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  • Immunocytochemistry  (104)
  • Auxin
  • Chromatographie, Dünnschicht
  • Springer  (184)
  • 1995-1999  (81)
  • 1980-1984
  • 1975-1979  (103)
  • 1925-1929
  • 1995  (81)
  • 1978  (55)
  • 1977  (48)
Collection
Keywords
Publisher
Years
  • 1995-1999  (81)
  • 1980-1984
  • 1975-1979  (103)
  • 1925-1929
Year
  • 1
    Electronic Resource
    Electronic Resource
    Infection and disintegration of vascular tissues of non-suberized roots of spruce byHeterobasidion annosum and use of antibodies for characterizing infection (1995)
    Springer
    Mycopathologia 129 (1995), S. 91-101 
    Details
    ISSN: 1573-0832
    Keywords: ELISA ; Endodermis ; H. annosum ; Immunocytochemistry ; Root rot ; Vascular tissues
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Vascular disintegration mainly of medulla rays of spruce roots is of major significance in root rot disease of spruce caused byH. annosum. Using seedling roots as an experimental model, the possible routes and initial host reactions preceding invasion of vascular tissues was investigated. Transmission electron microscopy showed that penetration through the endodermis was an obvious route but not without host resistance. Using antibodies againstH. annosum hyphal materials, some labelling of vascular tissues remote from sites of fungal colonization suggest the release of fungal secretory products partly active in tissue disintegration. Similarly, intense labelling was also observed in severely colonized host tissues at late stages of infection. Strong labelling recorded at 3 d p.i. mainly on fungal hyphae and scant gold particles on invaded host tissues could imply that induction of host antifungal metabolites may have been a late event. A correlation was found between total antigenic material in root homogenates measured by ELISA, density of tissue labelling by immunocytochemistry and severity of disease symptoms. The importance of this in relation to diagnosis of biotic root rot diseases in the field is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01103468
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  • 2
    Electronic Resource
    Electronic Resource
    Demonstration of enamel matrix proteins on root-analogue surfaces of rabbit permanent incisor teeth (1977)
    Springer
    Calcified tissue international 24 (1977), S. 223-229 
    Details
    ISSN: 1432-0827
    Keywords: Enamel-cementum-morphology ; Immunocytochemistry ; Biochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The continuously erupting rabbit incisor tooth is normally thought of as having an enamel covered “crown” on its labial surface and a cementum covered “root” on its lingual surface. We have examined both surfaces of continuously erupting rabbit incisor teeth taken from near term embryos by a variety of means, including transmission and scanning electron microscopy, biochemical fractionation, and immunohistochemistry. In all cases, we could detect no qualitative difference in the early extracellular matrices taken from the labial and lingual surfaces of the teeth. Both matrices were shown to be composed of dentin and enamel, although the thickness and geometry of the enamel matrix on the lingual surface was somewhat different from that on the labial surface.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02223320
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  • 3
    Electronic Resource
    Electronic Resource
    The role of cellulase in hormonal regulation of shoot morphogenesis in tobacco callus (1995)
    Springer
    Planta 196 (1995), S. 727-731 
    Details
    ISSN: 1432-2048
    Keywords: Antibody ; Auxin ; Callus ; Cellulase ; Morphogenesis ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A polyclonal antibody raised against cellulase (EC 3.2.1.4.) from callus of Nicotiana tabacum L. cv. Petit Havana SR1 reduced cellulase activity and induced shoot formation in tobacco callus in the presence of callus maintaining concentrations of auxin and cytokinin. Shoot induction as well as reduction of the cellulase activity was also obtained by withdrawing auxin from the callus medium. The effect of the two hormones on cellulase activity in the tobacco tissue was examined by varying the concentration of one of the hormones α-naphthylacetic acid (NAA) or benzylaminopurine (BAP) at a time while the other was kept at a level sufficient for either callus growth or shoot induction. While NAA stimulated the enzyme activity increasingly with concentration in the range 5 × 10−7 M to 5 × 10−5 M at both levels of BAP, BAP only stimulated the cellulase activity at an optimum concentration of 5 × 10−6 M when NAA was present at a level sufficient to induce callus growth. The results point to a pivotal role of the downward regulation of cellulase in the initiation of shoot induction. A series of events leading to oriented cell divisions as a result of the lowered cellulase level during the initial phase of the morphogenetic process is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00197338
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  • 4
    Electronic Resource
    Electronic Resource
    Affinity labels for auxin binding sites in corn coleoptile membranes (1977)
    Springer
    Planta 134 (1977), S. 145-149 
    Details
    ISSN: 1432-2048
    Keywords: Affinity labels ; Auxin ; Cell membranes ; Hormone Receptors ; Receptors ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two auxin analogues have been tested as affinity labels for auxin binding sites in coleoptile membranes of Zea mays L. Reacting the membranes at pH 8–9 with the diazonium salt of CAPA (2-chloro-4-aminophenoxyacetic acid) reduces their subsequent ability to bind NAA(1-naphthylacetic acid). Diazo-Chloramben (2,5-dichloro-3-aminobenzoic acid) is also effective in inhibiting NAA binding capacity and this inhibition is largely independent of reaction pH over the range pH 6–9. Similar experiments with sulphydryl reagents have shown that reaction of the membranes with p-mercuribenzoate (PMB) strongly inhibits subsequent auxin binding activity. Prior addition of NAA protects the binding sites against the action of diazo-Chloramben or PMB when the reactions are carried out at pH 6. From these results and from other considerations, several of the amino acid residues in the binding site environment have been tentatively assigned.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00384963
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  • 5
    Electronic Resource
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    Growth of Avena coleoptiles and pH drop of protoplast suspensions induced by chlorinated indoleacetic acids (1978)
    Springer
    Planta 140 (1978), S. 89-92 
    Details
    ISSN: 1432-2048
    Keywords: Auxin ; Avena ; Helianthus ; pH drop ; Pisum ; Protoplast suspension
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Several indoleacetic acids, substituted in the benzene ring, were compared in the Avena straight growth bioassay. 4-Chloroindoleacetic acid, a naturally occurring plant hormone, is one of the strongest hormones in this bioassay. With an optimum at 10-6 mol l-1, it is more active than indoleacetic acid, 2,4-dichlorphenoxyacetic acid and naphthaleneacetic acid. 5-Chloro- and 6-chloroindoleacetic acids are very strong auxins as well. Other derivatives tested have a lower activity. 5,7-Dichloro- and 5-hydroxyindoleacetic acids have very low auxin activity at 10-4 mol l-1 and may be anti-auxins. Some of the derivatives were compared for their effect on pH decline in stem protoplast suspensions of Helianthus annuus L. and Pisum sativum L. The change of pH occurs without a lag period or with only a very short one. Derivatives which are very active in the Avena straight growth assay cause a larger pH decline than indoleacetic acid, while inactive derivatives cause effectively no pH decline.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00389385
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  • 6
    Electronic Resource
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    Early time course and specificity of auxin effects on turgor movement of the bean pulvinus (1978)
    Springer
    Planta 140 (1978), S. 107-109 
    Details
    ISSN: 1432-2048
    Keywords: Auxin ; Leaf movement ; Phaseolus ; Pulvinus ; Turgor movement
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Auxin application to the upper side of the pulvinus of primary leaves of Phaseolus vulgaris L. promoted bending away from the place of application. The effect had a latency of less than 20 min and was specifically induced by substances known as active auxins in growth tests (indoleacetic and 1-naphthaleneacetic acid) but not by inactive auxin analogs (2-naphthaleneacetic, 3-indolepropionic and benzoic acid); 2,4-dichlorophenoxyacetic acid, and L-(-)-2,4-dichlorophenoxyisopropionic acid were of intermediate activity. Auxin-promoted bending was reversible and presumably caused by turgor increase in the treated cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00384908
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  • 7
    Electronic Resource
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    Continuous measurement of initial curvature of maize coleoptiles induced by lateral auxin application (1978)
    Springer
    Planta 140 (1978), S. 201-211 
    Details
    ISSN: 1432-2048
    Keywords: Auxin ; Cell elongation ; Coleoptiles ; Fusicoccin ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To analyze early effects of auxin application, an apparatus was developed which continuously and simultaneously registered the curvature of 10 individual maize (Zea mays L.) coleoptiles. Resolution was less than 5 μm over a range of ±0.5 mm. The data were evaluated and plotted via paper tape and Hewlett-Packard-computer. Unilateral application of 3×10-5 M indoleacetic acid (IAA) resulted in a transient inhibition of growth on the side of application for ca. 10 min (Phase I), followed by a strong stimulation (Phase II). The phytotoxin fusicoccin (FC) caused an immediate stimulation of elongation. The initial negative reaction of Phase I is auxin-specific. Only active auxins such as IAA and 1-naphtaleneacetic acid produced this initial inhibition; chemical analogs-inhibitory or neutral in long-term growth tests, e.g. phenylacetic acid-did not show any significant effects on Phase I. When the coleoptiles were symmetrically preloaded with different levels of auxin, only a large step-up of subsequent unilateral auxin application resulted in a negative phase I; a small step-up led to an immediate positive reaction. The results are discussed in context with the parallel kinetics for various other auxin-induced reactions of coleoptile cells which have already been published.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00390249
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  • 8
    Electronic Resource
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    Effects of validamycin A, a potent trehalase inhibitor, and phytohormones on trehalose metabolism in roots and root nodules of soybean and cowpea (1995)
    Springer
    Planta 197 (1995), S. 362-368 
    Details
    ISSN: 1432-2048
    Keywords: Auxin ; Carbohydrate metabolism ; Glycine (nodules) ; Symbiosis ; Trehalose ; Trehalase ; Validamycin A
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Trehalose, a common microbial disaccharide, has been reported to be toxic to plants, and plant trehalase has therefore been hypothesized to function as a detoxifying enzyme. To test this, aseptically grown soybean (Glycine max L. Merr.) plantlets were supplied with trehalose. The plants accumulated trehalose only when validamycin A, a potent trehalase inhibitor, was added as well. Under these conditions, they accumulated trehalose to up to 8% of the dry weight in their primary leaves without any detectable impairment of growth or health. We have previously shown that in soybean nodules, trehalose is generated by the symbiotic bacteria, and trehalase is strongly induced. However, direct exposure of plants to trehalose did not affect their trehalase activity, whereas a treatment with auxin strongly increased it, indicating that the enzyme level is regulated by hormones rather than by its substrate. Addition of validamycin A to nodules caused an increase in the amount of trehalose and a decrease in the sucrose and starch pools, but nitrogen fixation was not affected. Similar results were obtained with cowpea (Vigna unguiculata L.) plantlets and nodules. These results indicate that plant trehalase is functional in metabolizing trehalose from exogenous and endogenous sources, even though the disaccharide has no obvious toxic effects.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00202658
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  • 9
    Electronic Resource
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    Anions modify the response of guard-cell anion channels to auxin (1995)
    Springer
    Planta 197 (1995), S. 546-552 
    Details
    ISSN: 1432-2048
    Keywords: Auxin ; Anion channel (GCAC1) ; Guard cell ; Malate ; Vicia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The anion channel in the guard-cell plasma membrane of Vicia faba, GCAC1, possesses recognition sites for the plant growth hormone auxin at the extracellular mouth of the channel (Marten et al. 1991, Nature 353:759-762). Using the patch-clamp technique we could demonstrate that auxins induced a shift of the voltage dependence of the anion channel to hyperpolarized potentials; the shift was attenuated during an increase in the extracellular chloride concentration, indicating that chloride shields the hormone-binding site. The auxin-induced shift was concentration-dependent, characterized by a Michaelis-Menten type of behaviour with a half saturation constant (K m) of about 10 μM naphthalene-1-acetic acid (1-NAA) in the presence of 2 mM Cl− and 12 μM in 80 mM Cl−. In the presence of malate, another gating modulator of GCAC1, auxins were less effective, indicating that both ligands compete for common sites. Inactive auxins with respect to stomatal opening or stimulation of the plasma membrane H+-ATPase, such as 2-NAA, modulated the activation threshold and kinetics of GCAC1 similar to the active form 1-NAA. At a concentration of 100 μM 2-NAA the peak-current potential shifted by about 30 mV more negative.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00196677
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  • 10
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    Auxin inhibition of acid-and fusicoccin-induced elongation in lentil roots (1977)
    Springer
    Planta 136 (1977), S. 97-102 
    Details
    ISSN: 1432-2048
    Keywords: Acid growth ; Auxin ; Ethylene ; Fusicoccin ; Growth inhibition ; Lens ; Root growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Both acid pH (4.0) and fusicoccin (FC) strongly stimulate root elongation in intact lentil (Lens culinaris Med.) seedlings. FC-induced elongation is apparently mediated by FC-enhanced H+ secretion since the toxin induces massive secretion of H+ in these roots after a latent period of less than 5 min. Auxin (indole-3-acetic acid) strongly inhibits elongation in control roots as well as acid-induced and FC-induced root elongation. Treatment of apical root segments with auxin causes only a slight apparent uptake of H+ and has no inhibitory effect on FC-induced H+ secretion, whether the hormone is given before or after the toxin. Auxin induces ethylene production in excised roots of lentil but the latent period is at least 30 min while inhibition of root elongation by IAA is maximal within 30 min. It is concluded that the inhibitory action of auxin on acid-and fusicoccin-induced root elongation is a direct effect, independent of auxin-induced ethylene production or auxin-mediated modification of cell-wall pH.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00396184
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  • 11
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    Synergistic effect of gibberellic acid and indole-3-acetic acid on rooting in stem cuttings of Abelmoschus esculentus Moench (1978)
    Springer
    Planta 138 (1978), S. 111-112 
    Details
    ISSN: 1432-2048
    Keywords: Abelmoschus ; Auxin ; Gibberellin ; Root formation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Indole-3-acetic acid (IAA) and gibberellic acid (GA3) enhanced the formation of roots on the stem cuttings of Abelmoschus esculentus. The effect increased considerably when both IAA and GA3 were applied together.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00392926
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  • 12
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    Asymbiotic association of Rhizobium with pea epicotyls treated with a plant hormone (1978)
    Springer
    Planta 138 (1978), S. 107-110 
    Details
    ISSN: 1432-2048
    Keywords: Auxin ; Nitrogen fixation (asymbiotic) ; Pisum ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Treatment of epicotyls of dark-grown pea (Pisum sativum L.) seedlings with indole-3-acetic acid causes swelling of the tissue. Application of Rhizobium to the cut surface of the swollen tissue results in the development of an “infection”. The infection spreads in the cortical cells and proceeds 2–3 mm deep into the stem within 3–4 days. An acetylene reduction assay used for detecting nitrogen-fixation capacity of the infected tissue was negative at 10% [O2]; however, if [O2] was reduced to below 1%, some activity could be detected. Ultrastructural observations indicate that the cytoplasmic contents of the infected cells are destroyed and no membrane structure around the bacteria is formed during this infection. Rhizobium does not appear to have developed any symbiotic relationship with the host. Failure to develop symbiosis appears to result in a parasitic or saprophytic association and the nitrogen fixed under such conditions may not be of any use to the plant.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00392925
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  • 13
    Electronic Resource
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    The influence of growth regulators absorbed by the root on sex expression in hemp plants (1978)
    Springer
    Planta 138 (1978), S. 181-184 
    Details
    ISSN: 1432-2048
    Keywords: Abscisic acid ; Auxin ; Cannabis ; Cytokinin ; Flowers (sex) ; Gibberellin ; Sex expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Application, through the root system, of growth regulators to hemp (Cannabis sativa L.) plants having 2–3 pairs of visible leaves caused pronounced shifts of sex expression in the adult individuals. Treatment with gibberellic acid (25 mg/l) resulted in more than 80% of the plants being male, i.e. having staminate flowers (controls, ca. 30%). Treatment with 6-benzylaminopurine and with indole-3-acetic acid (in either case, 15 mg/l) resulted in all plants being either female (pistillate flowers) or intersexes (bisexual flowers); treatment with abscisic acid (10 mg/l) had a similar but somewhat less pronounced effect.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00391176
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  • 14
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    Ethylene and adventitious root formation in hypocotyl segments of etiolated mung-bean (Vigna radiata (L.) Wilczek) seedlings (1978)
    Springer
    Planta 138 (1978), S. 193-197 
    Details
    ISSN: 1432-2048
    Keywords: Auxin ; Ethylene ; Root formation ; Vigna
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Rooting responses and ethylene production by hypocotyl cuttings from etiolated mung-bean seedlings treated with the auxins α-naphthaleneacetic acid, γ-(indole-3)-n-butyric acid (IBA) and 2,4,5-trichloro-phenoxypropionic acid were determined. There was no relationship between the abilities of the auxins to induce root formation and their capacities for inducing ethylene production. Studies with mixtures of 3-indoleacetic acid, a poor stimulator of rooting but an effective inducer of ethylene production, and IBA, an effective rooting stimulator but a poor inducer of ethylene production, exposure of cuttings to ethylene or (2-chloroethyl) phosphonic acid (Ethephon), hypobaric storage (150 mb) of treated cuttings, and exposure of auxin-treated cuttings to 7% CO2 also indicated that ethylene is not directly involved in initiation of adventitious roots in this plant material.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00386810
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  • 15
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    3-O-Methyl glucose uptake stimulation by auxin and by fusicoccin in plant materials and its relationships with proton extrusion (1978)
    Springer
    Planta 138 (1978), S. 249-256 
    Details
    ISSN: 1432-2048
    Keywords: Auxin ; Fusicoccin ; Glucose transport ; Proton flux ; Roots ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Auxin and fusicoccin (FC) stimulate the active uptake of 3-O-methyl glucose (3-O-MG) in those materials in which they have been shown to activate an electrogenic proton extrusion (Pisum sativum L. stems, Zea mays L. coleoptiles and roots). In maize roots the curve relating 3-O-MG influx to external concentrations indicated that the values of the apparent Km increase in the 3-O-MG concentration range between 2×10-5 mol l-1 and 2×10-2 mol l-1. FC did not alter the Km values and its stimulating effect was nearly constant at all 3-O-MG concentrations tested. Basal and FC-induced uptake of 3-O-MG appeared associated with a transient proton influx suggesting that also in maize roots a sugar-proton contransport occurs. Diethyl stilbestrol, which inhibits proton extrusion, inhibited also basal and FC-induced 3-O-MG uptake. The data support the view that the stimulation by FC of 3-O-MG uptake is closely related to that of proton extrusion. The stimulation by FC of 3-O-MG uptake cannot be replaced by increasing extracellular proton concentration, nor may be explained only by the FC-induced hyperpolarization of transmembrane potential difference. The hypothesis is proposed that the effect of FC on 3-O-MG uptake depends on an increase of cytoplasmic pH, following the activation of the proton extruding system.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00386819
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  • 16
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    Phytochrome photoreversibility: Empirical test of the hypothesis that it varies as a consequence of pigment compartmentation (1978)
    Springer
    Planta 141 (1978), S. 129-134 
    Details
    ISSN: 1432-2048
    Keywords: Avena ; Immunocytochemistry ; Phytochrome
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phytochrome of oat (Avena sativa L., cv. Garry) coleoptile cells in the red-light-absorbing form, Pr, is diffusely distributed while after conversion to the far-red-light-absorbing form, Pfr, it is observed only in very small areas within the cell. Comparison of phytochrome photoversibility measurements to the distribution of the pigment within the cell indicates that the spectral assay is not influenced by the observed compartmentalization of the chromoprotein. However, the observed compartmentalization of phytochrome is correlated with a loss in spectrophotometrically detectable Pr.
