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  • Immunocytochemistry  (122)
  • Zea mays  (93)
  • Springer  (214)
  • American Geophysical Union
  • American Meteorological Society
  • Oxford University Press
  • 1995-1999  (120)
  • 1990-1994  (94)
  • 1995  (120)
  • 1992  (94)
Collection
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  • Springer  (214)
  • American Geophysical Union
  • American Meteorological Society
  • Oxford University Press
  • Wiley-Blackwell  (18)
Years
  • 1995-1999  (120)
  • 1990-1994  (94)
Year
  • 1
    Electronic Resource
    Electronic Resource
    Inheritance of resistance in maize silks to the corn earworm (1995)
    Springer
    Entomologia experimentalis et applicata 77 (1995), S. 315-321 
    Details
    ISSN: 1570-7458
    Keywords: Insecta ; Helicoverpa zea ; Zea mays ; resistance inheritance ; joint scaling test ; additive-dominance model
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The corn earworm,Helicoverpa zea (Boddie), is a perennial economic pest of field crops in the United States. Maize,Zea mays L., is the major host crop promoting the build-up of devastating corn earworm populations that limit full production of cotton, soybean, peanut, and grain sorghum. Resistance to the corn earworm in maize and in particular sweet maize, would provide an environmentally safe, economical method of control for this pest insect. Antibiotic effects of corn silks on this insect are: small larvae, extended developmental period, and reduced fecundity. Silks from individual maize plants of resistant and susceptible lines and progeny in six generations consisting of parents (P1, P2), F1, F2, and backcrosses BC1.1 (F1 × P1) and BC1.2 (F1 × P2) from each of four crosses were used to determine the genetic basis of the antibiotic resistance of silks to the corn earworm. In the cross of Zapalote Chico × PI340856, genes controlling resistance in the silks to the corn earworm larvae are dominant in PI340856 to those in Zapalote Chico. The cross of Zapalote Chico × GT114 involves parents differing in degree of resistance, and possibly differing for the genetic mechanism by which the resistance is inherited. The inheritance of resistance may involve non-additive (dominance and epistasis) genetic variance. A digenic 6-parameter model indicated (1) the resistance in this cross is controlled by more than one pair of genes and (2) some or all of the genes interact to cause non-allelic interaction. Thus, the resistance in this cross may be controlled by both dominant and recessive genes. The resistance of Zapalote Chico × CI64, an intermediate inbred, is influenced by additive gene effects. The digenic model adequately predicts all generation means of the cross of GT3 × PI340856 except for the F1. Thus, it appears that the additive-dominance model is not satisfactory for this cross involving susceptible and resistant parents. Generation mean analysis indicates that resistance to silk-feeding by corn earworm larvae is under genetic control, but gene action differs from one type of cross to another.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02383066
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  • 2
    Electronic Resource
    Electronic Resource
    Effects of a vesicular-arbuscular mycorrhizal fungus and other soil microorganisms on growth, mineral nutrient acquisition and root exudation of soil-grown maize plants (1995)
    Springer
    Mycorrhiza 5 (1995), S. 321-327 
    Details
    ISSN: 1432-1890
    Keywords: Glomus mosseae ; Zea mays ; Mineral uptake ; Root exudation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Maize (Zea mays L. cv. Alize) plants were grown in a calcareous soil in pots divided by 30-μm nylon nets into three compartments, the central one for root growth and the outer ones for hyphal growth. Sterle soil was inoculated with either (1) rhizosphere microorganisms other than vesicular-arbuscular mycorrhizal (VAM) fungi, (2) rhizosphere microorganisms together with a VAM fungus [Glomus mosseae (Nicol. and Gerd.) Gerdemann and Trappel], or (3) with a gamma-irradiated inoculum as control. Plants were grown under controlled-climate conditions and harvested after 3 or 6 weeks. VAM plants had higher shoot∶root ratios than non-VAM plants. After 6 weeks, the concentrations of P, Zn and Cu in roots and shoots had significantly increased with VAM colonization, whereas Mn concentrations had significantly decreased. Root exudates were collected on agar sheets placed on the interface between root and hyphal compartments. Six-week-old VAM and non-VAM plants had similar root exudate compositions of 72–73% reducing sugars, 17–18% phenolics, 7% organic acids and 3% amino acids. In another experiment in which root exudates were collected on agar sheets with or without antibiotics, the amounts of amino acids and carbohydrates recovered were similar in VAM and non-VAM plants. However, threeto sixfold higher amounts of carbohydrates, amino acids and phenolics were recovered when antibiotics were added to the agar sheets. Thus, the high microbial activity in the rhizosphere and on the rhizoplane limits the exudates recovered from roots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00207404
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  • 3
    Electronic Resource
    Electronic Resource
    Effects of a vesicular-arbuscular mycorrhizal fungus and other soil microorganisms on growth, mineral nutrient acquisition and root exudation of soil-grown maize plants (1995)
    Springer
    Mycorrhiza 5 (1995), S. 321-327 
    Details
    ISSN: 1432-1890
    Keywords: Key words Glomus mosseae ; Zea mays ; Mineral uptake ; Root exudation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Maize (Zea mays L. cv. Alize) plants were grown in a calcareous soil in pots divided by 30-μm nylon nets into three compartments, the central one for root growth and the outer ones for hyphal growth. Sterile soil was inoculated with either (1) rhizosphere microorganisms other than vesicular-arbuscular mycorrhizal (VAM) fungi, (2) rhizosphere microorganisms together with a VAM fungus [Glomus mosseae (Nicol. and Gerd.) Gerdemann and Trappel], or (3) with a gamma-irradiated inoculum as control. Plants were grown under controlled-climate conditions and harvested after 3 or 6 weeks. VAM plants had higher shoot : root ratios than non-VAM plants. After 6 weeks, the concentrations of P, Zn and Cu in roots and shoots had significantly increased with VAM colonization, whereas Mn concentrations had significantly decreased. Root exudates were collected on agar sheets placed on the interface between root and hyphal compartments. Six-week-old VAM and non-VAM plants had similar root exudate compositions of 72–73% reducing sugars, 17–18% phenolics, 7% organic acids and 3% amino acids. In another experiment in which root exudates were collected on agar sheets with or without antibiotics, the amounts of amino acids and carbohydrates recovered were similar in VAM and non-VAM plants. However, three- to sixfold higher amounts of carbohydrates, amino acids and phenolics were recovered when antibiotics were added to the agar sheets. Thus, the high microbial activity in the rhizosphere and on the rhizoplane limits the exudates recovered from roots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s005720050079
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  • 4
    Electronic Resource
    Electronic Resource
    Bioavailability of heavy metals and abundance of arbuscular mycorrhiza in a soil polluted by atmospheric deposition from a smelter (1995)
    Springer
    Biology and fertility of soils 19 (1995), S. 22-28 
    Details
    ISSN: 1432-0789
    Keywords: Arbuscular mycorrhiza ; Limed silty loam Heavy metals ; Pb-Zn smelter ; Root colonization Spore numbers ; Tolerance ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The bioavailability of heavy metals (Cd, Zn, Pb, Cu) and the abundance of arbuscular mycorrhiza (AM) were studied in two agricultural fields close to a Pb-Zn smelter and three fields outside the pollution zone all cultivated with maize (Zea mays L.). Metal extractability with ethylenediaminetetraacetic acid (EDTA)-NH4OAc and Ca(NO3)2, plant metal uptake, and mycorrhizal parameters (spore number, root colonization) were assessed at two growth stages (six-leaf and maturity). Despite regular liming, the availability of Cd, Zn, and Pb was markedly higher in the two metal-polluted fields than in the three uncontaminated fields. However, the AM abundance was not correlated with metal availability. Root colonization and spore numbers in the metal polluted fields were relatively high, though at plant maturity the former was significantly lower than in one of the uncontaminated fields. The very low AM abundance in the two other unpolluted fields was related to other factors, particular soil and plant P status and soil pH. AM root colonization did not substantially prevent plant metal accumulation, since the metal concentrations in maize grown on the polluted fields strongly exceeded normal values, and for Cd and Pb reached the limits of toxicity for animal feed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00336342
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  • 5
    Electronic Resource
    Electronic Resource
    Nitrogen use in maize-grain legume cropping systems in semi-arid Kenya (1995)
    Springer
    Biology and fertility of soils 20 (1995), S. 57-62 
    Details
    ISSN: 1432-0789
    Keywords: Nitrogen use ; Nitrogen fertilizer recovery ; Zea mays ; Phaseolus vulgaris ; Vigna unguiculata ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Locally suitable cultivars of maize, beans, and cowpeas were grown in field experiments for four seasons in semi-arid Kenya. For three seasons, the dry matter production and grain yield of maize and beans were not increased by N fertilizer additions up to 120 kg N ha-1. Fertilizer recoveries measured by 15N isotope dilution techniques were low, less than 20%. Inoculated and uninoculated beans failed to fix N2. By contrast the cowpea derived 50% of its N from fixation, equivalent to 197 kg N ha-1. The N content of the grain generally exceeded 40 kg N ha-1, and the N content of the seeds from the grain legumes were greater than those from the cereals. Large inputs of N fertilizer or N by fixation are required if maize-grain legume cropping system in semiarid Kenya are to be sustained in the long term.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00307842
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  • 6
    Electronic Resource
    Electronic Resource
    Sequential development of pathogens in the maize tarspot disease complex (1992)
    Springer
    Mycopathologia 117 (1992), S. 157-161 
    Details
    ISSN: 1573-0832
    Keywords: Phyllachora maydis ; Monographella maydis ; Coniothyrium phyllachorae ; Zea mays ; tarspot complex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The tarspot complex is caused by the interaction of Phyllachora maydis and Monographella maydis. Coniothyrium phyllachorae, possibly a mycoparasite, is found in older ascostromata of P. maydis, which always appears first causing tarspot. M. maydis follows and is responsible for the damaging “fisheye” symptom. The fisheye symptom is always associated with a tarspot in the center of the lesion, whereas 12 to 20% of the Phyllachora ascostromata remained free of M. maydis. Inoculations of maize leaves with the Microdochium anamorph of the Monographella (usually produced in lesions) failed to produce infections. Some infections with M. maydis were, however, obtained under unusual conditions in the field. Inoculations onto tarspots in the laboratory were unsuccessful, but in field experiments inoculations with conidia of M. maydis enhanced severity of the tarspot complex. Fisheye symptoms of the complex naturally appear 2 to 7 days after the manifestation of P. maydis. This is followed a week later by the appearance of M. maydis which became predominant in the lesions and is associated with empty perithecia of P. maydis. In the early stages of the tarspots pycnidia of the anamorph of P. maydis, Linochora sp., could occasionally be observed. Ascomata of M. maydis were rare in the field. Of the 36 genetic materials of CIMMYT tested, 30 developed the fisheye symptom, 4 tarspots only and 2 remained free of symptoms
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00442777
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  • 7
    Electronic Resource
    Electronic Resource
    The effects of aflatoxin B1 on immature germinating maize (Zea mays) embryos (1992)
    Springer
    Mycopathologia 119 (1992), S. 181-190 
    Details
    ISSN: 1573-0832
    Keywords: aflatoxin B1 ; electron microscopy ; in vitro ; immature maize embryo ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immature maize (Zea mays L.) embryos were treated with aflatoxin B1 concentrations, ranging from 0.1 μg ml−1 to 25 μg ml−1. Below 5 μg ml−1 aflatoxin B1, root and shoot elongation was not significantly inhibited. Ultrastructurally, root tip cells showed little deterioration, except a possible diffused clearing in mitochondria and plastids. As the toxin concentration was increased above 5 μgml−1, shoot, and particularly root elongation, was progressively inhibited. Associated with this, there was an apparent decrease in the ribosome population. Furthermore, membranes, particularly the vacuolar membrane, became abnormal and vacuolar distension occurred. At 20 and 25 μg ml−1, these effects were exacerbated, and mitochondria and plastid structure was disrupted. At these concentrations, there was evidence of a disruption in lipid metabolism. The results are discussed in the context of known aflatoxin effects on cellular control mechanisms and ultrastructure in animal systems.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00448817
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  • 8
    Electronic Resource
    Electronic Resource
    Infection and disintegration of vascular tissues of non-suberized roots of spruce byHeterobasidion annosum and use of antibodies for characterizing infection (1995)
    Springer
    Mycopathologia 129 (1995), S. 91-101 
    Details
    ISSN: 1573-0832
    Keywords: ELISA ; Endodermis ; H. annosum ; Immunocytochemistry ; Root rot ; Vascular tissues
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Vascular disintegration mainly of medulla rays of spruce roots is of major significance in root rot disease of spruce caused byH. annosum. Using seedling roots as an experimental model, the possible routes and initial host reactions preceding invasion of vascular tissues was investigated. Transmission electron microscopy showed that penetration through the endodermis was an obvious route but not without host resistance. Using antibodies againstH. annosum hyphal materials, some labelling of vascular tissues remote from sites of fungal colonization suggest the release of fungal secretory products partly active in tissue disintegration. Similarly, intense labelling was also observed in severely colonized host tissues at late stages of infection. Strong labelling recorded at 3 d p.i. mainly on fungal hyphae and scant gold particles on invaded host tissues could imply that induction of host antifungal metabolites may have been a late event. A correlation was found between total antigenic material in root homogenates measured by ELISA, density of tissue labelling by immunocytochemistry and severity of disease symptoms. The importance of this in relation to diagnosis of biotic root rot diseases in the field is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01103468
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  • 9
    Electronic Resource
    Electronic Resource
    Enterobacter cloacae is an endophytic symbiont of corn (1995)
    Springer
    Mycopathologia 129 (1995), S. 117-125 
    Details
    ISSN: 1573-0832
    Keywords: Biological control ; Corn seedling disease ; Enterobacter cloacae ; Fusarium moniliforme ; Maize ; Seedling blight ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The bacteriumEnterobacter cloacae is presently used for biocontrol of postharvest diseases of fruits and vegetables and as a preplant seed treatment for suppression of damping-off. This bacterium has apparent affinities for several grass species, but it is not considered to be an endophyte. While screening corn for fungi and bacteria with potential for biocontrol of various corn diseases, the surface-sterilized kernels of one unknown Italian corn cultivar produced fungus-free corn seedlings with roots endophytically infected byE. cloacae. This paper describes the microscopic nature ofE. cloacae RRC 101 with corn, and the in vitro control ofFusarium moniliforme and other fungi with this bacterium. Light and electron microscopy determined that this isolate ofE. cloacae was biologically associated with corn seedling roots, where it was distributed intercellularly within the cortex and stele. This is a first report of a strain of this bacterium as an endophytic symbiont of roots. Following a topical application ofE. cloacae to kernels, and upon germination this bacterium readily infected roots of two other corn cultivars. The bacterium was observed within the endosperm of germinating corn seedling, but germination was not affected. Further, the bacterium was isolated from leaves and stems of 3- to 6-week-old seedlings indicating that the above ground portions of corn were also colonized. There was no evidence of damage to cells of the root during a three to four week observation period. This bacterium was antagonistic to several isolates of the corn pathogenFusarium moniliforme, and to two other species of fungi, all of which produce mycotoxins on corn.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01103471
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  • 10
    Electronic Resource
    Electronic Resource
    The phytotoxicity of selected mycotoxins on mature, germinatingZea mays embryos (1995)
    Springer
    Mycopathologia 132 (1995), S. 173-183 
    Details
    ISSN: 1573-0832
    Keywords: Deoxynivalenol ; Embryo ; Mature ; Ochratoxin ; Plantlet ; Zearalenone ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Mature maize (Zea mays) embryos were exposed to 5, 10 and 25 µg ml−1 of deoxynivalenol (DON), zearalenone (ZEA), ochratoxin A (OA) and a mixture of zearalenone and deoxynivalenol (ZEA/DON) for 9 days. DON and the ZEA/DON combination were consistently more inhibitory of the measured parameters than either ZEA or OA. Based on the predicted additive values, it would appear that, in combination, ZEA and DON act synergistically to inhibit root and shoot growth. For ZEA alone, a concentration of 5 µg ml−1 ZEA was generally inhibitory of root and shoot elongation and fresh mass accumulation, while at 10 and 25 µg ml−1, this toxin had a stimulatory effect on these parameters. For OA, the measured effects on root and shoot growth at 5 and 25 µg ml−1 were stimulatory, while at 10 µg ml−1 OA, an inhibitory effect was observed. For all toxins, inhibitory/stimulatory effects were generally more marked for root parameters than for shoot elongation or mass.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01103984
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  • 11
    Electronic Resource
    Electronic Resource
    The egg-jelly macromolecule, a fucose sulphate glycoconjugate, originates from the accessory cells of the ovary in the sea urchin Hemicentrotus pulcherrimus (1992)
    Springer
    Development genes and evolution 201 (1992), S. 179-189 
    Details
    ISSN: 1432-041X
    Keywords: Sea urchin ; Jelly coat ; Accessory cell ; Oogenesis ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The immunocytochemical localization of the egg-jelly macromolecule, a fucose sulphate glycoconjugate (FSG) that induces the acrosome reaction in spermatozoa, was investigated in ovaries of the sea urchin Hemicentrotus pulcherrimus by use of a polyclonal antibody. The polyclonal antibody reacted with the accessory cells and oocytes in the ovarian lumen. In the accessory cells, evidence of an intense immunohistochemical reaction was observed in many globules of variable density. Products of the specific immunohistochemical reaction were frequently observed in the surface region of oocytes, at a distance from the ovarian wall. At the ultrastructural level, the polyclonal antibody was found to react with the material present in the vacuole-like structures of the globules in the accessory cells. Many gold particles, demonstrating specific immunolabelling, were associated with well-developed microvilli on the vitellogenic oocytes. In the mature oocytes, intense labelling was observed in the jelly coat but not in the vitelline coat. By contrast, oogonia and early oocytes were barely labelled. Quantitative data indicated that the extent of immunolabellings in the surface region of oocytes was very high in the vitellogenic and mature oocytes. In all cases, neither the oocyte cytoplasm nor the subcellular organelles were labelled. These results suggest that FSG is produced by the accessory cells and is deposited initially on the surface of vitellogenic oocytes for the formation of jelly. These findings may provide a new insight into the role of the accessory cells in the reproductive process of the sea urchin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00188717
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  • 12
    Electronic Resource
    Electronic Resource
    Expression of growth hormone receptor by immunocytochemistry in rat molar root formation and alveolar bone remodeling (1992)
    Springer
    Calcified tissue international 50 (1992), S. 541-546 
    Details
    ISSN: 1432-0827
    Keywords: Growth hormone ; Growth hormone receptor ; Odontogenesis ; Bone remodeling ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Growth hormone (GH) may regulate tooth formation and bone remodeling associated with tooth eruption. This study reports the distribution of growth hormone receptor/binding protein in developing rat molars and adjacent alveolar bone by immunocytochemistry using well-characterized anti-growth hormone receptor monoclonal antibodies. These tissues represent an excellent model for studying the ontogenic changes that occur in odontogenic and osteogenic cells, as these cells are found in linear arrays displaying the various stages of morphological and functional differention, and differentiated function. Immunoreactivity was first seen in precementoblasts in contact with the epithelial root sheath, and preodontoblasts. However, growth hormone receptor immunoreactivity was associated primarily with the cytoplasm of odontogenic and osteogenic cells forming their respective matrices. Thus, cementoblasts and odontoblasts at sites of new matrix formation showed intense immunoreactivity whereas cementocytes and mature odontoblasts at later stages of tooth development were nonreactive. Osteoblasts engaged in intramembranous ossification in the alveolar bone were positive, although osteocytes and endosteal cells were immunonegative. Osteoclasts at sites of alveolar bone remodeling resorption were also immunopositive. These patterns of receptor expression parallel the ontogenic sequences of odontogenic and osteogenic cells and suggest that GH promotes the functional state of these cells. Our results also imply that GH may influence differentiation or differentiated functions associated with odontogenesis, osteogenesis, and bone remodeling independent of systemic insulin-like GF-I.
