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  • Angiosperms  (60)
  • Immunocytochemistry  (42)
  • Nitrogen fixation  (36)
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  • Springer  (179)
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  • 1
    Electronic Resource
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    Springer
    Biology and fertility of soils 4 (1987), S. 61-66 
    ISSN: 1432-0789
    Keywords: Stem nodulation ; Aeschynomene afraspera ; Legume ; Nitrogen fixation ; Acetylene reduction assay (ARA)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Aeschynomene afraspera is a wild annual legume growing in periodically waterlogged soils in western Africa. This legume is characterized by a profuse stem nodulation. Nodules are formed on the stem at the emergence of lateral root primordia, called nodulation sites. These sites are irregularly distributed on vertical rows all along the stem and branches. Stem nodules are hemispherically shaped. Their outside is dark green and they contain a red-pigmented central zone. Stem nodules exhibit a high nitrogen-fixing potential. Acetylene reduction assays result in stem nodule activity of 309 μmol C2H4 g−1 dry nodule h−1. Field-grown stem nodulated Aeschynomene accumulated more N (51 g N m−2 in 10 weeks) than the root nodulated one. Because of this nitrogenfixing potential and its ability to grow in waterlogged conditions, A. afraspera could probably be introduced into tropical rice cropping systems.
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  • 2
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    Biology and fertility of soils 5 (1987), S. 83-87 
    ISSN: 1432-0789
    Keywords: Inoculation ; Inoculum dose ; Nitrogen fixation ; Chickpea ; Rhizobium spp. ; Cicer arietinum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The influence of three inoculum rates on the performance of three chickpea (Cicer arietinum L.) Rhizobium strains was examined in the field on a Mollisol soil. Increasing amounts of inoculum improved the performance of the strains. A normal dose (104 cells per seed) applied at different intervals gave non-significant increases in nodulation, nitrogenase activity (acetylene reduction assay), nitrogen uptake and grain yield. A ten-fold increase in inoculum increased nodule number, shoot dry weight, nitrogenase activity (ARA) and grain yield, but increases over the control were significant only for nodule dry weight and nitrogen uptake by shoot and grain. The highest level of inoculum (100 × normal) significantly increased nodule dry weight, grain yield, total nitrogenase activity (ARA) and nitrogen uptake by shoot and grain. Strain TAL 620 was more effective than the other two. Combined nitrogen (60 kg N ha−1) suppressed nodulation and nitrogenase activity (ARA).
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  • 3
    ISSN: 1432-0789
    Keywords: Triticum aestivum ; T. turgidum ; Nitrogen fixation ; Field inoculation ; Acetylene reduction assay (ARA)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Eight commercial Israeli spring wheat cultivars (six Triticum aestivum and two T. turgidum) grown with 40 and 120 kg N/ha were tested for responses to inoculation with Azospirillum brasilense. At the low level of N fertilization (40 kg/ha), five cultivars showed significant increases in plant dry weight measured at the milky ripe stage; however, by maturation only the cultivar “Miriam” showed a significant increase in grain yield. Two cultivars, which had shown a positive inoculation effect at the earlier stages, had a significant decrease in grain yield. No significant effect of inoculation was found at the high N level. To confirm those results, four wheat (T. aestivum) cultivars were tested separately over 4 years in 4 different locations under varying N levels. Only Miriam showed a consistently positive effect of Azospirillum inoculation on grain yield. Inoculation increased the number of roots per plant on Miriam compared with uninoculated plants. This effect was found at all N levels. Nutrient (N, P and K) accumulation and number of fertile tillers per unit area were also enhanced by Azospirillum, but these parameters were greatly affected by the level of applied N. It is suggested that the positive response of the spring wheat cultivar “Miriam” to Azospirillum inoculation is due to its capacity to escape water stresses at the end of the growth season.
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  • 4
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    Biology and fertility of soils 4 (1987), S. 15-19 
    ISSN: 1432-0789
    Keywords: Nitrogen fixation ; N-balance studies ; Azolla ; Blue-green algae ; Chemical N fertilization ; Rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A nitrogen balance study conducted in ceramic pots under net house conditions for four seasons showed that flooded rice soil leaves a positive nitrogen balance (N increase) in soil after rice cropping in both fertilized and unfertilized soil. Recovery of nitrogen from rice soil was more than its input in unfertilized soil, but it was reverse in fertilized soil. Incorporation of Azolla or BGA twice as basal and 20 days after transplanting (DAT) alone or in combination showed higher nitrogen balance and N2-fixation (N gain) in soil than in that where it was applied once either as basal or 20 DAT. Planted soil showed more N2-fixation than that of fallow rice, and flooded soil fixed more nitrogen in comparison to non-flooded soil in light but less in dark. Soil exposed to light fixed more nitrogen than that of unexposed soil in both flooded and non-flooded conditions.
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  • 5
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    Biology and fertility of soils 4 (1987), S. 9-14 
    ISSN: 1432-0789
    Keywords: Rhizosphere ; Nitrogen fixation ; Root exudates ; Soil bacteria ; Carbon budget ; Rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The association of rice seedlings (cv. Delta) with different strains of Azospirillum was studied under monoxenic conditions in the dark. Axenic 3-day-old seedlings were obtained on a C- and N-free medium and inoculated with 6 · 107 bacteria per plant in a closed vial. Seven days later, different components of a carbon budget were evaluated on them and on sterile controls: respired CO2, carbon of shoot and roots, bacterial and soluble carbon in the medium. Two strains (A. lipoferum 4B and A. brasilense A95) isolated from the rhizosphere of rice caused an increase in exudation, + 36% and + 17% respectively compared with sterile control. Shoot carbon incorporation and respiration were reduced by inoculation. A third strain (A. brasilense R07) caused no significant change in exudation. A. lipoferum B7C isolated from maize did not stimulate rice exudation either. We further investigated a possible effect of nitrogen fixation on this phenomenon: inhibition of nitrogen fixation by 10% C2H2 did not modify the extent of C exudation by rice associated with A. lipoferum 4B or with the non-motile A. lipoferum 4T.
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  • 6
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    Journal of molecular evolution 26 (1987), S. 347-357 
    ISSN: 1432-1432
    Keywords: Ribosome structure ; Electron microscopy ; Image analysis ; Evolutionary lineages
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Multivariate statistical analysis and classification techniques are powerful tools in sorting noisy electron micrographs of single particles according to their principal features, enabling one to form average images with an enhanced signal-to-noise ratio and a better reproducible resolution. We apply this methodology here to determining the characteristic views of the large (50S) ribosomal subunits from the eubacteriumEscherichia coli and the archaebacteriaMethanococcus vannielii, Sulfolobus solfataricus, andHalobacterium marismortui. Average images were obtained of the subunit in the common crown and kidney projections, but views of the particle in orientations intermediate between these two extremes were also elucidated for all species. These averages show reproducible detail of up to 2.0 nm resolution, thus enabling the visualization and interspecies comparison of many structural features as a first step toward comparing the actual three-dimensional structures. Our results disprove evolutionary lineages recently postulated on the basis of electron microscopical images of ribosomal subunits.
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  • 7
    ISSN: 1432-2285
    Keywords: Cambial activity ; Frost hardiness ; Phenology ; Salix ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The ultrastructure of cells in the cambial region of Salix dasyclados Wim. (clone 78056) was studied during the development of winter hardiness and the onset of cambial activity in spring. Plants were grown at relative growth rates (RG) of 8% and 12% respectively, resulting in different nitrogen content in the stems. Frost hardiness of the plants was estimated by standardized freezing tests. Plants with a higher nitrogen status ceased growth later and started re-growth earlier in spring than plants with lower nitrogen content. Differences in ability to withstand low temperatures during autumn and spring were found between plants grown in the two nutrient treatments. During the development of frost hardiness in the autumn, the number of meristematic cells in the cambial region decreased. The cessation of meristematic activity was accompanied by cell wall thickening and ultrastructural changes in the cells. Frost hardiness increased from the ability to survive -6° C in October to survival of -80° C at the beginning of December. From November to February the cambial region comprised a layer of 2–3 thick-walled cells with conspicuous ultrastructural features. Starch accumulated in plastids in September, decreased during November to March and then increased again in accordance with changes of frost hardiness. Onset of cambial activity began between the end of March and the beginning of April, as shown by increased vacuolization of meristematic cells and mitotic activity. By April, the starch content had increased and lipolysis was observed. Frost hardiness had decreased, and plants with low and high nitrogen content were able to survive -15° C and -10° C, respectively. After budburst, all axillary shoot parts were damaged at temperatures below-3° C.
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  • 8
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    Development genes and evolution 196 (1987), S. 367-371 
    ISSN: 1432-041X
    Keywords: Vitellogenesis ; Bufo marinus oocyte ; Yolk-platelet membrane ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Oocytes of the toad Bufo marinus have been studied by means of thin section and particularly freeze-fracture electron microscopy to characterize the cytoplasmic membranes around the yolk organelle, and the storage of yolk material in precursors and platelets. This appears to be a previously unknown type of yolk-platelet formation. During yolk-organelle development from the primordial precursor to the bi-partite fully grown yolk platelet, numerous lipoid droplets are attached to the periphery of the platelet, indicating an intense uptake of lipids. As is typical for amphibians, the fully grown yolk platelet has a crystalline internum covered by a dense osmiophilic externum, and the whole organelle is enveloped by a plasma membrane that shows no direct connection or fusion with endocytotic vesicles. The yolk membrane exhibits few intramembraneous particles (IMPs) at the core areas and some more where it borders fields of lipoid droplets. Here the IMPs show a net-like arrangement in the furrows between adjacent droplets.
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  • 9
    ISSN: 1432-1939
    Keywords: Lupinus succulentus ; Fabaceae ; Lupine ; Quinolizidine alkaloids ; Nitrogen fixation ; Defoliation ; Plant-herbivore interactions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We examined the effects of nitrogen nutrition and defoliation on the alkaloids, nitrogen levels, and growth of Lupinus succulentus by growing plants under five nitrogen/defoliation treatments: 1) fertilization with a high-nitrate nutrient solution, 2) fertilization with a low-nitrate solution, 3) inoculation with N-fixing bacteria but without available soil nitrogen, 4) high-nitrate solution plus periodic partial defoliation, and 5) low-nitrate plus defoliation. In the absence of defoliation, plants from both the N-fixing and high-N treatments had higher concentrations of alkaloids and nitrogen, and higher growth rates than the low-N plants. Periodic defoliation had little effect on the high-N plants, but defoliated N-fixing plants were severely stunted and had lower alkaloid and nitrogen levels. The experimental treatments also affected the relative concentrations of the alkaloids. Our results indicate that 1) alkaloid composition and concentration in L. succulentus are determined by both nitrogen availability and developmental state, 2) plants relying solely on N-fixation respond quite differently to defoliation than those with adequate soil nitrogen, and 3) the food value of the plant tissue can be affected by an interaction between the effects of defoliation and nitrogen status.
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  • 10
    ISSN: 1432-0983
    Keywords: Electron microscopy ; Replicative intermediates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Some physicochemical properties of the mitochondrial DNAs (mtDNA) from plants of flax, broad bean and mung bean, and from tissue culture cells of jimson weed, soybean, petunia and tobacco were determined. Circular molecules were observed in electron microscope preparations of each mtDNA. In soybean, petunia, broad bean and mung bean mtDNAs, the circular molecules had a continuous distribution of lengths (ranges between 1 to 36 kb, and 1 to 126 kb), heavily skewed toward smaller molecules. Eighty-six percent of the flax circular molecules were from 27 to 54 kb in size, and 78% of the jimson weed circular molecules were from 4 to 15 kb. Replicative forms of 1.2–1.6 kb circular molecules were observed in electron microscope preparations of broad bean mtDNA.
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  • 11
    ISSN: 1432-072X
    Keywords: Methanobacterium thermoautotrophicum ; Methyl-CoM reductase ; Immunocytochemistry ; Colloidal gold ; Energy conservation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cells of Methanobacterium thermoautotrophicum were fixed with glutaraldehyde, sectioned and labeled with antibodies against the β subunit of component C (=methyl-CoM reductase) of methyl-CoM reductase system and with colloidal gold-labeled protein A. It was found that the gold particles were located predominantly in the vicinity of the cytoplasmic membrane, when the cells were grown under conditions where methyl-CoM reductase was not overproduced. This finding confirms the recent data obtained with Methanococcus voltae showing via the same immunocytochemical localization technique that in this organism methyl-CoM reductase is membrane associated.
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  • 12
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    Archives of microbiology 148 (1987), S. 286-291 
    ISSN: 1432-072X
    Keywords: Sulfate-reducing bacteria ; Desulfobacter species ; Acetate ; Hydrogen ; Autotrophic growth ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Sulfate-reducing bacteria with oval to rod-shaped cells (strains AcRS1, AcRS2) and vibrio-shaped cells (strains AcRM3, AcRM4, AcRM5) differing by size were isolated from anaerobic marine sediment with acetate as the only electron donor. A vibrio-shaped type (strain AcKo) was also isolated from freshwater sediment. Two strains (AcRS1, AcRM3) used ethanol and pyruvate in addition to acetate, and one strain (AcRS1) grew autotrophically with H2, sulfate and CO2. Higher fatty acids or lactate were never utilized. All isolates were able to grow in ammonia-free medium in the presence of N2. Nitrogenase activity under such conditions was demonstrated by the acetylene reduction test. The facultatively lithoautotrophic strain (AcRS1), a strain (AcRS2) with unusually large cells (2×5 μm), and a vibrio-shaped strain (AcRM3) are described as new Desulfobacter species, D. hydrogenophilus, D. latus, and D. curvatus, respectively.