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    URL: http://dx.doi.org/10.1007/BF00387878
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  • 17
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    Auxin gradient along the root of the maize seedling (1978)
    Springer
    Planta 141 (1978), S. 179-181 
    Details
    ISSN: 1432-2048
    Keywords: Auxin ; Maize ; Root tip ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract [5-3H]Indol-3yl-acetic acid (IAA) applied to the shoot apices of intact 6-day-old maize (Zea mays L.) plants moved into the primary root and accumulated at the root apex. IAA from the shoot could partially satisfy the requirement of the primary root for IAA for growth.
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    URL: http://dx.doi.org/10.1007/BF00387886
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  • 18
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    Hormonal regulation of cotton ovule and fiber growth: Effects of bromodeoxyuridine, AMO-1618 and p-chlorophenoxyisobutyric acid (1978)
    Springer
    Planta 141 (1978), S. 269-272 
    Details
    ISSN: 1432-2048
    Keywords: AMO-1618 ; Antiauxin ; Auxin ; Bromodeoxyuridine ; Cell (fiber) growth ; Gibberellin ; Gossypium ; Ovules (in-vitro culture)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of 5-bromo-2-deoxyuridine (BUdR, thymidine analogue), AMO-1618 (2-isopropyl-4-dimethylamino-5-methylphenyl-1-piperidine carboxylate methyl chloride), a growth retardant, and p-chlorophenoxyisobutyric acid (PCIB, an antiauxin) on growth (dry weight increase) and fiber development in unfertilized cotton (Gossypium hirsutum L.) ovules grown in vitro have been studied. BUdR (5 μM) causes about 70% inhibition of fiber production, with little effect on ovule growth, if applied during the first 6 d of culture in the presence of GA3 and IAA. AMO-1618, when used with GA3 alone, causes only a small reduction in both dry weight and fiber production, but when used with IAA alone reduces both fiber production and dry weight, the effect on the latter being predominant. In the presence of both IAA and GA3, AMO-1618 causes a small decrease in fiber production but a major decrease in dry weight. PCIB completely inhibits fiber growth but has little effect on dry weight, especially when GA3 is present. These results indicate that GA3 mainly promotes ovule growth while IAA is largerly responsible for fiber growth.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00388342
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  • 19
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    Effect of 2,4-dichlorophenoxyacetic acid on the structure and function of developing chloroplasts (1977)
    Springer
    Planta 137 (1977), S. 65-69 
    Details
    ISSN: 1432-2048
    Keywords: Auxin ; Chlorophyll content ; Chloroplast ; Photosynthetic rate ; Raphanus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Dark-grown radish seedlings (Raphanus sativus L.) were sprayed with 10-3 mol·l-1 2,4-dichlorophenoxyacetic acid and then were exposed to a 14:10 light: dark cycle. Cotyledon samples from these seedlings and unsprayed controls were taken for electron microscopy, chlorophyll determinations, and photosynthetic rate measurements at regular intervals for 72 h. A normal development of etioplasts to chloroplasts with formation of typical grana-fret work system was observed in the control cotyledons. The chloroplasts in the 2,4-D-treated cotyledons showed changes in the organization of the grana thylakoids; these thylakoids being more appressed to each other than in the controls. The chlorophyll content of treated plants was less than that of controls but the rate of chlorophyll biosynthesis was unaffected. The photosynthetic rate/mg chlorophyll was considerably higher for treated plants suggesting that 2,4-D treatment resulted in decreased size of the photosynthetic unit.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00394437
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  • 20
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    The stimulation of cell extension by ethylene and auxin in aquatic plants (1978)
    Springer
    Planta 144 (1978), S. 39-47 
    Details
    ISSN: 1432-2048
    Keywords: Aquatic plants ; Auxin ; Cell elongation ; Ethylene ; Ethylene biosynthesis ; Silverions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Elongation of the shoots of three aquatic plants (Hydrocharis morsus-ranae, Regnellidium diphyllum and Ranunculus sceleratus) is stimulated by treatment with ethylene or IAA. The effects of the two hormones are additive, and experiments with an ethylene biosynthesis inhibitor and silver ions indicate that the mechanisms by which ethylene and IAA stimulate growth may be different. Hydrocharis and Ranunculus leaf discs synthesize [14C]ethylene from [14C]methionine, but no [14C]ethylene is formed by Regnellidium, suggesting the existence of an alternative pathway of ethylene biosynthesis in the fern.
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    A specific radioimmunoassay for nanogram quantities of the auxin, indole-3-acetic acid (1977)
    Springer
    Planta 136 (1977), S. 173-180 
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    ISSN: 1432-2048
    Keywords: Auxin ; Immunoassay ; Nicotiana ; Radioimmunoassay
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have developed a specific radioimmunoassay [RIA] for indole-3-acetic acid (IAA) in the 0.2 ng to 12 ng range which, in principle, can be extended to other indole auxins as well. Methods are presented for obtaining suitable antibody, for the RIA procedure, and for measuring IAA in methanolic extracts of plant tissues. Antibody specific for IAA was obtained from rabbits immunized with IAA bound to bovine serum albumin by formaldehyde treatment. In assays with this antibody, 2,4-dichlorophenoxyacetic acid and indoles structurally related to IAA reacted from 300- to 3000-fold less than did IAA itself. However, α-and β-naphthaleneacetic acid reacted significantly and hence interfered with the assay. Extracts of tobacco (Nicotiana tabacum L.) tissue were immunoassayed after partial purification by buffer-ether partition. Crown-gall tumor tissue, which is auxin-autotrophic, and pith tissue depleted of auxin by the diffusion method contained, respectively, 26.7 ng and 〈0.5 ng extractable IAA per gram fresh weight.
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    Short-term effects of plant hormones on membrane potential and membrane permeability of dwarf maize coleoptile cells (Zea mays L. d 1) in comparison with growth responses (1977)
    Springer
    Planta 137 (1977), S. 293-298 
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    ISSN: 1432-2048
    Keywords: Auxin ; Calcium ions ; Cell growth ; Gibberellin ; Membrane permeability ; Membrane potential ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The membrane potential difference of dwarf maize coleoptile cells is increased by both 10-5moll-1 gibberellic acid (GA3) and indoleacetic acid (IAA) a few minutes after application. A final level is reached after 10–20 min. The membrane permeability ratio P Na:P K is altered by both hormones during the first 15 min after application, indicating a rapid effect on the membrane. Elongation growth of coleoptile segments, however, is only stimulated by IAA. The auxin-induced growth as well as the auxin effect on membrane permeability depends on the calcium ion concentration of the medium. It is concluded that IAA acts via a proton extrusion pump that is electrically balanced by a potassium ion uptake, driven by the electromotive force of the pump. The mode of action of GA3 on elongation growth is assumed to involve a process that depends on the physiologic state of the tissue and/or metabolic energy.
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    Lateral electrical potential following asymmetric auxin application to maize coleoptiles (1978)
    Springer
    Planta 140 (1978), S. 31-35 
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    ISSN: 1432-2048
    Keywords: Auxin ; Coleoptiles ; Electrical Potential ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Following asymmetric application of indoleacetic acid to maize (Zea mays L.) coleoptiles the early time course of changes in lateral electrical potential was externally monitored with static-drop electrodes. First, an early negative potential change of ca.-1 mV was measured at the surface on the side of a strong auxin application. This negative auxin effect ended after ca. 15 min and was followed by a strong and lasting auxin stimulation of a positive lateral potential up to +12 mV at the auxin-treated side. The initial auxin effect appeared to depend on the size of the step-up in auxin concentration.
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    Does indoleacetic acid promote growth via cell wall acidification? (1978)
    Springer
    Planta 140 (1978), S. 137-142 
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    ISSN: 1432-2048
    Keywords: Auxin ; Cell elongation ; Proton secretion ; Triticum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Growth of Triticum aestivum L. cv. Cappelle Desprez coleoptiles is promoted by 5.7×10−5 M indole acetic acid (IAA) as effectively in pH 3.4 buffer as in water, but IAA is not effective in the presence of buffer at pH 3.0 or 3.2 A combination of 5.7×10−5 M IAA and pH 3.4 buffer promotes growth to a greater extent than pH 3.2 buffer alone, which is optimal for acid-induced growth. IAA employed at 10−7 M is still effective at promoting growth in the presence of pH 3.4 buffer, moreover, IAA at 10−7 M interacts synergistically with the acidic buffer to promote growth. It is concluded that IAA and acid promote growth via separate mechanisms, and that IAA does not promote cell wall loosening by rendering the cell wall more acid.
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    Lead toxicity effects on indole-3-ylacetic acid-induced cell elongation (1978)
    Springer
    Planta 144 (1978), S. 79-84 
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    ISSN: 1432-2048
    Keywords: Auxin ; Cell elongation ; Lead ; Triticum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In vitro studies of IAA-induced cell elongation in Triticum aestivum have demonstrated that lead causes a large reduction in elongation. Inhibition of elongation can be reduced by increasing the concentration of IAA, or by the addition of calcium. The inhibitory effect appears to be linked with changes in the properties of the cell walls. Experiments are described which show that lead becomes bound strongly to certain chemical substances involved in cell wall architecture.
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    Effect of growth regulators and role of roots in sex expression in spinach (1978)
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    Planta 142 (1978), S. 207-210 
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    ISSN: 1432-2048
    Keywords: Abscisic acid ; Auxin ; Cytokinin ; Flowers (sex) ; Gibberellin ; Sex expression ; Spinacia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When 7-d-old plantlets of spinach (Spinacia oleracea L.) were immersed with their roots for 24 h in 25 mg/l gibberellic acid (GA3), or 15 mg/l 6-benzylaminopurine (6-BAP), or 15 mg/l indole-3-acetic acid (IAA), or 10 mg/l abscisic acid (ABA) and subsequently grown on long (18-h) days, the ratio of plants with male and female flowers, which in the controls was almost 1:1 (48 and 52%, respectively), was greatly altered. The treatments with 6-BAP, IAA and ABA raised the percentage of female plants to 88, 76 and 71%, respectively; the GA3 treatment increased the percent of male plants to 79%. When young, vegetative spinach plants (3 visible leaves) grown in 18-h days were cut a the root neck, and the shoots grown with their bases in nutrient solution, with adventitious roots either being allowed to develop or being systematically removed, 85% of the plants without roots became males, 85% of those with roots became females. But if the cut shoots were first, for 28 h, placed in a 15-mg/l 6-BAP solution and then grown in the absence of roots, the percent of female plants was restored to 84. These results fully agree with those obtained previously with hemp, namely, that plant growth regulators exert a regulating effect on the sex expression of dioecious plants when applied through the roots in early stages of development; that the root system plays an important role in determining the sex of these plants, that this role of the roots is associated with the synthesis of cytokinins in them. Dioecious short- and long-day plants do not differ in these respects.
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    Naturally occuring modifiers of auxin-receptor interaction in corn: Identification as benzoxazolinones (1978)
    Springer
    Planta 142 (1978), S. 103-107 
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    ISSN: 1432-2048
    Keywords: Auxin ; Benzoxazolinones ; Receptors ; Supernatant factor ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A naturally-occurring material termed ‘supernatant factor’ [Ray, P.R., Dohrmann, U., Hertel, R.: Plant Physiol. 59. 357–364 (1977)], which has the property of modifying the binding affinity of auxins to receptor sites, has been isolated from corn (Zea mays L.) and characterised as a mixture of 6-methoxy-2-benzoxazolinone (MBOA) and 6,7-dimethoxy-2-benzoxazolinone (DMBOA). DMBOA is about 50 times more active than MBOA in inhibiting binding of the auxin 1-naphthylacetic acid to membrane-bound or solubilised receptors. The activity of these compounds and the parent analogue in inhibiting auxin binding is correlated with their ability to inhibit auxin-induced growth.
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    Fluxes and compartmentation of K+, Na+ and Cl−, and action of auxins in suspension-cultured Petroselinum cells (1978)
    Springer
    Planta 143 (1978), S. 213-223 
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    ISSN: 1432-2048
    Keywords: Auxin ; Compartmental analysis ; Ion fluxes ; Petroselinum ; Suspension culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transport of 86Rb+/K+, 22Na+, 36Cl−, and [3H]indole acetic acid (IAA) has been studied on suspension-cultured cells of the parsley, Petroselinum crispum (Mill) Nym. By compartmental analysis two intracellular compartments of K+, Na+, and Cl− have been identified and ascribed to the cytoplasm and vacuole; half-times of exchange were around 200 s and 5 h, respectively. According to the Ussing-Teorell flux equation, active transport is required for the influx into the cytoplasm at the plasmalemma (K+, Cl−) and the tonoplast (K+, Na+, Cl−). The plasmalemma permeability pattern, PK:PNa:PCl=1.00:0.24:0.38, features an increased chloride permeability compared with cells from higher plant tissues. IAA uptake showed an exponential timecourse, was half-maximal after 10 min, and a linear function of the IAA concentration from 10−9 to 10−5 M. IAA and 2,4-dichlorophenoxy acetic acid reduce the apparent influx of K+, Na+, Cl− during the initial 30 min after addition and subsequently accelerate both in- and efflux of these ions. We discuss that auxins could affect the ion fluxes in a complex way, e.g. by protonophorous activity and by control of the hypothetical proton pump.
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    Ribosome metabolism in hormone-treated Jerusalem artichoke tuber slices in the absence and presence of 5-fluorouracil (1977)
    Springer
    Planta 135 (1977), S. 267-273 
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    ISSN: 1432-2048
    Keywords: Auxin ; Cytokinin ; 5-Fluorouracil ; Helianthus ; Ribosome synthesis and activity ; Tuber
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In order to examine the relation of protein synthesis to the onset of growth, changes in ribosome content and activity were compared in aged, metabolically active Jerusalem artichoke (Helianthus tuberosus L.) slices incubated in water or 2,4-dichlorophenoxyacetic acid+kinetin. In water, cells do not grow or divide and rRNA and protein levels remain constant. The percentage membrane-bound (mb) ribosomes drops from 25% to 16% during 24h. At the same time the proportion of ribosomes active in protein synthesis in both free and mb populations declines from about 69% to 54%. In auxin+kinetin, cell expansion occurs and is accompanied by a 3-fold increase in rRNA and a 50% increase in total protein content. The percentage mb ribosomes remains at 25% throughout 48 h of growth. During the first 24h of growth 70% of ribosomes in both free and mb populations are active; this value declines to near water levels at 48 h. Considering the large increase in total ribosomes the number of synthetically active ribosomes is substantially increased during growth. 5-Fluorouracil (5-FU) does not inhibit hormone induced growth but does depress total rRNA content by about one-third. It also reduces [3H]uridine incorporation into ribosomes by 70% and the newly made ribosomes are mostly inactive in protein synthesis. On the other hand, the inhibitor does not significantly affect the proportion of total ribosomes active in protein synthesis and only partially reduces protein accumulation during the second 24 h of growth. It is suggested that while ribosome production is reduced in 5-FU, ribosome turnover is also retarded resulting in retention of near normal capacity for protein synthesis and growth.
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    Meristem differentiation in Riella helicophylla (Bory et Mont.) Mont. under the influence of auxin or antiauxin (1977)
    Springer
    Planta 135 (1977), S. 289-295 
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    ISSN: 1432-2048
    Keywords: Antiauxin ; Auxin ; Meristem differentiation ; Riella
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract During the development of the unistratose gemmae of Riella helicophylla, the single intercalary meristem of the very young gemmae is subdivided into two lateral meristems. The duration of the cell reproduction cycle increases from the margin to the median part of the gemmae. This polarization within the meristem disappears after addition of the antiauxin PCIB to the culture medium. PCIB leads to a retardation or blockage of the cell cycle during the light period of the culture. Under the influence of PCIB the amount of starch in the chloroplasts is strikingly increased, probably because of a reduction of starch degradation. Addition of sugars compensates the effect of PCIB on the cell cycle. The effects of PCIB are counteracted by auxin. The results are taken as evidence that auxin plays a role in directing the transport of substances needed for the continuation of the cell reproduction cycle between adjacent cells of the meristem.
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    Some quantitative effects of indoleacetic acid on the wood production and tracheid dimensions of Picea (1977)
    Springer
    Planta 134 (1977), S. 223-228 
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    ISSN: 1432-2048
    Keywords: Auxin ; Cambium ; Gibberellin ; Picea ; Tracheids-wood production
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The diameter and wall thickness of tracheids produced after indoleacetic acid treatment were not significantly different from those of the intact controls, for the first few weeks after treatment of disbudded shoots of Picea abies Karst. and Picea sitchensis (Bong.) Carr. However, lateral application of indoleacetic acid (IAA) to intact shoots increased both tracheid diameter and wall thickness; it is suggested that IAA acted synergistically with another endogenous growth regulator, which was also removed by disbudding. Increase in wall thickness after exogenous IAA was associated with increase in duration of the wall thickening phase of tracheid differentiation; this is discussed in relation to the seasonal change from early to latewood. Cambial dormancy was induced by disbudding during active wood production. Since this occurred with or without the presence of current leaves, it is concluded that in Picea continued cambial activity depends upon supply of auxin from the buds, and cannot be supplied from expanded leaves or from the internode itself. Neither indoleacetic acid nor gibberellic acid stimulated renewed cambial activity when applied after the cessation of wood production. With both disbudded and intact shoots, the effectiveness of exogenous IAA declined with time, probably due to decreasing penetration through callus developing at the wounded surface. It is suggested that this apparent change in IAA effectiveness may explain some discrepancies between the results of previous observers.