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    URL: http://dx.doi.org/10.1007/BF00582170
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  • 13
    Electronic Resource
    Electronic Resource
    Electroporation and PEG delivery of DNA into maize microspores (1992)
    Springer
    Plant cell reports 11 (1992), S. 567-570 
    Details
    ISSN: 1432-203X
    Keywords: Microspore ; Electroporation ; Transformation ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The ability to deliver and detect reporter gene activity in maize microspores was tested. Tested expression vectors contained the chloramphenicol acetyl transferase (CAT) gene and one of the following promoter-intron combinations: 1) cauliflower mosaic virus (CaMV 35S), 2) CaMV 35S + maize alcohol dehydrogenase 1 intron 6 (Adh1-I6), 3) maize alcohol dehydrogenase 1 + intron 1 (Adh1-I1), or 4) maize ubiquitin 1 + intron 1 (Ubiq 1-I1) promoter + intron. The expression vectors were delivered into maize microspores using electroporation or polyethylene glycol (PEG). Both methods were effective for delivering free DNA into microspores. Although all four promoters were active in maize protoplasts, only two promoters were active in maize microspores. The CaMV 35S and the Adh1 promoters did not promote gene expression in maize microspore. The CaMV 35S + Adh1-I6 and Ubiq1-I1 promoters produced high levels of CAT activity in maize microspores.
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    URL: http://dx.doi.org/10.1007/BF00233094
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  • 14
    Electronic Resource
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    Microfilaments (F-actin) in generative cells and sperm: an evaluation (1992)
    Springer
    Sexual plant reproduction 5 (1992), S. 89-100 
    Details
    ISSN: 1432-2145
    Keywords: Actin ; Cytoskeleton ; Generative cell ; Immunocytochemistry ; Microtubule ; Mitosis ; Phragmoplast ; Pollen ; Rhodamine phalloidin ; Sperm
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Disagreement has arisen over the presence of actin-containing microfilaments (Mfs) in angiosperm generative cells and sperm (GSP). In order to address this issue, we subjected GSP of Tradescantia virginiana, Nicotiana tabacum and Rhododendron laetum to a series of localizations using different antiactins, rhodamine phalloidin and antimyosin. Coordinate staining with antitubulin and Hoechst 33258 defined the status of the microtubule (Mt) cytoskeleton and stages of generative cell division. Additional experiments utilized cytochalasin D (CD). In no instance could Mfs be detected in GSP of the three species. Instead, Mfs seen at the periphery of GSP appear to be continuous with vegetative Mfs and thus are in the vegetative cytoplasm. Mfs are not seen in the constriction zone of dividing T. virginiana generative cells, nor are they indicated in the phragmoplast of N. tabacum and R. laetum. Myosin localizations reveal punctate staining in the vegetative cytoplasm and a thin line of fluorescence around the the outside of the generative cell. While CD seems to delay generative cell division, cytokinesis still takes place. CD-induced Mf fragments are evident in the vegetative cytoplasm but not in GSP. The weight of evidence therefore indicates that GSP do not contain Mfs. The implications of this conclusion for the behavior of GSP and the mechanism of cytokinesis in dividing generative cells are considerable.
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    URL: http://dx.doi.org/10.1007/BF00194867
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  • 15
    Electronic Resource
    Electronic Resource
    Cleavage polyembryony in maize (1992)
    Springer
    Sexual plant reproduction 5 (1992), S. 224-226 
    Details
    ISSN: 1432-2145
    Keywords: Zea mays ; Maize ; Polyembryony
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two types of cleavage polyembryony are described in the inbred line VIR 17 of maize. Suspensorial embryony was observed to occur spontaneously. Typical cleavage of the zygotic proembryo occurred spontaneously, but could also be induced by treating the developing caryopses with 2,4-Dichlorophenoxyacetic acid (2,4-D) on the second day after pollination. 2,4-D was active as a decorelative factor also evoking the expression of totipotency in individual proembryonal cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00189815
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  • 16
    Electronic Resource
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    Deleterious effect of minimal enzymatic treatments on the development of isolated maize embryo sacs in culture (1995)
    Springer
    Sexual plant reproduction 8 (1995), S. 313-317 
    Details
    ISSN: 1432-2145
    Keywords: Embryo sac ; Zea mays ; Enzymatic isolation ; Zygotic embryogenesis ; Microinjection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The long-term viability of isolated embryo sacs was studied in maize. Fertilised embryo sacs were digested in order to remove most of the nucellus cells present on their surfaces and then transferred to culture. Experiments on 161 embryo sacs showed that isolation treatments using even minimal enzymatic digestion affected the further development of the embryo sacs. Few embryo sacs survived in culture and those produced only abnormal embryos; they produced no plants. We concluded that embryo sacs isolated through enzymatic digestion may offer limited prospects for long-term studies where normal embryogenic development is required. Alternative strategies are discussed for maize.
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    URL: http://dx.doi.org/10.1007/BF00229389
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  • 17
    Electronic Resource
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    Analysis of pollen-tube growth in cultured maize silks (1992)
    Springer
    Sexual plant reproduction 5 (1992), S. 227-231 
    Details
    ISSN: 1432-2145
    Keywords: Zea mays ; Maize ; Pollen-tube growth regulation ; In vitro pollination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In vitro pollen-tube growth in maize was studied using an in vitro pollination system. In the ‘cut-silk’ method, ovaries with silks were placed on medium in vitro, whereafter the silk was cut and the upper part of the silk was pollinated. Pollen tubes were not able to bridge the space between the two silk parts. Even when silk parts were tightly connected, pollen tubes still were not able to pass the cut ends and reach the lower silk part. Pollen-tube growth rates and the direction of tube growth were not influenced by the presence or absence of an ovary. Prepollination did not have any influence on pollen-tube growth rate. Measurements of pollen-tube growth rate also showed that there was no ‘population effect’, i.e. growth rate was not stimulated by pollination with an excess of pollen grains. We found that the direction in which maize pollen grew was determined only by the positioning of the silk hairs.
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    URL: http://dx.doi.org/10.1007/BF00189816
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    Effects of calcium, magnesium, potassium and boron on sperm cells isolated from pollen of Zea mays L. (1995)
    Springer
    Sexual plant reproduction 8 (1995), S. 113-122 
    Details
    ISSN: 1432-2145
    Keywords: Zea mays ; Calcium ; Cell integrity ; Cell viability ; Sperm cell
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Our previous studies showed that Brewbaker and Kwack salts, which have been widely used in pollen germination and sperm isolation, are not appropriate for the maintenance of isolated maize (Zea mays L.) sperm cells. In the present study, we have characterized the effects of each BKS component salt on the integrity of isolated sperm cells using hemacytometry. At 0.01 and 0.1 mM, there were no differences in cell number between control and any salt-treated cells except a 22% decrease with 0.1 mM MgSO4 at 48 h. At the 1 mM level, cell number decreased with time in the presence of Ca(NO3)2 and MgSO4, with loss of integrity of most cells at 48 h, while KNO3 and H3BO3 had little or no effect. Further characterization of calcium-induced reduction in cell integrity using flow cytometry showed that depletion of possible residual free calcium by addition of EGTA to the suspension medium improved cell longevity and viability. Exposure of isolated sperm cells to 1 mM calcium had no effect on cell integrity and viability in 5 h; however, only 12% of cells remained intact at 24 h. The reduction in cell integrity was hastened when cells were pretreated with the calcium ionophore A23187 prior to exposure to 1 mM calcium, with a 54% reduction in cell number at 1 h and complete cell lysis at 24 h. However, depletion of cytosolic free calcium by pretreatment of cells with the calcium ionophore followed by resuspension in the presence of EGTA resulted in rapid reduction of cell integrity as well. These results collectively suggest that maize sperm cells are sensitive to exogenous free calcium; however, a certain level of cytosolic free calcium is necessary for maintenance of integrity. Mechanisms of calcium-induced reduction in cell integrity are discussed along with possible roles of the sensitivity of sperm cells to calcium in fertilization.
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    URL: http://dx.doi.org/10.1007/BF00230898
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  • 19
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    Maturation of maize pollen in vitro (1992)
    Springer
    Plant cell reports 11 (1992), S. 535-539 
    Details
    ISSN: 1432-203X
    Keywords: Zea mays ; in vitro culture ; in vitro pollen ; pollen germination ; fertilization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Maturation of maize pollen was obtained in male reproductive structures cultured in vitro. Immature tassels containing microspores at the mid-uninucleate to late-binucleate stage of development were excised and spikelets, anthers, and/or isolated microspores were cultured on a medium capable of supporting pollen maturation. Microspore mitosis, culminating in the production of starch-filled, trinucleate pollen capable of germination, was observed after 7–15 days, depending on the genotype and stage at which the cultures were initiated. Up to 100%, 70%, and 20% of the cultured spikelets, anthers, and isolated microspores, respectively, produced mature pollen, which germinated, however, at different frequencies (i.e., spikelets, 50–70%; anthers, 5–10%; microspores, 〈1%). Mature kernels were produced following fertilization with pollen from cultured spikelets and anthers. These procedures provide methods for the in vitro manipulation of a significant phase of the maize life cycle.
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    URL: http://dx.doi.org/10.1007/BF00236273
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    Matrix-associated DNA from maize is enriched in repetitive sequences (1992)
    Springer
    Plant cell reports 11 (1992), S. 355-358 
    Details
    ISSN: 1432-203X
    Keywords: Zea mays ; Matrix-associated ; DNA ; repetitive sequences ; DNA loops
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In order to elucidate some features of the topological organization of DNA within the plant nucleus, DNA fragments involved in the attachment of the DNA loops to the nuclear matrix in maize were studied. The matrix-associated DNA from dry embryo and meristematic cells after extensive digestion with DNase I and high salt treatment was about 2% of the total DNA, sized within the range of 50 and 250 bp. This DNA was found to be enriched in repetitive DNA sequences, both for nuclei from dry embryo and meristematic cells. The loop size of the DNA in cells of Zea mays appeared to be between 5 and 25 kbp.
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  • 21
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    Field evaluation of N2 fixation and N partitioning in climbing bean (Phaseolus vulgaris L.) using 15N (1992)
    Springer
    Biology and fertility of soils 13 (1992), S. 142-146 
    Details
    ISSN: 1432-0789
    Keywords: A value ; Common bean ; N remobilization ; Soil N balance ; Atom% 15N excess ; Phaseolus vulgaris ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The common bean (Phaseolus vulgaris L.) is generally regarded as a poor N2 fixer. This study assessed the sources of N (fertilizer, soil, and fixed N), N partitioning and mobilization, and soil N balance under field conditions in an indeterminate-type climbing bean (P. vulgaris L. cv. Cipro) at the vegetative, early pod-filling, and physiological maturity stages, using the A-value approach. This involved the application of 10 and 100 kg N ha-1 of 15N-labelled ammonium sulphate to the climbing bean and a reference crop, maize (Zea mays L.). At the late pod-filling stage (75 days after planting) the climbing bean had accumulated 119 kg N ha-1, 84% being derived from fixation, 16% from soil, and only 0.2% from the 15N fertilizer. N2 fixation was generally high at all stages of plant growth, but the maximum fixation (74% of the total N2 fixed) occurred during the interval between early (55 days after planting) and late podfilling. The N2 fixed between 55 and 75 days after planting bas a major source (88%) of the N demand of the developing pod, and only about 11% was contributed from the soil. There was essentially no mobilization of N from the shoots or roots for pod development. The cultivation of common bean cultivars that maintain a high N2-fixing capacity especially during pod filling, satisfying almost all the N needs of the developing pod and thus requiring little or no mobilization of N from the shoots for pod development, may lead to a net positive soil N balance.
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    URL: http://dx.doi.org/10.1007/BF00336269
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    Periplasmic location of nitrous oxide reductase and its apoform in denitrifying Pseudomonas stutzeri (1992)
    Springer
    Archives of microbiology 157 (1992), S. 218-222 
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    ISSN: 1432-072X
    Keywords: Denitrification ; N2O reductase ; Nitrite reductase ; Immunocytochemistry ; Localization ; Two-dimensional electrophoresis ; Cell fractionation ; Pseudomonas stutzeri
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Immunogold labelling techniques on ultrathin sections of low temperature embedded cells yielded evidence for the periplasmic location of the respiratory enzymes N2O reductase and nitrite reductase (cytochrome cd 1) in Pseudomonas stutzeri strain ZoBell. Cell fractionation by spheroplast preparation and two-dimensional electrophoresis showed the absence of a membrane association of these enzymes. Immunocytochemical localization of N2O reductase in a mutant strain deficient in the chromophore of N2O reductase showed the gold label at the cell periphery, indicating that the copper chromophore processing takes place after export of this protein's apoform.
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    Odors influence choice of oviposition sites byDiabrotica virgifera virgifera (Coleoptera: Chrysomelidae) (1992)
    Springer
    Journal of chemical ecology 18 (1992), S. 1227-1237 
    Details
    ISSN: 1573-1561
    Keywords: Western corn rootworm ; Diabrotica virgifera virgifera ; Coleoptera ; Chrysomelidae ; bacteria ; carbon dioxide ; pheromone ; semiochemicals ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract FemaleDiabrotica virgifera virgifera LeConte were allowed to choose between oviposition substrates that were and those that were not associated with potential sources of semiochemicals. Females deposited over five times more eggs on moist towelettes that were treated with homogenates of female abdomens than on towelettes treated with distilled water. Similar results were obtained when screening separated the homogenates from the towelettes, indicating that odors alone could elicit the response. In contrast, females did not choose towelettes that had previously been used for oviposition or towelettes containing eggs over unused towelettes. Further tests with homogenates of abdomens and a bacteriostatic agent (sorbate) indicated that the females were probably responding to bacterial odors rather than an oviposition-enhancing pheromone. Four strains of bacteria were isolated from a homogenate of female abdomens; females deposited 4 to 16 times more eggs on substrates with odors of the bacteria than on substrates with odors of uninoculated nutrient agar. In no-choice tests, bacterial odors did not increase the number of eggs deposited per female beetle; however, in choice tests with dishes that tended to retain any beetles that entered, there were more eggs per female (but not more beetles) after 24 hr in dishes with bacterial odors than in those without the odors. Females also chose dishes with odors of excised maize (Zea mays L.) roots or elevated levels of carbon dioxide over “control” dishes.
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    Mutational analysis of the ‘conserved region’ of maize streak virus suggests its involvement in replication (1992)
    Springer
    Plant molecular biology 19 (1992), S. 601-610 
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    ISSN: 1573-5028
    Keywords: geminivirus ; agroinfection ; Zea mays ; large intergenic region (LIR)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Maize streak virus as well as other geminiviruses contain a potential hairpin structure with the conserved sequence TAATATTAC in the loop. We assessed the possible involvement of this structure in replication and symptom induction of the virus. A series of insertion and deletion mutants were analyzed by agroinfection. Deletion of the hairpin or insertions in the conserved sequence abolished symptom development. Viral DNA could not be detected in the infected tissue. However, a mutant with a point mutation in the ‘conserved’ sequence, isolated after inoculation of maize plants with an insertion mutant, was able to replicate and to induce symptoms.
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    An mRNA that specifically accumulates in maize roots delineates a novel subset of developing cortical cells (1992)
    Springer
    Plant molecular biology 20 (1992), S. 821-831 
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    ISSN: 1573-5028
    Keywords: cortex ; developmental regulation ; in situ hybridization ; organ-specific gene expression ; roots ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A near full-length cDNA clone (pZRP3) corresponding to an mRNA that accumulates specifically in roots of maize was isolated. The ZRP3 mRNA is ca. 600 nucleotides in length. The amino acid sequence of the predicted polypeptide is rich in leucine (16%), proline (11%), and cysteine (8.5%). The zrp3 gene appears to be expressed exclusively in roots, whereas other ZRP3-related genes are expressed in additional organs of the maize plant. In situ hybridization shows that ZRP3 mRNA accumulation is largely confined to the cells of the cortical ground meristem. Furthermore, accumulation of this mRNA occurs within a distinct subset of cortical cells, the inner three to four cell layers.
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    A DNA polymerase from maize axes: its purification and possible role (1992)
    Springer
    Plant molecular biology 20 (1992), S. 1159-1168 
    Details
    ISSN: 1573-5028
    Keywords: DNA polymerase ; germination ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three different DNA polymerase activities can be resolved by passing a protein extract from 24 h imbibed maize axes through DEAE-cellulose. These activities have been numbered 1, 2 and 3, according to their elution order. One of them, DNA polymerase 2, elutes at 100–120 mM phosphates. This enzyme was further purified by passing it through Heparin-Sepharose, Sephacryl S-300 and DNA cellulose. Purification was nearly 5000-fold. The enzyme needs Mg2+, is stimulated by K+, has an optimum pH of 7.0 and its optimum temperature is 30–37 °C. Specific inhibitors for different types of polymerases, such as aphidicolin, dideoxythymidine triphosphate and N-ethyl maleimide, gave intermediate values of inhibition, making impossible the definition of the type of enzyme purified by its inhibitory pattern. SDS-PAGE indicated the presence of several bands of molecular masses of 28–40, 56 and 15 kDa. Most of these bands could be visualized when proteins from crude extracts were analyzed by western blot, using an antibody against calf thymus DNA polymerase α. A high molecular mass (around 500 kDa) was calculated by western blot of native gels using the same antibody. Finally, specific activity of this enzyme increased 100-fold during maize germination whereas polymerase 3 virtually did not increase. Furthermore, immunoprecipitation experiments with the antipolymerase α-antibody showed a decrease in DNA polymerase activity by 70%. The possibility that polymerase 2 is a replicative enzyme is discussed.