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  • 13
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    Archives of microbiology 149 (1987), S. 24-29 
    ISSN: 1432-072X
    Keywords: Frankia ; Nitrogen fixation ; Calcium ; Vesicle development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A calcium requirement was shown for both vesicle development and nitrogenase activity by Frankia strains EAN1pec and CpI1. Washing cells with EGTA or EDTA inhibited both vesicle development and nitrogenase activity. The inhibition of both was reversed by the addition of calcium. A variety of agents known to affect calcium-dependent biological processes, such as a Ca-ATPase inhibitor, Ca-channel blockers, Ca-ionophores, calmodulin antagonists and the local anaesthetics, tetracaine and dibucaine, inhibited nitrogenase activity. Respiratory studies showed that a CN-insensitive respiration process occurred only under nitrogen derepressing conditions. Respiration by NH4Cl-grown cells was completely inhibited by KCN while N2-grown cells were inhibited by only 70%. Removal of calcium ions by EGTA or by the addition of dibucaine or tetracaine blocked the CN-insensitive respiration. This CN-insensitive respiration may be involved in protecting nitrogenase inside the vesicles from oxygen.
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  • 14
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    Archives of microbiology 147 (1987), S. 383-388 
    ISSN: 1432-072X
    Keywords: Frankia ; Nitrogen fixation ; Vesicle development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The relationship between nitrogen fixation and development of a specialized cell structure, called the vesicle, was studied using four Frankia isolates. Nitrogenase activity was repressed in all four strains during growth with ammonia. Strain CpI1 formed no vesicles during NH4 growth. Strains ACN1 ag , EAN1pec and EUN1f produced low numbers of vesicles in the presence of ammonia. Following transfer to nitrogen-free media, a parallel increase in nitrogenase activity and vesicle numbers occurred with all four isolates. Appearance of nitrogenase activity was more rapid in those strains that possessed some vesicles at the time of shift to N2 as a nitrogen source. The ratio of vesicle numbers to level of nitrogenase activity varied widely among the four strains and in response to different growth conditions and culture age of the individual strains. Optimum conditions of temperature, carbon and energy source, nitrogen source and availability of iron and molybdenum were different for each of the four strains. Those conditions that significantly reduced nitrogenase activity were always associated with decreased numbers of vesicles.
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  • 15
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    Archives of microbiology 146 (1987), S. 327-331 
    ISSN: 1432-072X
    Keywords: Actinomycetes ; Nitrogen fixation ; Symbiosis ; Immunocytochemistry ; Ultracryotomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Immunogoldlabelling on ultrathin cryosections of Frankia sp. Cc1.17 showed specific labelling of nitrogenase in the spherical cells called vesicles. No label was found in the hyphae in any cells grown on a medium with combined nitrogen, nor in those to which no specific antiserum was added. Similar results were obtained with cultures grown under high (20%) and low (2%) oxygen tension in the gas phase.
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  • 16
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    Archives of microbiology 148 (1987), S. 150-154 
    ISSN: 1432-072X
    Keywords: Bacteroides ; Vesicles ; Ultrastructure ; Cellulolytic bacteria ; Rumen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In 3-day-old cultures of Bacteroides succinogenes grown on filter paper, no cell division was observed. When grown on cellulosic substrate, bacteria exhibited vesicles clustered within cell wall pockets. In 2 day-old filter paper cultures, cells adhered tightly to the substrate. Twenty to 30% of them were dividing. There were cell wall pockets in about 25% of the bacteria, but no vesicles. Whether they adhered to the cellulosic substrate or not, and irrespective of the age of the bacteria, storage polysaccharides were found in the form of dense granules in the cytoplasm. It would appear that vesicles are not essential for cellulose degradation, but are rather a sign of ageing of the cells.
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  • 17
    ISSN: 1476-5535
    Keywords: Sulfate-reducing bacteria ; Hydrogen metabolism ; Nitrogen fixation ; Deuterium-proton exchange
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Hydrogenase and nitrogenase activities of sulfate-reducing bacteria allow their adaptation to different nutritional habits even under adverse conditions. These exceptional capabilities of adaptation are important factors in the understanding of their predominant role in problems related to anaerobic metal corrosion. Although the D2−H+ exchange reaction indicated thatDesulfovibrio desulfuricans strain Berre-Sol andDesulfovibrio gigas hydrogenases were reversible, the predominant activity in vivo was hydrogen uptake. Hydrogen production was restricted to some particular conditions such as sulfate or nitrogen starvation. Under diazotrophic conditions, a transient hydrogen evolution was followed by uptake when dinitrogen was effectively fixed. In contrast, hydrogen evolution proceeded when acetylene was substituted as the nitrogenase substrate. Hydrogen can thus serve as an electron donor in sulfate reduction and nitrogen metabolism.
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  • 18
    ISSN: 1432-2048
    Keywords: Nitrogen fixation ; Peroxisome ; Root nodules ; Ureide biogenesis ; Uricase ; Vigna
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cowpea (Vigna unguiculata (L.) Walp.) nodules have been investigated by means of cytochemical and immunocytochemical procedures at the ultrastructural level in order to assess the role of the uninfected cells in ureide biogenesis. Uricase activity in the nodules was shown by cytochemical methods to be localized exclusively in the numberous large peroxisomes confined to the uninfected cells; the small peroxisomes in the infected cells did not stain for uricase. Uricase was also localized in the peroxisomes of uninfected cells by immunogold techniques employing polyclonal antibodies against nodule-specific uricase of soybean. There was no labeling above background of any structures in the infected cells. The results indicate that the uninfected cells are essential for ureide biogenesis in cowpea. Although tubular endoplasmic reticulum, the presumptive site of allantoinase, increases greatly in the uninfected cells during nodule development, it virtually disappears as the nodules mature. The inconsistency between the disappearance of the tubular endoplasmic reticulum from older nodules and the high allantoinase activity reported for older plants remains to be explained.
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  • 19
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    Theoretical and applied genetics 74 (1987), S. 177-187 
    ISSN: 1432-2242
    Keywords: Barley ; Grain development ; Mutants ; Ultrastructure ; Genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Eleven Na-azide induced barley shrunken endosperm mutants expressing xenia (sex) were characterized genetically and histologically. All mutants have reduced kernel size with kernel weights ranging from 11 to 57% of the wild type. With one exception, the mutant phenotypes are ascribable to single recessive mutant alleles, giving rise to a ratio of 3∶1 of normal and shrunken kernels on heterozygous plants. One mutant (B10), also monofactorially inherited, shows a gene dosage dependent pattern of expression in the endosperm. Among the 8 mutants tested for allelism, no allelic mutant genes were discovered. By means of translocation mapping, the mutant gene of B10 was localized to the short arm of chromosome 7, and that of B9 to the short arm of chromosome 1. Based on microscopy studies, the mutant kernel phenotypes fall into three classes, viz. mutants with both endosperm and embryo affected and with a non-viable embryo, mutants with both endosperm and embryo affected and with a viable embryo giving rise to plants with a clearly mutant phenotype, and finally mutants with only the endosperm affected and with a normal embryo giving rise to plants with normal phenotype. The mutant collection covers mutations in genes participating in all of the developmental phases of the endosperm, i.e. the passage from syncytial to the cellular endosperm, total lack of aleurone cell formation and disturbance in the pattern of aleurone cell formation. In the starchy endosperm, varying degrees of cell differentiation occur, ranging from slight deviations from wild type to complete loss of starchy endosperm traits. In the embryo, blocks in the major developmental phases are represented in the mutant collection, including arrest at the proembryo stage, continued cell divisions but no differentiation, and embryos deviating only slightly from the wild type.
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  • 20
    ISSN: 1432-0878
    Keywords: Ornithine decarboxylase ; Chick embryo ; Organogenesis A ; Autoradiography ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localization of ornithine decarboxylase (ODC), a key enzyme in polyamine biosynthesis and thus in cell growth, was determined in the 4.5-day-old chick embryo, using two independent methods of analysis. ODC protein was identified by indirect immunofluorescence with a monospecific ODC antibody, and catalytically active ODC was identified by autoradiography with α-(5-3H) difluoromethylornithine. Both methods revealed a basically similar distribution of ODC within the embryo. Among the organs, the brain exhibited the highest ODC levels. ODC levels were also high in spinal cord, mesonephric tubules and heart. Similar levels, but confined to limited areas, were found in liver tissue, head mesenchyme, and the oral and pharyngeal regions. Organs that exhibited high ODC levels are all engaged in rapid growth, as well as in extensive tissue remodeling and differentiation.
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  • 21
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Innervation ; Neu rophysins ; Mesotocin ; Immunocytochemistry ; Ultrastructure ; Snake, Natrix maura
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The subcommissural organ (SCO) of the snake Natrix maura was studied by use of the immunoperoxidase procedure. Primary antisera against bovine neurophysins (Nps I + II, OXY-Np), oxytocin (OXY), mesotocin (MST), arginine-vasotocin (AVT), somatostatin (SOM), β-endorphin (END) and bovine Reissner's fiber were used. A conventional ultrastructural study, with special emphasis on the nerve fibers present in the SCO, was also performed. Nerve fibers containing immunoreactive OXY-Np and MST were seen to reach the SCO. The staining of adjacent sections with the anti-Reissner's fiber serum showed that the OXY-Np- and MST-immunoreactive fibers were distributed among the cell bodies and processes of the ependymal secretory cells. No fibers containing immunoreactive OXY, AVT, SOM or END were found in the SCO. The ultrastructural analysis revealed in the SCO the presence of nerve fibers filled with electron-dense granules, 170–210 nm in diameter. Although a direct apposition between these fibers and the SCO cells was frequently seen, no synaptic differentiations were identified. Structures identical to the Herring bodies (found in the neurohypophysis) were seen in the SCO.
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  • 22
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    Cell & tissue research 249 (1987), S. 421-425 
    ISSN: 1432-0878
    Keywords: Cholinergic synapses ; Ultrastructure ; Exocytosis ; Non-synaptic release ; Neuropeptides ; Carassius auratus ; Rana pipiens ; Wistar white rat ; Hamster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Nerve terminals forming typical synapses with adrenal chromaffin tissues have been examined in the goldfish, frog (Rana pipiens), hamster and rat. Presumptive secretory inclusions present in the terminals are of two distinct types. Electron-lucent synaptic vesicles 30–50 nm in diameter are densely clustered adjacent to membrane thickenings and presumably discharge their contents into the synaptic clefts. Secretory granules (i.e. large dense-cored vesicles) 60–100 nm in diameter are more abundant in other parts of the terminals. Sites of granule exocytosis have been observed in each of the animals investigated. They are usually encountered within apparently undifferentiated areas of plasmalemma and only rarely occur within synaptic thickenings. Granule exocytosis from within synaptic terminals and chromaffin gland cells is most readily observed in specimens exposed, prior to fixation, to saline solutions containing both tannic acid, and 4-aminopyridine and/or elevated levels of K+. These findings show that the pattern of secretory discharge, involving both synaptic and non-synaptic release, which is widespread in invertebrate central nervous systems, is also characteristic of vertebrate, peripheral cholinergic terminals.
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  • 23
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    Cell & tissue research 249 (1987), S. 437-442 
    ISSN: 1432-0878
    Keywords: Angiotensin II ; Glomerulus ; Salmo gairdneri ; Seawater-adaptation ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effect of angiotensin infusion on the glomerular ultrastructure of freshwater- and seawater-adapted rainbow trout, Salmo gairdneri, has been examined by scanning and transmission electron microscopy. Adaptation of trout to seawater resulted in epithelial podocyte flattening, primary process broadening and apparent loss of foot processes in almost all glomeruli, features which were uncommon in freshwater-adapted trout. Similar changes were induced by infusion of freshwater-adapted animals with angiotensin, suggesting that the renin-angiotensin system plays a role in the modification of glomerular epithelial ultrastructure. Adaptation of trout to seawater also reduced glomerular diameter, but infusion of freshwater-adapted animals with angiotensin did not mirror this effect. Infusion of angiotensin into seawater-adapted animals increased the overall thickness of glomerular basement membrane by increasing the lamina rara interna and lamina densa. This did not occur when freshwater-adapted fish were either infused with angiotensin or adapted to seawater. These findings suggest that other humoral systems are involved in the control of glomerular diameter and basement membrane thickness as part of an integrated response to increased environmental salinity.
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  • 24
    ISSN: 1432-0878
    Keywords: Leydig cells ; Ethane dimethanesulphonate ; Ultrastructure ; Degeneration ; Resistance ; Rodents
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The morphological response of the testis of rats, guinea pigs, Syrian hamsters and mice to treatment with the cytotoxin ethane dimethanesulphonate was examined using light and electron microscopy. One to two days after a single administration of ethane dimethanesulphonate to adult rats, guinea pigs, and hamsters, the Leydig cells showed marked ultrastructural alterations suggestive of degeneration and cell death. The former alterations included karyopyknosis, cytoplasmic vesiculation and accumulation of lipid inclusions and large lipofuscin bodies. Fragments of necrotic Leydig cells were often engulfed by the interstitial tissue macrophages. The morphology of the seminiferous epithelium of these three species was unchanged from the morphology observed in vehicle-injected control animals. In contrast, multiple injections of ethane dimethanesulphonate given to mice produced no ultrastructural alterations to Leydig cells yet the seminiferous epithelium exhibited disruption of spermatogenesis. Although the Leydig cells of the mouse appear resistant to ethane dimethanesulphonate, this agent exerts a selective cytotoxic action upon Leydig cells of the rat, guinea pig and hamster thus identifying ethane dimethanesulphonate as a useful chemical for future endocrine and physiological studies of testicular function in three common laboratory species.
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  • 25
    ISSN: 1432-0878
    Keywords: Retina ; Pineal organ ; Hypothalamus ; Opsin ; α-Transducin ; Interstitial retinol-binding protein (IRBP) ; Immunocytochemistry ; Japanese quail
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The retinal proteins opsin,α-transducin, S-antigen and interstitial retinol-binding protein (IRBP) are essential for the processes of vision. By use of immunocyto-chemistry we have employed antibodies directed against these “photoreceptor proteins” in an attempt to identify the photoreceptor systems (retina, pineal and deep brain) of the Japanese quail. Opsin immunostaining was identified within many outer (basal portion) and inner segments of retinal photoreceptor cells and limited numbers of photoreceptor perikarya. Opsin immunostaining was also demonstrated in limited numbers of pinealocytes with all parts of these cells being immunoreactive. These results differ from previous observations. In contrast to the results obtained with the antibody against opsin, S-antigen andα-transducin immunostaining was seen throughout the entire outer segments and many photoreceptor perikarya of the retina. In the pineal organ immunostaining was seen in numerous pinealocytes in all follicles. These results conform to previous findings in birds. In addition, IRBP has been demonstrated for the first time in the avian retina and pineal organ. These findings underline the structural and functional similarities between the retina and pineal organ and provide additional support for a photoreceptive role of the avian pineal. No specific staining was detected in any other region of the brain in the Japanese quail; the hypothalamic photoreceptors of birds remain unidentified.