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    Abscisic acid and apical dominance in Phaseolus coccineus L. (1977)
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    Planta 134 (1977), S. 295-299 
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    ISSN: 1432-2048
    Keywords: Abscisic acid ; Apical dominance ; Auxin ; Gibberellin ; Hormone transport ; Phaseolus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Abscisic acid (ABA) in lanolin, applied to the internode of decapitated runner bean plants enhances the outgrowth of lateral buds. The optimum concentration of the paste is 10-5 M. The effect of ABA is counteracted by indoleacetic acid (IAA) but not by gibberellic acid (GA3). There is no effect when ABA is applied to the apical bud or lateral buds of intact plants. However, 13.2 ng given to the lateral buds of decapitated plants stimulate their growth, whereas higher concentrations are inhibitory. Consequently, ABA enhances growth of lateral buds directly, but only when apical dominance is already weakened. The growth of the decapitated 2nd internode was not affected by ABA. Radioactivity from [2-14C] ABA, applied to nonelongating 2nd internode stumps of decapitated runner bean plants moves to the lateral buds, whereas [1-14C]IAA-and [3H]GA1-translocation is much weaker. ABA transport is inhibited if IAA or [3H]GA1 is applied simultaneously. In elongating internodes [14C]ABA is almost completely immobile. [14C]IAA-and [3H]GA1-translocation is not affected by ABA. The amount of radioactivity from labelled ABA, translocated to the lateral buds, is highest during the early stages of bud outgrowth.
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    Rapid differentiation of tracheary elements in cultured explants of Jerusalem artichoke (1977)
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    Planta 135 (1977), S. 207-212 
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    ISSN: 1432-2048
    Keywords: Auxin ; Cytokinin ; Explants ; Helianthus ; Tuber (explants) ; Xylem differentiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract the culture of Jerusalem artichoke (Helianthus tuberosus L.) tuber explants on filter paper discs moistened with liquid medium resulted in rapid and consistent xylem differentiation. The number of tracheary elements increased in discrete steps, the first at 48 h with a second at 56–58 h, following partially synchronous mitoses at 20 and 30 h. Factors favouring xylem cell differentiation were optimum levels of both an auxin and a cytokinin, low medium nitrogen concentrations, small volumes of medium, and high culture temperatures. A cell counting method employing Feulgen-stained nuclei and suitable for quantifyings small numbers of immature tracheary elements is described.
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    The role of cellulase in hormonal regulation of shoot morphogenesis in tobacco callus (1995)
    Springer
    Planta 196 (1995), S. 727-731 
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    ISSN: 1432-2048
    Keywords: Antibody ; Auxin ; Callus ; Cellulase ; Morphogenesis ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A polyclonal antibody raised against cellulase (EC 3.2.1.4.) from callus ofNicotiana tabacum L. cv. Petit Havana SR1 reduced cellulase activity and induced shoot formation in tobacco callus in the presence of callus maintaining concentrations of auxin and cytokinin. Shoot induction as well as reduction of the cellulase activity was also obtained by withdrawing auxin from the callus medium. The effect of the two hormones on cellulase activity in the tobacco tissue was examined by varying the concentration of one of the hormones α-naphthylacetic acid (NAA) or benzylaminopurine (BAP) at a time while the other was kept at a level sufficient for either callus growth or shoot induction. While NAA stimulated the enzyme activity increasingly with concentration in the range 5 × 10−7 M to 5 × 10−5 M at both levels of BAP, BAP only stimulated the cellulase activity at an optimum concentration of 5 × 10−6 M when NAA was present at a level sufficient to induce callus growth. The results point to a pivotal role of the downward regulation of cellulase in the initiation of shoot induction. A series of events leading to oriented cell divisions as a result of the lowered cellulase level during the initial phase of the morphogenetic process is discussed.
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    The effect of auxin on cytokinin levels and metabolism in transgenic tobacco tissue expressing an ipt gene (1995)
    Springer
    Planta 196 (1995), S. 84-94 
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    ISSN: 1432-2048
    Keywords: Agrobacterium ; Auxin ; Cytokinin metabolism ; Cytokinin oxidase ; ipt gene ; Nicotiana (cytokinin)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The ipt gene from the T-DNA of Agrobacterium tumefaciens was transferred to tobacco (Nicotiana tabacum L.) in order to study the control which auxin appears to exert over levels of cytokinin generated by expression of this gene. The transgenic tissues contained elevated levels of cytokinins, exhibited cytokinin and auxin autonomy and grew as shooty calli on hormone-free media. Addition of 1-naphthylacetic acid to this culture medium reduced the total level of cytokinins by 84% while 6-benzylaminopurine elevated the cytokinin level when added to media containing auxin. The cytokinins in the transgenic tissue were labelled with 3H and auxin was found to promote conversion of zeatin-type cytokinins to 3H-labelled adenine derivatives. When the very rapid metabolism of exogenous [3H]zeatin riboside was suppressed by a phenylurea derivative, a noncompetitive inhibitor of cytokinin oxidase, auxin promoted metabolism to adenine-type compounds. Since these results indicated that auxin promoted cytokinin oxidase activity in the transformed tissue, this enzyme was purified from the tobacco tissue cultures. Auxin did not increase the level of the enzyme per unit tissue protein, but did enhance the activity of the enzyme in vitro and promoted the activity of both glycosylated and non-glycosylated forms. This enhancement could contribute to the decrease in cytokinin level induced by auxin. Studies of cytokinin biosynthesis in the transgenic tissues indicated that trans-hydroxylation of isopentenyladenine-type cytokinins to yield zeatin-type cytokinins occurred principally at the nucleotide level.
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    Developmental patterns during direct somatic embryogenesis in protoplast cultures of european larch (Larix decidua Mill.) (1995)
    Springer
    Plant cell reports 15 (1995), S. 242-247 
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    ISSN: 1432-203X
    Keywords: Auxin ; Larix decidua Mill ; Protoplasts ; Single cell monitoring ; Somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Protoplasts isolated from embryogenic suspension cultures of European larch (Larix decidua Mill.) were cultured in thin alginate layers using a nylon mesh to enable a monitoring of the development of single cells. The patterns of cell division and differentiation are characterized and compared with zygotic embryogenesis to which homologies can only be drawn to some extent when the protoplasts grow in an auxinfree environment. Already at 2.5 μM both 2,4-dichlorophenoxyacetic acid or indole-3-acetic acid cause vacuolation and elongation of individual cells, thus disturbing the process of somatic embryogenesis which generally lacks the precise quantitative patterns occurring in vivo. Prior to the formation of an embryo, a proembryonal mass develops. Oligonucleated products of spontaneous protoplast fusions are able to cellularize even without preceding karyokinesis and perform a normal embryogenic program.
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    Rapid auxin- and fusicoccin-enhanced Rb+ uptake and malate synthesis in Avena coleoptile sections (1978)
    Springer
    Planta 139 (1978), S. 35-41 
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    ISSN: 1432-2048
    Keywords: Auxin ; Avena ; Cell elongation ; Ion uptake ; Malate synthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The short-term effects of auxin (indole-3-acetic acid) and fusicoccin (FC) on Rb+ uptake and malate accumulation in Avena sativa L. coleoptile sections have been investigated. FC stimulates 86Rb+ uptake within 1 min while auxin-enhanced uptake begins after a 15–20-min lag period. Auxin has little or no effect on 86Rb+ uptake at external pHs of 6.0 or less, but substantial auxin effects can be observed in the range of pH 6.5 to 7.5. Competition studies indicate that the uptake mechanism is specific for Rb+ and K+. After 3 h of auxin treatment the total amount of malate in the coleoptile sections is doubled compared to control sections. FC causes a doubling of malate levels within 60 min of treatment. Auxin-induced malate accumulation exhibits a sensitivity to inhibitors and pH which is similar to that observed for the H+-extrusion and Rb+-uptake responses. Both auxin- and FC-enhanced malate accumulation are stimulated by monovalent cations but this effect is not specific for K+.
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    Identification and measurement of indoleacetic and abscisic acids in the cambial region of Picea sitchensis (Bong.) Carr. by combined gas chromatography-mass spectrometry (1978)
    Springer
    Planta 139 (1978), S. 133-138 
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    ISSN: 1432-2048
    Keywords: Abscisic acid ; Auxin ; Cambium ; Picea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Indole-3-acetic acid (IAA) and abscisic acid (ABA) were identified by combined gas chromatography-mass spectrometry (GCMS) in fractions obtained by diffusion and extraction from bark peelings of Sitka spruce. A procedure is described for the quantitative analysis of IAA and ABA levels in the same extract using the GCMS technique of single-ion current monitoring. This procedure was used to measure the diffusible, free, and bound fractions of IAA and ABA in the cambial region of Sitka spruce throughout one year; the range in concentration for these fractions was 0.06–0.30, 0.46–3.85, and 0.04–0.20 μg/g oven-dry weight, respectively, for IAA, and 0–0.08, 0.03–2.21, and 0.13–0.66 μg/g oven-dry weight, respectively, for ABA. Movement in the cambial region was found to be polar for endogenous IAA and nonpolar for endogenous ABA. Recoveries of [14C]IAA internal standards showed that 73–99.5% of the IAA was lost during purification, and that there could be up to 5-fold differences in recovery between purifications, indicating that IAA loss shold be measured in quantitative analyses.
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    Rapid-growth responses of corn root segments: Effect of auxin on elongation (1977)
    Springer
    Planta 135 (1977), S. 1-5 
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    ISSN: 1432-2048
    Keywords: Auxin ; Cell elongation ; Elongation ; Root growth ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of indole-3-acetic acid (IAA) on the elongation rates of 2 mm corn (Zea mays L.) root segments induced by citrate-phosphate buffer (or unbuffered) solutions of pH 4.0 and 7.0 was studied. At pH 7.0, auxin initially reduced the elongation rate in both buffered and unbuffered solutions. Only in buffer at pH 7.0 was auxin at a concentration of 0.1 μM found to promote the elongation rate though briefly. THis promoted rate represented only ca. 20% of the rate achieved with only buffer at pH 4.0. Auxin in pH 4.0 buffered and unbuffered solutions only served to reduce the elongation rates of root segments. Some comparative experiments were done using 2 mm corn coleoptile segments. Auxin (pH 6.8) promoted the elongation rate of coleoptile segments to a level equal or greater than the maximal H ion-induced rate. The two responses of root segments to auxin are compared to auxin action in coleoptile growth.
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    URL: http://dx.doi.org/10.1007/BF00387967
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    A canonical model for production and distribution of root mass in space and time (1995)
    Springer
    Journal of mathematical biology 33 (1995), S. 477-488 
    Details
    ISSN: 1432-1416
    Keywords: Root profile ; Auxin ; Mathematical model ; Diffusivity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Mathematics
    Notes: Abstract A mathematical model is presented for the production and distribution of plant root mass in a substrate in space and time. This model is intended to be used for the interpretation of data on root density profiles. It is assumed that a pulse of root apices appears at the surface of the substrate at zero time, emerging from seeds or stems, and that these apices migrate downwards in the substrate, leaving trails of root mass as they go, giving birth to daughter apices, and “dying” by terminal differentiation. The model has at least 3 parameters, representing, respectively, initial root generating capacity of root axes at zero time, net rate of root apex reproduction (or extinction), and rate of downwards migration of root apices. Two versions of the model are explored, a nondiffusive version and a diffusive version in which there is an additional parameter representing diffusivity of root apices. It is shown that both versions of this theoretical mechanistic model predict that the profile of root density with depth tends to a steady-state exponential form, parameterised by a density of root mass at zero depth and a logarithmic rate of change of root mass with respect to depth. This form has previously appeared in the literature as an empirical model. Data analyses are performed, using this model, on a set of data in which cultures of plants were subjected to treatment with auxin or by genetic manipulation, and the effects of the various treatments on the parameters of the model are described. Interpretations of the experimental results are proposed in the light of the theoretical results derived in this paper.
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    URL: http://dx.doi.org/10.1007/BF00163039
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    The AQP2 water channel: Effect of vasopressin treatment, microtubule disruption, and distribution in neonatal rats (1995)
    Springer
    The journal of membrane biology 143 (1995), S. 165-175 
    Details
    ISSN: 1432-1424
    Keywords: Water channels ; Vasopressin ; Rat kidney ; Immunocytochemistry ; Microtubules ; Cell polarity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Aquaporin 2 is a collecting duct water channel that is located in apical vesicles and in the apical plasma membrane of collecting duct principal cells. It shares 42% identity with the proximal tubule/thin descending limb water channel, CHIP28. The present study was aimed at addressing three questions concerning the location and behavior of the AQP2 protein under different conditions. First, does the AQP2 channel relocate to the apical membrane after vasopressin treatment? Our results show that AQP2 is diffusely distributed in cytoplasmic vesicles in collecting duct principal cells of homozygous Brattleboro rats that lack vasopressin. In rats injected with exogenous vasopressin, however, AQP2 became concentrated in the apical plasma membrane of principal cells, as determined by immunofluorescence and immunogold electron microscopy. This behavior is consistent with the idea that AQP2 is the vasopressin-sensitive water channel. Second, is the cellular location of AQP2 modified by microtubule disruption? In normal rats, AQP2 has a mainly apical and subapical location in principal cells, but in colchicine-treated rats, it is distributed on vesicles that are scattered throughout the entire cytoplasm. This is consistent with the dependence on microtubules of apical protein targeting in many cell types, and explains the inhibitory effect of microtubule disruption on the hydroosmotic response to vasopressin in sensitive epithelia, including the collecting duct. Third, is AQP2 present in neonatal rat kidneys? We show that AQP2 is abundant in principal cells from neonatal rats at all days after birth. The detection of AQP2 in early neonatal kidneys indicates that a lack of this protein is not responsible for the relatively weak urinary concentrating response to vasopressin seen in neonatal rats.
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    URL: http://dx.doi.org/10.1007/BF00233445
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    Autoradiographic studies of 3H-dexamethasone uptake by immunocytochemically characterized cells of the rat pituitary (1977)
    Springer
    Cell & tissue research 182 (1977), S. 347-356 
    Details
    ISSN: 1432-0878
    Keywords: Pituitary ; Dexamethasone ; ACTH ; Autoradiography ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary 3H-Dexamethasone (10 μg/kg) was injected intravenously in adrenalectomized rats and after survival times of 5, 30, 60, and 180 min its uptake within the pituitary was studied by autoradiography. Radioactivity was concentrated in cell nuclei in the pars nervosa and pars distalis. Within the pars intermedia, only cells of the marginal zone were labeled. In the pars distalis, some cells showed a weak nuclear accumulation of radioactivity as early as 5 min after injection. The tissue radioactivity was nearly maximal at 5 min, and the proportion of radioactivity in nuclei reached a maximum of 60–70% by 30 min. In competition experiments, non-radioactive steroids (1 mg/kg) were injected 5 min before 3H-dexamethasone and sacrifice was 30 min later. Dexamethasone markedly diminished the nuclear accumulation in the pars distalis, but corticosterone and progesterone did not. In the pars nervosa, corticosterone and progesterone competed for nuclear uptake of 3H-dexamethasone, although less effectively than dexamethasone itself. Different cell types in the pars distalis were characterized by treating autoradiograms with an immuno-peroxidase bridge procedure. Cells treated with anti-ACTH 17–39 had the greatest nuclear concentration of radioactivity, and those stained with anti-TSH were least heavily labeled. Cells treated with antisera to GH, PRL, and hCG were moderately labeled.
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    URL: http://dx.doi.org/10.1007/BF00219770
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    Isolation, characterization and localization of bovine adrenal medullary myosin (1977)
    Springer
    Cell & tissue research 178 (1977), S. 17-38 
    Details
    ISSN: 1432-0878
    Keywords: Myosin ; Immunocytochemistry ; Adrenal medulla ; Exocytosis ; Secretion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Myosin was isolated in high purity from the bovine adrenal medulla by gel filtration and ion exchange chromatography. The purified myosin was analyzed by electrophoresis in gels containing SDS and found to contain a 200,000 molecular weight heavy chain and major light chains of molecular weights 20,000 and 17,000 in a 1∶1∶1 molar ratio. At high ionic strength the myosin had high Ca-ATPase and K-EDTA-ATPase activities and low Mg-ATPase activity. At low ionic strength, the Mg-ATPase was activated to a low level by rabbit muscle actin. The myosin was found to decorate F-actin in the absence, but not the presence of ATP. In low ionic strength solutions, the myosin assembled into characteristic bipolar filaments. The distribution of this myosin in the adrenal medulla and of cross-reacting myosin in several other bovine tissues was determined with the use of antimedullary myosin immunoglobulin G as a specific stain that was detected by direct and indirect immunofluorescence. In the medulla strong staining was seen between the chords of chromaffin cells indicating the presence of a highly muscular vasculature that may perform functions analogous to those of the myoepithelium of exocrine glands. The chromaffin cells showed weak positive staining around the nuclei and in a pattern radiating toward adjacent blood vessels. Cells of the inner zone of the adrenal cortex showed strong staining in the peripheral cytoplasm while cells in the intermediate and outer zones did not stain. In a blood smear, platelets and the cytoplasm of leukocytes stained strongly while erythrocytes did not stain. In striated muscle and the gray and white matter of the cerebrum only the capillaries and larger vessels stained. In the liver the phagocytic cells bordering vascular sinuses stained strongly while the hepatocytes were separated from one another by a 2 micron trilaminar band possibly representing the microfilament web surrounding the bile canaliculi and associated with junctional complexes. The results suggest that myosin is present in several highly differentiated, non-motile tissue cells where it may play a role in secretion or other specialized functions.