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    Stress responses in maize: Sequence analysis of cDNAs encoding glycine-rich proteins (1992)
    Springer
    Plant molecular biology 18 (1992), S. 847-849 
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    ISSN: 1573-5028
    Keywords: cDNA ; nucleotide sequence ; glycine-rich proteins ; chemical stress ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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    URL: http://dx.doi.org/10.1007/BF00020034
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    Multiple ocs-like elements required for efficient transcription of the mannopine synthase gene of T-DNA in maize protoplasts (1992)
    Springer
    Plant molecular biology 20 (1992), S. 219-233 
    Details
    ISSN: 1573-5028
    Keywords: promoter ; electroporation ; protoplasts ; transient assay ; Agrobacterium ; Ti plasmid ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Regulatory elements controlling transcriptional activity of the mannopine synthase 2′ promoter (mas 2′) were defined by analysis of deletion mutants in transient expression assays in maize protoplasts. Deletion of the region between −305 and −290 containing sequence similarity to the octopine synthase (ocs) promoter element reduced activity by 67% compared to wild type activity. Less than 1% of the activity remained in 5′ deletions downstream of −153. Inclusion of various heterologous enhancer-like sequences immediately upstream of position −325 increased activity by up to 7.5-fold. Insertion of the −325 to −275 sequence alone, or in combination with heterologous enhancer-like elements, restored activity of some of the 5′-deletion mutants. Restoration of activity was not obtained with mutants deleted past position −127. Our results suggest that a single class of nuclear proteins from maize interact with high affinity at elements designated mas b (−306 to −275; mas 1′ element), d (−127 to −108), and e (−82 to −39; mas 2′ element) as well as the 20 bp element from the ocs promoter. Although the binding site at mas d only appears to accommodate a single protein, this element has the potential to make a weak, but positive, contribution to the activity of the mas 2′ promoter. The binding of nuclear proteins could not be demonstrated at mas a and c, both of which showed limited homology to the ocs element. Mutational evidence suggested that mas a and c may also contribute to mas 2′ transcription.
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    A Zea mays GTP-binding protein of the ARF family complements an Escherichia coli mutant with a temperature-sensitive malonyl-coenzyme A:acyl carrier protein transacylase (1995)
    Springer
    Plant molecular biology 27 (1995), S. 629-633 
    Details
    ISSN: 1573-5028
    Keywords: GTP binding ; ADP ribosylation ; Zea mays ; Escherichia coli ; fatty acid biosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In an attempt to isolate a plant malonyl-coenzyme A:acyl carrier protein transacylase cDNA clone, by direct genetic selection in an Escherichia coli fabD mutant (LA2-89) with a maize cDNA expression library, a Zea mays cDNA clone encoding a GTP-binding protein of the ARF family was isolated. Complementation of a mutation affecting bacterial membrane lipid biosynthesis by a plant ARF protein, could indicate the existence of as yet unidentified bacterial equivalents of this ubiquitous eucaryotic GTP-binding protein.
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    URL: http://dx.doi.org/10.1007/BF00019329
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    The involvement of Opaque 2 on β-prolamin gene regulation in maize and Coix suggests a more general role for this transcriptional activator (1995)
    Springer
    Plant molecular biology 27 (1995), S. 1015-1029 
    Details
    ISSN: 1573-5028
    Keywords: β-prolamin ; Coix lacryma-jobi ; different O2-binding sites ; Opaque 2 ; transcriptional regulation ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The maize opaque 2 (o2) mutation is known to have numerous pleiotropic effects. Some polypeptides have their expression depressed while others are enhanced. The best characterized effects of the o2 mutation are those exerted on endosperm genes encoding the storage protein class of the 22 kDa α-zeins and the ribosome inactivating protein b-32. The Opaque 2 (O2) locus encodes a basic domain-leucine zipper DNA-binding factor, O2, which transcriptionally regulates these genes. In the maize-related grass Coix lacryma-jobi, an O2-homologous protein regulates the 25 kDa α-coixin gene family. We show in this paper that O2 transcriptionally regulates the structurally and developmentally different class of the β-prolamins. A new O2-binding box was identified in β-prolamin genes from maize and Coix that, together with the boxes previously identified in other endosperm expressed genes, forms a curious collection of O2 cis elements. This may have regulatory implications on the role of O2 in the mechanism that controls coordinated gene expression in the developing endosperm. Considering that the O2 locus controls at least three distinct classes of genes in maize endosperm, we propose that the O2 protein may play a more general role in maize endosperm development than previously conceived.
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    Molecular cloning, characterization and expression of an elongation factor 1α gene in maize (1995)
    Springer
    Plant molecular biology 29 (1995), S. 611-615 
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    ISSN: 1573-5028
    Keywords: elongation factor 1α ; EF-1α ; Zea mays ; cDNA sequence ; gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA (zmEF1A) and the corresponding genomic clone (zmgEF1A) of a member of the gene family encoding the α subunit of translation elongation factor 1 (EF-1α) have been isolated from maize. The deduced amino acid sequence is 447 residues long interrupted by one intron. Southern blot analysis reveals that the cloned EF-1α gene is one member out of a family consisting of at least six genes. As shown by northern hybridizations in leaves the mRNA level increases at low temperature whereas time-course experiments over 24 h at 5°C show that in roots the overall mRNA level of EF-1α is transiently decreased. These results indicate that the expression of EF-1α is differently regulated in leaves and roots under cold stress.
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    Nucleotide sequence of a region of maize chloroplast DNA containing the 3′ end of clpP, exon 1 of rps12 and rpl20 and their cotranscription (1992)
    Springer
    Plant molecular biology 18 (1992), S. 415-418 
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    ISSN: 1573-5028
    Keywords: ribosomal protein ; rps12 ; rpl20 ; clpP ; chloroplast genome ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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    URL: http://dx.doi.org/10.1007/BF00034970
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    Nitrate reductase transcript is expressed in the primary response of maize to environmental nitrate (1992)
    Springer
    Plant molecular biology 18 (1992), S. 55-64 
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    ISSN: 1573-5028
    Keywords: cycloheximide ; leaf ; nitrate induction ; nitrate reductase transcript ; root ; scutellum ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nitrate induction of NADH:nitrate reductase mRNA in maize roots, scutella and leaves was investigated in the presence and absence of inhibitors of protein synthesis. In the absence of inhibitors, nitrate treatment caused a fairly rapid (2 to 3 h) increase in the level of the nitrate reductase transcript in all tissues. When cytoplasmic protein synthesis was inhibited by cycloheximide, nitrate reductase mRNA was induced by nitrate in all tissues to levels equal to or greater than those found with nitrate treatment alone. Treatment of maize tissues with cycloheximide in the absence of nitrate had only a small effect on the accumulation of the nitrate reductase mRNA. Inhibition of organellar protein synthesis with chloramphenicol also had little or no effect on nitrate-induced nitrate reductase mRNA accumuiation in roots and scutella, but did appear to partially inhibit appearance of transcript in leaves. Excision of scutella in the absence of nitrate was sufficient to cause some accumulation of the nitrate reductase transcript. Since cytoplasmic protein synthesis was not required for expression of nitrate reductase transcripts, induction of these transcripts by nitrate is a primary response of maize to this environmental signal. Thus, it appears that the signal transduction system mediating this response is constitutively expressed in roots, scutella and leaves of maize.
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    cDNA clones encoding Arabidopsis thaliana and Zea mays mitochondrial chaperonin HSP60 and gene expression during seed germination and heat shock (1992)
    Springer
    Plant molecular biology 18 (1992), S. 873-885 
    Details
    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; ATPase ; cpn60 ; developmental regulation ; molecular chaperones ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mitochondria contain a nuclear-encoded heat shock protein, HSP60, which functions as a chaperonin in the post-translational assembly of multimeric proteins encoded by both nuclear and mitochondrial genes. We have isolated and sequenced full-length complementary DNAs coding for this mitochondrial chaperonin in Arabidopsis thaliana and Zea mays. Southern-blot analysis indicates the presence of a single hsp60 gene in the genome of A. thaliana. There is a high degree of homology at the predicted amino acid levels (43 to 60%) between plant HSP60s and their homologues in prokaryotes and other eukaryotes which indicates that these proteins must have similar evolutionarily conserved functions in all organisms. Northern- and western-blot analyses indicate that the expression of the hsp60 gene is developmentally regulated during seed germination. It is also heat-inducible. Developmental regulation of the (β-subunit) of F1-ATPase, an enzyme complex that is involved in the cyanide-sensitive mitochondrial electron transport system, indicates that imbibed embryos undergo rapid mitochondrial biogenesis through the early stages of germination. Based on the functional role of HSP60 in macromolecular assembly, these data collectively suggest that the presence of higher levels of HSP60 is necessary during active mitochondrial biogenesis, when the need for this protein is greatest in assisting the rapid assembly of the oligomeric protein structures.
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    Dissection of a pollen-specific promoter from maize by transient transformation assays (1992)
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    Plant molecular biology 18 (1992), S. 211-218 
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    ISSN: 1573-5028
    Keywords: pollen-specific gene expression ; promoter analysis ; transient assays ; Tradescantia ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have previously reported the isolation and characterization of a gene (Zm 13) from Zea mays which shows a pollen-specific pattern of expression. Stably transformed tobacco plants containing a reporter gene linked to portions of the Zm 13 5′ flanking region show correct temporal and spatial expression of the gene. Here we present a more detailed analysis of the 5′ regions responsible for expression in pollen by utilizing a transient expression system. Constructs containing the β-glucuronidase (GUS) gene under the control of various sized fragments of the Zm13 5′ flanking region were introduced into Tradescantia and Zea mays pollen via high-velocity microprojectile bombardment, and monitored both visually and with a fluorescence assay. The results suggest that sequences necessary for expression in pollen are present in a region from −100 to −54, while other sequences which amplify that expression reside between −260 and −100. The replacement of the normal terminator with a portion of the Zm13 3′ region containing the putative polyadenylation signal and site also increased GUS expression. While the −260 to −100 region contains sequences similar to other protein-binding domains reported for plants, the −100 to −54 region appears to contain no significant homology to other known promoter fragments which direct pollen-specific expression. The microprojectile bombardment of Tradescantia pollen appears to be a good test system for assaying maize and possibly other monocot promoter constructs for pollen expression.
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    The maize GapC4 promoter confers anaerobic reporter gene expression and shows homology to the maize anthocyanin regulatory locus C1 (1995)
    Springer
    Plant molecular biology 29 (1995), S. 1293-1298 
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    ISSN: 1573-5028
    Keywords: anaerobiosis ; BMS cells ; glyceraldehyde-3-phosphate dehydrogenase ; transient gene expression ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The cytosolic glyceraldehyde-3-phosphate dehydrogenase (GapC) gene family of maize is differentially expressed in response to anaerobic stress. While GapC1 and GapC2 are downregulated, GapC3 and GapC4 are anaerobically induced. We have sequenced and analyzed a 3073 bp promoter fragment of GapC4. The promoter confers anaerobic induction of a reporter gene construct in a transient gene expression system in maize. Deletion analysis of the GapC4 promoter revealed a 270 bp long DNA region required for anaerobic induction. This region contains sequence motifs resembling the cis-acting sequences of the anaerobically induced maize Adh1 and Adh2 genes. Furthermore, the 3073 bp GapC4 promoter fragment displays homology to long terminal repeats of maize retrotransposons and to the 3′ region of the maize anthocyanin regulatory locus C1.
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    Molecular cloning and characterization of six cDNAs expressed during glucose starvation in excised maize (Zea mays L.) root tips (1995)
    Springer
    Plant molecular biology 28 (1995), S. 473-485 
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    ISSN: 1573-5028
    Keywords: carbon catabolite repression ; cDNA ; gene expression ; stress-induced genes ; glucose-starvation ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In order to isolate glucose-starvation-related cDNAs in maize (Zea mays L.) root tips, a cDNA library was constructed with poly(A)+ mRNA from 24 h starved root tips. After differential screening of the library, we isolated six different cDNAs (named pZSS2 and pZSS7) which were expressed during glucose starvation. Time course analysis revealed that maximum expression of five of these genes occurs 30 h after the onset of the starvation treatment. On the contrary, the expression of mRNAs corresponding to pZSS4 was maximal at an early stage of starvation and then dramatically decreased. The expression of this gene did not seem to be specific for glucose starvation. The pattern of induction of the genes corresponding to pZSS2, pZSS3, pZSS5, pZSS6 and pZSS7 revealed that non-metabolizable sugars such as L-glucose and mannitol induce mRNA transcription similarly to glucose starvation. When D-glucose or any other metabolizable sugar was supplied, the level of transcripts was reduced. Nucleotide sequence analyses of the six cDNAs allowed identification of five of them by comparison with sequence data bases. The protein encoded by clone pZSS2 is analogous to a wound-induced protein from barley. Clones pZSS4 to pZSS7 encode, respectively, a transmembrane protein, a cysteine protease, a metallothionein-like protein and a chymotrypsin/subtilisin-like protease inhibitor. Clone pZSS3 shares no significant homology with any known sequence.
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    URL: http://dx.doi.org/10.1007/BF00020395
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    Intron-dependent transient expression of the maize GapA1 gene (1995)
    Springer
    Plant molecular biology 28 (1995), S. 667-676 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; promoter ; glyceraldehyde-3-phosphate dehydrogenase ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transient expression experiments show that the maize GapA1 promoter exhibits a requirement for sequences contained within intron 1 and surrounding exon border regions for expression in maize Black Mexican Sweet cells. Maize GapA1-promoter constructs lacking intron 1 are inactive. Intron 1 and its exon border sequences, when reintroduced into constructs lacking introns, restore gene activity whereas intron 2 and its exon borders to not. The minimal promoter so defined encompasses roughly 250 bp upstream of the in vivo transcription start and appears also to include intron 1. An octameric sequence was identified in intron 1 of maize GapA1 which is similar to sequence motifs found in other maize introns known to increase transient expression. Partial restoration of gene expression in GapA1 constructs lacking intron 1 was achieved through insertion of the identified octameric sequence.
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    URL: http://dx.doi.org/10.1007/BF00021192
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    PCR-generated cDNA library of transition-stage maize embryos: cloning and expression of calmodulin genes during early embryogenesis (1995)
    Springer
    Plant molecular biology 27 (1995), S. 105-113 
    Details
    ISSN: 1573-5028
    Keywords: calmodulin ; cDNA library ; embryogenesis ; PCR ; transition stage ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract One hundred maize zygotic embryos microdissected at the transition stage were used to construct a cDNA library after non-selective PCR (NS-PCR) amplification of whole cDNA populations. The library contains 2.3 × 105 recombinants and two different calmodulin cDNAs were cloned using a heterologous probe from petunia. Calmodulin expression was confirmed throughout maize embryogenesis at the mRNA, amplified cDNA and protein levels. Sequence analysis suggests a maize origin for both clones and negligible nucleotide changes linked to PCR. This library is the first described for early plant embryos and represents a breakthrough to isolate genes involved in embryo differentiation.
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    URL: http://dx.doi.org/10.1007/BF00019182
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    Evidence for the thiamine biosynthetic pathway in higher-plant plastids and its developmental regulation (1995)
    Springer
    Plant molecular biology 29 (1995), S. 809-821 
    Details
    ISSN: 1573-5028
    Keywords: Zea mays ; thiamine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Thiamine or vitamin B-1, is an essential constituent of all cells since it is a cofactor for two enzyme complexes involved in the citric acid cycle, pyruvate dehydrogenase and α-ketoglutarate dehydrogenase. Thiamine is synthesized by plants, but it is a dietary requirement for humans and other animals. The biosynthetic pathway for thiamine in plants has not been well characterized and none of the enzymes involved have been isolated. Here we report the cloning and characterization of two cDNAs representing members of the maize thi1 gene family encoding an enzyme of the thiamine biosynthetic pathway. This assignment was made based on sequence homology to a yeast thiamine biosynthetic gene and by functional complementation of a yeast strain in which the endogenous gene was inactivated. Using immunoblot analysis, the thi1 gene product was found to be located in a plastid membrane fraction. RNA gel blot analysis of various tissues and developmental stages indicated thi1 expression was differentially regulated in a manner consistent with what is known about thiamine synthesis in plants. This is the first report of cDNAs encoding proteins involved in thiamine biosynthesis for any plant species.
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    URL: http://dx.doi.org/10.1007/BF00041170
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    Manipulating cytoplasmic pH under anoxia: A critical test of the role of pH in the switch from aerobic to anaerobic metabolism (1995)
    Springer
    Planta 195 (1995), S. 324-330 
    Details
    ISSN: 1432-2048
    Keywords: Anoxia ; Biochemical pH-stat ; Cytoplasmic pH ; Ethanol production ; Pyruvate decarboxylase ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Ethanol production by maize (Zea mays L.) root tips, measured by an enzymic assay of the suspending medium, was correlated with changes in the cytoplasmic pH, determined by in-vivo 31P nuclear magnetic resonance (NMR) spectroscopy, following the onset of anoxia. Strong evidence for the role of the cytoplasmic pH in triggering the switch to ethanol production under anoxia was obtained by: (i) varying the pH of the suspending medium between pH 4 and pH 10; and (ii) using the permeant weak base methylamine to combat the acidification of the cytoplasm induced by the anoxic conditions. Experimentally, it proved to be much easier to manipulate the cytoplasmic pH under anoxia after the pH had stabilised, rather than during the initial rapid acidification that occurred following the onset of anoxia, and in the presence of methylamine, it was possible to impose a normal aerobic cytoplasmic pH value on tissue that was metabolising anaerobically. By this means it was possible to demonstrate the reversibility of the pH effect on ethanol production under anoxia and thus to provide good evidence in support of the biochemical pH-stat model of the anoxic response. The NMR measurement of the cytoplasmic pH in the presence of methylamine was achieved by using a manganese pretreatment technique to eliminate interference between the cytoplasmic and vacuolar Pi signals, and it seems likely that the experimental approach used here will have further applications in studies of the metabolic response to anoxia.
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    URL: http://dx.doi.org/10.1007/BF00202588
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    Transport and dynamics of molecules dissolved in maize root cortex membranes (1995)
    Springer
    The journal of membrane biology 143 (1995), S. 19-28 
    Details
    ISSN: 1432-1424
    Keywords: Plant membrane ; Lipid domain ; Fluorescence photobleaching recovery ; Electron paramagnetic resonance ; Temperature stress ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Translational diffusion of a fluorescent sterol probe was measured in the plasma membranes of protoplasts isolated from cortical cells of the primary root of maize seedlings. The apparent lateral diffusion coefficient was typically observed to be nearly insensitive to temperature, while the mobile fraction increased with increasing temperature. These fluorescence photobleaching recovery (FPR) measurements were compared with the electron paramagnetic resonance (EPR) spectra of the methyl ester of 13-doxyl palmitic acid in membranes of corn root tissue in situ. The complex spectra observed with this probe were analyzed as weighted sums of simpler spectra of various order parameters and rotational correlation times. The reconstituted spectra calculated from the model show that EPR also detects a mobile (less ordered, fluid) fraction, distinguished by the order parameter S=0.1 to 0.2, which becomes more abundant as temperature increases and is qualitatively comparable to the mobile fraction determined by the FPR method. The observed results on the mobile fractions and the diffusion rates for translational (FPR) as well as rotational (EPR) motions are interpreted in terms of membrane organization, thus providing information on the population and structural patterns of the coexisting domains with a special emphasis on the response of the membrane to temperature changes.