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  • 26
    ISSN: 1432-0878
    Keywords: Endocrine pancreas ; Insulin ; Glucagon ; Somatostatin ; Immunocytochemistry ; Neoceratodus forsteri (Australian lungfish)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The endocrine pancreas of the Australian lungfish,Neoceratodus forsteri, was investigated immunocytochemically for the presence of polypeptide hormone-producing cells. Three cell types were identified, namely insulin-, glucagon-, and somatostatin-immunoreactive elements. The insulin cells are confined solely to the center of the islets. Glucagon and somatostatin cells are distributed peripherally around the central mass of the insulin cells. Isolated cells or clusters of glucagon and somatostatin cells are also dispersed within the exocrine parenchyma. The immunoreactive cell types are compared with those staining with standard histological procedures. The spatial relationships of the different cell populations are examined.
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  • 27
    ISSN: 1432-0878
    Keywords: Macula densa ; Intercellular spaces ; Ultrastructure ; Frusemide ; Tubulo-glomerular feedback ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In situ drip-fixation of superficial glomeruli and tubules in the rat kidney verified the results obtained from high pressure perfusion fixation studies indicating that distinct spaces normally exist between the cells of the macula densa. Following treatment with frusemide these intercellular spaces between the macula densa cells became closed in both drip-fixed and perfusion-fixed kidneys. These findings suggest that the variability of extracellular compartmentation found in the macula densa is unlikely to be an artifact but most likely represents changes in the in vivo status of the macula densa.
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  • 28
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    Cell & tissue research 248 (1987), S. 275-286 
    ISSN: 1432-0878
    Keywords: Nerve growth factor ; Salivary glands ; Antibodies ; Immunocytochemistry ; Affinity purification ; Specificity tests ; Testis ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A series of polyclonal affinity-purified antibodies against mouse submandibular-gland nerve growth factor (NGF) are described. Using the submandibular gland of the male mouse and indirect immunofluorescence, the specificity and sensitivity of affinity-purified immunoglobulins and various other fractions from the immunized animals have been tested. It will be shown that affinity-purification schemes, including pre-purification of protein A-fractionated immunoglobulins to remove antibodies that bind to unrelated hydrophilic and hydrophobic proteins, significantly enhance the signal-to-noise ratio and specificity of the antibodies. The antibodies effectively detect NGF-like immunoreactivity in both fresh and fixed glandular tissue. Optimal fixation procedures are described. Fluorescence intensities are linearly correlated to log antibody concentration. By use of the best antibody fractions and optimal fixation protocols, the distribution of NGF-like immunoreactivity is described in eight different salivary glands (rat and mouse, male and female, submandibular and sublingual glands). In addition to the well-known large numbers of immunoreactive cells in the submandibular gland of the male mouse, immunoreactive cells were found in the sublingual gland of male mice and in the submandibular and sublingual glands of female mice. One antibody revealed a weak specific fluorescence also in the submandibular gland of the male mouse. In a survey of genital organs of male mice, one antibody revealed fluorescence in the germ cell line. We conclude that several polyclonal affinity-purified antibodies have been characterized that show a strong NGF-dependent binding to the secretory granules of tubular cells in the submandibular gland of male mice. These antibodies should make it possible to locate endogenous and perturbed NGF levels immunocytochemically, e.g., in the peripheral and central nervous system, where NGF concentrations may be several orders of magnitude lower than in the salivary glands.
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  • 29
    ISSN: 1432-0878
    Keywords: Nervous system ; Immunocytochemistry ; FMRFamide ; Bovine pancreatic polypeptide ; Insects ; Schistocerca gregaria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of FMRFamide-irmunoreactive cell bodies in the brain and retrocerebral complex of the locust, Schistocerca gregaria, is described. Most of the immunoreactive cell bodies are found in the pars intercerebralis and in the optic lobes. Many, but not all, of the cell bodies also react with an antiserum raised against bovine pancreatic polypeptide, but this antiserum also reveals another population of cells that stain selectively with this antiserum. In addition to the cell bodies, numerous immunoreactive processes are revealed by both antisera in neuropilar regions of the brain. The results of blocking experiments suggest that a differential distribution of three locust antigens can be determined from the examination of alternate serial sections stained with the two antisera used.
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  • 30
    ISSN: 1432-0878
    Keywords: GABA (gamma-aminobutyric acid) ; Pituitary ; Neuroendocrine regulation ; Immunocytochemistry ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The GABAergic innervation of the mouse pituitary, including the median eminence, was studied at light microscopic and ultrastructural levels by use of a pre-embedding immunocytochemical technique with antibodies directed against GABA. In the median eminence, a high density of GABA-immunoreactive fibers was found in the external layer where the GABAergic varicosities were frequently observed surrounding the blood vessels of the primary capillary plexus. In the internal and subependymal layers, only few fibers were immunoreactive. The intense labeling of the external layer was observed in the entire rostro-caudal extent of the median eminence. In the pituitary proper, a dense network of GABA-immunoreactive fibers was revealed throughout the neural and intermediate lobes, entering via the hypophyseal stalk. The anterior and tuberal lobes were devoid of any immunoreactivity. The GABA-immunoreactive terminals were characterized in the median eminence, and in the intermediate and posterior lobes at the electron-microscopic level. They contained small clear vesicles, occasionally associated with dense-core vesicles or neurosecretory granules. In the intermediate lobe they were seen to be in contact with the glandular cells. In the posterior lobe and in the median eminence, GABA-immunoreactive terminals were frequently located in the vicinity of blood vessels. These results further support the concept of a role of GABA in the regulation of hypophyseal functions, via the portal blood for the anterior lobe, directly on the cells in the intermediate lobe, and via axo-axonic mechanisms in the median eminence and posterior lobe.
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  • 31
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Isoactin ; Muscle ; Fetus ; Ontogeny ; Rat (Wistar-Kyoto)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Actin, a cytoskeletal and contractile protein, is expressed in six different isoforms that exhibit striking specificity. No studies have considered the muscle-specific actin expression in multiple organ systems in the intact fetus. Using a monoclonal antibody (B4) which reacts specifically with the isoactins of the smooth and skeletal muscle our immunohistochemical study examined whole fetal body sections to follow the development of actin expression throughout the last third of gestation in the Wistar-Kyoto rat. B4 staining was exclusively localized to muscle, confirming its high specificity and its usefulness for studying the ontogeny of muscle-specific isoactins. At 15 days of gestation, B4 staining was detected in the heart, the thoracic aorta and the skeletal muscle of the chest wall. The distribution and intensity of staining in the heart were initially higher than in the aorta or skeletal muscle and remained unchanged throughout the remainder of gestation, suggesting that the maturation of cardiac actin expression is well developed, although not fully completed before birth. Expression of muscle-specific actins in skeletal muscle was age-dependent and correlated with the maturational changes of muscle cell precursors. B4 staining in the fetal kidney was not apparent until day 20 of gestation and was localized to the inner cortical vessels. in association with the most mature nephrons, suggesting a centrifugal maturation of the intrarenal vasculature. The intensity of B4 staining in most tissues including bronchi, bowel, diaphragm, chest wall muscle and peripheral and pulmonary arteries increased by the end of gestation.
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  • 32
    ISSN: 1432-0878
    Keywords: Reissner's fiber ; Massa caudalis ; Subcommissural organ ; Spinal cord ; Central canal ; Immunocytochemistry ; Lectin histochemistry ; Lamprey ; Geotria australis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The subcommissural organ (SCO), Reissner's fiber (RF) and its massa caudalis of lamprey larvae (Geotria australis) were investigated immunocytochemically by use of an antiserum raised against bovine RF as primary antibody. The affinities of RF and massa caudalis for Ricinus communis agglutinin I (RCA) with and without previous acid hydrolysis, concanavalin A (Con A), wheat-germ agglutinin (WGA), aldehyde fuchsin, and PAS reaction were also studied. SCO and massa caudalis were strongly immunoreactive, whereas RF proper was distinctly negative. RF did not react with Con A and RCA. Only the periphery of RF was WGA-positive. RCA showed affinity for RF only after acid hydrolysis. RF was homogeneously stained by the aldehyde-fuchsin and PAS-methods. At variance with RF proper, the periphery of the massa caudalis reacted with RCA without previous acid hydrolysis, but its core was WGA-positive and reacted with RCA only after hydrolysis. It is suggested that (i) RF has a coat of glycoproteins containing sialic acid as terminal residue, whereas the massa caudalis possesses a coat with galactose as terminal residue; (ii) in RF proper and the massa caudalis the spatial arrangement of glycoproteins might be different. Routine transmission electron-microscopic observations indicate that in larvae of Geotria australis an open communication exists between the ampulla caudalis and blood capillaries via large cavities or lacunae.
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  • 33
    ISSN: 1432-0878
    Keywords: Mammary gland ; Tissue culture ; Collagen gel ; Electron microscopy ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Fragments of human breast epithelium, devoid of all stromal and basal lamina components, which maintain their in vivo topological organisation can be cultured for up to 28 days within a reconstituted rat-tail-derived collagen matrix. These organoids initially undergo a loss of structural and 3-dimensional organisation, typified by loss of lumina formed by epithelial cells, and myosin from myoepithelial cells. Their subsequent reorganisation is dependent on the presence of serum, insulin, hydrocortisone, and cholera toxin in tissue culture medium. After this preliminary phase, a reduction in the concentration of serum, insulin, hydrocortisone, and cholera toxin is necessary to allow the structural differentiation of epithelial and myoepithelial cells. The myoepithelial cells also regain their ability to produce the basal lamina component laminin. The use of bovine-dermal collagen as the matrix, rather than rat-tail-derived collagen is shown to result in more stable organisation and differentiation of the organoids. The successful use of single-cell pellets (derived by trypsinisation of the organoids) in place of organoids in such cultures illustrates that there is no requirement for pre-existing cell/ cell contact or topological organisation of cells prior to embedding within the collagen matrix.
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  • 34
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    Cell & tissue research 247 (1987), S. 457-459 
    ISSN: 1432-0878
    Keywords: Vomeronasal glands ; Autonomic innervation ; Electron microscopy ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The morphological evidence for a direct autonomic innervation of the mouse vomeronasal glands is presented. Axonal varicosities containing a few densecore vesicles and numerous clear vesicles (36–60 nm in diameter) make synaptic contacts with the secretory cells at the base of the glandular acini. The axonal presynaptic membrane is associated with a distinct dense material and it is separated from the secretory cell by a synaptic cleft of about 12–14 nm. At the postsynaptical level, coated vesicles can be found. Additional postsynaptical specializations have not been observed.
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  • 35
    ISSN: 1432-0878
    Keywords: Nasal mucosa ; Odorant-binding protein ; 2-isobutyl-3-methoxypyrazine ; Immunocytochemistry ; Cow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Polyclonal antibodies have been raised against purified bovine pyrazine-binding protein, a protein that binds the odorant 2-isobutyl-3-methoxypyrazine. These antibodies have been utilized in immunocytochemical experiments to localize the pyrazine-binding protein in bovine nasal mucosa. Tissue fragments, macroscopically identified as olfactory and respiratory mucosa, were fixed in Bouin's fluid and embedded in paraffin. Consecutive serial sections were processed for immunofluorescence studies and restained either with haematoxylin-eosin or with periodic acid Schiff-Alcian Blue. In both olfactory and respiratory mucosa, only seromucous tubulo-acinar glands were specifically labelled. These glands are located in the lamina propria underlying typical respiratory epithelium, even in those tissues that are macroscopically defined as olfactory mucosa.
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  • 36
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    Cell & tissue research 247 (1987), S. 465-468 
    ISSN: 1432-0878
    Keywords: Adenylate cyclase ; Cytochemistry ; Gastrulation ; Ultrastructure ; Chick embryo
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Adenylate cyclase activity was localized in various tissues of the early chick embryo using an ultrastructural histochemical technique. Reaction product was deposited on the lateral plasma membrane of all cells, but with a preferential localization at the apical terminal complex in the epiblast. There was no activity associated with the free surfaces of these or other cells in the embryo. Intracellular deposits were found in all cells associated with the endoplasmic reticulum, nuclear envelope and Golgi bodies. In the last organelle, the deposit was sometimes observed to be distributed through the stack in a non-uniform way, with the heaviest deposits occurring at the forming face. No clear difference could be detected between the cytochemical activity associated with cells in various regions of the embryo, or with embryos at different stages of early development.
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  • 37
    ISSN: 1432-0878
    Keywords: Gonadotropic cells ; LHRH analogue ; Pimo-zide ; Ultrastructure ; Goldfish ; Carassius auratus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effect of pimozide and an LHRH-analogue (LHRH-A) on gonadotropic cells of the goldfish pituitary gland were described qualitatively and quantitatively. A scale of four categories was devised to reflect various ultrastructural appearances of the cells. Experimental animals were divided into a control group, a group injected with LHRH-A alone, pimozide alone, and groups receiving these two substances in combination. Fish injected with the single substance were killed 12 h after injection while the groups receiving the combined treatments were killed at 4, 12 and 48 h. Serum levels of gonadotropin measured by radioimmunoassay were used to indicate whether an increase in hormone release had occurred. An immunocytochemical technique, the protein A-gold procedure, assured that the cells studied were gonadotropes. The control group showed variation in the profiles of gonadotropic cells. The single treatment groups showed some increase in secretory inclusions. At 4 h after injection the combined treatment caused a significant increase in hormone granules; at 12 and 48 h there was a gradual decrease in content of secretory products, and an increase in vacuolization. The results indicate that the combined pimozide and LHRH-A treatment stimulated gonadotropin production as well as release.