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    URL: http://dx.doi.org/10.1007/BF00232821
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    Quantitative dünnschicht-chromatographische Bestimmung von aromatischen Aminen nach der Remissionsmethode (1978)
    Springer
    Fresenius' Zeitschrift für analytische Chemie 292 (1978), S. 381-384 
    Details
    ISSN: 1618-2650
    Keywords: Best. von Aminen, aromat. ; Chromatographie, Dünnschicht ; Remissionsmessung
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Description / Table of Contents: Zusammenfassung Es wird eine Methode zur quantitativen Bestimmung von aromatischen Aminen beschrieben. Diese werden dünnschicht-chromatographisch auf Kieselgel 60 F254 Fertigplatten in den Laufmitteln n-Propanol/Methanol/Wasser/Eisessig (65∶15∶15∶20; Vol.) oder n-Butanol/Wasser/Eisessig (66∶17∶17; Vol.) von den Begleitstoffen abgetrennt. Die getrennten Aminflecke werden entweder im Ultraviolett-Bereich oder nach Diazotierung im sichtbaren Bereich mit dem Chromatogramm-Spektralphotometer PMQ II nach der Remissionsmethode gemessen und anhand der auf derselben Platte aufgestellten Eichgerade ausgewertet. Die quantitative Bestimmung wird anhand der beiden Amine 5-Amino-2,4,6-trijod-isophthalsäurebis-(2,3-dihydroxy-propyl-N-methylamid) (I) und 2,4,6-Trijod-3-aminobenzoesäure (II) in Röntgenkontrastmittelpräparaten (RKP) demonstriert. Sowohl im UV- als auch im sichtbaren Bereich, läßt sich in 3000 μg RK-Säure (RKS) noch 0,1 μg freies aromatisches Amin, das sind 0,003%, bezogen auf die eingesetzte Menge RKS, mit einer maximalen relativen Standardabweichung von 4% bestimmen. Diese Methode ist spezifischer und empfindlicher als die der Diazotierung in Lösung.
    Notes: Summary The aromatic amines are separated from impurities by thin-layer chromatography on silica gel 60 F254 precoated plates in the mobile solvents n-propanol/methanol/water/glacial acetic acid (65∶15∶15∶20; vol.) or n-butanol/water/glacial acetic acid (66∶17∶17; vol.). The separated amine spots are measured by the remission method by means of the TLC densitometer PMQ II, either in the u.v. region or, after diazotization, in the visible region, and evaluated on the basis of the linear calibration curve set up on the same plate. Quantitative determination is demonstrated by means of the two amines, 5-amino-2,4,6-triiodoisophthalic acid-bis-(2,3-dihydroxypropyl-N-methylamide) (I) and 2,4,6-triiodo-3-aminobenzoic acid (II), in radiopaque contrast media (RCM). Both in the u.v. and visible regions, it is still possible to determine 0.1 μg of free aromatic amine in 3000 μg of radiopaque contrast medium acid (RCA), or 0.003% in terms of the amount of RCA used, with a maximum relative standard deviation of 4%. This method is more specific and more sensitive than that of diazotization in solution.
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    Analyse von Desodorantien in kosmetischen Präparaten (1978)
    Springer
    Fresenius' Zeitschrift für analytische Chemie 293 (1978), S. 135-137 
    Details
    ISSN: 1618-2650
    Keywords: Analyse von Desodorantien, Phenolderivaten in Kosmetika ; Chromatographie, Dünnschicht
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Description / Table of Contents: Zusammenfassung Phenolische Desodorantien in kosmetischen Präparaten lassen sich durch Umsetzung mit Echtrotsalz Al (Anthrachinondiazonium-chlorid-1) in wäßrig-alkalischer Lösung, Extraktion des gebildeten Azofarbstoffes mit einem organischen Lösungsmittel und anschließender Dünnschicht-Chromatographie schnell identifizieren. In den meisten Fällen ist die Abhängigkeit zwischen der Extinktion der Farbstofflösung und der eingesetzten Menge des betreffenden Desodorants linear, wodurch dann auch quantitative Bestimmungen möglich sind.
    Notes: Summary Phenolic compounds, used as deodorants in cosmetic products can be identified quickly by reaction with anthraquinonediazonium chloride-1 (Echtrotsalz Al) in aqueous alkaline medium, extraction of the azo dye with an organic solvent and separation by thinlayer chromatography. The relation between the absorbance of the coloured solution and the concentration of the deodorant concerned is generally linear; in these cases quantitative determinations are always possible.
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    Trennung und Nachweis von N-Heterocyclen auf CuSO4-imprägnierten Schichten (1978)
    Springer
    Fresenius' Zeitschrift für analytische Chemie 291 (1978), S. 366-368 
    Details
    ISSN: 1618-2650
    Keywords: Nachw. von N-Heterocycl. Verbindungen ; Chromatographie, Dünnschicht ; CuSO4-imprägnierte Schichten, Fluorescenz
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Description / Table of Contents: Zusammenfassung Durch die charakteristischen Fluorescenzfarben und die Fluorescenz-Thermochromie der CuJ-Komplexe lassen sich N-Heterocyclen auf Chromatogrammen meist einwandfrei identifizieren. Eine wesentliche Vereinfachung wird durch die Verwendung CuSO4-imprägnierter Kieselgelschichten erreicht. Die Nachweisempfindlichkeiten insbesondere bei tiefer Temperatur ändern sich im Vergleich zur ursprünglichen Technik kaum.
    Notes: Summary N-Heterocycles can usually be identified with certainty through the characteristic fluorescence colours and fluorescence-thermochromism of the CuI-complexes. The use of CuSO4-impregnated silica gel layers facilitates the procedure considerably, and scarcely reduces the sensitivity of detection, particularly at low temperature.
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    TLC separation of some inorganic ions on p-toluidine impregnated layers (1978)
    Springer
    Fresenius' Zeitschrift für analytische Chemie 292 (1978), S. 415-416 
    Details
    ISSN: 1618-2650
    Keywords: Trenn. von Anorgan. Ionen ; Chromatographie, Dünnschicht ; mit p-Toluidin imprägnierte Schichten
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00481570
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    Localization of thyrotropin (TSH) in the dog pituitary gland (1978)
    Springer
    Cell & tissue research 186 (1978), S. 399-412 
    Details
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Dog ; Pars distalis ; Thyrotropin (TSH) ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using the immunoperoxidase technique and antisera to the specific beta (β) subunits of bovine and rat TSH1, selective immunocytochemical staining was localized in a specific cell population in the pars distalis of the dog pituitary gland. These TSH cells were found to be positive to aldehyde fuchsin, alcian blue, periodic acid-Schiff (PAS) and aniline blue. With the performic acidalcian blue (pH 0.2) -PAS-orange G procedure these cells stained blue-purple, demonstrating FSH/LH cells (blue or turquoise), ACTH/MSH cells (redpurple) and PRL cells (orange-red). The TSH cells were further differentiated from other functional cell types of the pars distalis on the basis of their typical cytological features, intraglandular distribution and by immunocytochemical double staining. In the pars distalis of adult male dogs the TSH cells were mostly shown to be smaller in size and less numerous than in bitches in the anestrous phase of the sexual cycle. Moreover, cytological alterations in the immunoreactive thyrotrophs in the pituitary of male and female dogs generally paralleled the spontaneous changes in thyroid function associated with thyroid atrophy and/or pituitary insufficiency, and thyroid hyperplasia or goiter. In conclusion, because of their specificity and high potency, the antisera to the β-subunits of bovine and rat TSH represent an effective tool for the selective immunocytochemical localization of TSH in the dog pituitary. This allows the study of the morphology and function of TSH cells under different physiological, pathological and experimental conditions.
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    URL: http://dx.doi.org/10.1007/BF00224930
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    Immunocytochemical identification of an LHRH-producing system originating in the preoptic nucleus of the duck (1978)
    Springer
    Cell & tissue research 188 (1978), S. 99-106 
    Details
    ISSN: 1432-0878
    Keywords: LHRH-neurosecretion ; Avian hypothalamus ; Vasotocin neurosecretion ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A fluorescent technique applying specific LHRH and vasotocin antisera was used for the immunocytochemical localization of the respective neurosecretory systems in the hypothalamus of gonadectomized, testosteronetreated and/or serotonin injected male domestic ducks. An immunoreactive (IR) LHRH-producing system, with perikarya located in the preoptic nucleus, could be traced through the ventral hypothalamus down to the external layer of the rostral and caudal ME, in close vicinity to the hypophysial portal system. An IR-vasotocin system originating in the paraventricular and supraoptic nuclei ran through the ventral hypothalamus, but terminated in (i) the external layer of the rostral ME, and (ii) in the posterior lobe of the hypophysis.
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    URL: http://dx.doi.org/10.1007/BF00220517
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    Immunocytochemical study of the hypothalamo-neurohypophysial system (1978)
    Springer
    Cell & tissue research 188 (1978), S. 119-132 
    Details
    ISSN: 1432-0878
    Keywords: Neurophysin ; Paraventricular nucleus ; Supraoptic nucleus ; Sheep ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An antiserum cross-reactive against ovine neurophysins-I-II and -III has been used in conjunction with the immunoperoxidase histochemical procedure to localize the cells of the sheep paraventricular (PVN) and supraoptic nuclei (SON). In order to describe the topographical distribution of the SON and PVN a study was made on the serial sections cut (a) transversely from rostral to caudal positions and (b) sagittally from lateral to medial positions of the hypothalamus. The cells of the SON, when examined in the transverse aspect, extended approximately 1900 μ caudally and when examined in the sagittal plane were contained within a lateral-medial distance of 4830 μ. In each case the SON cells lay adjacent to the optic chiasm. As sections were cut transversely, the cells of the PVN first appeared in a rostral position defined as 0 μ and close to the ventral lining of the third ventricle. This general ventral and ventro-lateral distribution of cells maintained up to a caudal distance of approximately 840 μ. From positions 1260–2310 μ there was a dramatic dorsal shift of the PVN cells which by this time had also extended laterally. The total rostral-caudal distance occupied by the PVN cells was 3150 μ. That the lateral-medial distance occupied by the PVN was small (1050 μ) was determined on examining the magnocellular nuclei in sagittal section.
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    Presence of an oxytocin-like peptide in the hypothalamus and neurohypophysis of a turtle (Mauremys caspica) and a snake (Natrix maura) (1995)
    Springer
    Cell & tissue research 279 (1995), S. 75-84 
    Details
    ISSN: 1432-0878
    Keywords: Oxytocin ; Neurophysin ; Vasotocin ; Mesotocin ; Suprachiasmatic nucleus ; Medial nucleus of the infundibular recess ; Immunocytochemistry ; Natrix maura (Serpentes) ; Mauremys caspica (Chelonia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The probable presence of oxytocin in the hypothalamo-hypophysial system of two reptilian species, the snake Natrix maura and the turtle Mauremys caspica, was re-investigated. A high-pressure liquid chromatographic analysis of the turtle neural lobe revealed the existence of vasotocin, mesotocin, and a third compound co-eluting with oxytocin. Brains from both species were fixed by vascular perfusion with Bouin's fluid. Adjacent paraffin sections were immunostained using antisera against the following substances: (1) bovine oxytocin-neurophysin; (2) a mixture of bovine oxytocin-neurophysin and vasopressin-neurophysin; (3) dogfish neurophysins; (4) oxytocin; (5) arginine-vasotocin; (6) mesotocin; (7) somatostatin. Immunoreactivity against oxytocin was found in parvocellular neurons of the snake suprachiasmatic nucleus and cerebrospinal-fluid contacting neurons of the medial nucleus of the infundibular recess of both species, the latter immunoreactivity being much more conspicuous in the turtle. Numerous fibers containing immunoreactive oxytocin extended between the medial nucleus of the infundibular recess, and the internal region of the medium eminence and the neural lobe. The oxytocin-immunoreactivity in all locations was completely abolished by preabsorption of the anti-oxytocin serum with three different oxytocin preparations. None of the neurons of the suprachiasmatic and medial nucleus of the infundibular recess, including the oxytocin-immunoreactive elements, reacted with either the antineurophysin sera used, or the anti-vasotocin or anti-mesotocin antibodies. The possible existence of a reptilian oxytocin-neurophysin is discussed. The alternative that, in the reptilian hypothalamus, neurons synthesize a compound closely related to, but different from oxytocin is also considered.
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    The biased intracellular accumulation of the β-subunit of salmon gonadotropin (GTH II) in the pituitary of rainbow trout Oncorhynchus mykiss, during gametogenesis (1995)
    Springer
    Cell & tissue research 279 (1995), S. 93-99 
    Details
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Gonadotropin ; Subunits ; Gonadotropes ; Immunocytochemistry ; Immunoblotting ; Oncorhynchus mykiss (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Salmon gonadotropin (GTH II) is a heterodimeric glycoprotein hormone (α and IIβ subunits), serving as a maturational GTH, and is produced in a specific gonadotropic cell-type (GTH II-cells) containing small granules and large globules. In trout GTH II-cells, double immunolabeling for the α- and IIβ-subunits shows that colocalization of the α- and IIβ-immunolabeling is confined to the small granules, indicating storage of functional GTH II. On the other hand, α-immunolabeling is absent in the large globules, even though IIβ labeling is abundant throughout the period of seasonal gametogenesis. The α-specific antiserum recognizes the intact α-subunit as well as the reduced and deglycosylated α-subunits by immunoblotting. These results indicate that an accumulation of the IIβ-subunit is specifically generated in the large globules of these cells. In fact, with sexual maturity, the quantity of IIβ-subunits becomes elevated in the trout pituitary due to a marked increase in GTH II-cells containing many large globules. However, the derivation and function of the large globules and the fate of their contained IIβ-subunits remains unknown.
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    Regeneration and post-metamorphic development of the central nervous system in the protochordate Ciona intestinalis: a study with monoclonal antibodies (1995)
    Springer
    Cell & tissue research 279 (1995), S. 421-432 
    Details
    ISSN: 1432-0878
    Keywords: Trans-differentiation ; Proliferation ; Bromodeoxyuridine ; Immunocytochemistry ; Regeneration ; Ciona intestinalis (Tunicata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract In this study, we use three monoclonal antibodies that recognise antigens present in the central nervous system of the ascidian Ciona intestinalis to study regeneration and post-metamorphic development of the neural ganglion. We have also used bromodeoxyuridine labelling to study generation of the neuronal precursor cells. The first antibody, CiN 1, recognises all neurones in the ganglion, whereas the second, CiN 2, recognises only a subpopulation of the large cortical neurones. Western blotting studies show that CiN 2 recognises two membrane-bound glycoproteins of apparent Mr 129 and 100 kDa. CiN 1 is not reactive on Western blots. Immunocytochemical studies with these antibodies show that CiN 1-immunoreactive neurone-like cells are present at the site of regeneration as early as 5–7 days post-ablation, a sub-population of CiN 2-immunoreactive cells being detected by 9–12 days post-ablation. The third antibody, ECM 1, stains extracellular matrix components and recognises two diffuse bands on Western blots of whole-body and ganglion homogenates. The temporal and spatial pattern of appearance of CiN 1 and CiN 2 immunoreactivity both during post-metamorphic development and in regeneration occurs in the same sequence in both processes. Studies with bromodeoxyuridine show labelled nuclei in some neurones in the regenerating ganglion. Plausibly these originate from the dorsal strand, an epithelial tube that reforms by cell proliferation during the initial phases of regeneration. A second population of cells, the large cortical neurones, do not incorporate bromodeoxyuridine and thus must have been born prior to the onset of regeneration. This latter finding indicates a mechanism involving trans-differentiation of other cell types or differentiation of long-lived totipotent stem cells.
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    URL: http://dx.doi.org/10.1007/BF00318500
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    Presence of embryonic myosin in normal postural muscles of the adult rat (1995)
    Springer
    Cell & tissue research 280 (1995), S. 541-548 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Musle ; striated ; skeletal ; Regeneration ; Myosin ; Immunocytochemistry ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Indirect immunofluorescence was used to localize embryonic myosin heavy chains in soleus, adductor longus, tibialis anterior, plantaris, and extensor digitorum longus muscles of 6-month-old rats. A monoclonal antibody (2B6), specifically recognizing rat embryonic myosin, was applied to unfixed, transverse, frozen sections. The number of embryonic myosin-positive (EMP) extrafusal fibers was expressed as a percentage of the total number of fibers. EMP extrafusal fibers were only seen in the soleus and adductor longus muscles, both postural muscles. Approximately 1% of the soleus muscle fibers appeared positively stained for embryonic myosin. The majority of such fibers had a small diameter (〈500 μ2), appeared intensely fluorescent, and typically contained central nuclei. Re-expression of embryonic myosin due to spontaneous fiber denervation is not a likely factor in this study, since alpha-bungarotoxin and N-CAM localization were restricted to the motor end-plate region of EMP fibers. Since embryonic myosin was shown to disappear in all normal-sized myofibers by 2 to 3 months of age, the results suggest that the EMP extrafusal fibers seen in postural muscles of 6 to 12-month-old animals are regenerating myofibers. We speculate that a small number of muscle fibers may be regenerating in normal, adult postural muscles, in response to fiber damage possibly caused by excessive recruitment or overloading.
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    URL: http://dx.doi.org/10.1007/BF00318358
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    Development of melanin-concentrating hormone-immunoreactive elements in the brain of gilthead seabream (Sparus auratus) (1995)
    Springer
    Cell & tissue research 282 (1995), S. 523-526 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Melanin-concentrating hormone ; Immunocytochemistry ; Development ; ontogenetic ; Sparus auratus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The development of the hypothalamic melanin-concentrating hormone (MCH) system of the teleost Sparus auratus has been studied by immunocytochemistry using an anti-salmon MCH serum. Immunoreactive perikarya and fibers are found in embryos, larvae, and juvenile specimens. In juveniles, most labeled neurons are present in the nucleus lateralis tuberis; some are dispersed in the nucleus recessus lateralis and nucleus periventricularis posterior. From the nucleus lateralis tuberis, MCH neurons project a conspicuous tract of fibers to the ventral hypothalamus; this penetrates the pituitary stalk and reaches the neurohypophysis. Most fibers end close to the cells of the pars intermedia, and some reach the adenohypophysial rostral pars distalis. Immunoreactive fibers can also be seen in extrahypophysial localizations, such as the preoptic region and the nucleus sacci vasculosi. In embryos, MCH-immunoreactive neurons first appear at 36 h post-fertilization in the ventrolateral margin of the developing hypothalamus. In larvae, at 4 days post-hatching, perikarya can be observed in the ventrolateral border of the hypothalamus and in the mid-hypothalamus, near the ventricle. At 26 days post-hatching, MCH perikarya are restricted to the nucleus lateralis tuberis. The neurohypophysis possesses MCH-immunoreactive fibers from the second day post-hatching. The results indicate that MCH plays a role in larval development with respect to skin melanophores and cells that secrete melanocyte-stimulating hormone.