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    URL: http://dx.doi.org/10.1007/BF00232520
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    The AQP2 water channel: Effect of vasopressin treatment, microtubule disruption, and distribution in neonatal rats (1995)
    Springer
    The journal of membrane biology 143 (1995), S. 165-175 
    Details
    ISSN: 1432-1424
    Keywords: Water channels ; Vasopressin ; Rat kidney ; Immunocytochemistry ; Microtubules ; Cell polarity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Aquaporin 2 is a collecting duct water channel that is located in apical vesicles and in the apical plasma membrane of collecting duct principal cells. It shares 42% identity with the proximal tubule/thin descending limb water channel, CHIP28. The present study was aimed at addressing three questions concerning the location and behavior of the AQP2 protein under different conditions. First, does the AQP2 channel relocate to the apical membrane after vasopressin treatment? Our results show that AQP2 is diffusely distributed in cytoplasmic vesicles in collecting duct principal cells of homozygous Brattleboro rats that lack vasopressin. In rats injected with exogenous vasopressin, however, AQP2 became concentrated in the apical plasma membrane of principal cells, as determined by immunofluorescence and immunogold electron microscopy. This behavior is consistent with the idea that AQP2 is the vasopressin-sensitive water channel. Second, is the cellular location of AQP2 modified by microtubule disruption? In normal rats, AQP2 has a mainly apical and subapical location in principal cells, but in colchicine-treated rats, it is distributed on vesicles that are scattered throughout the entire cytoplasm. This is consistent with the dependence on microtubules of apical protein targeting in many cell types, and explains the inhibitory effect of microtubule disruption on the hydroosmotic response to vasopressin in sensitive epithelia, including the collecting duct. Third, is AQP2 present in neonatal rat kidneys? We show that AQP2 is abundant in principal cells from neonatal rats at all days after birth. The detection of AQP2 in early neonatal kidneys indicates that a lack of this protein is not responsible for the relatively weak urinary concentrating response to vasopressin seen in neonatal rats.
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    URL: http://dx.doi.org/10.1007/BF00233445
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    Occurrence and expression of acid phosphatase of Hymenoscyphus ericae (Read) Korf & Kernan, in isolation or associated with plant roots (1992)
    Springer
    Mycorrhiza 1 (1992), S. 137-146 
    Details
    ISSN: 1432-1890
    Keywords: Ericaceae ; Mycorrhizal fungi ; Acid phosphatase ; Protein expression ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The activity of acid phosphatase produced in pure culture by the endomycorrhizal fungus Hymenoscyphus ericae (Read) Korf & Kernan (H. ericae LPA 2) was inhibited by high phosphorus levels, alkaline pH, fluoride, molybdate and mannosidase, and activated by concanavalin A. Over 80% of the enzyme activity was due to two wall-bound acid phosphatase isozymes with the characteristics of mannose-rich glycoproteins. Antiserum was raised against the major, low-molecular-weight wall isozyme and its activity tested by immunoblotting and ELISA. The antiserum cross reacted 100% with exocellular (excreted) and 28% with cytoplasmic cellular fractions of H. ericae (LPA 2) cultures, and showed high reactivity with other strains of H. ericae but not with fungal isolates from Erica hispidula L. or E. mauritanica L. Ultrastructural localization of acid phosphatase by cytoenzymology and indirect immunogold labelling confirmed its association with the fungal wall in pure culture and showed that the influence of a high phosphorus level, fluoride and molybdate is through inactivation of the enzyme. Intense acid phosphatase activity, sensitive to the latter inhibitors, was also present on external hyphae growing over a host or non-host root but it was weak or absent from intracellular hyphae where these developed within a host root. Indirect immunolabelling confirmed that this acid phosphatase was of fungal origin and that the specific inhibitory effect of host cells is due to inactivation of the enzyme rather than repression of its synthesis. Possible implications of fungal acid phosphatase in ericoid endomycorrhizal infection processes are discussed together with mechanisms that may be regulating the enzyme activity.
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    Detection of the apomictic mode of reproduction in maize-Tripsacum hybrids using maize RFLP markers (1995)
    Springer
    Theoretical and applied genetics 90 (1995), S. 1198-1203 
    Details
    ISSN: 1432-2242
    Keywords: Diplospory ; RFLP ; Bulk-segregant analysis ; Genome similarity ; Intergeneric hybrids ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Polyploid plants in the genus Tripsacum, a wild relative of maize, reproduce through gametophytic apomixis of the diplosporous type, an asexual mode of reproduction through seed. Moving gene(s) responsible for the apomictic trait into crop plants would open new areas in plant breeding and agriculture. Efforts to transfer apomixis from Tripsacum into maize at CIMMYT resulted in numerou intergeneric F1 hybrids obtained from various Tripsacum species. A bulk-segregant analysis was carried out to identify molecular markers linked to diplospory in T. dactyloides. This was possible because of numerous genome similarities among related species in the Andropogoneae. On the basis of maize RFLP probes, three restriction fragments co-segregating with diplospory were identified in one maize-Tripsacum dactyloides F1 population that segregated 1∶1 for the mode of reproduction. The markers were also found to be linked in the maize RFLP map, on the distal end of the long arm of chromosome 6. These results support a simple inheritance of diplospory in Tripsacum. Manipulation of the mode of reproduction in maize-Tripsacum backcross generations, and implications for the transfer of apomixis into maize, are discussed.
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    URL: http://dx.doi.org/10.1007/BF00222943
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    Chromosomal and cell size analysis of cold tolerant maize (1992)
    Springer
    Theoretical and applied genetics 84 (1992), S. 798-802 
    Details
    ISSN: 1432-2242
    Keywords: Zea mays ; C-banding ; Cell size ; Chloroplast number ; Cold tolerance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary C-band number, guard cell length, and chloroplast number per guard cell were determined for eight maize populations. These populations consisted of maize selected for cold tolerance at the University of Nebraska as well as the original unselected populations. The genome size of these populations had previously been determined. C-band number fluctuated concertedly with the changes in genome size indicating that deletions and additions of constitutive heterochromatin occurred during selection, resulting in altered genome sizes. Guard cell size of all the cold tolerant populations was greater than the cell size of the respective nonselected populations. Chloroplast number per guard cell was also higher in all the cold tolerant populations than in their parental populations, but the increases were not statistically significant. The results indicate that changes in genome size that occurred during selection for cold tolerance are the result of changes in amounts of C-band heterochromatin and that the selection process results in an increase in cell size in the cold tolerant populations.
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    URL: http://dx.doi.org/10.1007/BF00227387
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    Immunocytochemical and electron-microscopic study of the elasmobranch nucleus sacci vasculosi (1992)
    Springer
    Cell & tissue research 270 (1992), S. 395-404 
    Details
    ISSN: 1432-0878
    Keywords: Nucleus sacci vasculosi ; Ultrastructure ; Immunocytochemistry ; Hypothalamus ; Tuberculum posterius ; Scyliorhinus caniculus, Raja undulata (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The elasmobranch nucleus sacci vasculosi was studied by means of electron microscopy (in the dogfish) and immunocytochemistry (in the dogfish and the skate) by using antibodies against tyrosine hydroxylase, alpha-melanocyte-stimulating hormone, somatostatin, serotonin, and substance P. Ultrastructural study of the dogfish nucleus sacci vasculosi shows the presence of medium-sized cells that possess numerous mitochondria but that have no dense-core vesicles in the cytoplasm or in cell processes. Fibres of the conspicuous tractus sacci vasculosi have a beaded appearance and form conventional synapses with dendrites and cell perikarya of the nucleus sacci vasculosi. The perikarya of this hypothalamic nucleus were not immunoreactive to any of the antibodies tested, and fibres immunopositive to tyrosine hydroxylase, alpha-melanocyte-stimulating hormone, somatostatin, serotonin, and substance P were scarce within this nucleus, in both the dogfish and the skate. Dorsal to the nucleus sacci vasculosi, there are numerous positive neuronal processes in addition to many small neurons that show immunoreactivity to alpha-melanocyte-stimulating hormone, somatostatin and tyrosine hydroxylase. Two types of neuron occur in this dorsal region, displaying dense-core vesicles of either 100–160 nm or 60–100 nm diameter in their cytoplasm; they were identified as peptide-containing and monoamine-containing neurons, respectively. The neuropil of this region has a significantly different ultrastructure from that of the nucleus sacci vasculosi, with many processes containing dense-core vesicles. This group of neurons, located dorsal to the nucleus sacci vasculosi and showing (a) immunoreactivity to neuropeptides or to monoamine-synthesizing enzyme, and (b) cytoplasm with dense-core vesicles, was considered not to be a part of the nucleus sacci vasculosi but rather part of the nucleus tuberculi posterioris. These results support the non-peptidergic and non-aminergic character of the nucleus sacci vasculosi.
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    Serotonin-like immunoreactivity in the ventral nerve cord of the Colorado potato beetle, Leptinotarsa decemlineata: Identification of five different neuron classes (1992)
    Springer
    Cell & tissue research 270 (1992), S. 405-413 
    Details
    ISSN: 1432-0878
    Keywords: Serotonin ; Whole-mount ; Immunocytochemistry ; Insect ventral nervous system ; Interneurons ; Efferent neurons ; Leptinotarsa decemlineata (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In an immunohistochemical study of the ventral nerve cord of L. decemlineata, five distinct neuron categories were distinguished: 1) Two paired segmental twin interneurons occur in each ganglion or neuromere; their axons distribute processes over almost the entire nerve cord and run to the cerebral ganglion complex. In contrast, other axons are distributed locally. 2) Four large frontal neurosecretory neurons occur in the suboesophageal ganglion (SOG), two of which have axons that run into the mandibular nerves to form a neurohemal plexus on the surface of cerebral nerves. 3) A pair of large caudal neurons occur in the terminal ganglion and innervate the hindgut. 4) Local miniature interneurons occur in the SOG. 5) Terminal neurons are present in the last abdominal ganglion. Segmental twin interneurons appear to be grouped into 3 ‘functional units’ spanning several ganglia. Their axons run to specific projection areas, which separate the functional units, and which mark the externally visible separation of condensed ganglion complexes. A possible role of the most caudal functional unit might be the synaptic control of caudal neurons innervating the hindgut.
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    Pericyte involvement in capillary sprouting during angiogenesis in situ (1992)
    Springer
    Cell & tissue research 270 (1992), S. 469-474 
    Details
    ISSN: 1432-0878
    Keywords: Pericytes ; Angiogenesis ; Capillaries ; Capillary sprouting ; Desmin ; Immunocytochemistry ; Rat Adenocarcinoma cells, rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary To investigate the participation of microvascular pericytes in the process of capillary sprouting, we examined whole-mount preparations of the rat mesentery by use of a double immunofluorescence approach. Angiogenesis was induced by intraperitoneal injections of either the mast cell-degranulating substance compound 48/80 or tumor cell-conditioned medium. Capillary sprouts were visualized by staining with rhodaminconjugated phalloidin and pericytes were simultaneosly stained by an antibody to the intermediate filament protein desmin. Developing pericytes were negative for the smooth-muscle isoform of α-actin, bbut were clearly reactive for desmin. Pericytes appear to be involved in the carliest stages of capillary sprouting. Pericytes were regularly found lying at and in front of the advancing tips of endothelial sprouts. At many sites pericytes were seen to bridge the gap between the leading edges of opposing endothelial sprouts, which were apparently preparing to merge, suggesting that pericytic processes may serve as guiding structures aiding outgrowth of endothelial cells.
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    Localization of basic fibroblast growth factor in a subpopulation of rat sensory neurons (1992)
    Springer
    Cell & tissue research 267 (1992), S. 125-130 
    Details
    ISSN: 1432-0878
    Keywords: Sensory ganglia ; Sympathetic ganglia ; Parasympathetic ganglia ; Basic fibroblast growth factor ; Substance P ; Somatostatin ; Bombesin ; Immunocytochemistry ; Rat (Wistar: Han)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of basic fibroblast growth factor (bFGF)-immunoreactivity (IR) was studied in rat sensory and autonomic ganglia. In postnatal and adult sympathetic superior cervical ganglia and in adult parasympathetic otic ganglia no bFGF-staining was found. Postnatal and adult neural crest-and placode-derived sensory ganglia displayed intensive bFGF-IR in a neuronal subpopulation. This subpopulation was characterized by use of consecutive sections of adult dorsal root ganglia stained with antibodies against substance P, somatostatin, bombesin, and bFGF. Basic FGF was colocalized with the somatostatin/bombesin subpopulation but not with substance P.
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    Aminergic neurons in the brain of blowflies and Drosophila: dopamine- and tyrosine hydroxylase-immunoreactive neurons and their relationship with putative histaminergic neurons (1992)
    Springer
    Cell & tissue research 267 (1992), S. 147-167 
    Details
    ISSN: 1432-0878
    Keywords: Dopamine ; Tyrosine hydroxylase ; Histamine ; Immunocytochemistry ; Insect nervous system ; Drosophila melanogaster, Phormia terraenovae, Calliphora erythrocephala (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution and morphology of neurons reacting with antisera against dopamine (DA), tyrosine hydroxylase (TH) and histamine (HA) were analyzed in the blowflies Calliphora erythrocephala and Phormia terraenovae. TH-immunoreactive (THIR) and HA-immunoreactive (HAIR) neurons were also mapped in the fruitfly Drosophila melanogaster. The antisera against DA and TH specifically labeled the same neurons in the blowflies. About 300 neurons displayed DA immunoreactivity (DAIR) and THIR in the brain and subesophageal ganglion of the blowflies. Most of these neurons were located in bilateral clusters; some were distributed as bilateral pairs, and two ventral unpaired median (VUM) neurons were seen in the subesophageal ganglion. Immunoreactive processes were found in all compartments of the mushroom bodies except the calyces, in all divisions of the central body complex, in the medulla, lobula and lobula plate of the optic lobe, and in non-glomerular neuropil of protocerebrum, tritocerebrum and the subesophageal ganglion. No DA or TH immunoreactivity was seen in the antennal lobes. In Drosophila, neurons homologous to the blowfly neurons were detected with the TH antiserum. In Phormia and Drosophila, 18 HA-immunoreactive neurons were located in the protocerebrum and 2 in the subesophageal ganglion. The HAIR neurons arborized extensively, but except for processes in the lobula, all HAIR processes were seen in non-glomerular neuropil. The deuto- and tritocerebrum was devoid of HAIR processes. Double labeling experiments demonstrated that TH and HA immunoreactivity was not colocalized in any neuron. In some regions there wasm however, substantial superposition between the two systems. The morphology of the extensively arborizing aminergic neurons described suggests that they have modulatory functions in the brain and subesophageal ganglion.
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    Differentiation of the melanotrophic cells of rat pituitary primordium in organotypic culture in defined medium (1992)
    Springer
    Cell & tissue research 267 (1992), S. 169-183 
    Details
    ISSN: 1432-0878
    Keywords: Fetal intermediate lobe ; Tissue culture ; Immunocytochemistry ; In situ hybridization ; Dopamine ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Organotypic cultures, in defined medium, of pituitary primordia obtained from 15-day-old rat fetuses were performed in order to study the in vitro differentiation of melanotrophic cells. The morphological and ultrastructural features of the transplants resembled those of the gland developing in vivo. In situ hybridization on semi-thin sections, using a 35S-labelled oligonucleotide probe, revealed pro-opiomelanocortin-mRNA-containing cells on the first day of culture in the anterior lobe and after 2–3 days in the intermediate lobe. Immunoperoxidase labelling of adjacent sections showed that the same cells reacted with antibodies against α-melanocyte-stimulating hormone (αMSH), γ3 and adrenocorticotropic hormone in both lobes. The pro-opiomelanocortin-mRNA-containing cells formed progressively conspicuous areas in the intermediate lobe, which was almost uniformly labelled after 6 days. In the anterior lobe, these cells remained scattered in small cell groups, and colloidal gold immunolabelling showed the progressive disappearance of αMSH labelling from the secretory vesicles in cells exhibiting morphological features of adult corticotrophic cells. Both the αMSH content of the explants and αMSH release into the culture medium increased with time. Treatment with the dopamine agonist bromocriptine induced a strong dose-dependent decrease in αMSH secretion, which was significant after 3 days in culture, indicating that dopamine D2 receptors are able to regulate hormonal release of melanotrophic cells at early stages. This system constitutes a suitable model for further studies of factors controlling cell differentiation and cellular interactions involved in histogenesis.
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    Light- and electron-microscopic immunocytochemistry of a molluscan insulin-related peptide in the central nervous system of Planorbarius corneus (1992)
    Springer
    Cell & tissue research 267 (1992), S. 473-481 
    Details
    ISSN: 1432-0878
    Keywords: Cerebral ganglia ; Neurohormones ; Molluscan insulin-related peptide ; Immunocytochemistry ; Tannic acid ; Planorbarius corneus (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Two groups of cerebral dorsal cells of the pulmonate snail Planorbarius corneus stain positively with antisera raised against synthetic fragments of the B- and C-chain of the molluscan pro-insulin-related prohormone, proMIP-I, of another pulmonate snail, Lymnaea stagnalis. At the light-microscopic level the somata of the dorsal cells and their axons and neurohemal axon terminals in the periphery of the paired median lip nerves are immunoreactive with both antisera. Furthermore, the canopy cells in the lateral lobes of the cerebral ganglia are positive. In addition, MIPB-immunoreactive neurons are found in most other ganglia of the central nervous system. At the ultrastructural level, pale and dark secretory granules are found in somata and axon terminals of the dorsal cells. Dark granules are about 4 times as immunoreactive to both antisera as pale granules. Release of anti-MIPB- and anti-MIPC-immunopositive contents of the secretory granules by exocytosis is apparent in material treated according to the tannic acid method. It is concluded that the dorsal and canopy cells synthesize a molluscan insulin-related peptide that is packed in the cell body into secretory granules and that is subsequently transported to the neurohemal axon terminals and released into the hemolymph by exocytosis. Thus, MIP seems to act as a neurohormone on peripheral targets. On the basis of the analogy between the dorsal cells and the MIP-producing cells in L. stagnalis, it is proposed that the dorsal cells of P. corneus are involved in the control of body growth and associated processes.
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    Culture and characterization of dental follicle cells from rat molars (1992)
    Springer
    Cell & tissue research 267 (1992), S. 483-492 
    Details
    ISSN: 1432-0878
    Keywords: Dental follicle ; Cell culture ; Fibroblasts ; Immunocytochemistry ; Ultrastructure ; Collagen ; Gel-electrophoresis ; Western blotting ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Because the dental follicle is necessary for the eruption of teeth of limited eruption, it was the objective of this study to determine if the cells of the follicle could be cultured in vitro. To achieve this, dental follicles and associated enamel organs were dissected from the first and second mandibular molars of 6–7-day-old rats (secretory stage of amelogenesis), and then cultured in a medium that promotes fibroblast growth — the predominant cell type of the dental follicle. The cultured cells grew to confluency and were kept through 3 passages before experimentation. The cultured cells were fibroblastic in shape, elongate with processes, and transmission electron microscopy revealed that they contained an abundant rough endoplasmic reticulum, but did not form desmosomes. Immunofluorescent staining for anti-vimentin showed that all the cells stained and electron-microscopic immunogold labeling indicated that the antibody was associated with intermediate filaments. As revealed by SDS-polyacrylamide gel electrophoresis and Western blotting, the cultured cells synthesized and secreted the extracellular matrix molecules fibronectin and procollagens. Subsequent immunofluorescence staining of permeabilized and non-permeabilized cells confirmed the presence of fibronectin and type I collagen both intra- and extracellularly. Thus, based on all the above characteristics, the cultured cells appeared to be fibroblasts derived from the dental follicle, although a few of the fibroblasts may be derived from undifferentiated mesenchymal cells interposed between the alveolar bone and follicle. Experiments now can be conducted to determine how these cultured cells respond directly to growth factors that alter the rates of tooth eruption.