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  • 38
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    Cell & tissue research 248 (1987), S. 49-54 
    ISSN: 1432-0878
    Keywords: Enterochromaffin cells ; Serotonin ; Duodenum ; Immunocytochemistry ; Electron microscopy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Enterochromaffin cells of the rat duodenum have been studied immunocytochemically by use of a specific antiserum to serotonin. At the light-microscopic level serotonin immunoreactivity was observed in enterochromaffin cells located in the epithelium of the duodenal mucosa. Most of the serotonin-immunoreactive material was localized to the basal portion of the enterochromaffin cells, but small amounts of immunoreactive material were regularly observed in the apical portion. At the electron-microscopic level serotonin immunoreactivity in enterochromaffin cells was found to be concentrated over the dense cores of the cytoplasmic granules. The majority of these granules was located in the basal cytoplasm of the enterochromaffin cells, but serotonin-immunoreactive granules were also observed in the apical cytoplasm immediately beneath the microvilli. These observations indicate that duodenal enterochromaffin cells are bipolar and that they secrete serotonin both basally, to the circulation, and apically, to the gut lumen. Rat duodenal enterochromaffin cells thus appear to have an exocrine as well as an endocrine function.
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  • 39
    ISSN: 1432-0878
    Keywords: LH-cells ; Prolactin cells ; Immunocytochemistry ; Estrogen ; Autoradiography ; Guinea pig ; Hamster ; Gerbil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Nuclear uptake and retention of3H-estradiol by luteinizing hormone (LH) and prolactin (PRL) cells was examined in three species of rodents (guinea pigs, hamsters and gerbils) using the combined techniques of immunocyto-chemistry and autoradiography. Castrated animals were injected with3H-estradiol and decapitated 1.5 h later. The pituitary glands were processed for thaw-mount autoradiography followed by conventional immunocytochemical staining for LH and PRL.3H-estradiol accumulated in more than 80% of the anterior pituitary cells in the gerbils, while only 33 and 22% of the cells accumulated3H-estradiol in the hamsters and guinea pigs, respectively. A varying percentage of immunoreactive LH and PRL cells in all three species were found also to contain binding sites for estradiol. Some LH and PRL cells in hamsters and guinea pigs and only some in PRL cells of gerbils were found to be devoid of grains. Quantitative analysis revealed that the number of grains per nucleus differed considerably from cell to cell. LH cells of guinea pigs accumulated much larger amounts of3H-estradiol than did the PRL cells, while the LH cells in the hamsters and gerbils accumulated only slightly more3H-estradiol than the PRL cells. These results confirm the previous observations in rats and baboons that demonstrated tremendous species differences in percentage of cells in the anterior pituitary gland that accumulated3H-estradiol. Also, these data suggest that there are functionally heterogeneous cell types among the LH and PRL cells in hamsters, guinea pigs and gerbils as has been previously demonstrated in rats and baboons.
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  • 40
    ISSN: 1432-0878
    Keywords: Retina ; Amacrine cells ; Immunocytochemistry ; Neurotensin ; Somatostatin ; Color vision ; Teleosts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neurotensin- and somatostatin-like immunoreactivities were localized by pre-embedding techniques in retinal whole-mounts and radial sections of a monochromatic glass catfish (Kryptopterus bicirrhis), a dichromatic cichlid species (Aequidens pulcher), and the tetrachromatic roach (Rutilus rutilus). Both neuropeptides were observed in perikarya and processes of amacrine cells. For a precise identification of cell types, tangential and radial views were correlated with Golgiimpregnated material. The dendritic pattern defining the morphological subtype of amacrine cells was determined by the given neuropeptide or by the species-specific degree of complexity of retinal structure and function. Neurotensin-like immunoreactivity was localized in amacrine cells of intermediate size, radial symmetry and dendrites with numerous varicosities; they were monostratified in sublayer 3 of the inner plexiform layer. This cell type was common to all three species. In the mono and dichromatic retinas, a single type of amacrine cell with somatostatinlike immunoreactivity was found with radially oriented, varicose dendrites in sublayer 5. In the tetrachromatic roach retina, two somatostatin-positive amacrine cell types were found with very different patterns of ramification; furthermore, both of these types occurred in more than one sublayer. Possible functional implications for color vision of neuropeptide-specific amacrine cells with uniform morphology in all three species and those with a more varied morphology in the tetrachromatic roach are discussed.
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  • 41
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    Cell & tissue research 248 (1987), S. 675-682 
    ISSN: 1432-0878
    Keywords: Molluscan muscles ; Innervation ; Chromatophores ; Cephalopods ; Electron microscopy ; Eledone cirrhosa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cephalopod chromatophores are made of a central pigment cell surrounded by 10 to 20 radially arranged muscle fibres under direct nervous control. Innervation of these muscle fibres was studied with anterograde cobalt fills of peripheral nerve bundles and light and electron microscopy. Individual axons branch repeatedly to innervate the muscles of chromatophores scattered over several millimeters. Axons contained in several dermal nerves converge to innervate the same chromatophores. Among the chromaophores, axons were found running either singly or in small bundles, often accompanied by sheath cells. Single chromatophore muscles were innervated by at least one axon running across or along its length. Since nerves terminating on chromatophore muscles are very rare, neuromuscular contact seems to be made “en passant”. Varicosities of the axons apposed to the muscles are thought to be presynaptic sites. However, morphological differentiations of the pre-or post-synaptic membranes were not visible. Two types of innervating processes were found containing either electron-clear or a mixture of electron-clear and dark-core synaptic vesicles.
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  • 42
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    Cell & tissue research 248 (1987), S. 699-702 
    ISSN: 1432-0878
    Keywords: Albumin ; Ovarian follicles ; Granulosa cells ; Immunocytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The purpose of this study was to investigate whether albumin (Alb) can be detected in ovarian rat granulosa cells. Using immunocytochemistry and morphometrics, the percentages of Alb-positive follicles (follicle-index), of Alb-positive granulosa cells (granulosa-index), and of strongly reacting follicles (intensity-index) were evaluated in intact and regressing follicles of different diameter groups during different stages of the estrous cycle. In intact follicles, the follicle- and the granulosa-index increased from small-sized to large-sized follicles. Although the follicle-index did not change in any group during the stages of the estrous cycle, the granulosa-index was higher during proestrus than during the other stages. Intact follicles showed a stronger immunoreactivity than regressing follicles throughout the stages of the estrous cycle. Thus, Alb may be a requirement for the control of follicle growth in fertile rats. This Alb function may be attributable to Alb binding to specific cell-membrane components followed by the intracellular uptake of Alb-bound substances.
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  • 43
    ISSN: 1432-0878
    Keywords: Endolymphatic sac ; Calcium carbonate ; Otoconia ; Ultrastructure ; Frog (Hyla arborea japonica)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The endolymphatic sac of the tree frog and its crystals were observed by light- and electron microscopy. Scanning electron microscopy revealed that the crystals have a faceted body and two pointed ends. Light- and transmission electron microscopy revealed that the endolymphatic sac is composed of many small chambers. In their lumina, numerous “ghosts” of crystals that resulted from decalcification were observed. The ghosts were demarcated by a linear dense material or embedded in a flocculent substance. The epithelium of the endolymphatic sac is simple squamous or cuboidal and peculiar cytoplasmic granules are found in most cells. The granules are surrounded by a limiting membrane and have varying electron density. Some granules contain a core and/or tubular structures. Vacuoles containing large ghosts are also found in the epithelial cells. These ghosts were quite similar to those in the lumen and sometimes coexist with cell debris. The fine structure of the endolymphatic sac and its crystals is discussed.
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  • 44
    ISSN: 1432-0878
    Keywords: Epididymis ; Immunogold technique ; Ultrastructure ; Protein secretion ; Antagglutinin ; Boar
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Antagglutinin, a specific protein synthesized by the boar epididymis, was localized by an ultrastructural immunogold-labeling procedure in the principal cells of the three regions of the caput epididymidis, most notably at the sites of synthesis and secretion. The intensity of the reaction was variable in the three epididymal zones. Labeling was of low intensity in the proximal and middle caput, except in the granules of the latter. These granular “storage sites” did not correspond to typical secretory granules but appeared to be intracellular sites of degradation of this protein. In the distal caput, which was devoid of these granules, intense secretory activity for antagglutinin was detected. Few gold particles were localized in the RER profiles but labeling was detected in the Golgi zone, in numerous dense vesicles, in structures distributed between the Golgi zone and the apex of the cell, and in the epididymal lumen. This study has enabled us to visualize immunocytochemically antagglutinin along its intracellular secretory pathway, i.e. at the site of its synthesis, during its passage via the Golgi zone, and its intracellular transport to the lumen.
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  • 45
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    Cell & tissue research 249 (1987), S. 111-116 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Serotonin ; Mytilus galloprovincialis ; pedal ganglion ; Invertebrate nervous system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Serotonergic cell bodies and fibers were identified in the pedal ganglia of Mytilus gattoprovincialis with a serum raised against serotonin and the unlabelled peroxidase-antiperoxidase pre- and post-embedding methods. Examination of Vibratome and serial semithin sections showed that most reactive perikarya are located in the ganglionic cortex, being mainly concentrated at the medial aspect of the postero-dorsal portion of the ganglia. Immunoreactive fibers form a dense network in the neuropil, extend throughout the commissure and run parallel in the nerves and connective tracts. The morphology of serotonin-positive cells compared with that of Golgi-impregnated neurons allows the identification of a main population of unipolar, probably projecting neurons and of smaller multipolar cells likely representing local circuit elements. The ultrastructure of labelled neurons is comparable to that of serotonergic cells described in both vertebrate and invertebrate nervous systems.
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  • 46
    ISSN: 1432-0878
    Keywords: Weibel-Palade body ; Human umbilical vein ; Factor VIII-related antigen ; Histamine ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The present study was done to elucidate the biological significance of the Weibel-Palade body of human umbilical vein endothelial cells. Quantitative determinations of these endothelial-specific granules throughout pregnancy revealed that their numbers and size per cell profile were maintained at low levels from 12 to 19 weeks of gestation; then both rapidly increased from 33 weeks to full term. This increase coincided with the development of the rough endoplasmic reticulum and an increase in the number of endothelial cell pinocytotic vesicles. Light-microscopic peroxidase anti-peroxidase and electron-microscopic protein A-gold techniques provided evidence that factor VIII-related antigen was localized in the Weibel-Palade bodies. Furthermore, in vitro treatment of incubated umbilical vein tissue with compound 48/80, a histamine releaser, induced degranulation of Weibel-Palade bodies from the endothelium. The present study indicates that Weibel-Palade bodies are storage sites of both histamine and factor VIII-related antigen and have an important role in the obliteration of this vessel.
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  • 47
    ISSN: 1432-0878
    Keywords: Pigment-dispersing hormone ; Ultrastructural histochemistry ; Immunocytochemistry ; Neurosecretion ; Crustaceans (Carcinus maenas, Cancer pagurus, Uca pugilator, Orconectes limosus)
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary A high-liter antiserum has been obtained from two rabbits immunized with a glutaraldehyde conjugate of synthetic pigment-dispersing hormone (PDH) from Uca pugilator and bovine thyroglobulin. The antiserum blocked melanophore-dispersing activity of the peptide in vivo. In sinus glands (SG) of Carcinus maenas, Cancer pagurus, Uca pugilator and Orconectes limosus, electron-microscopic immunocytochemistry revealed sparsely distributed axon endings containing a distinct PDH-immunoreactive type of neurosecretory granules (diameter 90–130 nm). Exocytotic figures indicating release of the content of these granules into hemolymph lacunae were occasionally observed. Preservation of fine structure and antigenicity of the PDH granules were markedly dependent on the fixation procedure used. A preliminary experiment with C. maenas showed that preterminal axon dilatations near the basal lamina seemed to accumulate PDH-granules when animals were kept in complete darkness for three days. Immunodot blotting of fractions after high pressure liquid chromatography (HPLC) of extracts from SGs of C. maenas and O. limosus revealed a strongly immunoreactive substance at a retention time very similar to those of synthetic PDHs of Uca pugilator and Pandalus borealis. It is also coincident with a zone of biological activity. Thus, the antigen demonstrated by immunocytochemistry is identical or very similar to one of the known PDHs.
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  • 48
    ISSN: 1432-0878
    Keywords: Pigment-dispersing hormone (crustaceans) ; FMRFamide ; Immunocytochemistry ; Neurosecretion ; Crustaceans: Carcinus maenas, Orconectes limosus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By use of a new antiserum, raised against synthetic pigment-dispersing hormone (PDH) from Uca pugilator, immunoreactive structures were studied at the light-microscopic level in the eyestalk ganglia of Carcinus maenas and Orconectes limosus. PDH-reactivity was mainly found in two types of neurons that were located between the medulla interna (MI) and the medulla terminalis (MT) in both species. Several additional perikarya were located in the distal part of the MI in O. limosus. In C. maenas, two to three PDH-positive perikarya were found in the region of the X-organ (XO) in the MT. Processes from single and clustered cells could be traced into all medullae of the eyestalk. Axons from the immunoreactive perikarya running between MI and MT form a larger tract that traverses the MT. Fibers from this tract give rise to extensive arborizations and plexuses throughout the proximal MT. A plexus containing very fine fibers is located at the surface of the MT in a position distal to the XO-area of C. maenas only. The proximal plexus also receives PDH-positive fibers through the optic nerve. PDH-perikarya in the cerebral ganglion may also project into the more distal regions of the eyestalk. Distal projections of the perikarya between the MI and MT consist of several branches. Most of these are directed toward the MI and ME (medulla externa) wherein they form highly organized, layered plexuses. One branch was traced into the principal neurohemal organ, the sinus gland (SG). In the SG, the tract gives off arborizations and neurosecretory terminals. It then proceeds in a proximal direction out of the SG, adjacent to the MT. Its further course could not be elucidated. The lamina ganglionaris (LG) receives PDH-fibers from the ME and fine processes from small perikarya located in close association with the LG in the distal part of the first optic chiasma. The architecture of PDH-positive elements was similar in both C. maenas and O. limosus. The distribution of these structures suggests that PDH is not only a neurohormone but may, in addition, have a role as a neurotransmitter or modulator. Immunostaining of successive sections with an FMRF-amide antiserum revealed co-localization of FMRFamideand PDH-immunoreactivities in most, but not all PDH-containing perikarya and fibers. The axonal branch leading to the SG and the SG proper were devoid of FMRFamide immunoreactivity.