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    Ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the dogfish (1995)
    Springer
    Cell & tissue research 282 (1995), S. 33-40 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Neuropeptide Y ; Gastroenteropancreatic (GEP) endocrine system ; Development ; ontogenetic ; Vitellointestinal duct ; Pancreas ; exocrine ; Pancreas ; endocrine ; Immunocytochemistry ; Scyliorhinus torazame (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. This immunocytochemical study was carried out to elucidate the ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the cloudy dogfish, Scyliorhinus torazame. Immunostained cells first appeared in the pancreas of the embryo at the 15-mm stage, and were also detected in the vitellointestinal duct of the yolk stalk at the 20-mm stage. These cells were polymorphic, with occasional processes that were sometimes directed toward the vascular wall or into the cavity of the vitellointestinal duct. At the 34-mm stage, immunostained cells could also be found in the proximal part of the spiral intestine and, by the 74-mm stage, immunopositive cells were present in the gastric mucosa. In the gut and pancreas, the cells gradually increased in number with development, whereas in the vitellointestinal duct and internal yolk sac, they decreased and seemed to disappear following hatching. Thus, in juveniles, the distribution of the neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system had attained that of adults. Electron-microscopic immunocytochemistry demonstrated that, in the labeled cells of the vitellointestinal duct, the neuropeptide Y-like antigen was located in cytoplasmic granules, as in the cells of the gut and pancreas.
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    URL: http://dx.doi.org/10.1007/s004410050456
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    Rat intestinal galactoside-binding lectin L-36 functions as a structural protein in the superficial squamous cells of the esophageal epithelium (1995)
    Springer
    Cell & tissue research 281 (1995), S. 77-83 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Esophagus ; Epithelial cells ; Intestinal lectin ; L-36 ; RI-H fragment ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Using an affinity purified antibody raised against the RI-H fragment of rat intestinal lectin L-36, the latter protein has been identified within the esophageal epithelium by means of ultracryotomy followed by immunogold labeling. The epithelium consists of 4 morphologically distinct cell-types, namely, the basal, spiny, granular and squamous cells, and each of these exhibits a different immunolabeling pattern. The basal cells form a layer on the basal lamina, and in these a diffuse cytoplasmic staining is observed. This basal cell layer is overlaid by spiny cells that extend many cell processes into wide intercellular spaces. In these cells, immunogold particles are found only on small granular inclusions consisting of an electron-lucent homogeneous substance. The granular cells form a third layer over the spiny cells, and are characterized by a number of large granular inclusions with an electron-dense core rimmed by a less electron-dense substance. Immunogold labeling is found on these granules, both on the core and peripheral region. Squamous cell-types constitute the most superficial layer of the epithelium. They are without granular inclusions, and immunogold labeling is confined to the cytoplasmic surface of the thickened plasma membrane. These findings suggest that L-36 is produced in the basal cells as free cytosolic protein, then becomes progressively aggregated into the granular inclusions of the spiny and granular cells, and is eventually transferred onto the cytoplasmic surface of the squamous cell plasma membrane where it may interact with complementary glycoconjugate(s) located at this site. The membrane lining substance thus formed may play a role in stabilizing the squamous cell membranes, thereby maintaining the structural integrity of the epithelium against mechanical stress coming from the esophageal lumen.
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    URL: http://dx.doi.org/10.1007/BF00307960
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    Immunocytochemical localisation of some lysosomal hydrolases, their presence in luminal fluid and their directional secretion by human epididymal cells in culture (1995)
    Springer
    Cell & tissue research 280 (1995), S. 415-425 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Epididymis ; Efferent ducts ; Cell culture ; Immunocytochemistry ; Immunoprecipitation ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The way in which the human epididymis modifies spermatozoa during their sojourn in this structure might be clarified by knowledge of the nature of its secretions. We have examined the presence of several lysosomal hydrolases in human epididymal tissue and fluids, and their synthesis and secretion by monolayer cultures. Tissues were obtained from men undergoing orchidectomy for prostatic carcinoma. The enzymes cathepsin D and acid α-glucosidase were localised in the lysosomes of epithelial cells from the corpus epididymidis, by an immunocytochemical technique. Cathepsin D was also found in epithelial cells of the efferent ducts within lysosomes, apical vesicles and multivesicular bodies. No immunolocalisation of acid glucosidase in the efferent ducts or on the microvilli of the corpus was demonstrable. Cathepsin D, β-hexosaminidase (N-acetylglucosaminidase) and α-glucosidase were measurable in the luminal fluid from the human corpus epididymidis; β-hexosaminidase was secreted into the culture medium by confluent monolayers of epididymal and efferent duct cells. Immunoprecipitation of cell extracts and culture medium of these cultures incubated with 35S-methionine revealed that the precursors of cathepsin D and β-hexosaminidase were synthesized and secreted by such monolayers. Thus, active lytic enzymes are secreted by the human epididymis and could modify sperm membranes.
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    URL: http://dx.doi.org/10.1007/BF00307815
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    Immunolocalization of the Na,K-ATPase in photoreceptor cells of the moth Manduca sexta-evidence for Na,K-ATPase on both the nonmicrovillar and the microvillar domains of the plasma membrane (1995)
    Springer
    Cell & tissue research 281 (1995), S. 119-126 
    Details
    ISSN: 1432-0878
    Keywords: Compound eye ; Photoreceptor cells ; Ion pumps ; Polarity ; Immunocytochemistry ; Manduca sexta (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunohistochemical and physiological studies on various insect photoreceptors have demonstrated that the Na,K-ATPase (sodium pump) is restricted to the nonreceptive nonmicrovillar area of the plasma membrane. Here, we examined the distribution of the Na,K-ATPase in photoreceptor cells of the superposition-type compound eye in the moth Manduca sexta. Using immunofluorescent and immunogold cytochemistry, we show that the Na,K-ATPase is localized to both the nonmicrovillar and the microvillar parts of the plasma membrane. Manduca photoreceptors thus deviate from the common concept that the sodium pump and the molecular components of the photoreceptive machinery reside on different domains of the plasma membrane.
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    Putative neurohemal areas in the peripheral nervous system of an insect, Gryllus bimaculatus, revealed by immunocytochemistry (1995)
    Springer
    Cell & tissue research 281 (1995), S. 43-61 
    Details
    ISSN: 1432-0878
    Keywords: Neurohemal areas ; Neuropeptides ; Monoamines ; Immunocytochemistry ; Nervous system, insect ; Gryllus bimaculatus (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The morphology and position of putative neurohemal areas in the peripheral nervous system (ventral nerve cord and retrocerebral complex) of the cricket Gryllus bimaculatus are described. By using antisera to the amines dopamine, histamine, octopamine, and serotonin, and the neuropeptides crustacean cardioactive peptide, FMRFamide, leucokinin 1, and proctolin, an extensive system of varicose fibers has been detected throughout the nerves of all neuromeres, except for nerve 2 of the prothoracic ganglion. Immunoreactive varicose fibers occur mainly in a superficial position at the neurilemma, indicating neurosecretory storage and release of neuroactive compounds. The varicose fibers are projections from central or peripheral neurons that may extend over more than one segment. The peripheral fiber varicosities show segment-specific arrangements for each of the substances investigated. Immunoreactivity to histamine and octopamine is mainly found in the nerves of abdominal segments, whereas serotonin immunoreactivity is concentrated in subesophageal and terminal ganglion nerves. Immunoreactivity to FMRFamide and crustacean cardioactive peptide is widespread throughout all segments. Structures immunoreactive to leucokinin 1 are present in abdominal nerves, and proctolin immunostaining is found in the terminal ganglion and thoracic nerves. Codistribution of peripheral varicose fiber plexuses is regularly seen for amines and peptides, whereas the colocalization of substances in neurons has not been detected for any of the neuroactive compounds investigated. The varicose fiber system is regarded as complementary to the classical neurohemal organs.
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    URL: http://dx.doi.org/10.1007/BF00307957
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    Immunolocalization of interleukin-1α in rat mandibular molars and its enhancement after in vivo injection of epidermal growth factor (1995)
    Springer
    Cell & tissue research 280 (1995), S. 21-26 
    Details
    ISSN: 1432-0878
    Keywords: Interleukin ; Stellate reticulum ; Immunocytochemistry ; Epidermal growth factor ; Interleukin-1 receptor type I messenger RNA ; Tooth eruption ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunolocalization of interleukin-1α in the first mandibular molars of rats from day 0–12 postnatally showed that the protein was localized in the epithelial stellate reticulum adjacent to the dental follicle. Staining of the stellate reticulum was most prominent in the early days postnatally and was absent by postnatal day 11. Injection of epidermal growth factor into rats at day 0 greatly increased the intensity of the staining for interleukin-1α in the stellate reticulum. Epidermal growth factor (EGF) enhanced the gene expression of interleukin-1α in stellate reticulum cells in vitro, and this study suggests there is enhanced translation of interleukin-1α messenger RNA in the stellate reticulum following EGF injection. In turn, the interleukin-1α may exert its effect on the dental follicle cells adjacent to the stellate reticulum because EGF also enhanced expression of the interleukin-1 receptor type I messenger RNA in cultured dental follicle cells as well as enhancing its expression in vivo. In view of the fact that injection of EGF will stimulate precocious eruption of teeth, its stimulus of interleukin-1α synthesis in the stellate reticulum may be the mechanism by which EGF initiates a cascade of molecular events to signal the onset of tooth eruption.
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    Expression of the endogenous 14-kDa β-galactoside-binding lectin galectin in normal human skin (1995)
    Springer
    Cell & tissue research 280 (1995), S. 1-10 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Galectin ; β-Galactoside-binding lectin ; Human ; Skin ; Immunocytochemistry ; Immunohistochemistry ; Hybridization ; in situ ; Langerhans cell ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The localization of an endogenous 14-kDa β-galactoside-binding lectin (galectin) and its pattern of gene expression were examined in normal human skin by light- and electron microscopy. Under the light microscope, immunostaining of 14-kDa galectin was observed in the cell membrane of cells in the basal and spinous layers of the epidermis. Galectin was also found in the Langerhans cells, as shown by double labeling using anti-14-kDa galectin and anti-CD1a antibodi es. In the dermis, immunostaining for the 14-kDa galectin was positive in the extracellular matrix and fibroblasts. At the electron-microscopic level of resolution, galectin was located primarily along the plasma membrane of keratinocytes, and in both the cytoplasm and nucleus of Langerhans cells in the epidermis, whereas in the dermis it was detected in the extracellular matrix and in both the nucleus and cytoplasm of fibroblasts. The gene expression of 14-kDa galectin was visualized by the HRP-staining me thod following in situ hybridization techniques. The expression was detected in the cytoplasm of cells in the basal and spinous layers of the epidermis; whereas, in the dermis, it was detected in the cytoplasm of fibroblasts. Moreover, SDS-polyacrylamide gel electrophoresis and lectin-blot analysis revealed that this galectin bound to glycoproteins of approximately 17, 62, and 72 kDa in the epidermis and to those of 29, 54, and 220 kDa in the dermis. The present study indicates that 1) normal human skin produces the β-galactoside-binding 14-kDa galectin, and 2) this galectin is located in both the epidermis, particularly in the keratinocytes and Langerhans cells, and in the dermis. These results suggest that galectin is important for cell-cell contact and/or adhesion in the epidermis and for cell-extracellular matrix interaction in the dermis.
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    Expression of the endogenous 14-kDa β-galactoside-binding lectin galectin in normal human skin (1995)
    Springer
    Cell & tissue research 280 (1995), S. 1-10 
    Details
    ISSN: 1432-0878
    Keywords: Galectin ; β-Galactoside-binding lectin ; Human ; Skin ; Immunocytochemistry ; Immunohistochemistry ; Hybridization, in situ ; Langerhans cell ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localization of an endogenous 14-kDa β-galactoside-binding lectin (galectin) and its pattern of gene expression were examined in normal human skin by light- and electron microscopy. Under the light microscope, immunostaining of 14-kDa galectin was observed in the cell membrane of cells in the basal and spinous layers of the epidermis. Galectin was also found in the Langerhans cells, as shown by double labeling using anti-14-kDa galectin and anti-CD1a antibodies. In the dermis, immunostaining for the 14-kDa galectin was positive in the extracellular matrix and fibroblasts. At the electron-microscopic level of resolution, galectin was located primarily along the plasma membrane of keratinocytes, and in both the cytoplasm and nucleus of Langerhans cells in the epidermis, whereas in the dermis it was detected in the extracellular matrix and in both the nucleus and cytoplasm of fibroblasts. The gene expression of 14-kDa galectin was visualized by the HRP-staining method following in situ hybridization techniques. The expression was detected in the cytoplasm of cells in the basal and spinous layers of the epidermis; whereas, in the dermis, it was detected in the cytoplasm of fibroblasts. Moreover, SDS-polyacrylamide gel electrophoresis and lectin-blot analysis revealed that this galectin bound to glycoproteins of approximately 17, 62, and 72 kDa in the epidermis and to those of 29, 54, and 220 kDa in the dermis. The present study indicates that 1) normal human skin produces the β-galactoside-binding 14-kDa galectin, and 2) this galectin is located in both the epidermis, particularly in the keratinocytes and Langerhans cells, and in the dermis. These results suggest that galectin is important for cell-cell contact and/or adhesion in the epidermis and for cell-extracellular matrix interaction in the dermis.
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    Characterization and localization of an Mr 225 kDa polypeptide specifically involved in the defence mechanisms of the clam Tapes semidecussatus (1995)
    Springer
    Cell & tissue research 280 (1995), S. 27-37 
    Details
    ISSN: 1432-0878
    Keywords: Defence mechanisms ; Encapsulation ; Granulocytes ; Immunocytochemistry ; Parasitism ; Perkinsus sp. (Protozoa) ; Tapes semidecussatus (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Parasitosis by the trophozoite protozoan Perkinsus sp. (Apicomplexa, Perkinsea) induces in the gill filaments of the clam Tapes semidecussatus (Mollusca, Bivalvia) a cellular reaction, which is constituted by infiltrated granulocytes. This cellular reaction has characteristics of those of a holocrine gland, since the parasites are encapsulated by the secretion product of the granulocytes after cell death. An enriched fraction of prezoosporangia and their associated capsule was obtained after culture of the parasitized gills in fluid thioglycollate medium. Specific polypeptides from this fraction were separated by SDS-PAGE and isolated for rabbit immunizations. The serum obtained against an Mr 225 kDa polypeptide, revealed its exclusive localization in the capsule and in the granules of the infiltrated granulocytes, thus indicating that this polypeptide is synthesized by these cells and secreted, in a polarized way, around the trophozoites resulting in their encapsulation. Selective deglycosylation of the polypeptide, by Endo H and alkaline β-elimination, did not show an effect on its molecular weight or antibody recognition. Furthermore, the absence of the 225 kDa band in the Western-blots of non-parasitized gills indicated the specific association of this polypeptide with the parasitosis. Finally, this is the first tissue-specific factor described in molluscs in relation to defence mechanisms.
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    Immunocytochemical localization of ribonuclease in yolk granules of adult Rana cttesbeiana oocytes (1995)
    Springer
    Cell & tissue research 280 (1995), S. 259-265 
    Details
    ISSN: 1432-0878
    Keywords: Oocyte ; Yolk granules ; Ribonuclease ; Immunocytochemistry ; Bullfrog, Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract To determine the localization of the pyrimidine-guanine sequence-specific ribonuclease in Rana catesbeiana (bullfrog) oocytes, the RNase was first isolated and used to prepare a specific rabbit antiserum. Only one protein of similar molecular size to the RNase was immunoprecipitated from ovary homogenate by the antiserum, but two bands were observed by Western blotting analysis. These two proteins were shown by further purification of antibody and Western blotting analysis to have similar antigenicity. Immunoprecipitation and Western blotting of tissue homogenates showed that the RNase was found predominantly in the ovary, but not in other tissues. The specific localization of the RNase was determined by immuno-electron microscopy of oocyte sections incubated with the specific antiserum; the yolk granules, but not other organelles, were found to contain the RNase. Most of the RNase was evenly distributed in the lateral amorphous area of the yolk granule but not in the central yolk crystal area which contains stored vitellogenin proteins. Our results indicate that the RNase is compartmentalized in the yolk granules of oocytes, which might prevent damage to cellular RNAs.
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    Rat intestinal galactoside-binding lectin L-36 functions as a structural protein in the superficial squamous cells of the esophageal epithelium (1995)
    Springer
    Cell & tissue research 281 (1995), S. 77-83 
    Details
    ISSN: 1432-0878
    Keywords: Esophagus ; Epithelial cells ; Intestinal lectin, L-36 ; RI-H fragment ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Using an affinity purified antibody raised against the RI-H fragment of rat intestinal lectin L-36, the latter protein has been identified within the esophageal epithelium by means of ultracryotomy followed by immunogold labeling. The epithelium consists of 4 morphologically distinct cell-types, namely, the basal, spiny, granular and squamous cells, and each of these exhibits a different immunolabeling pattern. The basal cells form a layer on the basal lamina, and in these a diffuse cytoplasmic staining is observed. This basal cell layer is overlaid by spiny cells that extend many cell processes into wide intercellular spaces. In these cells, immunogold particles are found only on small granular inclusions consisting of an electron-lucent homogeneous substance. The granular cells from a third layer over the spiny cells, and are characterized by a number of large granular inclusions with an electron-dense core rimmed by a less electron-dense substance. Immunogold labeling is found on these granules, both on the core and peripheral region. Squamous cell-types constitute the most superficial layer of the epithelium. They are without granular inclusions, and immunogold labeling is confined to the cytoplasmic surface of the thickened plasma membrane. These findings suggest that L-36 is produced in the basal cells as free cytosolic protein, then becomes progressively aggregated into the granular inclusions of the spiny and granular cells, and is eventually transferred onto the cytoplasmic surface of the squamous cell plasma membrane where it may interact with complementary glycoconjugate(s) located at this site. The membrane lining substance thus formed may play a role in stabilizing the squamous cell membranes, thereby maintaining the structural integrity of the epithelium against mechanical stress coming from the esophageal lumen.
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    Localization of integrin subunits α6 and β1 during somitogenesis in the long-tailed macaque (M. fascicularis) (1995)
    Springer
    Cell & tissue research 281 (1995), S. 101-108 
    Details
    ISSN: 1432-0878
    Keywords: Somite ; Intergin ; Extracellular matrix, structures ; Embryo ; Laminin ; Immunocytochemistry ; Macaca fascicularis (Primates)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of integrin subunits α6 and β1, and the α6β1 integrin ligand, laminin, was examined during somitogenesis in developmental stages 11, 13, and 16 in the long-tailed macaque, using peroxidase immunocytochemistry. Within differentiating somites in stage 11, α6 expression was observed in the sclerotome, basal surface of dermamyotomal cells adjacent to the basal lamina and on scattered cells throughout the dermamyotome. In further advanced somites in stages 13 and 16, α6 immunoreactivity become restricted to the myotome, α6 was expressed on mesenchymal core cells within the myocele of undifferentiated epitheliod somites and the ventromedial wall of somites commencing differentiation at each stage. β1 distribution resembled that of α6 in stage 11 somitic tissue, however, it remained present on myotome and sclerotome cells in the later stages, and was also expressed on dermatomal cells in stage 16. Laminin immunoreactivity, while more intense and prevalent than α6 and β1 in each stage examined, occurred on the same somite cell populations as the 2 integrin subunits. These results show a defined distribution of α6 on somitic tissue, and suggest this integrin is involved in somite differentiation. They also support a possible role for α6 in myoblast formation and migration. Overlapping of β1 and laminin immunoreactivity with that of α6 further suggests that α6 paris with β1 as a functional heterodimer for laminin in defined somitic regions.