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    URL: http://dx.doi.org/10.1007/BF00319370
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    Characterization and fine-structural localization of actin-and fibronectin-like proteins in planaria (Dugesia lugubris s.l.) (1992)
    Springer
    Cell & tissue research 267 (1992), S. 499-506 
    Details
    ISSN: 1432-0878
    Keywords: Actin-like protein ; Fibronectin-like protein ; Regeneration ; Cell migration ; Immunocytochemistry ; Dugesia lugubris (Tricladida)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Actin- and fibronectin-like proteins were characterized in the planarian, Dugesia lugubris s.l., by sodium dodecyl sulphate polyacrylamide gel electrophoresis and immunoblotting analysis using antisera to vertebrate actin and fibronectin. These antisera recognized protein bands of 42 kDa and 220 kDa, respectively. In addition, the immunohistochemical distribution of both actin- and fibronectin-like material was examined by using immuno-electron microscopy. Actin-like protein was localized in myofibrils in various differentiation stages, and in the peripheral cytoplasm and lamellipodia of cells that were migrating. The fibronectin-like component was associated with the extracellular matrix in the fibrillar structures and with the surface of the migrating cells. Our data suggest that similar cellular and molecular mechanisms are involved in cell-matrix interactions and in the morphogenesis of living organisms at different evolutionary levels.
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    URL: http://dx.doi.org/10.1007/BF00319372
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    Synaptic contacts of tyrosine hydroxylase-immunoreactive elements in the inner plexiform layer of the retina of Bufo marinus (1992)
    Springer
    Cell & tissue research 267 (1992), S. 525-534 
    Details
    ISSN: 1432-0878
    Keywords: Retina ; Dopaminergic neurons ; Synapses ; Inner plexiform layer ; Immunocytochemistry ; Electron microscopy ; Bufo marinus (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Tyrosine hydroxylase (TH) immunocytochemistry was utilized to quantify dopaminergic synapses in the inner plexiform layer of the retina of Bufo marinus. Since dopaminergic cells have bistratified dendritic arborisation in the inner plexiform layer, attention was given to the segregation of synapses between the scleral and the vitreal sublaminae. Light-microscopically, a more elaborate dendritic branching was observed in the scleral than in the vitreal sublamina. In contrast, about 55% of synapses occurred in the vitreal one fifth of the inner plexiform layer, 30% in the scleral fifth, and 15% in the intermediate laminae. Input sources and output targets showed only minor quantitative differences between sublaminae 1 and 5. TH-immunoreactive processes were found in presynaptic (62.8%) and postsynaptic (37.2%) positions. Synapses to the stained dendrites derived from bipolar (40.4%) and amacrine (59.6%) cells, whereas outputs from the TH-positive processes were directed to amacrine cells (56.8%) and to small and medium-sized dendrites (35.4%); at least some of these can be considered as ganglion cell dendrites. TH-positive profiles neither formed synapses with each other nor were presynaptic to bipolar cell terminals. Junctional appositions of the immunoreactive profiles were occasionally seen on non-stained amacrine and ganglion cell dendrites in the scleral sublamina of the inner plexiform layer and on optic axons in the optic fibre layer. Although dopaminergic cells are mainly involved in amacrine-amacrine interactions, inputs from bipolar terminals and outputs to ganglion cell dendrites were also substantial, suggestive of a role also in vertical information processing.
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    Embryonic origin and development of the corpuscles of Stannius in chum salmon (Oncorhynchus keta) (1992)
    Springer
    Cell & tissue research 268 (1992), S. 65-70 
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    ISSN: 1432-0878
    Keywords: Stanniocalcin ; Corpuscles of Stannius ; Embryology ; Immunocytochemistry ; Pronephros ; Oncorhynchus keta (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An immunocytochemical technique was used to follow the embryological origin and development of the corpuscles of Stannius (CS) in the chum salmon, Oncorhynchus keta. Stanniocalcin immunoreactive (ir-) cells can be observed as early as 13 days before hatching. The ir-CS cells appear in clusters of variable size in close association with nephric ducts. In addition, individual ir-cells also occur at this stage amoung epithelial cells of the nephric ducts. these individual cells may give rise to clusters which subsequently increase in size, the largest reaching 100 μm in diameter by the time of hatching. During this period, dispersed CS cells become evident and develop into secondary clusters in the vicinity of the primary clusters. These clusters appear to fuse to form larger clusters with a lobular structure. Transfer of the larvae (20 days after hatching) from fresh water to 50% seawater, accelerates the development of the CS tissue, suggesting an important role of the CS in seawater adaptation.
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    Distribution of neuropeptide Y-like immunoreactivity in the brain and hypophysis of the cloudy dogfish, Scyliorhinus torazame (1992)
    Springer
    Cell & tissue research 268 (1992), S. 453-461 
    Details
    ISSN: 1432-0878
    Keywords: Neuropeptide Y ; Brain, vertebrate ; Hypothalamus ; Pituitary gland, pars intermedia ; Nervus terminalis ; Immunocytochemistry ; Scyliorhinus torazame (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using a specific antiserum raised against synthetic neuropeptide Y, we examined the localization of immunoreactivity in the brain and hypophysis of the cloudy dogfish, Scyliorhinus torazame, by the peroxidase-antiperoxidase method. Immunoreactive perikarya were demonstrated in the ganglion of the nervus terminalis, the dorsocaudal portions of the pallium dorsale, the basal telencephalon, and the nucleus lateralis tuberis and the nucleus lobi lateralis in the hypothalamus. Labeled perikarya were also found in the tegmentum mesencephali, the corpus cerebelli, and the medulla oblongata. Some of the immunoreactive neurons in the hypothalamus were of the CSF-contacting type. The bulk of the labeled fibers in the nervus terminalis ran toward the basal telencephalon, showing radial projections and ramifications. Large numbers of these fibers coursed into the nucleus septi caudoventralis and the nucleus interstitialis commissurae anterioris, where they became varicose and occasionally formed fine networks or invested immunonegative perikarya. In the diencephalon, immunoreactive fibers were observed throughout the hypothalamus, e.g., in the pars neurointermedia of the hypophysis, the subependymal layer of the lobus inferior hypothalami, and in the neuropil of the posterior (mammillary) recess organ. Labeled fibers were scattered throughout the rest of the brain stem and were also seen in the granular layer of the cerebellum. These results suggest that, in the dogfish brain, neuropeptide Y or a related substance is involved in a variety of physiological processes in the brain, including the neuroendocrine control of the hypophysis.
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    URL: http://dx.doi.org/10.1007/BF00319152
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    Comparative study of the olfactory epithelium of the three-spined stickleback (Gasterosteus aculeatus) and the nine-spined stickleback (Pungitius pungitius) (1992)
    Springer
    Cell & tissue research 269 (1992), S. 267-273 
    Details
    ISSN: 1432-0878
    Keywords: Olfactory epithelium ; Comparative study ; Histochemistry ; Immunocytochemistry ; Non-specific label ; Microsmatic fish ; Three-spined stickleback, Gasterosteus aculeatus (Teleostei) ; Nine-spined stickleback, Pungitius pungitius (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The olfactory epithelium of the three-spined stickleback (Gasterosteus aculeatus) and the nine-spined stickleback (Pungitius pungitius) has been studied with a conventional histochemical and a novel immunological staining technique. In both species, the sensory epithelium is arranged in folds separated by non-sensory epithelial tissue. In the nine-spined stickleback, intrinsic folds consisting of non-sensory cells are found in the apical part of the sensory epithelium where they divide the surface of the sensory epithelium into small islets. These non-sensory cells are non-ciliated, flattened and piled on top of each other; they contain numerous electron-translucent vesicles. The intrinsic folds are absent from the sensory epithelium of the three-spined stickleback. In both species, axons of receptor cells form a layer of fibers in the sensory epithelium immediately above the basal cells. In the three-spined stickleback, thick branches of the olfactory nerve are frequently found in this layer. These branches are only occasionally observed in the sensory epithelium of the nine-spined stickleback. Thus, the three-spined stickleback and the nine-spined stickleback show considerable differences in the organization of the sensory regions of the olfactory epithelium.
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    Pituitary adenylate cyclase-activating polypeptide (PACAP): occurrence in rodent pancreas and effects on insulin and glucagon secretion in the mouse (1992)
    Springer
    Cell & tissue research 269 (1992), S. 275-279 
    Details
    ISSN: 1432-0878
    Keywords: Pituitary adenylate cyclase-activating peptide (PACAP) ; Immunocytochemistry ; Pancreas, endocrine, exocrine ; Insulin secretion ; Glucagon secretion ; Mouse (NMRI) ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide that occurs in several tissues, e.g., in the gut. We have studied PACAP-like immunoreactivity in the pancreas of rat and mouse, and the effects of PACAP-38 on basal and stimulated insulin and glucagon secretion in the mouse. Immunofluorescence staining demonstrated the presence of PACAP-like immunoreactivity in nerve fibers in both the rat and mouse pancreas. The nerve fibers were seen in the exocrine pancreas and surrounding the islets. Occasionally, the nerve fibers occurred within the islets. Most PACAP-positive nerve fibers innervated the intrapancreatic ganglia, although no nerve cell bodies contained PACAP-like immunoreactivity. In-vivo experiments in mice revealed that basal plasma glucagon levels were increased by PACAP-39 injected intravenously at dose levels exceeding 1.8 nmol/kg. Furthermore, PACAP-38 (7 nmol/kg) potentiated the plasma glucagon response to the cholinergic agonist carbachol (0.16 μmol/kg). This potentiation was reduced to simple addition by pretreatment with a combined α- and β-adrenergic blockade by phentolamine (35 μmol/kg) and propranolol (8.5 μmol/kg). Moreover, PACAP-38 inhibited a carbachol-induced increase in the level of plasma insulin in the absence but not in the presence of adrenergic blockade. PACAP-38 increased basal plasma insulin levels and increased basal plasma glucose levels 6 min and 10 min, respectively, after injection of the peptide. We conclude that PACAP-like immunoreactivity exists in nerve fibers innervating the mouse and rat pancreas, particularly the intrapancreatic ganglia, and that PACAP-38 augments both basal and carbachol-stimulated glucagon secretion in the mouse.
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    Localization of calcitonin gene-related peptide and islet amyloid polypeptide in the rat and mouse pancreas (1992)
    Springer
    Cell & tissue research 269 (1992), S. 315-322 
    Details
    ISSN: 1432-0878
    Keywords: Calcitonin gene-related peptide ; Islet amyloid polypeptide ; Immunocytochemistry ; Pancreas endocrine, exocrine ; Rat (Sprague-Dawley) ; Mouse (NMRI)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary It was previously demonstrated that the two chemically related peptides calcitonin gene-related peptide (CGRP) and islet amyloid polypeptide (IAPP) both occur in the pancreas. We have now examined the cellular localization of CGRP and IAPP in the rat and the mouse pancreas. We found, in both the rat and the mouse pancreas, CGRP-immunoreactive nerve fibers throughout the parenchyma, including the islets, with particular association with blood vessels. CGRP-immunoreactive nerve fibers were regularly seen within the islets. In contrast, no IAPP-immunoreactive nerve fibers were demonstrated in this location. Furthermore, in rat islets, CGRP immunoreactivity was demonstrated in peripherally located cells, constituting a major subpopulation of the somatostatin cells. Such cells were lacking in the mouse islets. IAPP-like immunoreactivity was demonstrated in rat and mouse islet insulin cells, and, in the rat, also in a few non-insulin cells in the islet periphery. These cells seemed to be identical with somatostatin/CGRP-immunoreactive elements. In summary, the study shows (1) that CGRP, but not IAPP, is a pancreati neuropeptide both in the mouse and the rat; (2) that a subpopulation of rat somatostatin cells contain CGRP; (3) that mouse islet endocrine cells do not contain CGRP; (4) that insulin cells in both the rat and the mouse contain IAPP; and (5) that in the rat, a non-insulin cell population apparently composed of somatostatin cells stores immunoreactive IAPP. We conclude that CGRP is a pancreatic neuropeptide and IAPP is an islet endocrine peptide in both the rat and the mouse, whereas CGRP is an islet endocrine peptide in the rat.
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    Immunocytochemistry of somatotrophs, gonadotrophs, prolactin and adrenocorticotropin cells in larval sea bream (Sparus auratus) pituitaries (1992)
    Springer
    Cell & tissue research 269 (1992), S. 341-346 
    Details
    ISSN: 1432-0878
    Keywords: Growth hormone ; Prolactin ; Gonadotropin ; Adrenocorticotropin ; Immunocytochemistry ; Pituitary gland ; Sparus auratus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The chronological appearance of endocrine cells in the pituitary of sea-bream (Sparus auratus) larvae was studied using antisera against salmon prolactin, trout growth hormone, salmon gonadotropin and N-terminal human adrenocorticotropin. The larval pituitary (1–12 days after hatching) was oval in shape and was composed of a dense mass of cells with few neurohypophysial fibres. By 60 days after hatching it began to resemble the adult and was divisible into a distinct rostral pars distalis containing prolactin and adrenocorticotropin cells; a proximal pars distalis containing somatotrophs and gonadotrophs and a pars intermedia. Cells immunoreactive with antisera against growth hormone were observed immediately after hatching (2 days post-fertilization). Weakly staining prolactin cells were observed 2 days later in the region corresponding to the rostral pars distalis. Cells immunoreactive with anti-gonadotropin and anti-adrenocorticotropin sera were observed in the pituitary 6 and 8 days after hatching, respectively. All the cell-types studied were immunoreactive from the time they were first identified until the final samples 90 days after hatching.
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    Immuno-electron-microscopic localization of laminin and collagen type IV in normal and denervated tooth pulp of the cat (1992)
    Springer
    Cell & tissue research 270 (1992), S. 157-164 
    Details
    ISSN: 1432-0878
    Keywords: Dental pulp ; Laminin ; Collagen IV ; Odontoblast ; Nerve regeneration ; Immunocytochemistry ; Cat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of laminin-like immunoreactivity in adult normal and denervated cat mandibular tooth pulps was studied by the use of fluorescence microscopy and pre-embedding immunogold electron microscopy. Immunoreactivity to collagen IV was also assessed in order to distinguish basement membranes. In normal pulps, light-microscope laminin-like immunoreactivity was strong along blood vessels and Schwann cell sheaths, and a faint immunoreactivity was seen also in the odontoblast layer. Electron microscopy confirmed the laminin-like immunoreactivity of endothelial and Schwann cell basement membranes at all pulpal levels. In the odontoblast layer and the predentine, nerve-like structures lacking basement membranes but possessing strong membrane laminin-like immunoreactivity were encountered. In addition, a clear-cut laminin-like immunoreactivity of plasma membranes of the somata and processes of odontoblasts was seen. Observations on denervated pulps as well as pulps in which nerve regeneration had taken place did not reveal any changes in the pattern of laminin-immunoreactivity in basement membranes or odontoblasts. Distribution of collagen IV-like immunoreactivity was very similar to laminin-like immunoreactivity in basement membranes of blood vessels and Schwann cells, and appeared unaffected by denervation. The odontoblasts and nerve-like profiles in the odontoblast layer were devoid of collagen IV-like immunoreactivity. We propose that odontoblast-associated laminin could be of significance as guidance for regenerating terminal pulpal nerve fibers to appropriate targets.
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    Presence of an oxytocin-like peptide in the hypothalamus and neurohypophysis of a turtle (Mauremys caspica) and a snake (Natrix maura) (1995)
    Springer
    Cell & tissue research 279 (1995), S. 75-84 
    Details
    ISSN: 1432-0878
    Keywords: Oxytocin ; Neurophysin ; Vasotocin ; Mesotocin ; Suprachiasmatic nucleus ; Medial nucleus of the infundibular recess ; Immunocytochemistry ; Natrix maura (Serpentes) ; Mauremys caspica (Chelonia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The probable presence of oxytocin in the hypothalamo-hypophysial system of two reptilian species, the snake Natrix maura and the turtle Mauremys caspica, was re-investigated. A high-pressure liquid chromatographic analysis of the turtle neural lobe revealed the existence of vasotocin, mesotocin, and a third compound co-eluting with oxytocin. Brains from both species were fixed by vascular perfusion with Bouin's fluid. Adjacent paraffin sections were immunostained using antisera against the following substances: (1) bovine oxytocin-neurophysin; (2) a mixture of bovine oxytocin-neurophysin and vasopressin-neurophysin; (3) dogfish neurophysins; (4) oxytocin; (5) arginine-vasotocin; (6) mesotocin; (7) somatostatin. Immunoreactivity against oxytocin was found in parvocellular neurons of the snake suprachiasmatic nucleus and cerebrospinal-fluid contacting neurons of the medial nucleus of the infundibular recess of both species, the latter immunoreactivity being much more conspicuous in the turtle. Numerous fibers containing immunoreactive oxytocin extended between the medial nucleus of the infundibular recess, and the internal region of the medium eminence and the neural lobe. The oxytocin-immunoreactivity in all locations was completely abolished by preabsorption of the anti-oxytocin serum with three different oxytocin preparations. None of the neurons of the suprachiasmatic and medial nucleus of the infundibular recess, including the oxytocin-immunoreactive elements, reacted with either the antineurophysin sera used, or the anti-vasotocin or anti-mesotocin antibodies. The possible existence of a reptilian oxytocin-neurophysin is discussed. The alternative that, in the reptilian hypothalamus, neurons synthesize a compound closely related to, but different from oxytocin is also considered.
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    The biased intracellular accumulation of the β-subunit of salmon gonadotropin (GTH II) in the pituitary of rainbow trout Oncorhynchus mykiss, during gametogenesis (1995)
    Springer
    Cell & tissue research 279 (1995), S. 93-99 
    Details
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Gonadotropin ; Subunits ; Gonadotropes ; Immunocytochemistry ; Immunoblotting ; Oncorhynchus mykiss (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Salmon gonadotropin (GTH II) is a heterodimeric glycoprotein hormone (α and IIβ subunits), serving as a maturational GTH, and is produced in a specific gonadotropic cell-type (GTH II-cells) containing small granules and large globules. In trout GTH II-cells, double immunolabeling for the α- and IIβ-subunits shows that colocalization of the α- and IIβ-immunolabeling is confined to the small granules, indicating storage of functional GTH II. On the other hand, α-immunolabeling is absent in the large globules, even though IIβ labeling is abundant throughout the period of seasonal gametogenesis. The α-specific antiserum recognizes the intact α-subunit as well as the reduced and deglycosylated α-subunits by immunoblotting. These results indicate that an accumulation of the IIβ-subunit is specifically generated in the large globules of these cells. In fact, with sexual maturity, the quantity of IIβ-subunits becomes elevated in the trout pituitary due to a marked increase in GTH II-cells containing many large globules. However, the derivation and function of the large globules and the fate of their contained IIβ-subunits remains unknown.