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  • 49
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    Cell & tissue research 247 (1987), S. 215-225 
    ISSN: 1432-0878
    Keywords: Kidney ; Endoplasmic reticulum ; Ultrastructure ; Membrane transport ; Metabolism ; Rat
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    Topics: Biology , Medicine
    Notes: Summary The spatial organization of endoplasmic reticulum (ER) was examined in all segments of rat nephron. Tissues were fixed with glutaraldehyde, impregnated “en bloc” with osmium tetroxide, prepared for and examined by standard (80–100 kV) and high voltage (1 mEV) transmission electron microscopy. In all proximal tubule cells, ER forms a continuous and extensive network of canaliculi and abundant fenestrated saccules which surround mitochondria and cytoplasmic bodies; the cage-like structure of the fenestrated saccules was most evident around the spherical mitochondria of the S3 segment. In the cells of the distal straight and convoluted tubules, the network consists mostly of canaliculi with rare non-fenestrated saccules. The ER network of canaliculi is particularly rich in intercalated cells, in contrast with its rudimentary appearance in the adjacent principal cells of the collecting tubule. In fact, in these cells there are few isolated ER cisternae and they are rarely impregnated. The nuclear envelope is well impregnated in most cells throughout the various segments. Segmental variations in ER organization and its relative abundance are most likely related to the well, established functional heterogeneity of the nephron segments. Moreover, the extensive and unique organization among mitochondria, ER and the basolateral membrane suggests that these three organelles function as a unit which is related to active electrolyte transport. In addition, because of its transepithelial organization, ER may well constitute a transcellular pathway for molecules.
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  • 50
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    Cell & tissue research 247 (1987), S. 249-258 
    ISSN: 1432-0878
    Keywords: Lactoferrin ; Electron microscope ; Immunocytochemistry ; Neutrophils ; Morphometry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The subcellular localization of lactoferrin in human neutrophils was studied by an electron-microscopic immunoperoxidase method. This molecule was detected in small granules of blood polymorphonuclear leukocytes. A morphometrical analysis showed that there was no significant difference in the mean size between lactoferrin-positive and myeloperoxidase-negative granules. In contrast, the mean size of myeloperoxidase-positive granules was significantly larger than that of lactoferrin-positive granules. This indicates that lactoferrin is contained in the myeloperoxidase-negative, secondary, granules of human neutrophils. In immature bone marrow mononuclear neutrophils, lactoferrin was present in cytoplasmic granules of somewhat larger size than lactoferrin-positive granules of polymorphonuclear leucocytes. A morphometrical study showed that the mean size of lactoferrin-positive granules was significantly greater in immature bone marrow cells than in polymorphonuclear leucocytes. This indicates that lactoferrin-positive granules decrease in size as the cells mature. Besides cytoplasmic granules, lactoferrin was demonstrated in the Golgi complex and a part of the rough endoplasmic reticulum of immature bone marrow neutrophils, probably myelocytes and early metamyelocytes. These results show that lactoferrin is synthesized and packed into secondary granules in immature bone marrow neutrophils and therefore that the secondary granules are a type of secretory granule.
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  • 51
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    Cell & tissue research 247 (1987), S. 259-265 
    ISSN: 1432-0878
    Keywords: Epidermis ; Acidophilic cell ; Histochemistry ; Ultrastructure ; Salmo trutta ; Salvelinus alpinus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Sacciform cells containing an acidophilic, proteinaceous secretion, were identified in the epidermis of the brown trout and Arctic char. This cell type increased in number during the chronic stages of infestation by the ectoparasitic flagellate, Ichthyobodo sp., in immature brown trout, and decreased during sexual maturation in male brown trout and char. It is suggested that the salmonid sacciform cell produces a secretion which protects the fish against infestation or damage by skin parasites.
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  • 52
    ISSN: 1432-0878
    Keywords: Nucleus preopticus ; Nucleus lateralis tuberis ; Corticotropin-releasing factor (CRF) ; Arginine vasotocin (AVT) ; Immunocytochemistry ; Catostomus commersoni (Teleostei)
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    Notes: Summary The peroxidase-antiperoxidase immunocytochemical procedure was used to study the distribution of ovine corticotropin-releasing factor (CRF) and arginine vasotocin (AVT) immunoreactivities sequentially in the same sections or in adjacent sections of the brain and pituitary of Catostomus commersoni. It was found that all CRF-immunoreactive (IR) neurons in the nucleus preopticus (NPO) also contained AVT immunoreactivity. Co-localization of both immunoreactivities was also observed in fibres forming the preoptic-pituitary tract and in the neurohypophyseal digitations, the IR-CRF and IR-AVT fibres projecting mainly to the neurointermediate lobe (NIL) of the pituitary. An additional population of exclusively IR-AVT neurons and fibres in the NPO, preoptic-pituitary tract and NIL was also observed. Exclusive CRF-immunostaining was found in neurons of the nucleus lateralis tuberis (NLT), in fibres distributed in some diencephalic nuclei and in the neurohypophyseal digitations in the region of the rostral pars distalis (RPD). These results suggest (i) that CRF- and AVT-like substances, present in NIL fibres (probably originating in the NPO), may have an integrated role in the release of the cell products from the pars intermedia, and (ii) that the control of corticotrops in the rostral pars distalis, innervated exclusively by IR-CRF fibres (probably originating in the NLT), does not require a simultaneous presence of CRF- and AVT-like substances.
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  • 53
    ISSN: 1432-0878
    Keywords: Pancreatic polypeptide ; FMRFamide ; Retina ; Immunocytochemistry ; Carassius auratus
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    Notes: Summary Immunocytochemical studies were conducted on goldfish to determine whether a retinal efferent fiber system, immunoreactive to the tetrapeptide Phe-Met-Arg-Phe-NH2 (FMRFamide), might contain instead a substance similar to one of the 36-amino acid pancreatic polypeptides, the C-terminus of which is similar to FMRFamide. Our results demonstrate the presence of two separate peptidergic systems, one containing FMRFamide-like, and the other pancreatic polypeptide-like peptides. Antisera to FMRFamide reveal the efferent fibers, whose axons exit the optic nerve and terminate in layer 1 of the inner plexiform layer, as previously described. Antisera to porcine neuropeptide Y, and to avian and bovine pancreatic polypeptides label a sparse population of putative amacrine cell bodies and a dense fiber plexus in layers 1, 3, and 5 of the inner plexiform layer. Based on intensity of staining, this amacrine cell peptide appears to be most similar to neuropeptide-Y. Radioimmunoassay and immunocytochemical staining of retinas in which the efferent fiber peptide was depleted by optic nerve crush confirm in large part the observation that the two peptide systems are distinct. However, there is some cross-recognition of the FMRFamide-like tissue antigen by pancreatic polypeptide antibodies. Double-label studies with antisera to tyrosine hydroxylase and neuropeptide-Y indicate that the pancreatic polypeptide antigen is not co-localized with catecholamines.
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  • 54
    ISSN: 1432-0878
    Keywords: Calcergy ; Calciphylaxis ; Cutaneous calcinosis ; Experimental calcification ; Ultrastructure ; Rat
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    Notes: Summary Skin calcification induced by topical calciphylaxis was provoked by a subcutaneous injection of iron chloride in rats previously sensitized by dihydrotachysterol. A cutaneous topical calcergy was induced by an injection of potassium permanganate. An electron-microscopical study of the long-term evolution of both these models of calcification was made. After the initial stages, mineralization of the connective tissue continued by a secondary nucleation process without matrix vesicles. The mineral composed of needle-like structures, apatite in nature, was mainly deposited between and around collagen fibrils, and showed various arrangements in calcified plaques. Intrafibrillar calcification was rarely observed and appeared only in the later stages. The extension of calcified deposits then stopped. Finally, there was a fragmentation of the mineralized area which was progressively surrounded by uncalcified collagen fibrils. A demineralization process, caused by cells such as macrophages and multinucleated giant cells, rather than a resorption of the calcified deposits, was noted. It is important to emphasize that, in both models of ectopic calcification, an evolution toward ectopic ossification was never observed, which is perhaps due to the absence of extensive resorption mechanisms.
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  • 55
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    Cell & tissue research 248 (1987), S. 1-24 
    ISSN: 1432-0878
    Keywords: Insect nervous system ; Protocerebrum ; Suboesophageal ganglion ; GABA ; Immunocytochemistry ; Manduca sexta
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    Notes: Summary We have used specific antisera against protein-conjugatedγ-aminobutyric acid (GABA) in immunocytochemical preparations to investigate the distribution of putatively GABAergic neurons in the brain and suboesophageal ganglion of the sphinx mothManduca sexta. About 20000 neurons per brain hemisphere exhibit GABA-immunoreactivity. Most of these are optic-lobe interneurons, especially morphologically centrifugal neurons of the lamina and tangential neurons that innervate the medulla or the lobula complex. Many GABA-immunoreactive neurons, among them giant fibers of the lobula plate, project into the median protocerebrum. Among prominent GABA-immunoreactive neurons of the median protocerebrum are about 150 putatively negative-feedback fibers of the mushroom body, innervating both the calyces and lobes, and a group of large, fan-shaped neurons of the lower division of the central body. Several commissures in the supra- and suboesophageal ganglion exhibit GABA-immunoreactivity. In the suboesophageal ganglion, a group of contralaterally descending neurons shows GABA-like immunoreactivity. The frontal ganglion is innervated by immunoreactive processes from the tritocerebrum but does not contain GABA-immunoreactive somata. With few exceptions the brain nerves do not contain GABA-immunoreactive fibers.
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  • 56
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    Cell & tissue research 248 (1987), S. 393-398 
    ISSN: 1432-0878
    Keywords: Adrenaline ; Phenylethanolamine-N-methyl transferase ; Immunocytochemistry ; Electron microscopy ; Central nervous system ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The fine structure of adrenergic axon terminals was examined in the paraventricular nucleus of the thalamus (PNT) and in the hypothalamic arcuate nucleus-median eminence (ARC-ME) complex by use of phenylethanolamine-N-methyl transferase (PNMT) immunocytochemistry. In the PNT, immunoreactive terminals formed a dense and well-circumscribed plexus. In the ARC, labeled varicosities were less numerous and more evenly distributed. In the ME, they were scarce and confined to the inner zone. In all these areas, the diameter of immunoreactive varicosities ranged between 0.2 and 1.3 μm; in the ME and in the transitional zone between the ARC and the ME, a population of larger boutons (〉2 μm) was also visible. All immunoreactive varicosities exhibited densely packed small, clear vesicles associated with a few large granular vesicles. In the PNT and the ARC, but not in the ME, they formed synaptic contacts with dendritic elements and were occasionally apposed to neuronal cell bodies. These axo-somatic appositions showed no junctional specializations. In the ME and transitional zone, immunoreactive terminals were frequently juxtaposed to, and occasionally established differentiated synaptic contacts with, tanycytes. These data support a transmitter role for adrenaline in the diencephalon and suggest that adrenaline plays a role in hypothalamo-hypophysiotropic regulation through interactions with neural and glial elements.
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  • 57
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    Cell & tissue research 248 (1987), S. 409-415 
    ISSN: 1432-0878
    Keywords: Bone matrix ; Osteonectin ; Osteoblasts ; Immunocytochemistry ; Differentiation ; Human
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    Topics: Biology , Medicine
    Notes: Summary Bone matrix consists of type-I collagen and noncollagenous proteins. The latter represent only 10% of its total protein content. Since type-I collagen is also present in various other connective tissue sites (e.g., skin) it cannot be considered as bone specific. Among the non-collagenous components osteonectin — a 32 kilodalton (KD) glycoprotein linking mineral to collagen fibrils — is thought to be bone specific due to its biochemical properties. In the present study various skeletal and non-skeletal tissues were investigated for the presence of osteonectin by means of immunocytochemical methods. Two polyclonal antibodies against human and bovine osteonectin were applied. Immunocytochemically, osteonectin could be demonstrated in active osteoblasts and osteoprogenitor cells as well as in young osteocytes, while aged, quiescent osteocytes did not contain the protein, suggesting that the protein is a marker of the osteoblastic functional differentiation of bone cells. Osteonectin was absent in all non-skeletal tissues with the exception of chondrocytes in so-called mineralizing chondroid bone.
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  • 58
    ISSN: 1432-0878
    Keywords: Serotonin ; Immunocytochemistry ; Central nervous system ; Annelids ; Lumbricus terrestris
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    Topics: Biology , Medicine
    Notes: Summary The distribution patterns of serotonin-immunoreactive somata in the cerebral and subpharyngeal ganglion, and in the head and tail ganglia of the nerve cord of Lumbricus terrestris are described from whole-mount preparations. A small number of serotonin-immunoreactive neurons occurs in the cerebral ganglion, in contrast to the large population of serotonin-immunoreactive neurons that exists in all parts of the ventral nerve cord. From the arrangement of serotonin-immunoreactive somata in the subpharyngeal ganglion, we suggest that this ganglion arises from the fusion of two primordial ganglia. In head and tail ganglia, the distribution of serotonin-immunoreactive somata resembles that in midbody segments. Segmental variations in the pattern and number of serotonin-immunoreactive somata in the different body regions are discussed on the background of known developmental mechanisms that result in metameric neuronal populations in annelids and arthropods.
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  • 59
    ISSN: 1432-0878
    Keywords: Kidney ; Collecting duct ; Immunocytochemistry ; Intercalated cell ; Rabbit
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    Notes: Summary A monoclonal antibody against an antigen (PCD2) derived from the rabbit renal papilla recognized principal and intercalated cells of the collecting duct system in the adult rabbit kidney. Intercalated cells were heterogeneous in the connecting tubule and the cortical collecting duct, where immunoreactive and unreactive cells were shown to coexist. In the outer medullary collecting duct, all intercalated cells exhibited PCD2-immunoreactivity. Connecting tubule cells proper were not recognized by the antibody, whereas all principal cells of the collecting duct revealed specific immunoreactivity. The immunocytochemical heterogeneity of the intercalated cells is discussed in terms of a functional heterogeneity. Cytologically, the immunogold labeling of principal and intercalated cells was shown to occur along the plasmalemma, in the intracellular membrane structures and along the Golgi transport route. This pattern suggests that the antigenic determinant, which is ubiquitous in both principal and reactive intercalated cells, belongs to a membrane protein.