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    Immunocytochemical, electrophoresis, and immunoblot analysis of Heliothis virescens gap junctions isolated in the presence and absence of protease inhibitors (1995)
    Springer
    Cell & tissue research 281 (1995), S. 179-186 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Gap junction ; Intercellular junction ; Insect ; Arthropod ; Immunocytochemistry ; Heliothis virescens (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Gap junction-enriched fractions were prepared from larvae of the tobacco budworm Heliothis virescens using the NaOH procedure in the presence or absence of protease inhibitors and were analyzed by SDS-PAGE, immunoblotting and EM immunocytochemistry. Protease inhibitor fractions contained a 48-kDa protein in addition to the ∼10 proteins in fractions with and without inhibitors. Three polyclonal antibodies were used as probes for gap junction plaques and proteins: R16, against an ∼40-kDa candidate gap junction protein from Drosophila melanogaster; R17, against the 40-kDa candidate gap junction protein from H. virescens; and R18AP, an affinity purified antibody against a consensus sequence of N-terminal amino acids 2–21 of the H. virescens 40-kDa protein. R16, R17, and R18AP stain the 40- and 48-kDa proteins, R16 and R18AP stain a 64-kDa protein, and R16 stains an ∼30-kDa protein in the absence of inhibitors. Inclusion of protease inhibitors had no effect on gap junction ultrastructure. R16 and R17 label gap junction plaques in crude membrane and NaOH fractions, whereas R18AP exhibits only a low level of reactivity with gap junctions in crude membrane fractions and none with gap junctions in NaOH fractions. The results show that the 30-, 40-, 48- and 64-kDa proteins are immunologically related and are associated with gap junctions in H. virescens, the N-terminus of the 40-kDa protein is relatively inaccessible or easily lost, and the 48-kDa protein is protease-sensitive.
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    Unusual distribution of tubulin isoforms in the snail Lymnaea stagnalis (1995)
    Springer
    Cell & tissue research 281 (1995), S. 507-515 
    Details
    ISSN: 1432-0878
    Keywords: Microtubules ; Isoforms ; Nervous system ; Locomotion ; Cilia ; Immunocytochemistry ; Western blotting ; Lymnaea stagnalis (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunocytochemistry and Western blotting techniques demonstrated that the nervous system and foot of the pond snail Lymnaea stagnalis are rich sources of tubulin, which can be extracted and assembled in vitro in the presence of taxol. Various broad-spectrum antibodies raised against α-tubulin and β-tubulin yielded qualitatively similar results. One monoclonal antibody to trypanosome α-tubulin, however, labelled α-tubulin more strongly on both probed sections and Western blots. Cytochemistry and immunoblotting revealed that tyrosinated tubulin constitutes a large proportion of total α-tubulin in locomotor cilia of the foot and in axons of the nervous system. Detyrosinated tubulin also appeared to be abundant in the foot cilia but only a very faint band of detyrosinated tubulin was found on protein blots extracted from the central ganglia, and staining was barely detectable in central ganglia or peripheral nerves. Similarly, acetylated tubulin appeared to be abundant in foot cilia, but Western blotting indicated only low levels of acetylated tubulin in the nervous system. Immunocytochemistry indicated that, while most neurons possessed little or no acetylated tubulin, a small number of axons contained significant amounts of this isoform. Thus, while a large amount of tubulin was expected in the nervous system and locomotor cilia of L. stagnalis, the observed distribution of isoforms was unanticipated. Specifically, neurons of other organisms have generally been reported to contain substantial amounts of both detyrosinated α-tubulin and acetylated α-tubulin. Our results indicate that such findings cannot be generalized across all species. L. stagnalis, with its well studied nervous system and unusual distribution of tubulin isoforms, may prove to be particularly useful for studying the roles of tubulin isoforms in microtubule function and cell activity.
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    Immunocytochemical characterization of the accessory medulla in the cockroach Leucophaea maderae (1995)
    Springer
    Cell & tissue research 282 (1995), S. 3-19 
    Details
    ISSN: 1432-0878
    Keywords: Circadian rhythm ; Colocalization ; Immunocytochemistry ; Brain (CNS), invertebrate ; Optic lobe ; Pigment-dispersing hormone, insect ; Leucophaea maderae (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Several lines of evidence suggest that pigment-dispersing hormone-immunoreactive neurons with ramifications in the accessory medulla are involved in the circadian system of insects. The present study provides a detailed analysis of the anatomical and neurochemical organization of the accessory medulla in the brain of the cockroach Leucophaea maderae. We show that the accessory medulla is compartmentalized into central dense nodular neuropil surrounded by a shell of coarse fibers. It is innervated by neurons immunoreactive to antisera against serotonin and the neuropeptides allatostatin 7, allatotropin, corazonin, gastrin/cholecystokinin, FMRFamide, leucokinin I, and pigment-dispersing hormone. Some of the immunostained neurons appear to be local neurons of the accessory medulla, whereas others connect this neuropil to various brain areas, including the lamina, the contralateral optic lobe, the posterior optic tubercles, and the superior protocerebrum. Double-label experiments show the colocalization of immunoreactivity against pigment-dispersing hormone with compounds related to FMRFamide, serotonin, and leucokinin I. The neuronal and neurochemical organization of the accessory medulla is consistent with the current hypothesis for a role of this brain area as a circadian pacemaking center in the insect brain.
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    URL: http://dx.doi.org/10.1007/BF00319128
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    Ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the dogfish (1995)
    Springer
    Cell & tissue research 282 (1995), S. 33-40 
    Details
    ISSN: 1432-0878
    Keywords: Neuropeptide Y ; Gastroenteropancreatic (GEP) endocrine system ; Development, ontogenetic ; Vitellointestinal duct ; Pancreas, exocrine ; Pancreas, endocrine ; Immunocytochemistry ; Scyliorhinus torazame (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract This immunocytochemical study was carried out to elucidate the ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the cloudy dogfish, Scyliorhinus torazame. Immunostained cells first appeared in the pancreas of the embryo at the 15-mm stage, and were also detected in the vitellointestinal duct of the yolk stalk at the 20-mm stage. These cells were polymorphic, with occasional processes that were sometimes directed toward the vascular wall or into the cavity of the vitellointestinal duct. At the 34-mm stage, immunostained cells could also be found in the proximal part of the spiral intestine and, by the 74-mm stage, immunopositive cells were present in the gastric mucosa. In the gut and pancreas, the cells gradually increased in number with development, whereas in the vitellointestinal duct and internal yolk sac, they decreased and seemed to disappear following hatching. Thus, in juveniles, the distribution of the neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system had attained that of adults. Electron-microscopic immunocytochemistry demonstrated that, in the labeled cells of the vitellointestinal duct, the neuropeptide Y-like antigen was located in cytoplasmic granules, as in the cells of the gut and pancreas.
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    URL: http://dx.doi.org/10.1007/BF00319130
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    Immunocytochemical characterization of the accessory medulla in the cockroach Leucophaea maderae (1995)
    Springer
    Cell & tissue research 282 (1995), S. 3-19 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Circadian rhythm ; Colocalization ; Immunocytochemistry ; Brain (CNS) ; invertebrate ; Optic lobe ; Pigment-dispersing hormone ; insect ; Leucophaea maderae (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Several lines of evidence suggest that pigment-dispersing hormone-immunoreactive neurons with ramifications in the accessory medulla are involved in the circadian system of insects. The present study provides a detailed analysis of the anatomical and neurochemical organization of the accessory medulla in the brain of the cockroach Leucophaea maderae. We show that the accessory medulla is compartmentalized into central dense nodular neuropil surrounded by a shell of coarse fibers. It is innervated by neurons immunoreactive to antisera against serotonin and the neuropeptides allatostatin 7, allatotropin, corazonin, gastrin/cholecystokinin, FMRFamide, leucokinin I, and pigment-dispersing hormone. Some of the immunostained neurons appear to be local neurons of the accessory medulla, whereas others connect this neuropil to various brain areas, including the lamina, the contralateral optic lobe, the posterior optic tubercles, and the superior protocerebrum. Double-label experiments show the colocalization of immunoreactivity against pigment-dispersing hormone with compounds related to FMRFamide, serotonin, and leucokinin I. The neuronal and neurochemical organization of the accessory medulla is consistent with the current hypothesis for a role of this brain area as a circadian pacemaking center in the insect brain.
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    URL: http://dx.doi.org/10.1007/s004410050454
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    Immunocytological evidence for oxytocin neurons in the human fetal hypothalamus (1978)
    Springer
    Cell & tissue research 188 (1978), S. 259-264 
    Details
    ISSN: 1432-0878
    Keywords: Hypothalamus ; Human fetus ; Oxytocin ; Neurophysin ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The use of antibodies against oxytocin or neurophysin enabled the detection by immunocytochemistry of oxytocin-neurophysin neurons in the hypothalamus in the human fetus. The perikarya of these neurons are located in the paraventricular and supraoptic nuclei. Immunoreactive neurons occur in the median eminence. The neurophysin immunoreactive neurons were more numerous than the oxytocin immunoreactive neurons. The specificity of the immunocytological reaction was controlled. The first oxytocin-neurophysin neurons are seen as early as the 14th week of gestation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00222635
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    Immunocytochemical localisation of some lysosomal hydrolases, their presence in luminal fluid and their directional secretion by human epididymal cells in culture (1995)
    Springer
    Cell & tissue research 280 (1995), S. 415-425 
    Details
    ISSN: 1432-0878
    Keywords: Epididymis ; Efferent ducts ; Cell culture ; Immunocytochemistry ; Immunoprecipitation ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The way in which the human epididymis modifies spermatozoa during their sojourn in this structure might be clarified by knowledge of the nature of its secretions. We have examined the presence of several lysosomal hydrolases in human epididymal tissue and fluids, and their synthesis and secretion by monolayer cultures. Tissues were obtained from men undergoing orchidectomy for prostatic carcinoma. The enzymes cathepsin D and acid α-glucosidase were localised in the lysosomes of epithelial cells from the corpus epididymidis, by an immunocytochemical technique. Cathepsin D was also found in epithelial cells of the efferent ducts within lysosomes, apical vesicles and multivesicular bodies. No immunolocalisation of acid glucosidase in the efferent ducts or on the microvilli of the corpus was demonstrable. Cathepsin D, β-hexosaminidase (N-acetylglucosaminidase) and α-glucosidase were measurable in the luminal fluid from the human corpus epididymidis; β-hexosaminidase was secreted into the culture medium by confluent monolayers of epididymal and efferent duct cells. Immunoprecipitation of cell extracts and culture medium of these cultures incubated with 35S-methionine revealed that the precursors of cathepsin D and β-hexosaminidase were synthesized and secreted by such monolayers. Thus, active lytic enzymes are secreted by the human epididymis and could modify sperm membranes.
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    URL: http://dx.doi.org/10.1007/BF00307815
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    Presence of embryonic myosin in normal postural muscles of the adult rat (1995)
    Springer
    Cell & tissue research 280 (1995), S. 541-548 
    Details
    ISSN: 1432-0878
    Keywords: Musle, striated, skeletal ; Regeneration ; Myosin ; Immunocytochemistry ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Indirect immunofluorescence was used to localize embryonic myosin heavy chains in soleus, adductor longus, tibialis anterior, plantaris, and extensor digitorum longus muscles of 6-month-old rats. A monoclonal antibody (2B6), specifically recognizing rat embryonic myosin, was applied to unfixed, transverse, frozen sections. The number of embryonic myosin-positive (EMP) extrafusal fibers was expressed as a percentage of the total number of fibers. EMP extrafusal fibers were only seen in the soleus and adductor longus muscles, both postural muscles. Approximately 1% of the soleus muscle fibers appeared positively stained for embryonic myosin. The majority of such fibers had a small diameter (〈500 ν), appeared intensely fluorescent, and typically contained central nuclei. Re-expression of embryonic myosin due to spontaneous fiber denervation is not a likely factor in this study, since alpha-bungarotoxin and N-CAM localization were restricted to the motor end-plate region of EMP fibers. Since embryonic myosin was shown to disappear in all normal-sized myofibers by 2 to 3 months of age, the results suggest that the EMP extrafusal fibers seen in postural muscles of 6 to 12-month-old animals are regenerating myofibers. We speculate that a small number of muscle fibers may be regenerating in normal, adult postural muscles, in response to fiber damage possibly caused by excessive recruitment or overloading.
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    URL: http://dx.doi.org/10.1007/BF00318358
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    Sources of inputs to longitudinal muscle motor neurons and ascending interneurons in the guinea-pig small intestine (1995)
    Springer
    Cell & tissue research 280 (1995), S. 549-560 
    Details
    ISSN: 1432-0878
    Keywords: Enteric nervous system ; Immunocytochemistry ; Calretinin ; Calbindin ; Bombesin ; Small intestine ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Light- and electron-microscopic studies were used to investigate connections between specific subgroups of neurons in the myenteric plexus of the guineapig small intestine. Inputs to two classes of calretinin-immunoreactive (IR) nerve cells, longitudinal muscle motor neurons and ascending interneurons, were examined. Inputs from calbindin-IR primary sensory neurons and from three classes of descending interneurons were studied. Electron-microscopic analysis showed that calbindin-IR axons formed two types of inputs, synapses and close contacts, on calretinin-IR neurons. About 40% of inputs to the longitudinal muscle motor neurons and 70% to ascending interneurons were calbindin-IR. Approximately 50% of longitudinal muscle motor neurons were surrounded by bombesin-IR dense pericellular baskets and 40% by closely apposed varicosities. At the electron-microscope level, the bombesin-IR varicosities were found to form synapses and close contacts with the motor neurons. Dense pericellular baskets with bombesin-IR surrounded 36% of all ascending interneurons, and a further 17% had closely apposed varicosities. Somatostatin-and 5-HT-IR descending interneurons provided no dense pericellular baskets to calretinin-IR nerve cells. Thus, calretinin-IR, longitudinal muscle motor neurons and ascending interneurons receive direct synaptic inputs from intrinsic primary sensory neurons and from non-cholinergic, bombesin-IR, descending interneurons.
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    URL: http://dx.doi.org/10.1007/BF00318359
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    Localization of the 110 kDa receptor for laminin in brains of embryonic and postnatal mice (1995)
    Springer
    Cell & tissue research 279 (1995), S. 371-377 
    Details
    ISSN: 1432-0878
    Keywords: Laminin ; Nerve tracts ; Ontogenetic development ; Brain ; Immunocytochemistry ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Laminin, a large glycoprotein of the basement membrane that promotes the growth of nerve cell processes in vitro has also been detected in the brains of developing embryos in situ where it is postulated to promote or guide neural outgrowth. We have investigated the histological and developmental patterns of a receptor to a specific pentapeptide sequence in the A chain of the laminin molecule (PA22-2 or IKVAV) that has been identified as a neuron growth-promoting sequence. Standard immunocytochemical procedures were used to localize the receptor by means of a polyclonal antibody to affinity-purified receptor (MR=110 kDa) from mouse brains. Results for postnatal stages (P) stages (P 1,7,8,25,30,and adult) show that the 110 kDa receptor is localized in fibers in the cortex and hippocampus, in astroglial cells at the surface of the cortex, and in neuronal cell bodies in the hippocampus. In contrast, the A-chain ligand is localized in cell bodies in the same regions at P stages. For embryonic stages (E) (E 14 and E 16) the receptor is localized in bundles of fibers in the superficial and deep cortical layers, and in cell bodies in these regions at E 14 only. Staining for the A chain ligand of the receptor was first seen postnatally. We speculate that the inverse histological pattern of receptor and ligand with respect to cell bodies and fibers may reflect a role in controlling axon guidance during development or repair during regeneration.
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    URL: http://dx.doi.org/10.1007/BF00318494
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    The distribution of neurones immunoreactive for β-tyrosine hydroxylase, dopamine and serotonin in the ventral nerve cord of the cricket, Gryllus bimaculatus (1995)
    Springer
    Cell & tissue research 280 (1995), S. 583-604 
    Details
    ISSN: 1432-0878
    Keywords: Dopamine ; Serotonin ; Tyrosine hydroxylase ; Immunocytochemistry ; Nervous system, insect-Gryllus bimaculatus (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The cellular localization of the biogenic amines dopamine and serotonin was investigated in the ventral nerve cord of the cricket, Gryllus bimaculatus, using antisera raised against dopamine, β-tyrosine hydroxylase and serotonin. Dopamine-(n〈-70) and serotonin-immunoreactive (n〈-120) neurones showed a segmental arrangement in the ventral nerve cord. Some neuromeres, however, did not contain dopamine-immunoreactive cell bodies. The small number of stained cells allowed complete identification of brain and thoracic cells, including intersegmentally projecting axons and terminal arborizations. Dopamine-like immunostaining was found primarily in plurisegmental interneurones with axons descending to the soma-ipsilateral hemispheres of the thoracic and abdominal ganglia. In contrast, serotonin-immunostaining occurred predominantly in interneurones projecting via soma-contralaterally ascending axons to the thorax and brain. In addition, serotonin-immunoreactivity was also present in efferent cells and afferent elements. Serotonin-immunoreactive, but no dopamine-immunoreactive, varicose fibres were observed on the surface of some peripheral nerves. Varicose endings of both dopamine-and serotonin-immunoreactive neurones occurred in each neuromere and showed overlapping neuropilar projections in dorsal and medial regions of the thoracic ganglia. Ventral associative neuropiles lacked dopamine-like immunostaining but were innervated by serotonin-immunoreactive elements. A colocalization of the two amines was not observed. The topographic representation of neurone types immunoreactive for serotonin and dopamine is discussed with respect to possible modulatory functions of these biogenic amines in the central nervous system of the cricket.