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    Ultrastructural evidence for multiple mucous domains in frog olfactory epithelium (1992)
    Springer
    Cell & tissue research 270 (1992), S. 47-56 
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    ISSN: 1432-0878
    Keywords: Cilia ; Microvilli ; Secretory granules ; Mucus ; Freeze-substitution ; Immunocytochemistry ; Rana pipiens (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary This study showed that the olfactory mucus is a highly structured extracellular matrix. Several olfactory epithelial glycoconjugates in the frog Rana pipiens were localized ultrastructurally using rapid-freeze, freeze-substitution and post-embedding (Lowicryl K11M) immunocytochemistry. Two of these conjugates were obtained from membrane preparations of olfactory cilia, the glycoproteins gp95 and olfactomedin. The other conjugates have a carbohydrate group which in the olfactory bulb appears to be mostly on neural cell-adhesion molecules (N-CAMs); in the olfactory epithelium this carbohydrate is present on more molecules. Localization of the latter conjugates was determined with monoclonal antibodies 9-OE and 5-OE. Ultrastructurally all antigens localized in secretory granules of apical regions of frog olfactory supporting cells and in the mucus overlying the epithelial surface, where they all had different, but partly overlapping, distributions. Monoclonal antibody 18.1, to gp95, labeled the mucus throughout, whereas poly- and monoclonal anti-olfactomedin labeled a deep mucous layer surrounding dendritic endings, proximal parts of cilia, and supporting cell microvilli. Labeling was absent in the superficial mucous layer, which contained the distal parts of the olfactory cilia. Monoclonal antibody 9-OE labeled rather distinct areas of mucus. These areas sometimes surrounded dendritic endings and olfactory cilia. Monoclonal antibody 5-OE labeled membranes of dendritic endings and cilia, and their glycocalyces, and also dendritic membranes.
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    Regeneration and post-metamorphic development of the central nervous system in the protochordate Ciona intestinalis: a study with monoclonal antibodies (1995)
    Springer
    Cell & tissue research 279 (1995), S. 421-432 
    Details
    ISSN: 1432-0878
    Keywords: Trans-differentiation ; Proliferation ; Bromodeoxyuridine ; Immunocytochemistry ; Regeneration ; Ciona intestinalis (Tunicata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract In this study, we use three monoclonal antibodies that recognise antigens present in the central nervous system of the ascidian Ciona intestinalis to study regeneration and post-metamorphic development of the neural ganglion. We have also used bromodeoxyuridine labelling to study generation of the neuronal precursor cells. The first antibody, CiN 1, recognises all neurones in the ganglion, whereas the second, CiN 2, recognises only a subpopulation of the large cortical neurones. Western blotting studies show that CiN 2 recognises two membrane-bound glycoproteins of apparent Mr 129 and 100 kDa. CiN 1 is not reactive on Western blots. Immunocytochemical studies with these antibodies show that CiN 1-immunoreactive neurone-like cells are present at the site of regeneration as early as 5–7 days post-ablation, a sub-population of CiN 2-immunoreactive cells being detected by 9–12 days post-ablation. The third antibody, ECM 1, stains extracellular matrix components and recognises two diffuse bands on Western blots of whole-body and ganglion homogenates. The temporal and spatial pattern of appearance of CiN 1 and CiN 2 immunoreactivity both during post-metamorphic development and in regeneration occurs in the same sequence in both processes. Studies with bromodeoxyuridine show labelled nuclei in some neurones in the regenerating ganglion. Plausibly these originate from the dorsal strand, an epithelial tube that reforms by cell proliferation during the initial phases of regeneration. A second population of cells, the large cortical neurones, do not incorporate bromodeoxyuridine and thus must have been born prior to the onset of regeneration. This latter finding indicates a mechanism involving trans-differentiation of other cell types or differentiation of long-lived totipotent stem cells.
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    Presence of embryonic myosin in normal postural muscles of the adult rat (1995)
    Springer
    Cell & tissue research 280 (1995), S. 541-548 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Musle ; striated ; skeletal ; Regeneration ; Myosin ; Immunocytochemistry ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Indirect immunofluorescence was used to localize embryonic myosin heavy chains in soleus, adductor longus, tibialis anterior, plantaris, and extensor digitorum longus muscles of 6-month-old rats. A monoclonal antibody (2B6), specifically recognizing rat embryonic myosin, was applied to unfixed, transverse, frozen sections. The number of embryonic myosin-positive (EMP) extrafusal fibers was expressed as a percentage of the total number of fibers. EMP extrafusal fibers were only seen in the soleus and adductor longus muscles, both postural muscles. Approximately 1% of the soleus muscle fibers appeared positively stained for embryonic myosin. The majority of such fibers had a small diameter (〈500 μ2), appeared intensely fluorescent, and typically contained central nuclei. Re-expression of embryonic myosin due to spontaneous fiber denervation is not a likely factor in this study, since alpha-bungarotoxin and N-CAM localization were restricted to the motor end-plate region of EMP fibers. Since embryonic myosin was shown to disappear in all normal-sized myofibers by 2 to 3 months of age, the results suggest that the EMP extrafusal fibers seen in postural muscles of 6 to 12-month-old animals are regenerating myofibers. We speculate that a small number of muscle fibers may be regenerating in normal, adult postural muscles, in response to fiber damage possibly caused by excessive recruitment or overloading.
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    Development of melanin-concentrating hormone-immunoreactive elements in the brain of gilthead seabream (Sparus auratus) (1995)
    Springer
    Cell & tissue research 282 (1995), S. 523-526 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Melanin-concentrating hormone ; Immunocytochemistry ; Development ; ontogenetic ; Sparus auratus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The development of the hypothalamic melanin-concentrating hormone (MCH) system of the teleost Sparus auratus has been studied by immunocytochemistry using an anti-salmon MCH serum. Immunoreactive perikarya and fibers are found in embryos, larvae, and juvenile specimens. In juveniles, most labeled neurons are present in the nucleus lateralis tuberis; some are dispersed in the nucleus recessus lateralis and nucleus periventricularis posterior. From the nucleus lateralis tuberis, MCH neurons project a conspicuous tract of fibers to the ventral hypothalamus; this penetrates the pituitary stalk and reaches the neurohypophysis. Most fibers end close to the cells of the pars intermedia, and some reach the adenohypophysial rostral pars distalis. Immunoreactive fibers can also be seen in extrahypophysial localizations, such as the preoptic region and the nucleus sacci vasculosi. In embryos, MCH-immunoreactive neurons first appear at 36 h post-fertilization in the ventrolateral margin of the developing hypothalamus. In larvae, at 4 days post-hatching, perikarya can be observed in the ventrolateral border of the hypothalamus and in the mid-hypothalamus, near the ventricle. At 26 days post-hatching, MCH perikarya are restricted to the nucleus lateralis tuberis. The neurohypophysis possesses MCH-immunoreactive fibers from the second day post-hatching. The results indicate that MCH plays a role in larval development with respect to skin melanophores and cells that secrete melanocyte-stimulating hormone.
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    Ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the dogfish (1995)
    Springer
    Cell & tissue research 282 (1995), S. 33-40 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Neuropeptide Y ; Gastroenteropancreatic (GEP) endocrine system ; Development ; ontogenetic ; Vitellointestinal duct ; Pancreas ; exocrine ; Pancreas ; endocrine ; Immunocytochemistry ; Scyliorhinus torazame (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. This immunocytochemical study was carried out to elucidate the ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the cloudy dogfish, Scyliorhinus torazame. Immunostained cells first appeared in the pancreas of the embryo at the 15-mm stage, and were also detected in the vitellointestinal duct of the yolk stalk at the 20-mm stage. These cells were polymorphic, with occasional processes that were sometimes directed toward the vascular wall or into the cavity of the vitellointestinal duct. At the 34-mm stage, immunostained cells could also be found in the proximal part of the spiral intestine and, by the 74-mm stage, immunopositive cells were present in the gastric mucosa. In the gut and pancreas, the cells gradually increased in number with development, whereas in the vitellointestinal duct and internal yolk sac, they decreased and seemed to disappear following hatching. Thus, in juveniles, the distribution of the neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system had attained that of adults. Electron-microscopic immunocytochemistry demonstrated that, in the labeled cells of the vitellointestinal duct, the neuropeptide Y-like antigen was located in cytoplasmic granules, as in the cells of the gut and pancreas.
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    URL: http://dx.doi.org/10.1007/s004410050456
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    Rat intestinal galactoside-binding lectin L-36 functions as a structural protein in the superficial squamous cells of the esophageal epithelium (1995)
    Springer
    Cell & tissue research 281 (1995), S. 77-83 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Esophagus ; Epithelial cells ; Intestinal lectin ; L-36 ; RI-H fragment ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Using an affinity purified antibody raised against the RI-H fragment of rat intestinal lectin L-36, the latter protein has been identified within the esophageal epithelium by means of ultracryotomy followed by immunogold labeling. The epithelium consists of 4 morphologically distinct cell-types, namely, the basal, spiny, granular and squamous cells, and each of these exhibits a different immunolabeling pattern. The basal cells form a layer on the basal lamina, and in these a diffuse cytoplasmic staining is observed. This basal cell layer is overlaid by spiny cells that extend many cell processes into wide intercellular spaces. In these cells, immunogold particles are found only on small granular inclusions consisting of an electron-lucent homogeneous substance. The granular cells form a third layer over the spiny cells, and are characterized by a number of large granular inclusions with an electron-dense core rimmed by a less electron-dense substance. Immunogold labeling is found on these granules, both on the core and peripheral region. Squamous cell-types constitute the most superficial layer of the epithelium. They are without granular inclusions, and immunogold labeling is confined to the cytoplasmic surface of the thickened plasma membrane. These findings suggest that L-36 is produced in the basal cells as free cytosolic protein, then becomes progressively aggregated into the granular inclusions of the spiny and granular cells, and is eventually transferred onto the cytoplasmic surface of the squamous cell plasma membrane where it may interact with complementary glycoconjugate(s) located at this site. The membrane lining substance thus formed may play a role in stabilizing the squamous cell membranes, thereby maintaining the structural integrity of the epithelium against mechanical stress coming from the esophageal lumen.
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    URL: http://dx.doi.org/10.1007/BF00307960
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    Localization of dopamine in the freshwater hydrozoanHydra attenuata (1992)
    Springer
    Cell & tissue research 270 (1992), S. 601-607 
    Details
    ISSN: 1432-0878
    Keywords: Dopamine ; HPLC ; Immunocytochemistry ; Hydra attenuata (Cnidaria)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary High performance liquid chromatography (HPLC), with electrochemical detection, is an analytical method sensitive enough to permit quantification of dopamine, dihydroxyphenylalanine (DOPA) and 5-S-cysteinyl DOPA in single or hemisected specimens ofHydra attenuata. Dopamine and 5-S-cysteinylDOPA appear to be the quantitatively predominant catechol compounds inH. attenuata, whereas DOPA is present in minor amounts. The presence of DOPA and 5-S-cysteinylDOPA, and the quantitative correlation between dopamine and these compounds in many specimens, suggests that dopamine inH. attenuata, as in higher animals, is formed through decarboxylation of DOPA. Contrary to the dopaminergic nerves in higher animals, DOPA inHydra seems to be oxidized and 5-S-cysteinyl DOPA is formed as a by-product. The oxidation of DOPA indicates that the hydroxylation of tyrosine into DOPA in the tissues ofH. attenuata is mediated by a tyrosinase rather than a tyrosine hydroxylase. Immunocytochemical methods demonstrate a highly variable distribution of dopamine in the tissues of different specimens ofH. attenuata. Dopamine immunoreactivity is confined to ectodermal tissue and can be found in several different cell types including nerve cells, battery cells, nematocytes, epithelial cells and interstitial undifferentiated cells. The large amounts of dopamine found in some specimens ofH. attenuata indicate some biological function, although its sporadic occurrence in neurites makes it less plausible as a generally utilized neurotransmitter in this animal.
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    URL: http://dx.doi.org/10.1007/BF00645064
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    Immunocytochemical identification of some regulatory peptides (gastrin, gastrin-releasing peptide, neurotensin and vasoactive intestinal polypeptide) in the Harderian gland of the green frog,Rana esculenta (1992)
    Springer
    Cell & tissue research 270 (1992), S. 609-611 
    Details
    ISSN: 1432-0878
    Keywords: Gastrin ; Gastrin-releasing peptide ; Neurotensin ; Vasoactive intestinal peptide ; Immunocytochemistry ; Harderian gland ; Rana esculenta (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence and distribution of gastrin-, gastrin-releasing peptide-, neurotensin-and vasoactive intestinal polypeptide-like immunoreactivity in the Harderian gland ofRana esculenta were studied at different times of the annual cycle. Gastrin-releasing peptide, neurotensin and vasoactive intestinal polypeptide-like substances were found either in the glandular cells, or in the nerve fibers surrounding the glandular acini. Gastrin-like immunoreactivity was confined to the glandular cells. The immunoreactivity varied during the annual cycle, with the greatest concentration being noted during the recovery phase of glandular secretory activity.
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    Distribution and development of the serotonin-and RFamide-like immunoreactive neurons in the arrowworm, Paraspadella gotoi (Chaetognatha) (1992)
    Springer
    Cell & tissue research 267 (1992), S. 215-222 
    Details
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Serotonin ; RFamide ; Development ; Paraspadella gotoi (Chaetognatha)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution and development of serotonin-and RFamide-like immunoreactivities in the nervous system of Chaetognatha, Paraspadella gotoi, were examined in whole-mount preparations. In adults, a single serotonin-like immunoreactive (5HTLI) neuron and numerous RFamide-like immunoreactive (RFaLI) neurons were found in the central nervous system. Based on the structure of the fins, hooks, and eyes, seven postembryonic developmental stages were recognized. The most obvious features of the stages are: stage 1, newly hatched young; stage 2, elongation of a continuous lateral tail fin; stage 3, separation of the lateral and tail fins; stage 4, appearance of hooks; stage 5, pigmentation of eyes, stage 6, attachment by tail adhesive fins; stage 7, prey capture. Stage 1 did not show any immunoreactivity. The 5HTLI neuron first appeared at stage 4 and its axonal pathway became similar to the adult at stage 6. On the other hand, the RFaLI neurons appeared at stage 3 in the ventral ganglion. Some of their somata disappeared at stage 5 and the neuronal architecture resembled the adult at stage 7 although the RFaLI neurons in the cerebral ganglion were complete at the juvenile stage.
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    URL: http://dx.doi.org/10.1007/BF00302958
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    Sulfhydryl oxidase immunoreactivity in seminiferous tubules of infertile men (1992)
    Springer
    Cell & tissue research 267 (1992), S. 209-214 
    Details
    ISSN: 1432-0878
    Keywords: Testis ; Sulfhydryl oxidase ; Hypospermatogenesis ; Sertoli cell integrity ; Immunocytochemistry ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Sulfhydryl oxidase (SOx) immunoreactivity was investigated in the seminiferous epithelium of human biopsy material from the testes of 33 adult men with disturbed fertility. SOx immunoreactivity was expressed in normal seminiferous epithelium in type-A spermatogonia (27±4% of all spermatogonia) (n=4), in spermatocytes and round spermatids. Mature spermatozoa as well as Sertoli cells were unlabelled. within the interstitium, Leydig cells were immunopositive. In biopsies of oligozoospermic men showing hypospermatogenesis (n=24), an increase in labelled spermatogonia up to more than 90% was observed in biopsies, where seminiferous epithelia revealed only spermatogonia and Sertoli cells. Within the group of oligozoospermic patients there was a significant increase of labelled spermatogonia from 43±13% (〉20 mill/ejaculate) (n=7) to 55±16% ( 20 and 〉20 mill/ejaculate) (n=6) to 68±8% (〈5 mill/ejaculate) (n=11) and a significant (P=0.01) decrease of score count from 7.0±2.7 to 2.0±1.8. In this group the increase of labelled spermatogonia was correlated with sperm concentrations in the ajaculate (correlation coefficient: r=-0.6). In biopsies of azoospermic patients showing maturation arrest at the level of spermatocytes or spermatids (n=5) the percentage of labelled spermatogonia was within the range of 24% to 59%. Immunoreactivity in Sertoli cells was only found in single degenerating cells and in tubules showing Sertoli Cell Only Syndrome (SCO) without lumen formation. Sertoli cells within immature seminiferous cords were immunonegative, indicating that Sertoli cell SOx immunoreactivity is rather a sign of physiological alterations in degenerating cells than dependent on the stage of differentiation. Leydig cells did not show changes of immunoreactivity in any biopsy. It is concluded that SOx expression in spermatogonia may serve as a marker for spermatogenic efficiency.
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    The alpha-subunit of glycoprotein hormones exists in the prolactin secretory granules of the bullforg (Rana catesbeiana) pituitary gland (1992)
    Springer
    Cell & tissue research 267 (1992), S. 223-231 
    Details
    ISSN: 1432-0878
    Keywords: α-Subunit ; Pituitary glycoprotein hormone ; PRL cell ; Pars distalis ; Colocalization ; Immunocytochemistry ; Bullfrog, Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Our recent finding that the number of immunoreactive α-subunit cells was invariably greater than the total number of immunoreactive gonadotropin (GTH) and thyrotropin (TSH) cells in the bullfrog (Rana catesbeiana) pituitary gland raises the possibility that the α-subunit also exists in pituitary cells other than GTH and TSH cells. The present study demonstrates that there are a considerable number of immunoreactive prolactin (PRL) cells that are also stained with antibody against the α-subunit when adjacent sections are immunocytochemically examined. Neither immunoreactive growth hormone nor adrenocorticotropin cells are stained with the antibody against the α-subunit. The specificity of the antibody against the α-subunit and of that against PRL was demonstrated by preabsorption test, non-competitive binding test, and immunoblot analysis. Double-immunolabeling with gold particles of different sizes for the α-subunit and PRL revealed that most of the immunolabeled PRL-secretory granules are also labeled with the α-subunit antibody. The gold particles indicating the presence of the α-subunit were mostly found in the peripheral zone of the secretory granules.
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    URL: http://dx.doi.org/10.1007/BF00302959
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    The VD1/RPD2 neuronal system in the central nervous system of the pond snail Lymnaea stagnalis studied by in situ hybridization and immunocytochemistry (1992)
    Springer
    Cell & tissue research 267 (1992), S. 551-559 
    Details
    ISSN: 1432-0878
    Keywords: Nervous system, central ; VD1/RPD2 system ; Neuropeptide immunocytochemistry ; Hybridization, in situ ; Immunocytochemistry ; Lymnaea stagnalis (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary VD1 and RPD2 are two giant neuropeptidergic neurons in the central nervous system (CNS) of the pond snail Lymnaea stagnalis. We wished to determine whether other central neurons in the CNS of L. stagnalis express the VD1/RPD2 gene. To this end, in situ hybridization with the cDNA probe of the VD1/RPD2 gene and immunocytochemistry with antisera specific to VD1 and RPD2 (the α1-antiserum, Mab4H5 and ALMA 6) and to R15 (the α1 and 16-mer antisera) were performed on alternate tissue sections. A VD1/RPD2 neuronal system comprising three classes of neurons (A1–A3) was found. All neurons of the system express the gene. Division into classes is based on immunocytochemical characteristics. Class A1 neurons (VD1 and RPD2) immunoreact with the α1-antiserum, Mab4H5 and ALMA 6. Class A2 neurons (1–5 small and 1–5 medium sized neurons in the visceral and right parietal ganglion, and two clusters of small neurons and 5 medium-sized neurons in the cerebral ganglia) immunoreact with the α1-antiserum and Mab4H5, but not with ALMA 6. Class A3 neurons (3–4 medium-sized neurons and a cluster of 4–5 small neurons located in the pedal ganglion) immunoreact with the α1-antiserum only. All neurons of the system are immunonegative to the R15 antisera. The observations suggest that the neurons of the VD1/RPD2 system produce different sets of neuropeptides. A group of approximately 15 neurons (class B), scattered in the ganglia, immunostained with one or more of the antisera, but did not react with the cDNA probe in in situ hybridization.