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  • 60
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    Cell & tissue research 249 (1987), S. 641-646 
    ISSN: 1432-0878
    Keywords: Pineal organ ; Development ; Photoreceptors ; Retinal S-antigen ; α-Transducin ; Opsin ; Serotonin ; Immunocytochemistry ; Teleosts (Salmo salar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The initial appearance of S-antigen, α-transducin, opsin and 5-HT during embryogenesis of the pineal organ and retina was studied by means of immunocytochemistry in the Atlantic salmon, Salmo salar L. The presence of these substances may be taken as a good indication of photoreceptor differentiation; α-transducin and S-antigen are involved in the phototransduction process, opsin is the proteinaceous component of the photopigment rhodopsin, and 5-HT is a neurotransmitter or neurohormone produced by pineal photoreceptors. Two days after the retinal pigment layer became visible in the eggs, the outer segments of a few pineal photosensory cells showed immunoreactivity to opsin and α-transducin. At the same time S-antigen and serotonin were present in pineal cells of the photoreceptor type. The number of immunoreactive cells in the pineal organ increased up to hatching. In the differentiating retina of the salmon, no immunoreactivity to antibodies raised against the mentioned substances was detectable until after hatching. These results indicate that in ontogeny the developing pineal organ of the salmon embryo has the ability to perceive light information much earlier than the retina.
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  • 61
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    Cell & tissue research 248 (1987), S. 169-180 
    ISSN: 1432-0878
    Keywords: M-band structure ; Fibre type ; Skeletal muscle ; Cryo ultramicrotomy ; Electron microscopy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The fine structure of the M-band in soleus (SOL) and extensor digitorum longus (EDL) muscles in newborn and four-week-old rats was studied using electron-microscopic techniques. In newborn rats, all myotubes and fibres in both muscles had an identical myofibrillar appearance. A five-line M-band pattern was seen in longitudinal sections and distinct M-bridges in cross-sections. The Z-discs were of medium width. On the other hand, in four-week-old rats, different muscle fibre types were observed on the basis of their myofibrillar pattern. In SOL two fibre types were distinguished in longitudinal sections. One had a four-line M-band pattern and very broad Z-discs, whereas the other type had five lines in the M-band and broad Z-discs. In EDL, three different myofibrillar patterns were observed. The M-bands were composed of three, four or five lines. Fibres had either thin, broad or medium Z-disc widths, respectively. In cross-sections of the SOL muscle one group of fibres showed indistinct M-bridges, whereas distinct M-bridges were seen in the other fibres and in all observed EDL muscle fibres. We conclude that initially there seems to be a single intrinsic program for M-band genesis; this program becomes modified upon the induction of functionally differentiated fibres.
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  • 62
    ISSN: 1432-0878
    Keywords: Human growth hormone-releasing factor (hGRF) ; Paraventricular nucleus ; Immunocytochemistry ; Electron microscopy ; Synapses ; Guinea pig
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    Notes: Summary By means of a preembedding immuno-electronmicroscopic technique, a large number of nerve endings containing a substance related to human growth hormonereleasing factor (hGRF) have been demonstrated in the paraventricular nucleus of the guinea pig. They made synaptic contacts primarily with dendritic shafts: 80% of these contacts were symmetrical. The immunoprecipitate was located mainly in large granules and around small clear vesicles. These findings suggest that a peptide related to hGRF may play a role in neural communication in the paraventricular nucleus.
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  • 63
    ISSN: 1432-0878
    Keywords: Serotonin ; Dopamin ; Immunocytochemistry ; Terminal ganglion ; Acheta domestica
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary The distribution and ultrastructure of serotonin- and dopamine-immunoreactive (5-HTi and DAi) neurones have been investigated in the terminal ganglion of the cricket, Acheta domestica, using a pre-embedding chopper technique. Special attention has been paid to the immunoreactive structures in the neuropil. 5-HTi structures are extensively distributed and densely packed throughout the 5 neuromeres of the terminal ganglion and originate from several interneurones and efferent neurones. In contrast, DAi fibres are distributed sparsely although they extend to all neuromeres of the ganglion and originate from 6 interneurons only. For both 5-HTi and DAi neurones characteristic axonal projections and branching patterns can be distinguished. The 5-HTi axons exhibit rich varicose arborizations, whereas DAi neurones possess fewer varicosities in the neuropil. Electron microscopy shows that 5-HTi varicosities contain small (∼ 60 nm) and large (∼ 100 nm) agranular vesicles, and large (∼ 100 nm) granular vesicles, whereas in DAi varicosities small (∼ 60 nm) agranular and large (∼ 100 nm) granular vesicles are seen. Both 5-HTi and DAi varicosities form synaptic contacts. We conclude that both serotonin and dopamine may be used as neurotransmitters in the terminal ganglion of the cricket.
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  • 64
    ISSN: 1432-0878
    Keywords: Sublingual gland ; Ultrastructure ; Golgi apparatus ; Rapid-freezing ; Freeze-substitution ; Gerbil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We compare the ultrastructure of the gerbil sublingual gland as seen after cryofixation followed by substitution with osmium tetroxide, with the more familiar appearance of material processed by glutaraldehyde-osmium chemical fixation. After primary cryofixation of fresh salivary glands, the nuclei of the mucous cells are found to be spherical in shape and, rather than being displaced toward the cell base, occupy a nearly central position in the cytoplasm, even in the storage phase of the secretory cycle. The mucous secretory granules are seen as membrane-limited inclusions, only rarely partially fused to each other. In both mucous and serous cells the Golgi cisterns have numerous large fenestrae which are aligned to form cytoplasmic channels which extend across the stack.
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  • 65
    ISSN: 1432-0878
    Keywords: Serotonin ; Neuropeptide Y ; Suprachiasmatic nucleus ; Radioautography ; Immunocytochemistry ; Rat
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    Notes: Summary The morphological substrate of putative serotonin (5-HT)/neuropeptide Y (NPY) interactions in thé suprachiasmatic nucleus (SCN) was investigated by combined radioautography and immunocytochemistry after intraventricular administration of (3H)5-HT in the rat. In the ventral portion of the SCN, the distribution of (3H)5-HT uptake sites overlapped closely the NPY-immunoreactive terminals. Previous investigations have shown that the dense 5-HT and NPY innervations of the SCN originate in different structures, i.e., the midbrain raphe nuclei and the ventral lateral geniculate nucleus, respectively. Accordingly, in the present study, destruction of 5-HT afferents by 5,7-dihydroxytryptamine was not found to induce any modification in NPY staining and, in ultrastructural immuno-radioautographic preparations, two distinct pools of axonal varicosities could be identified. Both 5-HT and NPY terminals established morphologically defined synaptic junctions, sometimes on the same neuronal target. Some cases of direct axo-axonic appositions between the two types of terminals were also encountered. These data constitute additional criteria for characterizing the cytological basis of the multiple transmitter interactions presumably involved in the function of the SCN as a central regulator of circadian biological rhythms.
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  • 66
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    Cell & tissue research 247 (1987), S. 167-177 
    ISSN: 1432-0878
    Keywords: Arteriovenous anastomoses ; Autonomic innervation ; Synaptic vesicles ; Histochemistry ; Electron microscopy ; Tongue ; Dog
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The innervation of the arteriovenous anastomoses in the dog tongue has been investigated. At the lightmicroscopic level, the vessels were found to be densely supplied with adrenergic and AChE-positive nerve plexuses and less densely with the quinacrine-binding nerve plexus. At the electron-microscopic level, at least two apparently different types of axon profiles were identified: 1) Small vesicle-containing axons, characterized by many small granular vesicles, variable numbers of small clear vesicles and large granular vesicles. Storage of endogenous amines and uptake of exogenous amines into most small granular vesicles and many large granular vesicles was demonstrated. These axons stained only lightly with reaction products for AChE activity and thus seemed to be adrenergic in nature. Some axons contained numerous large granular vesicles, whose cores occasionally stained with uranyl ions; this suggests a co-localization of ATP or peptides as neurotransmitters. 2) Small granular vesicle-free axons, containing small clear vesicles and large granular vesicles in variable ratio. Most cores of these large granular vesicles were heavily stained with uranyl ions. No storage or uptake of amine into the synaptic vesicles was detected. Some axons appeared to be typically cholinergic, some, typically non-adrenergic, noncholinergic, and the rest, intermediate between the two. All axons stained heavily with reaction products for AChE activity, suggesting their cholinergic nature.
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    Cell & tissue research 247 (1987), S. 573-582 
    ISSN: 1432-0878
    Keywords: Seminal vesicle ; Histology ; Histochemistry ; Electron microscopy ; Physiology ; Clarias gariepinus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The seminal vesicle of the African catfish, Clarias gariepinus, consists of 36–44 fingerlike lobes built up of tubules in which a fluid is secreted containing acid polysaccharides, acid-, neutral- and basic proteins, and phospholipids. In this fluid sperm cells are stored. The seminal vesicle fluid immobilizes the sperm cells. After ejaculation, it prolongs the period of sperm activity. The seminal vesicle fluid is secreted by the epithelium lining the tubules. The tubules in the proximal part of the lobes are predominantly lined by a simple cylindrical and those of the distal part by a simple squamous epithelium. These epithelial cells contain enzymes involved in energy-liberating processes, the enzyme activites being proportional to the height of the cells. Interstitial cells between the tubules have enzyme-histochemical and ultrastructural features indicative of steroid biosynthesis. Similar characteristics are found in testicular interstitial cells. The most rostral seminal vesicle lobes and the most caudal testicular efferent tubules form a network of tubules that opens at the point where the paired parts of the sperm ducts fuse with each other. The tubules of most seminal vesicle lobes, however, form a complex system that fuses with the unpaired part of the sperm duct.
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  • 68
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    Cell & tissue research 247 (1987), S. 667-673 
    ISSN: 1432-0878
    Keywords: Secretory granules ; Cell fusion ; Cell migration ; Immunocytochemistry ; Immunogold localisation ; Placenta ; Placental lactogen ; Trinucleate cell ; Cow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Bovine placental lactogen activity is shown by immunogold electron microscopy to be restricted to (a) the granules and the Golgi body from which they form in the bovine fetal trophectodermal binucleate cell, and (b) granules of similar size and staining reaction in trinucleate “giant” cells found in the maternal uterine epithelium throughout pregnancy. These results support the hypothesis that a fetal binucleate cell forms a maternal giant cell by migration to and fusion with a uterine epithelial cell.
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  • 69
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    Cell & tissue research 248 (1987), S. 559-563 
    ISSN: 1432-0878
    Keywords: α-MSH ; α-MSH-like peptide ; Hypothalamus ; Pituitary ; Immunocytochemistry ; Frog (Rana esculenta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of immunoreactive α-melanocyte-stimulating hormone (α-MSH) within the brain of the frog, Rana esculenta, has been studied on adjacent serial sections using an indirect immunofluorescence technique. Immunoreactive cell bodies are found in the anterior part of the preoptic nucleus and in some ventral subependymal cerebrospinal fluid-contacting elements, and in the nucleus infundibularis ventralis. Numerous α-MSH-like immunoreactive fibers are present in the preoptic area, in the pars ventralis of the tuber cinereum, and in the outer layer of the median eminence. This staining pattern is completely eliminated after preabsorbing the antiserum with the corresponding antigen, but blocking tests with α-MSH-related peptides do not lead to any change in the immunoreaction. From these results it may be inferred that an α-MSH-like system is present in the hypothalamic neurosecretory area of R. esculenta, and is probably related to its hypophysiotropic functions. The results are compared to the distribution of α-MSH within the hypothalamus of reptiles and mammals.
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  • 70
    ISSN: 1432-0878
    Keywords: Gastrin/CCK ; Immunocytochemistry ; Corpus cardiacum-corpus allatum ; Insect ; Leucophaea maderae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By use of immunocytochemistry, a gastrin/CCK-like material has been demonstrated in the corpus cardiacum-corpus allatum complex of the cockroach Leucophaea maderae. Reactivity toward gastrin and CCK with region-specific antisera suggests that the gastrin/CCK-like peptide of this insect contains the COOH-terminal tetrapeptide sequence which is common to gastrin and CCK, and that the material is more gastrin-like than CCK-like. The results indicate that, like other neuropeptides, the gastrin/CCK peptide family appeared early in evolution within neuronal elements, and that the COOH-terminal region of gastrin has been conserved during phylogeny.
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  • 71
    ISSN: 1432-0878
    Keywords: Microtubules ; Terminal web ; Small intestine ; Electron microscopy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The terminal web (TW) region of mature absorptive cells in the small intestine of the rat contains an elaborate cytoskeleton which supports the apical microvillus membrane. In studies regarding the structural organization of the cytoskeleton and associated proteins in the small intestine, microtubules have not been mentioned as components of the TW. By transmission electron microscopy of conventional resin-embedded sections of rat small intestine, we observe many microtubule profiles in the TW of mature absorptive cells. These microtubules are found in various orientations, although most course parallel to the long axis of the cell, and many microtubule profiles are seen in close association with smooth-surfaced vesicles.
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  • 72
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    Cell & tissue research 249 (1987), S. 1-5 
    ISSN: 1432-0878
    Keywords: Somatostatin ; Sympathetic ganglion ; Co-existence ; Tyrosine hydroxylase ; Immunocytochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localization of somatostatin-like immunore-activity (SOM-LI) was examined in human lumbar sympathetic ganglia using the peroxidase-antiperoxidase method. Few of the principal neurons showed immunolabelling for somatostatin and sparse networks of nerve terminals were unevenly associated with ganglion cells. Using light microscopy, the most intense SOM-LI was seen in the perinuclear zone of the neurons. Electron-microscopically, the staining was localized on the membranes of the Golgi apparatuses. In the nerve terminals, SOM-LI was seen inside the small vesicles (40–60 nm diameter). All neurons with SOM-LI were also found to be tyrosine-hydroxylase immunoreactive when excamined with a double-staining technique. These results provide evidence that somatostatin and noradrenaline co-exist in human sympathetic neurons.