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    URL: http://dx.doi.org/10.1007/BF00318362
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    Specific immunocytochemical localization of cathepsin E at the ruffled border membrane of active osteoclasts (1995)
    Springer
    Cell & tissue research 281 (1995), S. 85-91 
    Details
    ISSN: 1432-0878
    Keywords: Cathepsin E ; Aspartic proteinase ; Osteoclasts ; Immunocytochemistry ; Rat (WKA)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The immunocytochemical localization of cathepsin E, a non-lysosomal aspartic proteinase, was investigated in rat osteoclasts using the monospecific antibody to this protein. At the light-microscopic level, the preferential immunoreactivity for cathepsin E was found at high levels in active osteoclasts in the physiological bone modeling process. Neighboring osteoblastic cells were devoid of its immunoreactivity. At the electron-microscopic level, cathepsin E was exclusively confined to the apical plasma membrane at the ruffled border of active osteoclasts and the eroded bone surface. Cathepsin E was also concentrated in some endocytotic vacuoles of various sizes in the vicinity of the ruffled border membrane, some of which appeared to be secondary lysosomes containing the phagocytosed materials. These results strongly suggest that this enzyme is involved both in the extracellular degradation of the bone organic matrix and in the intracellular breakdown of the ingested substances in osteoclasts.
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    URL: http://dx.doi.org/10.1007/BF00307961
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    Localization of the 110 kDa receptor for laminin in brains of embryonic and postnatal mice (1995)
    Springer
    Cell & tissue research 279 (1995), S. 371-377 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Laminin ; Nerve tracts ; Ontogenetic development ; Brain ; Immunocytochemistry ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Laminin, a large glycoprotein of the basement membrane that promotes the growth of nerve cell processes in vitro has also been detected in the brains of developing embryos in situ where it is postulated to promote or guide neural outgrowth. We have investigated the histological and developmental patterns of a receptor to a specific pentapeptide sequence in the A chain of the laminin molecule (PA22-2 or IKVAV) that has been identified as a neuron growth-promoting sequence. Standard immunocytochemical procedures were used to localize the receptor by means of a polyclonal antibody to affinity-purified receptor (MR=110 kDa) from mouse brains. Results for postnatal stages (P) stages (P 1,7,8,25,30,and adult) show that the 110 kDa receptor is localized in fibers in the cortex and hippocampus, in astroglial cells at the surface of the cortex, and in neuronal cell bodies in the hippocampus. In contrast, the A-chain ligand is localized in cell bodies in the same regions at P stages. For embryonic stages (E) (E 14 and E 16) the receptor is localized in bundles of fibers in the superficial and deep cortical layers, and in cell bodies in these regions at E 14 only. Staining for the A chain ligand of the receptor was first seen postnatally. We speculate that the inverse histological pattern of receptor and ligand with respect to cell bodies and fibers may reflect a role in controlling axon guidance during development or repair during regeneration.
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    URL: http://dx.doi.org/10.1007/s004410050293
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    Regeneration and post-metamorphic development of the central nervous system in the protochordate Ciona intestinalis: a study with monoclonal antibodies (1995)
    Springer
    Cell & tissue research 279 (1995), S. 421-432 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Trans-differentiation ; Proliferation ; Bromodeoxyuridine ; Immunocytochemistry ; Regeneration ; Ciona intestinalis (Tunicata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. In this study, we use three monoclonal antibodies that recognise antigens present in the central nervous system of the ascidian Ciona intestinalis to study regeneration and post-metamorphic development of the neural ganglion. We have also used bromodeoxyuridine labelling to study generation of the neuronal precursor cells. The first antibody, CiN 1, recognises all neurones in the ganglion, whereas the second, CiN 2, recognises only a subpopulation of the large cortical neurones. Western blotting studies show that CiN 2 recognises two membrane-bound glycoproteins of apparent Mr 129 and 100 kDa. CiN 1 is not reactive on Western blots. Immunocytochemical studies with these antibodies show that CiN 1-immunoreactive neurone-like cells are present at the site of regeneration as early as 5–7 days post-ablation, a sub-population of CiN 2-immunoreactive cells being detected by 9–12 days post-ablation. The third antibody, ECM 1, stains extracellular matrix components and recognises two diffuse bands on Western blots of whole-body and ganglion homogenates. The temporal and spatial pattern of appearance of CiN 1 and CiN 2 immunoreactivity both during post-metamorphic development and in regeneration occurs in the same sequence in both processes. Studies with bromodeoxyuridine show labelled nuclei in some neurones in the regenerating ganglion. Plausibly these originate from the dorsal strand, an epithelial tube that reforms by cell proliferation during the initial phases of regeneration. A second population of cells, the large cortical neurones, do not incorporate bromodeoxyuridine and thus must have been born prior to the onset of regeneration. This latter finding indicates a mechanism involving trans-differentiation of other cell types or differentiation of long-lived totipotent stem cells.
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    URL: http://dx.doi.org/10.1007/s004410050299
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    Localization of choline acetyltransferase-expressing neurons in the larval visual system of Drosophila melanogaster (1995)
    Springer
    Cell & tissue research 282 (1995), S. 193-202 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Choline acetyltransferase ; Cholinergic neuron ; Visual system ; Bolwig’s organ ; Immunocytochemistry ; In situ hybridization ; Drosophila melanogaster (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Choline acetyltransferase (ChAT) is the enzyme catalyzing the biosynthesis of acetylcholine and is considered to be a phenotypically specific marker for cholinergic neurons. We have examined the distribution of ChAT-expressing neurons in the larval nervous system of Drosophila melanogaster by three different but complementary techniques: in situ hybridization with a cRNA probe to ChAT messenger RNA, immunocytochemistry using a monoclonal anti-ChAT antibody, and X-gal staining of transformed animals carrying a reporter gene composed of 7.4  kb of 5′ flanking DNA from the ChAT gene fused to a lacZ reporter gene. All three techniques demonstrated ChAT-expressing neurons in the larval visual system. In embryos, the photoreceptor organ (Bolwig’s organ) exhibited strong cRNA hybridization signals. The optic lobe of late third-instar larvae displayed ChAT immunoreactivity in Bolwig’s nerve and a neuron close to the insertion site of the optic stalk. This neuron’s axon ran in parallel with Bolwig’s nerve to the larval optic neuropil. This neuron is likely to be a first-order interneuron of the larval visual system. Expression of the lacZ reporter gene was also detected in Bolwig’s organ and the neuron stained by anti-ChAT antibody. Our observations indicate that acetylcholine may be a neurotransmitter in the larval photoreceptor cells as well as in a first-order interneuron in the larval visual system of Drosophila melanogaster.
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    URL: http://dx.doi.org/10.1007/s004410050472
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    Origin and destination of globules and irregular masses in the gonadotropin cells from the pituitary of the African catfish, Clarias gariepinus: a morphological study (1995)
    Springer
    Cell & tissue research 280 (1995), S. 113-122 
    Details
    ISSN: 1432-0878
    Keywords: Pituitary ; Gonadotrops ; Crinophagy ; Electron microscopy ; Enzyme cytochemistry ; Immunocytochemistry ; Autoradiography ; Catfish, Clarias gariepinus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The possible function of globules and irregular membrane-bound masses in the gonadotropin cells of the pituitary of Clarias gariepinus was studied. Strong secretory stimulation led to the disappearance of the secretory granules from gonadotropin cells but globules and irregular masses remained present. Acid phosphatase was detected enzyme-cytochemically in both globules and irregular masses. Radiolabelling with tritiated amino acids followed by autoradiography demonstrated that globules received radioactive material after secretory granules. The latter received radioactive material within 75 min of administration of radioactive amino acids but globules and irregular masses did not. Although some globules became radioactively labelled within 24 h of the administration of radioactive amino acids, irregular masses remained unlabelled during this period. Secretory granules reacted positively with antisera against α and β gonadotropin subunits, whereas globules and irregular masses only reacted with the antiserum against the β subunit. A moderate anti-7B2 immunoreactivity was demonstrated in secretory granules and globules, whereas irregular masses labelled strongly. The combined cytological results indicate that globules and irregular masses are degradative, possibly crinophagic structures which develop by fusional events from secretory granules to globules and then to irregular masses.
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    URL: http://dx.doi.org/10.1007/BF00304516
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    Immunolocalization of interleukin-1α in rat mandibular molars and its enhancement after in vivo injection of epidermal growth factor (1995)
    Springer
    Cell & tissue research 280 (1995), S. 21-26 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Interleukin ; Stellate reticulum ; Immunocytochemistry ; Epidermal growth factor ; Interleukin-1 receptor type I messenger RNA ; Tooth eruption ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunolocalization of interleukin-1α in the first mandibular molars of rats from day 0–12 postnatally showed that the protein was localized in the epithelial stellate reticulum adjacent to the dental follicle. Staining of the stellate reticulum was most prominent in the early days postnatally and was absent by postnatal day 11. Injection of epidermal growth factor into rats at day 0 greatly increased the intensity of the staining for interleukin-1α in the stellate reticulum. Epidermal growth factor (EGF) enhanced the gene expression of interleukin-1α in stellate reticulum cells in vitro, and this study suggests there is enhanced translation of interleukin-1α messenger RNA in the stellate reticulum following EGF injection. In turn, the interleukin-1α may exert its effect on the dental follicle cells adjacent to the stellate reticulum because EGF also enhanced expression of the interleukin-1 receptor type I messenger RNA in cultured dental follicle cells as well as enhancing its expression in vivo. In view of the fact that injection of EGF will stimulate precocious eruption of teeth, its stimulus of interleukin-1α synthesis in the stellate reticulum may be the mechanism by which EGF initiates a cascade of molecular events to signal the onset of tooth eruption.
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    URL: http://dx.doi.org/10.1007/BF00304507
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    Characterization and localization of an Mr 225 kDa polypeptide specifically involved in the defence mechanisms of the clam Tapes semidecussatus (1995)
    Springer
    Cell & tissue research 280 (1995), S. 27-37 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Defence mechanisms ; Encapsulation ; Granulocytes ; Immunocytochemistry ; Parasitism ; Perkinsus sp. (Protozoa) ; Tapes semidecussatus (Mollusca
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Parasitosis by the trophozoite protozoan Perkinsus sp. (Apicomplexa, Perkinsea) induces in the gill filaments of the clam Tapes semidecussatus (Mollusca, Bivalvia) a cellular reaction, which is constituted by infiltrated granulocytes. This cellular reaction has characteristics of those of a holocrine gland, since the parasites are encapsulated by the secretion product of the granulocytes after cell death. An enriched fraction of prezoosporangia and their ass ociated capsule was obtained after culture of the parasitized gills in fluid thioglycollate medium. Specific polypeptides from this fraction were separated by SDS-PAGE and isolated for rabbit immunizations. The serum obtained against an Mr 225 kDa polypeptide, revealed its exclusive localization in the capsule and in the granules of the infiltrated granulocytes, thus indicating that this polypeptide is synthesized by these cells and secreted, in a polarized way, around the trophozoites resulting in t heir encapsulation. Selective deglycosylation of the polypeptide, by Endo H and alkaline β-elimination, did not show an effect on its molecular weight or antibody recognition. Furthermore, the absence of the 225 kDa band in the Western-blots of non-parasitized gills indicated the specific association of this polypeptide with the parasitosis. Finally, this is the first tissue-specific factor described in molluscs in relation to defence mechanisms.
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    URL: http://dx.doi.org/10.1007/BF00304508
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    Origin and destination of globules and irregular masses in the gonadotropin cells from the pituitary of the African catfish, Clarias gariepinus:a morphological study (1995)
    Springer
    Cell & tissue research 280 (1995), S. 113-122 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Pituitary ; Gonadotrops ; Crinophagy ; Electron microscopy ; Enzyme cytochemistry ; Immunocytochemistry ; Autoradiography ; Catfish ; Clarias gariepinus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The possible function of globules and irregular membrane-bound masses in the gonadotropin cells of the pituitary of Clarias gariepinus was studied. Strong secretory stimulation led to the disappearance of the secretory granules from gonadotropin cells but globules and irregular masses remained present. Acid phosphatase was detected enzyme-cytochemically in both globules and irregular masses. Radiolabelling with tritiated amino acids followed by autoradiography demons trated that globules received radioactive material after secretory granules. The latter received radioactive material within 75 min of administration of radioactive amino acids but globules and irregular masses did not. Although some globules became radioactively labelled within 24 h of the administration of radioactive amino acids, irregular masses remained unlabelled during this period. Secretory granules reacted positively with antisera against α and β gonadotropin subunits, whereas globules and irregular masses only reacted with the antiserum against the β subunit. A moderate anti-7B2 immunoreactivity was demonstrated in secretory granules and globules, whereas irregular masses labelled strongly. The combined cytological results indicate that globules and irregular masses are degradative, possibly crinophagic structures which develop by fusional events from secretory granules to globules and then to irregular masses.
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    URL: http://dx.doi.org/10.1007/BF00304516
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    Development of melanin-concentrating hormone-immunoreactive elements in the brain of gilthead seabream (Sparus auratus) (1995)
    Springer
    Cell & tissue research 282 (1995), S. 523-526 
    Details
    ISSN: 1432-0878
    Keywords: Melanin-concentrating hormone ; Immunocytochemistry ; Development, ontogenetic ; Sparus auratus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The development of the hypothalamic melanin-concentrating hormone (MCH) system of the teleost Sparus auratus has been studied by immunocytochemistry using an anti-salmon MCH serum. Immunoreactive perikarya and fibers are found in embryos, larvae, and juvenile specimens. In juveniles, most labeled neurons are present in the nucleus lateralis tuberis; some are dispersed in the nucleus recessus lateralis and nucleus periventricularis posterior. From the nucleus lateralis tuberis, MCH neurons project a conspicuous tract of fibers to the ventral hypothalamus; this penetrates the pituitary stalk and reaches the neurohypophysis. Most fibers end close to the cells of the pars intermedia, and some reach the adenohypophysial rostral pars distalis. Immunoreactive fibers can also be seen in extrahypophysial localizations, such as the preoptic region and the nucleus sacci vasculosi. In embryos, MCH-immunoreactive neurons first appear at 36 h post-fertilization in the ventrolateral margin of the developing hypothalamus. In larvae, at 4 days post-hatching, perikarya can be observed in the ventrolateral border of the hypothalamus and in the mid-hypothalamus, near the ventricle. At 26 days post-hatching, MCH perikarya are restricted to the nucleus lateralis tuberis. The neurohypophysis possesses MCH-immunoreactive fibers from the second day post-hatching. The results indicate that MCH plays a role in larval development with respect to skin melanophores and cells that secrete melanocyte-stimulating hormone.
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    URL: http://dx.doi.org/10.1007/BF00318885
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  • 88
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    5′-Nucleotidase in PC12 cells as revealed by immunocytochemistry (1995)
    Springer
    Cell & tissue research 280 (1995), S. 123-131 
    Details
    ISSN: 1432-0878
    Keywords: Differentiation ; 5′-Nucleotidase ; Immunocytochemistry ; PC12 cells ; Synaptophysin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract 5′-Nucleotidase hydrolyzes 5′-mononucleotides to their nucleosides but is also thought to have a function in neuronal differentiation and synapse formation. The distribution of the enzyme, a glycosyl-phosphatidylinositol-anchored sialoglycoprotein, was investigated in PC12 cells using immunofluorescence microscopy. 5′-Nucleotidase was located both in intracellular compartments and at the cell surface. There was no principal difference in the cellular distribution between undifferentiated cells and after neuritogenic differentiation by nerve growth factor. Intracellularly, 5′-nucleotidase often revealed a sickle-shaped perinuclear distribution and a dotted pattern throughout the cytoplasm, including that of neurites and growth cones. The intracellular distribution was clearly different from that of the synaptic vesicle protein synaptophysin. However, the dotted fluorescence resembled that obtained after uptake of the endosomal marker acridine orange. 5′-Nucleotidase was present on the entire cell surface including all neurites formed after differentiation. There was no increase in 5′-nucleotidase fluorescence at synapse-like contacts between the tips of neurites and other PC12 cells. Surfacelocated 5′-nucleotidase could no longer be detected after the application of glycosyl-phosphatidylinositol-specific phospholipase C to cultured cells. This treatment did not affect PC12 cell differentiation. Our results thus reveal 5′-nucleotidase both at the surface and within organelles and suggest that PC12 cells may be used as a model system for the study of the physiological function of 5′-nucleotidase in neural cells.
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    URL: http://dx.doi.org/10.1007/BF00304517
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  • 89
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    5′-Nucleotidase in PC12 cells as revealed by immunocytochemistry (1995)
    Springer
    Cell & tissue research 280 (1995), S. 123-131 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Differentiation ; 5′-Nucleotidase ; Immunocytochemistry ; PC12 cells ; Synaptophysin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. 5′-Nucleotidase hydrolyzes 5′-mononucleotides to their nucleosides but is also thought to have a function in neuronal differentiation and synapse formation. The distribution of the enzyme, a glycosyl-phosphatidylinositol-anchored sialoglycoprotein, was investigated in PC12 cells using immunofluorescence microscopy. 5′-Nucleotidase was located both in intracellular compartments and at the cell surface. There was no principal difference in the cellular distrib ution between undifferentiated cells and after neuritogenic differentiation by nerve growth factor. Intracellularly, 5′-nucleotidase often revealed a sickle-shaped perinuclear distribution and a dotted pattern throughout the cytoplasm, including that of neurites and growth cones. The intracellular distribution was clearly different from that of the synaptic vesicle protein synaptophysin. However, the dotted fluorescence resembled that obtained after uptake of the endosomal marker acridine orange. 5′-Nucleotidase was present on the entire cell surface including all neurites formed after differentiation. There was no increase in 5′-nucleotidase fluorescence at synapse-like contacts between the tips of neurites and other PC12 cells. Surface-located 5′-nucleotidase could no longer be detected after the application of glycosyl-phosphatidylinositol-specific phospholipase C to cultured cells. This treatment did not affect PC12 cell differentiation. Our results thus reveal 5′ -nucleotidase both at the surface and within organelles and suggest that PC12 cells may be used as a model system for the study of the physiological function of 5′-nucleotidase in neural cells.