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    Octopamine-immunoreactive neurons in the central nervous system of the cricket, Gryllus bimaculatus (1992)
    Springer
    Cell & tissue research 268 (1992), S. 287-304 
    Details
    ISSN: 1432-0878
    Keywords: Nervous system, central ; Ganglia, invertebrate ; Octopamine ; Immunocytochemistry ; DUM neurone ; Neurosecretion ; Gryllus bimaculatus (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of octopamine-immunoreactive neurons is described using whole-mount preparations of all central ganglia of the cricket, Gryllus bimaculatus. Up to 160 octopamine-immunoreactive somata were mapped per animal. Medial unpaired octopamine-immunoreactive neurons occur in all but the cerebral ganglia and show segment-specific differences in number. The position and form of these cells are in accordance with well-known, segmentally-organized clusters of large dorsal and ventral unpaired medial neurons demonstrated by other techniques. In addition, bilaterally arranged groups of immunoreactive somata have been labelled in the cerebral, suboesophageal and terminal ganglia. A detailed histological description of octopamine-immunoreactive elements in the prothoracic ganglion is given. Octopamine-immunoreactive somata and axons correspond to the different dorsal unpaired medial cell types identified by intracellular single-cell staining. In the prothoracic ganglion, all efferent neurons whose primary neurites are found in the fibre bundle of dorsal unpaired cells are immunoreactive. Intersegmental octopamine-immunoreactive neurons are also present. Collaterals originating from dorsal intersegmental fibres terminate in different neuropils and fibre tracts. Fine varicose fibres have been located in several fibre tracts, motor and sensory neuropils. Peripheral varicose octopamine-immunoreactive fibres found on several nerves are discussed in terms of possible neurohemal releasing sites for octopamine.
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    URL: http://dx.doi.org/10.1007/BF00318798
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    Species variability in the expression of met- and leu-enkephalin-like immunoreactivity in mammalian Merkel cell dense-core granules (1992)
    Springer
    Cell & tissue research 269 (1992), S. 347-351 
    Details
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Enkephalin ; Skin ; Touch dome ; Merkel cells ; Dense-core granules ; Mammalian species
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunogold staining failed to show met-enkephalin immunoreactivity in the Merkel cell dense-core granules of rats when examined by electron microscopy, but showed gold particle staining in the Merkel cell dense-core granules of mice and nude mice. Merkel cells of hamster, guinea pig, rabbit, cat and dog were also examined using a similar method, and different antisera dilutions. Immunogold particles were consistently found in the dense-core granules of mice and nude mice at all antisera dilutions, but not in the other species, except in the dog, where a very low labelling response was encountered. Merkel cells from skin touch domes or sinus hair follicles, did not exhibit any difference in peptide expression as far as met-enkephalin immunoreactivity was concerned. In addition, all species studied, including mice and nude mice, did not show leu-enkephalin immunoreactivity in their Merkel cell dense-core granules. It is concluded that species variability in peptide expression occurs in the Merkel cell dense-core granules, and may be closely related to the different methodologies used.
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    URL: http://dx.doi.org/10.1007/BF00319627
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    The distribution of pheromone-biosynthesis-activating neuropeptide (PBAN) immunoreactivity in the central nervous system of the corn earworm moth, Helicoverpa zea (1992)
    Springer
    Cell & tissue research 270 (1992), S. 229-240 
    Details
    ISSN: 1432-0878
    Keywords: Pheromone-biosynthesis-activating neuropeptide ; Immunocytochemistry ; Helicoverpa zea (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Production of sex pheromone in several species of moths has been shown to be under the control of a neuropeptide termed pheromone-biosynthesis-activating neuropeptide (PBAN). We have produced an antiserum to PBAN from Helicoverpa zea (Lepidoptera: Noctuidae) and used it to investigate the distribution of immunoreactive peptide in the brain-suboesophageal ganglion complex and its associated neurohemal structures, and the segmental ganglia of the ventral nerve cord. Immunocytochemical methods reveal three clusters of cells along the ventral midline in the suboesophageal ganglion (SOG), one cluster each in the presumptive mandibular (4 cells), maxillary (12–14 cells), and labial neuromeres (4 cells). The proximal neurites of these cells are similar in their dorsal and lateral patterns of projection, indicating a serial homology among the three clusters. Members of the mandibular and maxillary clusters have axons projecting into the maxillary nerve, while two additional pairs of axons from the maxillary cluster project into the ventral nerve cord. Members of the labial cluster project to the retrocerebral complex (corpora cardiaca and cephalic aorta) via the nervus corpus cardiaci III (NCC III). The axons projecting into the ventral nerve cord appear to arborize principally in the dorsolateral region of each segmental ganglion; the terminal abdominal ganglion is distinct in containing an additional ventromedial arborization in the posterior third of the ganglion. Quantification of the extractable immunoreactive peptide in the retrocerebral complex by ELISA indicates that PBAN is gradually depleted during the scotophase, then restored to maximal levels in the photophase. Taken together, our findings provide anatomical evidence for both neurohormonal release of PBAN as well as axonal transport via the ventral nerve cord to release sites within the segmental ganglia.
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    URL: http://dx.doi.org/10.1007/BF00328008
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    Immunocytochemical localization of collagen types I, III, IV, and fibronectin in the human dermis (1992)
    Springer
    Cell & tissue research 268 (1992), S. 505-511 
    Details
    ISSN: 1432-0878
    Keywords: Dermis ; Collagen fibers ; Extracellular matrix ; Fibronectin ; Immunocytochemistry ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of collagen types I, III, IV, and of fibronectin has been studied in the human dermis by light and electron-microscopic immunocytochemistry, using affinity purified primary antibodies and tetramethylrhodamine isothiocyanate-conjugated secondary antibodies. Type I collagen was present in all collagen fibers of both papillary and reticular dermis, but collagen fibrils, which could be resolved as discrete entities, were labeled with different intensity. Type III collagen codistributed with type I in the collagen fibers, besides being concentrated around blood vessels and skin appendages. Coexistence of type I and type III collagens in the collagen fibrils of the whole dermis was confirmed by ultrastructural double-labelling experiments using colloidal immunogold as a probe. Type IV collagen was detected in all basement membranes. Fibronectin was distributed in patches among collagen fibers and was associated with all basement membranes, while a weaker positive reaction was observed in collagen fibers. Ageing caused the thinning of collagen fibers, chiefly in the recticular dermis. The labeling pattern of both type I and III collagens did not change in skin samples from patients of up to 79 years of age, but immunoreactivity for type III collagen increased in comparison to younger skins. A loss of fibronectin, likely related to the decreased morphogenetic activity of tissues, was observed with age.
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    Immunocytochemical localisation of some lysosomal hydrolases, their presence in luminal fluid and their directional secretion by human epididymal cells in culture (1995)
    Springer
    Cell & tissue research 280 (1995), S. 415-425 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Epididymis ; Efferent ducts ; Cell culture ; Immunocytochemistry ; Immunoprecipitation ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The way in which the human epididymis modifies spermatozoa during their sojourn in this structure might be clarified by knowledge of the nature of its secretions. We have examined the presence of several lysosomal hydrolases in human epididymal tissue and fluids, and their synthesis and secretion by monolayer cultures. Tissues were obtained from men undergoing orchidectomy for prostatic carcinoma. The enzymes cathepsin D and acid α-glucosidase were localised in the lysosomes of epithelial cells from the corpus epididymidis, by an immunocytochemical technique. Cathepsin D was also found in epithelial cells of the efferent ducts within lysosomes, apical vesicles and multivesicular bodies. No immunolocalisation of acid glucosidase in the efferent ducts or on the microvilli of the corpus was demonstrable. Cathepsin D, β-hexosaminidase (N-acetylglucosaminidase) and α-glucosidase were measurable in the luminal fluid from the human corpus epididymidis; β-hexosaminidase was secreted into the culture medium by confluent monolayers of epididymal and efferent duct cells. Immunoprecipitation of cell extracts and culture medium of these cultures incubated with 35S-methionine revealed that the precursors of cathepsin D and β-hexosaminidase were synthesized and secreted by such monolayers. Thus, active lytic enzymes are secreted by the human epididymis and could modify sperm membranes.
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    Immunolocalization of the Na,K-ATPase in photoreceptor cells of the moth Manduca sexta-evidence for Na,K-ATPase on both the nonmicrovillar and the microvillar domains of the plasma membrane (1995)
    Springer
    Cell & tissue research 281 (1995), S. 119-126 
    Details
    ISSN: 1432-0878
    Keywords: Compound eye ; Photoreceptor cells ; Ion pumps ; Polarity ; Immunocytochemistry ; Manduca sexta (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunohistochemical and physiological studies on various insect photoreceptors have demonstrated that the Na,K-ATPase (sodium pump) is restricted to the nonreceptive nonmicrovillar area of the plasma membrane. Here, we examined the distribution of the Na,K-ATPase in photoreceptor cells of the superposition-type compound eye in the moth Manduca sexta. Using immunofluorescent and immunogold cytochemistry, we show that the Na,K-ATPase is localized to both the nonmicrovillar and the microvillar parts of the plasma membrane. Manduca photoreceptors thus deviate from the common concept that the sodium pump and the molecular components of the photoreceptive machinery reside on different domains of the plasma membrane.
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    Putative neurohemal areas in the peripheral nervous system of an insect, Gryllus bimaculatus, revealed by immunocytochemistry (1995)
    Springer
    Cell & tissue research 281 (1995), S. 43-61 
    Details
    ISSN: 1432-0878
    Keywords: Neurohemal areas ; Neuropeptides ; Monoamines ; Immunocytochemistry ; Nervous system, insect ; Gryllus bimaculatus (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The morphology and position of putative neurohemal areas in the peripheral nervous system (ventral nerve cord and retrocerebral complex) of the cricket Gryllus bimaculatus are described. By using antisera to the amines dopamine, histamine, octopamine, and serotonin, and the neuropeptides crustacean cardioactive peptide, FMRFamide, leucokinin 1, and proctolin, an extensive system of varicose fibers has been detected throughout the nerves of all neuromeres, except for nerve 2 of the prothoracic ganglion. Immunoreactive varicose fibers occur mainly in a superficial position at the neurilemma, indicating neurosecretory storage and release of neuroactive compounds. The varicose fibers are projections from central or peripheral neurons that may extend over more than one segment. The peripheral fiber varicosities show segment-specific arrangements for each of the substances investigated. Immunoreactivity to histamine and octopamine is mainly found in the nerves of abdominal segments, whereas serotonin immunoreactivity is concentrated in subesophageal and terminal ganglion nerves. Immunoreactivity to FMRFamide and crustacean cardioactive peptide is widespread throughout all segments. Structures immunoreactive to leucokinin 1 are present in abdominal nerves, and proctolin immunostaining is found in the terminal ganglion and thoracic nerves. Codistribution of peripheral varicose fiber plexuses is regularly seen for amines and peptides, whereas the colocalization of substances in neurons has not been detected for any of the neuroactive compounds investigated. The varicose fiber system is regarded as complementary to the classical neurohemal organs.
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    URL: http://dx.doi.org/10.1007/BF00307957
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    Immunolocalization of interleukin-1α in rat mandibular molars and its enhancement after in vivo injection of epidermal growth factor (1995)
    Springer
    Cell & tissue research 280 (1995), S. 21-26 
    Details
    ISSN: 1432-0878
    Keywords: Interleukin ; Stellate reticulum ; Immunocytochemistry ; Epidermal growth factor ; Interleukin-1 receptor type I messenger RNA ; Tooth eruption ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunolocalization of interleukin-1α in the first mandibular molars of rats from day 0–12 postnatally showed that the protein was localized in the epithelial stellate reticulum adjacent to the dental follicle. Staining of the stellate reticulum was most prominent in the early days postnatally and was absent by postnatal day 11. Injection of epidermal growth factor into rats at day 0 greatly increased the intensity of the staining for interleukin-1α in the stellate reticulum. Epidermal growth factor (EGF) enhanced the gene expression of interleukin-1α in stellate reticulum cells in vitro, and this study suggests there is enhanced translation of interleukin-1α messenger RNA in the stellate reticulum following EGF injection. In turn, the interleukin-1α may exert its effect on the dental follicle cells adjacent to the stellate reticulum because EGF also enhanced expression of the interleukin-1 receptor type I messenger RNA in cultured dental follicle cells as well as enhancing its expression in vivo. In view of the fact that injection of EGF will stimulate precocious eruption of teeth, its stimulus of interleukin-1α synthesis in the stellate reticulum may be the mechanism by which EGF initiates a cascade of molecular events to signal the onset of tooth eruption.
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    URL: http://dx.doi.org/10.1007/BF00304507
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    Expression of the endogenous 14-kDa β-galactoside-binding lectin galectin in normal human skin (1995)
    Springer
    Cell & tissue research 280 (1995), S. 1-10 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Galectin ; β-Galactoside-binding lectin ; Human ; Skin ; Immunocytochemistry ; Immunohistochemistry ; Hybridization ; in situ ; Langerhans cell ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The localization of an endogenous 14-kDa β-galactoside-binding lectin (galectin) and its pattern of gene expression were examined in normal human skin by light- and electron microscopy. Under the light microscope, immunostaining of 14-kDa galectin was observed in the cell membrane of cells in the basal and spinous layers of the epidermis. Galectin was also found in the Langerhans cells, as shown by double labeling using anti-14-kDa galectin and anti-CD1a antibodi es. In the dermis, immunostaining for the 14-kDa galectin was positive in the extracellular matrix and fibroblasts. At the electron-microscopic level of resolution, galectin was located primarily along the plasma membrane of keratinocytes, and in both the cytoplasm and nucleus of Langerhans cells in the epidermis, whereas in the dermis it was detected in the extracellular matrix and in both the nucleus and cytoplasm of fibroblasts. The gene expression of 14-kDa galectin was visualized by the HRP-staining me thod following in situ hybridization techniques. The expression was detected in the cytoplasm of cells in the basal and spinous layers of the epidermis; whereas, in the dermis, it was detected in the cytoplasm of fibroblasts. Moreover, SDS-polyacrylamide gel electrophoresis and lectin-blot analysis revealed that this galectin bound to glycoproteins of approximately 17, 62, and 72 kDa in the epidermis and to those of 29, 54, and 220 kDa in the dermis. The present study indicates that 1) normal human skin produces the β-galactoside-binding 14-kDa galectin, and 2) this galectin is located in both the epidermis, particularly in the keratinocytes and Langerhans cells, and in the dermis. These results suggest that galectin is important for cell-cell contact and/or adhesion in the epidermis and for cell-extracellular matrix interaction in the dermis.
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    URL: http://dx.doi.org/10.1007/BF00304505
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  • 87
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    Expression of the endogenous 14-kDa β-galactoside-binding lectin galectin in normal human skin (1995)
    Springer
    Cell & tissue research 280 (1995), S. 1-10 
    Details
    ISSN: 1432-0878
    Keywords: Galectin ; β-Galactoside-binding lectin ; Human ; Skin ; Immunocytochemistry ; Immunohistochemistry ; Hybridization, in situ ; Langerhans cell ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localization of an endogenous 14-kDa β-galactoside-binding lectin (galectin) and its pattern of gene expression were examined in normal human skin by light- and electron microscopy. Under the light microscope, immunostaining of 14-kDa galectin was observed in the cell membrane of cells in the basal and spinous layers of the epidermis. Galectin was also found in the Langerhans cells, as shown by double labeling using anti-14-kDa galectin and anti-CD1a antibodies. In the dermis, immunostaining for the 14-kDa galectin was positive in the extracellular matrix and fibroblasts. At the electron-microscopic level of resolution, galectin was located primarily along the plasma membrane of keratinocytes, and in both the cytoplasm and nucleus of Langerhans cells in the epidermis, whereas in the dermis it was detected in the extracellular matrix and in both the nucleus and cytoplasm of fibroblasts. The gene expression of 14-kDa galectin was visualized by the HRP-staining method following in situ hybridization techniques. The expression was detected in the cytoplasm of cells in the basal and spinous layers of the epidermis; whereas, in the dermis, it was detected in the cytoplasm of fibroblasts. Moreover, SDS-polyacrylamide gel electrophoresis and lectin-blot analysis revealed that this galectin bound to glycoproteins of approximately 17, 62, and 72 kDa in the epidermis and to those of 29, 54, and 220 kDa in the dermis. The present study indicates that 1) normal human skin produces the β-galactoside-binding 14-kDa galectin, and 2) this galectin is located in both the epidermis, particularly in the keratinocytes and Langerhans cells, and in the dermis. These results suggest that galectin is important for cell-cell contact and/or adhesion in the epidermis and for cell-extracellular matrix interaction in the dermis.
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    URL: http://dx.doi.org/10.1007/BF00304505
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  • 88
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    Characterization and localization of an Mr 225 kDa polypeptide specifically involved in the defence mechanisms of the clam Tapes semidecussatus (1995)
    Springer
    Cell & tissue research 280 (1995), S. 27-37 
    Details
    ISSN: 1432-0878
    Keywords: Defence mechanisms ; Encapsulation ; Granulocytes ; Immunocytochemistry ; Parasitism ; Perkinsus sp. (Protozoa) ; Tapes semidecussatus (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Parasitosis by the trophozoite protozoan Perkinsus sp. (Apicomplexa, Perkinsea) induces in the gill filaments of the clam Tapes semidecussatus (Mollusca, Bivalvia) a cellular reaction, which is constituted by infiltrated granulocytes. This cellular reaction has characteristics of those of a holocrine gland, since the parasites are encapsulated by the secretion product of the granulocytes after cell death. An enriched fraction of prezoosporangia and their associated capsule was obtained after culture of the parasitized gills in fluid thioglycollate medium. Specific polypeptides from this fraction were separated by SDS-PAGE and isolated for rabbit immunizations. The serum obtained against an Mr 225 kDa polypeptide, revealed its exclusive localization in the capsule and in the granules of the infiltrated granulocytes, thus indicating that this polypeptide is synthesized by these cells and secreted, in a polarized way, around the trophozoites resulting in their encapsulation. Selective deglycosylation of the polypeptide, by Endo H and alkaline β-elimination, did not show an effect on its molecular weight or antibody recognition. Furthermore, the absence of the 225 kDa band in the Western-blots of non-parasitized gills indicated the specific association of this polypeptide with the parasitosis. Finally, this is the first tissue-specific factor described in molluscs in relation to defence mechanisms.
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    URL: http://dx.doi.org/10.1007/BF00304508
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  • 89
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    Immunocytochemical localization of ribonuclease in yolk granules of adult Rana cttesbeiana oocytes (1995)
    Springer
    Cell & tissue research 280 (1995), S. 259-265 
    Details
    ISSN: 1432-0878
    Keywords: Oocyte ; Yolk granules ; Ribonuclease ; Immunocytochemistry ; Bullfrog, Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract To determine the localization of the pyrimidine-guanine sequence-specific ribonuclease in Rana catesbeiana (bullfrog) oocytes, the RNase was first isolated and used to prepare a specific rabbit antiserum. Only one protein of similar molecular size to the RNase was immunoprecipitated from ovary homogenate by the antiserum, but two bands were observed by Western blotting analysis. These two proteins were shown by further purification of antibody and Western blotting analysis to have similar antigenicity. Immunoprecipitation and Western blotting of tissue homogenates showed that the RNase was found predominantly in the ovary, but not in other tissues. The specific localization of the RNase was determined by immuno-electron microscopy of oocyte sections incubated with the specific antiserum; the yolk granules, but not other organelles, were found to contain the RNase. Most of the RNase was evenly distributed in the lateral amorphous area of the yolk granule but not in the central yolk crystal area which contains stored vitellogenin proteins. Our results indicate that the RNase is compartmentalized in the yolk granules of oocytes, which might prevent damage to cellular RNAs.