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  • 73
    ISSN: 1432-0878
    Keywords: Retina ; Photoreceptor cells ; Freeze-fracturing ; Ultrastructure ; Immunocytochemistry ; Lampetra japonica
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary In common with other cyclostomata, the Japanese river lamprey (Lampetra japonica) has a retina consisting of distinct types of photoreceptor cells called long and short photoreceptor cells. After freeze-fracture, disc membranes of these photoreceptor cells were characterized in common by a homogeneous distribution of intramembrane particles on the protoplasmic fracture faces, in contrast to those of the myeloid bodies bearing scattering particles. Immunofluorescent examination was applied to the retina with monoclonal antibodies raised against bovine and chicken rhodopsins. Positive immunoreactivity was found to be limited to outer segments of the short cell, leaving the entire body of the long cell and all other components of the retina negative. The results suggest that the short cell is more closely related to a rod-type photoreceptor cell characterized by rhodopsin as its visual pigment.
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  • 74
    ISSN: 1432-0878
    Keywords: Serotonin ; Immunocytochemistry ; Ventral nerve cord ; Annelids ; Lumbricus terrestris
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary The distribution of serotonin-immunoreactive cell bodies and fibers in the ventral nerve cord of the earthworm has been investigated from whole-mount preparations and serial sections. Serotonin-immunoreactive neurons are organized in seven soma groups per ganglion; these are defined by cell number, soma shape, diameter and fiber projections. Positional variations of this pattern have been studied quantitatively. The number of labeled perikarya is constant in midbody ganglia, but increases markedly rostral to the posterior margin of the clitellum. Variability of position and cell number differs between the cell groups. Stained nerve fibers could only be partially traced; their distribution is described in relation to defined neuronal fiber bundles and segmental nerves. The distribution and morphology of serotonin-immunoreactive cells are compared with previous observations, based on pattern analysis of primary catecholamines and indolamines with the use of a formaldehyde-induced fluorescence technique. The possible role of serotonin-immunoreactive nervous elements in different nerve cord compartments is discussed with respect to physiological effects of serotonin in earthworms and other invertebrates.
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  • 75
    ISSN: 1432-0878
    Keywords: Pancreas ; Exocrine glands ; Islets of Langerhans ; Differentiation ; Immunocytochemistry ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The islet cells of the mammalian pancreas are comprised of four different endocrine cell types, each containing a specific hormone. Islet cells also contain two enzymes of the catecholamine biosynthetic pathway: tyrosine hydroxylase (TH) and aromatic L-amino acid decarboxylase (AADC). The cell lineage relationships of these different cell types have not been examined and it is not known whether, during development, they originate from the same or from different precursor populations. In this study we used immunocytochemical procedures to determine whether developing pancreatic cells express markers common to endocrine and exocrine cell types. We found that acinar cell precursors express AADC prior to the appearance of an exocrine marker and that the expression of AADC in acinar cells persists throughout embryogenesis to the first month of postnatal life. At this time, acinar cells do not contain AADC. We also found that exocrine cells containing AADC never express other islet-cell markers. These findings suggest that while acinar and islet cells both arise from precursor cells containing AADC, these progenitor cells do not express a combined endocrine-exocrine phenotype.
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  • 76
    ISSN: 1432-0878
    Keywords: Tentacles ; Ultrastructure ; Adhesion ; Echinodermata ; Dendrochirodita (11 species)
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    Notes: Summary The tentacles of eleven species of dendrochirote holothurians have been studied. The water vascular system, deep fibre system, ectoneural nerve ring and superficial fibre system are described and are similar to those of other holothurian tentacles. A conspicuous fuzzy coat covers the entire tentacular surface except for the tips of cilia. On the basis of its structure it is thought to be an attenuated glycocalyx. Its function is discussed in relation to anti-fouling and surface adhesion. The two surface coats underlying the fuzzy coat are termed the cuticle. Bacteria are found both within the surface coats and in the sub-cuticular space. Primary fixatives lacking osmium give poor preservation of the surface coats. The adhesive papillae of the apices of the tentacles contain elements of support cells and two other cells named Type-1 and Type-2 papillar cells. The secretions of Type-1 papillar cells are dense-cored vesicles and may contain a proteinaceous adhesive. The vesicles fuse with the cuticle and release their products which are apparently disseminated along the fuzzy coat filaments. The secretions of Type-2 papillar cells may have a neurosecretory function. The different models of food capture by dendrochirote tentacles are discussed as are duo-glandular adhesive systems in relation to dendrochirote tentacles.
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  • 77
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Alkaline phosphatase ; Small intestine ; Colchicine ; Rat
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary The purpose of this study was to investigate the effect of colchicine and vinblastine on the localization of alkaline phosphatase (AlPase) in rat duodenum in relation to structural changes. AlPase was localized on the membranes of rough endoplasmic reticulum, Golgi stacks, cytoplasmic vesicles, microvilli, on lateral plasma membranes, and in some lysosomes of the duodenal epithelial cells of rats treated with either lumicolchicine or 0.9% NaCl alone. Microvilli were most intensely stained, and AlPase-positive Golgi stacks were regularly distributed in the supranuclear regions. After colchicine treatment, microvilli were shortened and the staining intensity became weaker, whereas basal as well as lateral plasma membranes showed stronger staining. The AlPase-positive microvilli appeared not only on the luminal surfaces, but also on the baso-lateral plasma membranes and even on the surfaces of characteristic intracytoplasmic cysts. Golgi stacks became smaller and their distribution became less localized, and the staining intensity of the Golgi stacks became weaker. AlPase localization in rats treated with vinblastine was almost identical with that of rats treated with colchicine. Thus, colchicine and vinblastine appeared to have elicited a disorientation of intracellular transport of intestinal AlPase by inhibiting microtubule organization.
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  • 78
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    Cell & tissue research 250 (1987), S. 513-519 
    ISSN: 1432-0878
    Keywords: Oocytes ; Structural patterns ; Pore canals ; Ultrastructure ; Salmonids ; Onchorhynchus tshawytscha, O. kisutch, O. gorbuscha, Salmo trutta, S. gairdneri, Salvelinus namaycush
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary The zona radiata from unactivated and activated eggs from chinook salmon (Oncorhynchus tshawytscha), chum salmon (O. kisutch), pink salmon (O. gorbuscha), brown trout (Salmo trutta), rainbow trout (S. gairdneri) and lake trout (Salvelinus namaycush) were examined using scanning and transmission microscopy. The zona radiata in all species examined consisted of an outer adhesive coating, a thin densely staining zona radiata externa with pore canal plugs and a thick, fibrous zona radiata interna with a fibrous network on the inner surface. There was a two layer adhesive coating over the zona radiata externa in all species except pink salmon in which only one layer was observed. There were structural differences among species in the adhesive layer, zona radiata externa and plugs in the pore-canal openings.
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  • 79
    ISSN: 1432-0878
    Keywords: Alpha-bungarotoxin ; Autoradiography ; Cholinergic receptors ; Electron microscopy ; Suprachiasmatic nucleus ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of (125I) alpha bungarotoxin (α-BTX) binding sites in the suprachiasmatic nucleus (SCN) of the adult female rat was examined by electron-microscopic autoradiography. The ultrastructural distribution of silver grains was analysed by line source, direct point count, and 50% probability circle methods. Real grain distribution was significantly different from that of randomly generated hypothetical grains. Line source analysis demonstrated two populations of sources: one associated with membranes, and one inside neuronal structures. Probability circle analysis of shared grains indicated that membrane-bound-radioactive sources were mainly asssociated with axo-dendritic appositions. Only a small proportion of labeled neuronal interfaces exhibited synaptic differentiations in the plane of section. However, the compartment containing synaptic terminals was the most enriched when comparing real to hypothetical grains. Probability circle analysis of exclusive grains demonstrated that sources that were not associated with neuronal plasma membranes were likely to be within nerve cell bodies and dendrites. It is concluded that the majority of specifically labeled α-BTX binding sites in the SCN is membrane bound, and may be associated with axodendritic synaptic transmission. The presence of a significant proportion of the label in the soma and dendrites of suprachiasmatic neurons 24 h after ventricular infusion suggests that some of the labeled binding sites (junctional or nonjunctional) may be internalized within these two compartments.
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  • 80
    ISSN: 1432-0878
    Keywords: Polypeptide 7B2 ; Adenohypophysis ; Secretion ; Immunocytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A highly conserved polypeptide termed “7B2”, isolated from human and porcine pituitaries, has been reported by immunoreactivity to be distributed in various organs. However, the highest concentration has been found in the pituitary as demonstrated by a specific radioimmunoassay. In order to determine the type of cells within the pituitary that contain 7B2 and to analyse its intracellular localization, specific immunocytochemistry techniques (unlabeled antibody, peroxidase-antiperoxidase) were used both for light and electron microscopy. Immunocytochemistry of both expiants and monolayer-cell cultures of the adenohypophysis was studied. Immunoreactivity to 7B2 has been found in 21.9% of the total number of cells. After simultaneous staining of serial sections with appropriate antibodies, 7B2 was found to be colocalized with β-LH/β-FSH in gonadotrophs and with β-TSH in thyrotrophs. In situ immunocytochemistry at the electron-microscopic level showed that immunoreactive 7B2 is compartmentalized within secretory granules. The small (130 to 250 nm) but not the large granules (400 to 700 nm) were labeled in gonadotroph-like cells and small granules (90 to 150 nm) were also labeled in thyrothrophlike cells. Study of the gonadotrophs in cell culture after Zamboni's fixation revealed weak to moderate immunoreaction in rough endoplasmic reticulum. The current findings as well as previous data indicate that 7B2 is synthesized, stored and possibly released from the adenohypophysis similarly to many other secretory products.
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  • 81
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    Molecular genetics and genomics 208 (1987), S. 481-484 
    ISSN: 1617-4623
    Keywords: PEP carboxylase ; Azotobacter chroococcum ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Azotobacter chroococcum Fos 189 is a Tn1-induced mutant which, unlike the parent strain MCD1, does not fix nitrogen in air when provided with glucose or pyruvate as sole carbon sources. Fos 189 showed 5% of parental activity for phosphoenolpyruvate carboxylase though PEP synthetase activity was normal. The A. chroococcum phosphoenolpyruvate carboxylase (ppc) gene was isolated after complementation of an appropriate Escherichia coli mutant using a broad host range gene bank prepared from A. chroococcum genomic DNA. The gene was localised by transposon mutagenesis and subcloning on a minimum DNA fragment of 6.6 kb. Broad host range plasmids containing the A. chroococcum ppc gene complemented the mutation in Fos 189 thereby restoring aerotolerant nitrogen fixation.
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  • 82
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    Molecular genetics and genomics 207 (1987), S. 503-508 
    ISSN: 1617-4623
    Keywords: Bradyrhizobium ; Nif genes ; Nitrogen fixation ; Root nodule symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The 17 kb region between the Bradyrhizobium japonicum nitrogenase genes (nifDK and nifH) was investigated for the presence of further nif or fix genes by site-directed insertion or deletion/replacement mutagenesis and interspecies hybridization. Mutant strains were tested for their ability to reduce acetylene in free-living, microaerobic culture (Nif phenotype) and in soybean root nodules (Fix phenotype). The presence of a gene, previously identified by hybridization with the Klebsiella pneumoniae nifB gene, was proved by isolation of a nifB insertion mutant which was completely Nif- and Fix-. Three other regions were found to be homologous to the K. pneumoniae genes nifE, nifN, and nifS, NifE and nifN insertion mutants were completely Nif-/Fix- whereas nifS mutants were leaky with 30% residual Fix activity. Taken together, the data show that the B. japonicum genome harbours a cluster of closely adjacent genes which are directly concerned with nitrogenase function.
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  • 83
    ISSN: 1617-4623
    Keywords: Azorhizobium sesbaniae ORS571 ; Nitrogen fixation ; Regulation ; Tn5 mutagenesis ; lacZ fusions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A cosmid bank of ORS571, a diazotrophic bacterium capable of inducing aerial stem and root nodules on Sesbania rostrata, was constructed in the vector pLAFR1. A DNA probe carrying the Klebsiella pneumoniae nifA gene was used to identify nifA-and ntrC-like regions of ORS571 in the cosmid bank by colony hybridization. Cosmids carrying these regions were mapped by restriction endonuclease analysis, Southern blotting and transposon Tn5 mutagenesis. Selected Tn5 insertion mutations in the nifA/ntrC homologous regions were used for gene-replacement experiments and the resulting ORS571 mutants were examined for Nif, Fix and Ntr phenotypes. Two clearly distinct regulatory loci were thus identified and named nifA and ntrC. Plasmids carrying gene fusions of the ORS571 nifH and nifD genes to lacZ were constructed and the regulation of the ORS571 nifHDK promoter, and of the Rhizobium meliloti nifHDK promoter, was studied under varying physiological conditions in ORS571, ORS571 nifA::Tn5 and ORS571 nitrC::Tn5 strains. A model for the role of nifA and ntrC in the regulation of ORS571 nif and other nitrogen assimilation genes is proposed.
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  • 84
    ISSN: 1617-4623
    Keywords: Gene regulation ; Melanin synthesis ; Nitrogen fixation ; Phaseolus beans ; Rhizobium phaseoli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The symbiotic plasmid pRP2JI of Rhizobium phaseoli strain 8002 was shown to contain two separate regions of DNA which are required and sufficient for the synthesis of the pigment melanin. One of these regions containing the class II mel gene(s) was located to other genes involved in nodulation and in nitrogen fixation. Mutations in this region abolished both the ability to synthesize melanin and to fix nitrogen in Phaseolus bean root nodules. Mutations in the other, unlinked region, containing class I mel gene(s), also abolished melanin synthesis but did not affect symbiotic nitrogen fixation. Transcriptional fusions between the class I mel gene and the Escherichia coli lacZ gene were constructed and it was demonstrated that the class II mel gene(s) activated their transcription in free-living culture. Further, strains containing the cloned regulatory class II gene(s) synthesized melanin when growing in minimal medium, in contrast to wild-type strains which became pigmented only in complete medium containing yeast extract and tryptone. It was shown by hybridization experiments that the regulatory mel gene was closely linked to or may correspond to the regulatory nifA gene; a fragment of R. phaseoli DNA which included the class II gene(s) of R. phaseoli hybridized to a previously identified nifA-like gene of R. leguminosarum, the species that nodulates peas.