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    URL: http://dx.doi.org/10.1007/BF00304517
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  • 90
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    Secretory glycoproteins of the subcommissural organ of the dogfish (Scyliorhinus canicula): evidence for the existence of precursor and processed forms (1995)
    Springer
    Cell & tissue research 283 (1995), S. 75-84 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Subcommissural organ ; Secretory glycoproteins ; Antibodies ; Immunochemistry ; Immunocytochemistry ; Dogfish ; Scyliorhinus canicula (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The subcommissural organ of the dogfish, Scyliorhinus canicula (L), has been investigated by use of antibodies and lectins applied to blots and tissue sections processed for light and electron microscopy. Antibodies have been raised against each of the bands that have previously been identified in immunoblots by the use of antisera raised against secretory glycoproteins extracted from the dogfish subcommissural organ, viz., the 600-kDa band and two gel regions including the 475 to 400-kDa and the 145-kDa bands obtained from preparative gels; they are referred to as Ab-600, Ab-475/400, and Ab-145. These antisera and the lectins concanavalin A and wheat germ agglutinin have been used for the staining of: (1) blots of extracts of the dogfish subcommissural organ and optic tectum; (2) tissue sections of the dogfish brain. The findings indicate that the bands of 600, 475 and 400 kDa contain compounds that should be regarded as secretory glycoproteins of the dogfish subcommissural organ. The 600-kDa and 400-kDa bands are labeled by concanavalin A; wheat germ agglutinin labels the 475-kDa band strongly and the other two weakly. Ab-600 reacts with the bands at 600, 475 and 400 kDa and stains materials stored in the rough endoplasmic reticulum and secretory granules of 200–600 nm in diameter. The 600-kDa compound is probably a precursor form. Ab-475/400 stains the same three bands revealed by Ab-600; immunocytochemically, it reacts with two types of secretory granules (200–600 and 800–1200 nm in diameter) but it does not label the rough endoplasmic reticulum. Ab-145 reveals the bands at 600, 475 and 400 kDa and a diffuse zone in the region of 145 kDa; in light-microscopic immunocytochemistry, it behaves as Ab-475/400. The 475-kDa and 400-kDa glycoproteins, and a compound of approximately 145 kDa thus probably correspond to processed forms. Ab-475/400 stains granules present in cell processes ending on local blood vessels and at the leptomeninges. Since this antiserum selectively labels secretory granules, this finding may be taken as evidence for a basal route of secretion.
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    URL: http://dx.doi.org/10.1007/s004410050514
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  • 91
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    Presence of sauvagine-like epitopes in the interrenal gland of the bullfrog Rana catesbeiana (1995)
    Springer
    Cell & tissue research 283 (1995), S. 117-123 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Sauvagine ; Corticotropin-releasing factor ; Immunocytochemistry ; Interrenal gland ; Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunocytochemistry was used to investigate the presence of corticotropin-releasing factor-like peptides in the interrenal (adrenal) glands of the bullfrog Rana catesbeiana by using specific antisera raised against synthetic nonconjugated rat/human corticotropin-releasing factor, urotensin I, and sauvagine. From these three antisera, covering a broad range of corticotropin-releasing factor-like immunoreactivities, only the sauvagine antiserum gave positive immunoreactivity. Sauvagine immunoreactivity was found in cortical cells grouped into cords in the renal zone of the interrenal gland. The central and subcapsular cords were less stained. Tyrosine hydroxylase-positive chromaffin cells were not sauvagine-immunoreactive. The immunoreactivity was abolished, in all cases, by previous immunoabsorption of the sauvagine antiserum with synthetic sauvagine (0.1 μM), but it was not eliminated by sucker (Catostomus commersoni) urotensin I, sole (Hippoglossoides elassodon) urotensin I, sucker corticotropin-releasing factor, rat/human corticotropin-releasing factor, or ovine corticotropin-releasing factor (0.1–10 μM). In a sauvagine radioimmunoassay, interrenal extracts displaced 125I-sauvagine from antiserum only partially, and not in parallel with the sauvagine standard curve. The results suggest that the sauvagine immunoreactivity in the R. catesbeiana interrenal gland may represent a novel sauvagine-like peptide.
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    URL: http://dx.doi.org/10.1007/s004410050519
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  • 92
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    Immunocytochemical and morphometric study on the changes of TSH, PRL, GH and ACTH cells during the development of Bufo arenarum (1995)
    Springer
    Cell & tissue research 283 (1995), S. 125-132 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Pituitary hormones ; Immunocytochemistry ; Morphometry ; Metamorphosis ; Bufo arenarum (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The development and dynamics of thyrotropin (TSH), adrenocorticotropic hormone (ACTH), prolactin (PRL), and growth hormone (GH) cells have been studied using immunocytochemical techniques and rabbit antisera, raised against the relevant human hormone, in the pars distalis of Bufo arenarum larvae at different stages of development. The four types of cells studied were identified in different zones of the pars distalis: TSH cells occurred mainly in the centro-ventral zone, ACTH cells in the rostral and dorsal zones, GH cells in the central and caudal zones, and PRL cells in the anterior two-thirds of the gland. This distribution pattern does not show significant changes with development. Morphometry and stereology were used to evaluate the changes observed in the volume of the pars distalis and the immunoreactive cells during development. The former increased during larval growth and decreased throughout the metamorphic climax. The results obtained on cell number, volume density, and total volume suggest that, during larval growth (pre-prometamorphosis) of B. arenarum, TSH, PRL, GH and ACTH cells show a proliferative period with storage of their hormones; a second period involving hormone release occurs at the metamorphic climax.
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    URL: http://dx.doi.org/10.1007/s004410050520
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  • 93
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    Ultrastructural distribution of endogenous immunoglobulin-G in human term amniochorion (1995)
    Springer
    Cell & tissue research 281 (1995), S. 367-374 
    Details
    ISSN: 1432-0878
    Keywords: Placenta ; Amniochorion ; Cytotrophoblast cells ; Immunoglobulin G ; Immunocytochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Maternal immunoglobulin-G (IgG) is known to be transported across the placental syncytiotrophoblast during the period when the human fetus is incapable of manufacturing these defensive molecules. In this study we investigated the possible role of the amniochorion, that surrounds the amniotic cavity in which the fetus lies, in the transfer of immunoglobulin. Endogenous IgG was localised in the amniochorion by confocal immunofluorescence microscopy and by ultrastructural labelling of ultrathin frozen tissue sections using the protein A-gold technique. Immunoreactivity was identified in the extracellular matrix tissues and necrotic amniotic epithelial cells. Healthy amniotic epithelial cells and cytotrophoblast cells of the chorion laeve were devoid o endogenous IgG. These results suggest a possible non-specific paracellular transport pathway between cytotrophoblast cells, which may conceivably contribute to the acquisition of passive immunity by the fetus, and offer a rational explanation for the presence of small quantities of maternal IgG in the amniotic fluid.
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    URL: http://dx.doi.org/10.1007/BF00583405
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  • 94
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    Unusual distribution of tubulin isoforms in the snail Lymnaea stagnalis (1995)
    Springer
    Cell & tissue research 281 (1995), S. 507-515 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Microtubules ; Isoforms ; Nervous system ; Locomotion ; Cilia ; Immunocytochemistry ; Western blotting ; Lymnaea stagnalis (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunocytochemistry and Western blotting techniques demonstrated that the nervous system and foot of the pond snail Lymnaea stagnalis are rich sources of tubulin, which can be extracted and assembled in vi- tro in the presence of taxol. Various broad-spectrum antibodies raised against α-tubulin and β-tubulin yielded qualitatively similar results. One monoclonal antibody to trypanosome α−tubulin, however, labelled α-tubulin more strongly on both probed sections and Western blots. Cytochemistry and immunoblotting revealed that tyrosinated tubulin constitutes a large proportion of total α-tubulin in locomotor cilia of the foot and in axons of the nervous system. Detyrosinated tubulin also appeared to be abundant in the foot cilia but only a very faint band of detyrosinated tubulin was found on protein blots extracted from the central ganglia, and staining was barely detectable in central ganglia or peripheral nerves. Similarly, acetylated tubulin appeared to be abundant in foot cilia, but Western blotting indicated only low levels of acetylated tubulin in the nervous system. Immunocytochemistry indicated that, while most neurons possessed little or no acetylated tubulin, a small number of axons contained significant amounts of this isoform. Thus, while a large amount of tubulin was expected in the nervous system and locomotor cilia of L. stagnalis, the observed distribution of isoforms was unanticipated. Specifically, neurons of other organisms have generally been reported to contain substantial amounts of both detyrosinated α-tubulin and acetylated α-tubulin. Our results indicate that such findings cannot be generalized across all species. L. stagnalis, with its well studied nervous system and unusual distribution of tubulin isoforms, may prove to be particularly useful for studying the roles of tubulin isoforms in microtubule function and cell activity.
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    URL: http://dx.doi.org/10.1007/s004410050446
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  • 95
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    Distribution, ontogeny and ultrastructure of somatostatin immunoreactive cells in the pancreas and gut (1977)
    Springer
    Cell & tissue research 185 (1977), S. 465-479 
    Details
    ISSN: 1432-0878
    Keywords: Somatostatin cells ; Pancreas ; Gut ; Immunocytochemistry ; Comparative study
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Somatostatin cells are numerous in the pancreas and digestive tract of mammals as well as birds. In the pancreas of chicken, cat and dog they occur in both the exocrine parenchyma and in the islets. In the rat and rabbit, somatostatin cells have a peripheral location in the islets, whereas in the cat, dog and man the cells are usually more randomly distributed. In the stomach of rabbits and pigs, somatostatin cells are more numerous in the oxyntic gland area than in the pyloric gland area, whereas the reverse is true for the cat, dog and man. In the cat, pig and man, somatostatin cells are fairly numerous in the duodenum, whereas in the rat, rabbit and dog they are few in this location. In the remainder of the intestines somatostatin cells are few but regularly observed. Somatostatin cells are numerous in the human fetal pancreas and gut. In the fetal rat, somatostatin cells first appear in the pancreas and duodenum (at about the 16–17th day of gestation) and subsequently in the remainder of the intestine. Somatostatin cells do not appear in the gastric mucosa until after birth. Three weeks after birth, somatostatin cells show the adult frequency of occurrence and pattern of distribution. In the chicken, somatostatin cells are numerous in the proventriculus, absent from the gizzard, abundant in the gizzard-duodenal junction (antrum), infrequent in the duodenum and virtually absent from the remainder of the intestines. No immunoreactive cells can be observed in the thyroid of any species nor in the ultimobranchial gland of the chicken. In the chick embryo, somatostatin cells are first detected in the pancreas and proventriculus (at about the 12th day of incubation). They appear in the remainder of the gut much later, in the duodenum at the 16th day, in the antrum at about the 19th day and still later in the lower small intestine. The ultrastructure of the somatostatin cells was studied in the chicken, rat, cat and man; the cells were identified by the consecutive semithin/ultrathin section technique. The somatostatin cells display the properties of the D cell. There was no difference in granule ultrastructure between somatostatin cells in the gut and the pancreas. The granules, which are the storage site of the peptide, are round, supplied with a tightly fitting membrane and have a moderately electron-dense, fine-granulated core. The mean diameter of the somatostatin granules is smallest in rat (155–170 nm) and largest in the chicken (270–290 nm).
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    URL: http://dx.doi.org/10.1007/BF00220651
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  • 96
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    GABA and glutamate-like immunoreactivity at synapses received by dorsal unpaired median neurones in the abdominal nerve cord of the locust (1995)
    Springer
    Cell & tissue research 280 (1995), S. 325-333 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Nervous system ; insect ; DUM neuron ; Synapses ; Immunocytochemistry ; GABA ; Glutamate ; Locusta migratoria (Insecta) ; Schistocerca gregaria (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Dorsal unpaired median (DUM) neurones in the abdominal ganglia of the locust were impaled with microelectrodes and some were injected intracellularly with horseradish peroxidase so that their synapses could be identified in the electron microscope. Simultaneous recordings from DUM neurones in different abdominal ganglia revealed that they received common postsynaptic potentials from descending interneurones. Post-embedding immunocytochemistry using antibodies against GABA and glutamate was carried out on ganglia containing HRP-stained neurones. GABA-like immunoreactivity was found in 39% (n=82) of processes presynaptic to abdominal DUM neurones and glutamate-like immunoreactivity in 21% (n=42) of presynaptic processes. Output synapses from the DUM neurites were rarely observed within the neuropile. Structures resembling presynaptic dense bars but not associated with synaptic vesicles, were seen in some large diameter neurites.
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    URL: http://dx.doi.org/10.1007/BF00307805
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  • 97
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    Leu-callatostatin gene expression in the blowflies Calliphora vomitoria and Lucilia cuprina studied by in situ hybridisation: comparison with Leu-callatostatin confocal laser scanning immunocytochemistry (1995)
    Springer
    Cell & tissue research 280 (1995), S. 355-364 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Leu-callatostatin ; Allatostatins ; Neuropeptides ; In situ hybridisation ; Immunocytochemistry ; Hindgut innervation ; Midgut endocrine cells ; Calliphora vomitoria ; Lucilia cuprina (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. In situ hybridisation studies using a digoxigenin-labelled DNA probe encoding the Leu-callatostatin prohormone of the blowflies Calliphora vomitoria and Lucilia cuprina have revealed a variety of neurones in the brain and thoracico-abdominal ganglion, peripheral neurosecretory neurones, and endocrine cells of the midgut. With two exceptions, the hybridising cells are the same as those previously identified in immunocytochemical studies of sections and whole-mounts using Leu-callatostatin COOH-terminal-specific antisera. Within the brain and suboesophageal ganglion, there is a variety of neurones ranging from a single pair of large cells situated in the dorsal protocerebrum, to the several pairs of neurones in the tritocerebrum, some of which, in immunocytochemical preparations, can be seen to project via axons in the cervical connective to the thoracico-abdominal ganglion. In the medulla of the optic lobes, numerous small interneurones hybridise with the probe, as do clusters of similar-sized neurones close to the roots of the ocellar nerves. These results indicate that the Leu-callatostatin neuropeptides of the brain play a variety of roles in neurotransmission and neuromodulation. There are only three pairs of Leu-callatostatin-immunoreactive neurones in the thoracico-abdominal ganglion, at least two pairs of which project axons along the median abdominal nerve to provide extensive innervation of the hindgut. The Leu-callatostatin peripheral neurosecretory cells are located in close association with both nerve and muscle fibres in the thorax. In addition to neuronal Leu-callatostatin, the presence of the peptide and its mRNA has been demonstrated in endocrine cells in the posterior part of the midgut. These observations provide an example of a named brain/gut peptide in an insect.
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    URL: http://dx.doi.org/10.1007/BF00307808
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    Immunocytochemical localization of ribonuclease in yolk granules of adult Rana catesbeiana oocytes (1995)
    Springer
    Cell & tissue research 280 (1995), S. 259-265 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Oocyte ; Yolk granules ; Ribonuclease ; Immunocytochemistry ; Bullfrog ; Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. To determine the localization of the pyrimidine-guanine sequence-specific ribonuclease in Rana catesbeiana (bullfrog) oocytes, the RNase was first isolated and used to prepare a specific rabbit antiserum. Only one protein of similar molecular size to the RNase was immunoprecipitated from ovary homogenate by the antiserum, but two bands were observed by Western blotting analysis. These two proteins were shown by further purification of antibody and Western blotting analysis to have similar antigenicity. Immunoprecipitation and Western blotting of tissue homogenates showed that the RNase was found predominantly in the ovary, but not in other tissues. The specific localization of the RNase was determined by immuno-electron microscopy of oocyte sections incubated with the specific antiserum; the yolk granules, but not other organelles, were found to contain the RNase. Most of the RNase was evenly distributed in the lateral amorphous area of the yolk granule but not in the central yolk crystal area which contains stored vitellogenin proteins. Our results indicate that the RNase is compartmentalized in the yolk granules of oocytes, which might prevent damage to cellular RNAs.
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    URL: http://dx.doi.org/10.1007/BF00307797
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  • 99
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    Sources of inputs to longitudinal muscle motor neurons and ascending interneurons in the guinea-pig small intestine (1995)
    Springer
    Cell & tissue research 280 (1995), S. 549-560 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Enteric nervous system ; Immunocytochemistry ; Calretinin ; Calbindin ; Bombesin ; Small intestine ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Light- and electron-microscopic studies were used to investigate connections between specific subgroups of neurons in the myenteric plexus of the guinea-pig small intestine. Inputs to two classes of calretinin-immunoreactive (IR) nerve cells, longitudinal muscle motor neurons and ascending interneurons, were examined. Inputs from calbindin-IR primary sensory neurons and from three classes of descending interneurons were studied. Electron-microscopic analysis showed that calbindin-IR axons formed two types of inputs, synapses and close contacts, on calretinin-IR neurons. About 40% of inputs to the longitudinal muscle motor neurons and 70% to ascending interneurons were calbindin-IR. Approximately 50% of longitudinal muscle motor neurons were surrounded by bombesin-IR dense pericellular baskets and 40% by closely apposed varicosities. At the electron-microscope level, the bombesin-IR varicosities were found to form synapses and close contacts with the motor neurons. Dense pericellular baskets with bombesin-IR surrounded 36% of all ascending interneurons, and a further 17% had closely apposed varicosities. Somatostatin- and 5-HT-IR descending interneurons provided no dense pericellular baskets to calretinin-IR nerve cells. Thus, calretinin-IR, longitudinal muscle motor neurons and ascending interneurons receive direct synaptic inputs from intrinsic primary sensory neurons and from non-cholinergic, bombesin-IR, descending interneurons.
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    URL: http://dx.doi.org/10.1007/BF00318359
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  • 100
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    Ultrastructural investigation of nitric oxide synthase-immunoreactive nerves associated with coronary blood vessels of rat and guinea-pig (1995)
    Springer
    Cell & tissue research 280 (1995), S. 575-582 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Nitric oxide synthase ; Coronary vasculature ; Electron microscopy ; Immunocytochemistry ; Rat (Sprague Dawley) ; Guinea-pig (Dunkin Hartley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Ultrastructural investigation of nitric oxide synthase-immunoreactive nerves closely associated with blood vessels in rat and guinea-pig hearts revealed many labelled nerve fibres in the walls of the main branches of the coronary arteries, and in arterioles, capillaries and post-capillary venules. The number of nitric oxide synthase-containing nerve fibres associated with different vessels, even those of the same calibre, varied. Terminal regions of nitric oxide synthase-immunoreactive fibres were observed in the endocardium and myocardium. Nitric oxide synthase-labelled fibres displayed electron-dense immunoproduct in both varicose and intervaricose regions. Immunoreactive axonal varicosities contained both small and large synaptic vesicles. The characteristics of the nitric oxide synthase-immunoreactive nerve fibres observed in the heart and the possibility that these fibres represent the processes of intracardiac neurones and/or sensory neurones of extrinsic origin are discussed.
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    URL: http://dx.doi.org/10.1007/BF00318361
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