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    URL: http://dx.doi.org/10.1007/BF00307797
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  • 90
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    Rat intestinal galactoside-binding lectin L-36 functions as a structural protein in the superficial squamous cells of the esophageal epithelium (1995)
    Springer
    Cell & tissue research 281 (1995), S. 77-83 
    Details
    ISSN: 1432-0878
    Keywords: Esophagus ; Epithelial cells ; Intestinal lectin, L-36 ; RI-H fragment ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Using an affinity purified antibody raised against the RI-H fragment of rat intestinal lectin L-36, the latter protein has been identified within the esophageal epithelium by means of ultracryotomy followed by immunogold labeling. The epithelium consists of 4 morphologically distinct cell-types, namely, the basal, spiny, granular and squamous cells, and each of these exhibits a different immunolabeling pattern. The basal cells form a layer on the basal lamina, and in these a diffuse cytoplasmic staining is observed. This basal cell layer is overlaid by spiny cells that extend many cell processes into wide intercellular spaces. In these cells, immunogold particles are found only on small granular inclusions consisting of an electron-lucent homogeneous substance. The granular cells from a third layer over the spiny cells, and are characterized by a number of large granular inclusions with an electron-dense core rimmed by a less electron-dense substance. Immunogold labeling is found on these granules, both on the core and peripheral region. Squamous cell-types constitute the most superficial layer of the epithelium. They are without granular inclusions, and immunogold labeling is confined to the cytoplasmic surface of the thickened plasma membrane. These findings suggest that L-36 is produced in the basal cells as free cytosolic protein, then becomes progressively aggregated into the granular inclusions of the spiny and granular cells, and is eventually transferred onto the cytoplasmic surface of the squamous cell plasma membrane where it may interact with complementary glycoconjugate(s) located at this site. The membrane lining substance thus formed may play a role in stabilizing the squamous cell membranes, thereby maintaining the structural integrity of the epithelium against mechanical stress coming from the esophageal lumen.
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    URL: http://dx.doi.org/10.1007/BF00307960
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  • 91
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    Localization of integrin subunits α6 and β1 during somitogenesis in the long-tailed macaque (M. fascicularis) (1995)
    Springer
    Cell & tissue research 281 (1995), S. 101-108 
    Details
    ISSN: 1432-0878
    Keywords: Somite ; Intergin ; Extracellular matrix, structures ; Embryo ; Laminin ; Immunocytochemistry ; Macaca fascicularis (Primates)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of integrin subunits α6 and β1, and the α6β1 integrin ligand, laminin, was examined during somitogenesis in developmental stages 11, 13, and 16 in the long-tailed macaque, using peroxidase immunocytochemistry. Within differentiating somites in stage 11, α6 expression was observed in the sclerotome, basal surface of dermamyotomal cells adjacent to the basal lamina and on scattered cells throughout the dermamyotome. In further advanced somites in stages 13 and 16, α6 immunoreactivity become restricted to the myotome, α6 was expressed on mesenchymal core cells within the myocele of undifferentiated epitheliod somites and the ventromedial wall of somites commencing differentiation at each stage. β1 distribution resembled that of α6 in stage 11 somitic tissue, however, it remained present on myotome and sclerotome cells in the later stages, and was also expressed on dermatomal cells in stage 16. Laminin immunoreactivity, while more intense and prevalent than α6 and β1 in each stage examined, occurred on the same somite cell populations as the 2 integrin subunits. These results show a defined distribution of α6 on somitic tissue, and suggest this integrin is involved in somite differentiation. They also support a possible role for α6 in myoblast formation and migration. Overlapping of β1 and laminin immunoreactivity with that of α6 further suggests that α6 paris with β1 as a functional heterodimer for laminin in defined somitic regions.
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    URL: http://dx.doi.org/10.1007/BF00307963
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  • 92
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    Immunocytochemical, electrophoresis, and immunoblot analysis of Heliothis virescens gap junctions isolated in the presence and absence of protease inhibitors (1995)
    Springer
    Cell & tissue research 281 (1995), S. 179-186 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Gap junction ; Intercellular junction ; Insect ; Arthropod ; Immunocytochemistry ; Heliothis virescens (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Gap junction-enriched fractions were prepared from larvae of the tobacco budworm Heliothis virescens using the NaOH procedure in the presence or absence of protease inhibitors and were analyzed by SDS-PAGE, immunoblotting and EM immunocytochemistry. Protease inhibitor fractions contained a 48-kDa protein in addition to the ∼10 proteins in fractions with and without inhibitors. Three polyclonal antibodies were used as probes for gap junction plaques and proteins: R16, against an ∼40-kDa candidate gap junction protein from Drosophila melanogaster; R17, against the 40-kDa candidate gap junction protein from H. virescens; and R18AP, an affinity purified antibody against a consensus sequence of N-terminal amino acids 2–21 of the H. virescens 40-kDa protein. R16, R17, and R18AP stain the 40- and 48-kDa proteins, R16 and R18AP stain a 64-kDa protein, and R16 stains an ∼30-kDa protein in the absence of inhibitors. Inclusion of protease inhibitors had no effect on gap junction ultrastructure. R16 and R17 label gap junction plaques in crude membrane and NaOH fractions, whereas R18AP exhibits only a low level of reactivity with gap junctions in crude membrane fractions and none with gap junctions in NaOH fractions. The results show that the 30-, 40-, 48- and 64-kDa proteins are immunologically related and are associated with gap junctions in H. virescens, the N-terminus of the 40-kDa protein is relatively inaccessible or easily lost, and the 48-kDa protein is protease-sensitive.
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    URL: http://dx.doi.org/10.1007/BF00307972
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    Unusual distribution of tubulin isoforms in the snail Lymnaea stagnalis (1995)
    Springer
    Cell & tissue research 281 (1995), S. 507-515 
    Details
    ISSN: 1432-0878
    Keywords: Microtubules ; Isoforms ; Nervous system ; Locomotion ; Cilia ; Immunocytochemistry ; Western blotting ; Lymnaea stagnalis (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunocytochemistry and Western blotting techniques demonstrated that the nervous system and foot of the pond snail Lymnaea stagnalis are rich sources of tubulin, which can be extracted and assembled in vitro in the presence of taxol. Various broad-spectrum antibodies raised against α-tubulin and β-tubulin yielded qualitatively similar results. One monoclonal antibody to trypanosome α-tubulin, however, labelled α-tubulin more strongly on both probed sections and Western blots. Cytochemistry and immunoblotting revealed that tyrosinated tubulin constitutes a large proportion of total α-tubulin in locomotor cilia of the foot and in axons of the nervous system. Detyrosinated tubulin also appeared to be abundant in the foot cilia but only a very faint band of detyrosinated tubulin was found on protein blots extracted from the central ganglia, and staining was barely detectable in central ganglia or peripheral nerves. Similarly, acetylated tubulin appeared to be abundant in foot cilia, but Western blotting indicated only low levels of acetylated tubulin in the nervous system. Immunocytochemistry indicated that, while most neurons possessed little or no acetylated tubulin, a small number of axons contained significant amounts of this isoform. Thus, while a large amount of tubulin was expected in the nervous system and locomotor cilia of L. stagnalis, the observed distribution of isoforms was unanticipated. Specifically, neurons of other organisms have generally been reported to contain substantial amounts of both detyrosinated α-tubulin and acetylated α-tubulin. Our results indicate that such findings cannot be generalized across all species. L. stagnalis, with its well studied nervous system and unusual distribution of tubulin isoforms, may prove to be particularly useful for studying the roles of tubulin isoforms in microtubule function and cell activity.
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    URL: http://dx.doi.org/10.1007/BF00417868
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    Immunocytochemical characterization of the accessory medulla in the cockroach Leucophaea maderae (1995)
    Springer
    Cell & tissue research 282 (1995), S. 3-19 
    Details
    ISSN: 1432-0878
    Keywords: Circadian rhythm ; Colocalization ; Immunocytochemistry ; Brain (CNS), invertebrate ; Optic lobe ; Pigment-dispersing hormone, insect ; Leucophaea maderae (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Several lines of evidence suggest that pigment-dispersing hormone-immunoreactive neurons with ramifications in the accessory medulla are involved in the circadian system of insects. The present study provides a detailed analysis of the anatomical and neurochemical organization of the accessory medulla in the brain of the cockroach Leucophaea maderae. We show that the accessory medulla is compartmentalized into central dense nodular neuropil surrounded by a shell of coarse fibers. It is innervated by neurons immunoreactive to antisera against serotonin and the neuropeptides allatostatin 7, allatotropin, corazonin, gastrin/cholecystokinin, FMRFamide, leucokinin I, and pigment-dispersing hormone. Some of the immunostained neurons appear to be local neurons of the accessory medulla, whereas others connect this neuropil to various brain areas, including the lamina, the contralateral optic lobe, the posterior optic tubercles, and the superior protocerebrum. Double-label experiments show the colocalization of immunoreactivity against pigment-dispersing hormone with compounds related to FMRFamide, serotonin, and leucokinin I. The neuronal and neurochemical organization of the accessory medulla is consistent with the current hypothesis for a role of this brain area as a circadian pacemaking center in the insect brain.
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    URL: http://dx.doi.org/10.1007/BF00319128
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    Ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the dogfish (1995)
    Springer
    Cell & tissue research 282 (1995), S. 33-40 
    Details
    ISSN: 1432-0878
    Keywords: Neuropeptide Y ; Gastroenteropancreatic (GEP) endocrine system ; Development, ontogenetic ; Vitellointestinal duct ; Pancreas, exocrine ; Pancreas, endocrine ; Immunocytochemistry ; Scyliorhinus torazame (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract This immunocytochemical study was carried out to elucidate the ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the cloudy dogfish, Scyliorhinus torazame. Immunostained cells first appeared in the pancreas of the embryo at the 15-mm stage, and were also detected in the vitellointestinal duct of the yolk stalk at the 20-mm stage. These cells were polymorphic, with occasional processes that were sometimes directed toward the vascular wall or into the cavity of the vitellointestinal duct. At the 34-mm stage, immunostained cells could also be found in the proximal part of the spiral intestine and, by the 74-mm stage, immunopositive cells were present in the gastric mucosa. In the gut and pancreas, the cells gradually increased in number with development, whereas in the vitellointestinal duct and internal yolk sac, they decreased and seemed to disappear following hatching. Thus, in juveniles, the distribution of the neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system had attained that of adults. Electron-microscopic immunocytochemistry demonstrated that, in the labeled cells of the vitellointestinal duct, the neuropeptide Y-like antigen was located in cytoplasmic granules, as in the cells of the gut and pancreas.
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    URL: http://dx.doi.org/10.1007/BF00319130
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  • 96
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    Immunocytochemical characterization of the accessory medulla in the cockroach Leucophaea maderae (1995)
    Springer
    Cell & tissue research 282 (1995), S. 3-19 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Circadian rhythm ; Colocalization ; Immunocytochemistry ; Brain (CNS) ; invertebrate ; Optic lobe ; Pigment-dispersing hormone ; insect ; Leucophaea maderae (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Several lines of evidence suggest that pigment-dispersing hormone-immunoreactive neurons with ramifications in the accessory medulla are involved in the circadian system of insects. The present study provides a detailed analysis of the anatomical and neurochemical organization of the accessory medulla in the brain of the cockroach Leucophaea maderae. We show that the accessory medulla is compartmentalized into central dense nodular neuropil surrounded by a shell of coarse fibers. It is innervated by neurons immunoreactive to antisera against serotonin and the neuropeptides allatostatin 7, allatotropin, corazonin, gastrin/cholecystokinin, FMRFamide, leucokinin I, and pigment-dispersing hormone. Some of the immunostained neurons appear to be local neurons of the accessory medulla, whereas others connect this neuropil to various brain areas, including the lamina, the contralateral optic lobe, the posterior optic tubercles, and the superior protocerebrum. Double-label experiments show the colocalization of immunoreactivity against pigment-dispersing hormone with compounds related to FMRFamide, serotonin, and leucokinin I. The neuronal and neurochemical organization of the accessory medulla is consistent with the current hypothesis for a role of this brain area as a circadian pacemaking center in the insect brain.
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    URL: http://dx.doi.org/10.1007/s004410050454
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    The distribution of dopamine-like immunoreactivity in the thoracic and abdominal ganglia of the locust (Schistocerca gregaria) (1992)
    Springer
    Cell & tissue research 270 (1992), S. 113-124 
    Details
    ISSN: 1432-0878
    Keywords: Dopamine ; Nervous sytem, insect ; Ganglia, invertebrate ; Immunocytochemistry ; Schistocerca gregaria (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of dopamine-like immunoreactivity in somata and neurites within the thoracic and abdominal nervous system of the locust Schistocerca gregaria was mapped using two polyclonal antibodies. The prothoracic ganglion contains three bilateral pairs of immunoreactive somata. Two of these lie close to the root of the anterior connective while the third lies between the somata of anterior and common inhibitory motor neurones. The primary neurites of the third pair have a distinctive branching pattern that can be followed through the neuropile and gives rise to a large descending axon. The mesothoracic ganglion has a single pair of antero-lateral immunoreactive somata but the metathoracic and fourth abdominal ganglia have none. The fifth, sixth, and seventh abdominal ganglia each have one pair of somata which are situated at the root of the sternal nerve while the terminal ganglion has a similar pair in the eighth neuromere only. Seven to ten immunoreactive axons enter each ganglion from each of the connectives. Some of these can be traced along the longitudinal tracts that traverse the neuropile. In each ganglion there is one axon in the median dorsal tract and two in ventral median tract. In the meso- and metathoracic ganglia an additional large axon is seen in lateral dorsal tract and dorsal intermediate tract. The axons in median dorsal, and dorsal intermediate tracts send out long ventro-lateral and ventro-medial branches which extend throughout much of the neuropile. Those in median ventral tract send markedly varicose branches to the ventral and dorsal edges of the medial neuropile.
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    URL: http://dx.doi.org/10.1007/BF00381886
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    Ontogeny of the endocrine pancreas in sea bass (Dicentrarchus labrax) (1992)
    Springer
    Cell & tissue research 270 (1992), S. 339-352 
    Details
    ISSN: 1432-0878
    Keywords: Endocrine pancreas ; Ontogeny ; Regulatory peptides ; Immunocytochemistry ; Dicentrarchus labrax (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The development of the endocrine pancreas of the teleost sea bass (Dicentrarchus labrax, L.) was examined from hatching to 61 days, using the peroxidase-antiperoxidase technique for light microscopy. Mammalian and bonito insulin (mI and bI)-, salmo somatostatin-25 (SST-25)-, somatostatin-14 (SST-14a and b)-, glucagon-, bovine pancreatic polypeptide (PP)-, peptide tyrosine-tyrosine (PYY)- and salmo neuropeptide Y (NPY)-like immunoreactivity was demonstrated. Four ontogenetic stages were established according to the organization and immunostaining of the endocrine cells. One cell strand or primordial cord showing mI/bI- and SST-25/SST-14a-like immunoreactivity was first found at hatching in the dorsal epithelium of the anterior zone of the midgut (stage 1). One primitive islet, comprising outer SST-25/SST-14a- and inner mI/bI- and SST-14a/ SST-14b-immunoreactive cells, was found in 2- to 5-day-old larvae (stage 2). One single islet, in which glucagon-immunoreactive cells appear in the periphery, was found in larvae from 9 to 20 days after hatching (stage 3). One big islet containing, in addition, PP-immunoreactive cells in the outer region and slender cell processes which showed PYY-like immunoreactivity, was found from 25 to 61 days after hatching. During this period, primordial islets, composed of SST-25- and bI-immunoreactive cells, and clustered or isolated pancreatic endocrine cells, close to the pancreatic duct, as well as small and intermediate islets (secondary islets), in which glucagon, PP, PYY and NPY seem to be co-localized, were progressively found (stage 4). The origin of the endocrine pancreas of sea bass, and the ontogenetic and phylogenetic significance, are discussed.
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    URL: http://dx.doi.org/10.1007/BF00328018
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    Gonadotropin-releasing hormone neurons and pathways in the brain of the female mink (Mustela vison) (1992)
    Springer
    Cell & tissue research 270 (1992), S. 383-393 
    Details
    ISSN: 1432-0878
    Keywords: Gonadotropin-releasing hormone ; Immunocytochemistry ; Annual cycle-Mink ; Mustela vison
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of gonadotropin-releasing hormone-immunoreactive neurons and processes was mapped in the female mink brain using coronal, horizontal and sagittal sections. Perikarya were found along a ventral continuum including the olfactory tubercle, the diagonal band of Broca, the lateral septum, the preoptic and anterior hypothalamic area and the mediobasal hypothalamus; 80% of the perikarya were counted in the mediobasal hypothalamus. Fibres were mainly observed in the organum vasculosum of the lamina terminalis and the median eminence. A few processes terminated in the ependymal cells lining the third and lateral ventricles. The total number of immunoreactive perikarya was the highest in the brains of females sacrificed in July; it then significantly decreased until December. This variation is discussed in relation to the annual breeding cycle.
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    URL: http://dx.doi.org/10.1007/BF00328022
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  • 100
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    Immunocytochemical localisation of some lysosomal hydrolases, their presence in luminal fluid and their directional secretion by human epididymal cells in culture (1995)
    Springer
    Cell & tissue research 280 (1995), S. 415-425 
    Details
    ISSN: 1432-0878
    Keywords: Epididymis ; Efferent ducts ; Cell culture ; Immunocytochemistry ; Immunoprecipitation ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The way in which the human epididymis modifies spermatozoa during their sojourn in this structure might be clarified by knowledge of the nature of its secretions. We have examined the presence of several lysosomal hydrolases in human epididymal tissue and fluids, and their synthesis and secretion by monolayer cultures. Tissues were obtained from men undergoing orchidectomy for prostatic carcinoma. The enzymes cathepsin D and acid α-glucosidase were localised in the lysosomes of epithelial cells from the corpus epididymidis, by an immunocytochemical technique. Cathepsin D was also found in epithelial cells of the efferent ducts within lysosomes, apical vesicles and multivesicular bodies. No immunolocalisation of acid glucosidase in the efferent ducts or on the microvilli of the corpus was demonstrable. Cathepsin D, β-hexosaminidase (N-acetylglucosaminidase) and α-glucosidase were measurable in the luminal fluid from the human corpus epididymidis; β-hexosaminidase was secreted into the culture medium by confluent monolayers of epididymal and efferent duct cells. Immunoprecipitation of cell extracts and culture medium of these cultures incubated with 35S-methionine revealed that the precursors of cathepsin D and β-hexosaminidase were synthesized and secreted by such monolayers. Thus, active lytic enzymes are secreted by the human epididymis and could modify sperm membranes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00307815
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