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  • 85
    ISSN: 1617-4623
    Keywords: Agrobacterium rhizogenes ; Plant transformation ; Transgenic legumes ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A procedure for transformation and regeneration of the legume species Lotus corniculatus (Bird's-foot trefoil) has been developed. The Agrobacterium rhizogenes 15834 and 8196 strains were used to transform plant cells in wound site infections and transformed roots were propagated in vitro. Transformation was monitored by hybridization with pRi T-DNA sequences and by detection of agropine and mannopine. Transformation frequencies of up to 90% were obtained. Shoots spontaneously formed on hairy root cultures were excised, rooted and inoculated with Rhizobium. Root nodules formed on transformed plants had nitrogenase activities comparable to untransformed nodules. Transcript levels from the nodule-specific leghemoglobin genes and the constitutive ubiquitin genes were similar in transformed and untransformed root nodules. Transformed plants responded to R. loti and Bradyrhizobium sp. (Lotus) strains with phenotypes identical to phenotypes for untransformed plants.
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    Molecular genetics and genomics 206 (1987), S. 291-299 
    ISSN: 1617-4623
    Keywords: Nitrogen fixation ; Stem nodulation ; Tn5 mutagenesis ; nod genes ; nodC homology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary After random Tn5 mutagenesis of the stem-nodulating Sesbania rostrata symbiont strain ORS571, Nif-, Fix- and Nod- mutants were isolated. The Nif- mutants had lost both free-living and symbiotic N2 fixation capacity. The Fix- mutants normally fixed N2 in the free-living state but induced ineffective nodules on S. rostrata. They were defective in functions exclusively required for symbiotic N2 fixation. A further analysis of the Nod- mutants allowed the identification of two nod loci. A Tn5 insertion in nod locus 1 completely abolished both root and stem nodulation capacity. Root hair curling, which is an initial event in S. rostrata root nodulation, was no longer observed. A 400 bp region showing weak homology to the nodC gene of Rhizobium meliloti was located 1.5 kb away from this nod Tn5 insertion. A Tn5 insertion in nod locus 2 caused the loss of stem and root nodulation capacity but root hair curling still occurred. The physical maps of a 20.5 kb DNA region of nod locus 1 and of a 40 kb DNA region of nod locus 2 showed no overlaps. The two nod loci are not closely linked to nif locus 1, containing the structural genes for the nitrogenase complex (Elmerich et al. 1982).
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    Plant systematics and evolution 155 (1987), S. 45-48 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Asteraceae ; Inuleae ; Phagnalon ; Ph. rechingeri spec. nova ; Taxonomy ; Flora of Iran
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phagnalon rechingeri spec. nova from S Baluchestan (Iran) is described as a species new to science and illustrated; its relationships to other species of the genus, in particular to thePh. woodii group from S Arabia, are discussed.
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  • 88
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    Plant systematics and evolution 155 (1987), S. 49-54 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Chenopodiaceae ; Noaea griffithii ; Salsola griffithii ; Taxonomy ; sand vegetation ; psammophytes ; Flora SW Asia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Noaea griffithii, which hitherto was known from the type collection only, is transferred toSalsola by reason of its horizontal embryo and other fruit characters. The description is completed; ecological, phytocoenological and distributional data are given. The species is a common endemic psammophyte of SE Iran, S Afghanistan and adjacent Pakistan Baluchistan.
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  • 89
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    Plant systematics and evolution 155 (1987), S. 27-43 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Cyperaceae ; Carex ; Endemics ; floristical connections ; Flora Iranica area ; Flora of: Iran ; Afghanistan ; Iraq ; Turkemenia ; Pakistan
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 88 species and 20 subspecies ofCarex, altogether 94 taxa, occur in the area ofRechinger's Flora Iranica. Of these, 48 taxa, have an Euro-Siberian, 11 an Irano-Turanian and 35 a Central Asiatic distribution, the last including five species with connections to E. and tropical SE. Asia. The Saharo-Sindian element is missing. Endemic taxa are found in all groups. Eight taxa, endemic to the NW. Himalayas occur in the E. part of the area. The Euro-Siberian taxa are concentrated in the NW. of the area, but have connections to the east; similarly, Central-Asiatic taxa are concentrated in E. Afghanistan and N. Pakistan, but have connections to the west. Some Euro-Siberian taxa are widely distributed in the area, but avoid the most arid regions. The Irano-Turanian taxa are few and extend from Egypt through the area of Flora Iranica to Kazakhstan SSR and Central China.
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    Plant systematics and evolution 155 (1987), S. 15-25 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Compositae ; Cousinia ; Generic distribution map ; centres of diversity ; distribution patterns ; Flora of the Irano-Turanian Region ; Flora of SW. Asia ; of Iran ; Iraq ; Afghanistan ; Pakistan
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract On the basis of new floristic works a distribution map of the genusCousinia is given and discussed. The generic distribution area ofCousinia is nearly identical with the Irano-Turanian Region, but most of the 662 species are concentrated on the Iranian and Turkestanian mountain regions. Here, both in the eastern and in the western parts of the distribution area, four centres of diversity with high numbers of—mostly endemic—species are found: The western Tienshan (61 species), Pamiro-Alaj (169), NE-Afghanistan (80), NW-Afghanistan (44); Kopetdagh (66 in Iran, 33 in Turkmenia), Elburz (66), the northern part of Zagros (44), Azerbaijan (36). The outlines of the generic area are formed by genetic-historical and by climatic factors. The distribution patterns ofCousinia species confirm the close connections between the Iranian and the Turkestanian mountains as centres of origin and conservation of palaeo-xeromorphic mountain floras, contrasting with the Turanian lowland flora.
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  • 91
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    Plant systematics and evolution 155 (1987), S. 67-69 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Caryophyllaceae ; Acanthophyllum ; Diaphanoptera ; Ochotonophila ; Scleranthopsis ; Capsule dehiscence ; Flora of Iran ; Afghanistan
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The shape and the mode of dehiscence of the capsule had been regarded as good differential characters betweenAcanthophyllum and related genera.—Studies of these characters, including the shape of the ovary, in species ofAcanthophyllum, Diaphanoptera, Ochotonophila andScleranthopsis show, however, that they cannot be used as differential characters for the genusAcanthophyllum.
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  • 92
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    Plant systematics and evolution 155 (1987), S. 71-75 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Rubiaceae ; Rubieae ; Galium ; G. subfalcatum ; spec. nova ; G. campylotrichum ; spec. nova ; G. tetraphyllum ; spec. nova ; Flora of Pakistan
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three new species ofGalium from the NW. Himalaya in Pakistan are described and illustrated.
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    Plant systematics and evolution 155 (1987), S. 77-87 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Rubiaceae—Rubieae:Galium gymnopetalum ; spec. nova ; G. lahulense ; spec. nova ; G. saipalense ; spec. nova ; G. nepalense ; spec. nova ; G. serpylloides Royle exHook. f. typified ; Alpine flora of the Himalaya
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Galium serpylloides agg. comprises a group of locally endemic, disjunct and vicarious taxa in the (sub)alpine zone of the W. Himalaya, from W to E:G. gymnopetalum, G. lahulense, G. serpylloides s. str.,G. saipalense, andG. nepalense. Four of these are new to science;G. serpylloides is newly typified. The main characters of the five species are compared and illustrated, their affinities and evolution are discussed, and a map of distribution is presented.
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    Plant systematics and evolution 155 (1987), S. 93-103 
    ISSN: 1615-6110
    Keywords: Angiosperms ; new taxa ; Arenaria ; Campanula ; Scutellaria ; Stachys ; Calamintha ; Aristolochia ; Allium ; Flora of Turkey
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 10 new Turkish taxa are described:Arenaria eliasiana, A. sivasica, A. monscragus, A. angustifolioides; Campanula lycica; Scutellaria orientalis subsp.tortumensis; Stachys choruhensis, S. tundjeliensis; Calamintha caroli-henricana; Aristolochia rechingeriana, the latter two species named in honour ofKarl Heinz Rechinger;Allium vuralii.
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    Plant systematics and evolution 155 (1987), S. 89-92 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Hypericaceae ; Hypericum ; H. dogonbadanicum ; H. socotranum subsp.smithii ; Chorology ; phytogeography ; Flora of Iran ; Socotra
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Hypericum dogonbadanicum Assadi, a local species from theQuercus brandtii forests of the Zagros Mountains in south-western Iran, belongs to sect.Campylosporus (Spach)R. Keller and is most closely related to a Socotran endemic,H. socotranum Good subsp.smithii N. Robson. Despite the inclusion of these forests in the Irano-Turanian Element of the Iran flora byZohary (1963),H. dogonbadanicum appears to belong rather to the Mediterranean dwarf-shrub maquis type of community and therefore to be a member ofZohary's (1963) Mediterranean/Irano-Turanian group of Iranian species.—The key position of the Socotran endemics to the understanding of the inter-sectional relationships is discussed.
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  • 96
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    Plant systematics and evolution 155 (1987), S. 105-132 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Scrophulariaceae ; Veronica ; Veronica agrestis group ; V. bungei ; V. ceratocarpa ; V. persica ; V. polita ; V. siaretensis ; V. francispetae ; V. agrestis ; V. opaca ; V. filiformis ; Allopolyploidy ; significance of polyploidy ; neophytes ; anthropochory ; migration ; evolution of weeds ; Flora of Iran ; SW. Asia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A character analysis reveals a clearly intermediate position of the tetraploidV. persica (2n = 28) between the two diploid speciesV. polita andV. ceratocarpa (both 2n = 14) which are morphologically rather different and have been placed by several authors in different sections of the genus.V. ceratocarpa is native to subhumid deciduous forests of the Caucasus and of the Elburz mountains (N. Iran);V. polita has its centre of variation in the Elburz range where it grows in therophyte habitats. Three other closely related species,V. bungei, V. siaretensis, andV. francispetae, are endemic to the Elburz range which is the main centre of diversity and variability of theV. agrestis group. This comprises all the above mentioned species and also two more European weeds:V. agrestis andV. opaca. Veronica polita, was probably originally native to open places in deciduous mountain forests, before becoming a weed in neolithic times and migrating to Europe; nowadays it has an almost world-wide distribution. The allotetraploidV. persica combines the ecological characters of its parents, the slightly xerophyticV. polita and the more mesophyticV. ceratocarpa, thus being preadapted to become a highly successful weed with a large ecological range. It has spread rapidly almost all over the world since the early 19th century.
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    Plant systematics and evolution 155 (1987), S. 165-187 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Guttiferae ; Hypericaceae ; Growth form ; vicarious taxa ; nomenclature ; Flora of the Mediterranean ; of Greece ; of Crete
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The morphology, altitudinal distribution and phytocoenology ofHypericum empetrifolium Willd., var.oliganthum Rech. fil. and var.tortuosum Rech. fil. are reassessed and as a result, the taxa are treated at subspecific rank. Essential characters proved to be growth form, number of flowers, foliage and epidermal surface characters, shape of seeds and testa sculpturing. The different growth forms can be interpreted as adaptations to the local habitats.Hypericum empetrifolium Willd. subsp.empetrifolium is a low subshrub of various vegetation types known as phrygana whereasHypericum empetrifolium Willd. subsp.oliganthum (Rech. f.)Hagemann grows as a cushion-like dwarf shrub in chasmophyte associations rich in relic species.Hypericum empetrifolium Willd. subsp.tortuosum (Rech. f.)Hagemann is confined in the high mountain regions and grows as a true dwarf espalier shrub with low spreading shoots.
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    Plant systematics and evolution 155 (1987), S. 189-195 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Anacardiaceae ; Pistacia lentiscus var.chia. ; Taxonomy ; mastic production ; Flora of Greece
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The author discusses the history of an unusual form ofPistacia lentiscus L. (referred to as var.chia (Desf. exPoiret) DC. on the basis of literature data and personal field observations on Chios island. As a result of incisions made on the trunk and stems the tree exudes a specific resinous gum called mastic. The production of mastic currently amounts to 160–170 tons per annum and plays an important role in the economy of the island constituting the main source of income for approx. 20 villages in the south of Chios. The history of mastic dates back 2500 years to the time ofHerodotus. The author points out that on the plantation only male individuals are cultivated; these have been selected over a long period based on mastic yield. As they represent a group of cultivated clones it would be preferable to consider them as a collective cultivar ‘Chia’.
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    Plant systematics and evolution 155 (1987), S. 197-213 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Lamiaceae (Labiatae) ; Thymus sect.Teucrioides ; Th. teucrioides ; Th. rechingeri spec. nov. ; Th. leucospermus spec. nov. ; Taxonomy ; Mountain flora of Greece
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract TheThymus teucrioides Boiss. & Spruner aggregate is revised and the following new taxa, all from the alpine zone in the Greek mountains, are described:Th. leucospermus Hartvig from the calcareous mountains of Pindhos and Mt Parnassos in Sterea Ellas,Th. rechingeri Hartvig with the subsp.macrocalyx Hartvig from calcareous mountains in Sterea Ellas and N Peloponnissos, andTh. teucrioides subsp.alpinus Hartvig from the serpentine areas of N Pindhos. In the variableTh. teucrioides s. str. many characters have turned out to be markedly geographically correlated and many local populations can be distinguished by a particular combination of characters.
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    Plant systematics and evolution 155 (1987), S. 215-217 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Leguminosae ; Fabaceae ; Onobrychis aliacmonia ; Endemism ; threatened plants ; rediscovery ; Flora of Greece (Makedhonia, Peloponnisos)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Onobrychis aliacmonia was discovered in 1956 and described in 1973, the year when it was last collected. Its single known locality was flooded soon afterward by an artificial lake now extending over the middle portion of the Aliakmon Valley (western Greek Makedhonia), and the species was believed extinct. It has now been rediscovered in the southern Peloponnisos, in an area far south of the locus classicus and in a widely different ecological context.
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