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  • 1
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Compared with neurons of the CNS, the organization of the peripheral adrenergic axon and nerve terminal is more complex because two types of neurotrarismitter-containing vesicles, i.e., large (LDVs) and small densecore vesicles, coexist with the axonal reticulum (AR) and the well-characterized small synaptic vesicles. The AR, which is still poorly examined, is assumed to play some role in neurosecretion. We have studied the subcellular localization of noradrenaline, cytochrome b561, and synaptophysin in control and ligated dog splenic nerve using both biochemical and ultrastructural approaches. Noradrenaline and cytochrome b561 coaccumulated proximal to a ligation, whereas distally only the latter was found. Despite a codistribution with noradrenaline at high densities in sucrose gradients, Synaptophysin did not accumulate on either side of the ligation. At the ultrastructural level, cytochrome b561 immunoreactivity was found on LDVs and AR elements, both accumulating proximal to the ligation. Distally, the multivesicular bodies (MVBs), immunolabeled for cytochrome b561, account for the retrograde transport of LDVs and AR membranes retrieved at the nerve terminal. No Synaptophysin immunoreactivity could be detected on LDVs, AR, or MVBs. The results obtained from the ligation experiments together with the ultrastructural data Clearly illustrate that Synaptophysin is absent from LDVs and AR elements in adrenergic axons.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 228 (1970), S. 1203-1204 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Subcellular fractionation studies were done on the spleens of young (6-12 month) mongrel dogs. The spleens were decapsulated and homogenized in four volumes of ice cold 0-25 M sucrose in a glass Potter-Elvehjem homo-genizer fitted with a conical 'Teflon' pestle. A pellet sediment ing between 105 ...
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-6903
    Keywords: Sympathetic neurons ; large dense cored vesicle ; small dense cored vesicle ; exocytosis ; noradrenaline
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract More than 25 years have passed since the original demonstration that proteins such as chromogranin A and dopamine-β-hydroxylase, which are co-stored together with noradrenaline in large dense cored vesicles in adrenergic nerves, are released by exocytosis. Despite much evidence in favour, it was for a long time thought that large dense cored vesicles were not eminently involved in the release of noradrenaline. The present review attempts to demonstrate, making use of evidence from different approaches, that the release of noradrenaline from sympathetic neurons occurs ultimately from large dense cored vesicles. A model of the secretory cycle is proposed.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 0887-6134
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The technique of isotope dilution mass spectrometry has been used for the measurement of biogenic amine metabolites in cerebrospinal fluid (CSF). CSF samples were collected from rabbits treated with α2-antagonists. The aim of our study was to determine the specificity of these drugs on the central nervous noradrenergic, dopaminergic and serotonergic activity as measured by the release of corresponding monoamine metabolites. 3-Methoxy-4-hydroxyphenylethylene glycol (MHPG) and vanilmandelic acid (VMA) were used as parameters for the noradrenergic activity, whereas homovanillic acid (HVA) and 5-hydroxyindole-3-acetic acid (5-HIAA) were employed to follow the dopaminergic and serotonergic activity, respectively. For the measurement of the biogenic amine metabolites a published GCMS method has been adapted. Samples of 200 μl CSF were processed. Following addition of deuterated internal standards and acidification, extraction was carried out with ethyl acetate. Preliminary experiments with the analysis of MHPG using diethyl ether for extraction gave rise to emulsion formation and resulted in poor recoveries for MHPG and in irreproducibility problems due to a preferential extraction of non-labelled MHPG, effects which were not observed with ethyl acetate extraction. Derivatization was done with a mixture of pentafluoropropionic anhydride/pentafluoropropanol (or hexafluoroisopropanol) in order to derivatize both hydroxyl and carboxylic acid groups. The derivatization procedure was optimized for the analysis of 5-HIAA by carrying out a second reaction step with pentafluoropropionic anhydride alone in order to complete the derivatization for the indolic NH moiety. The molecular ions of the derivatized products were selected for detection. The methodology has been applied to evaluate the effects of two α2-antagonists, i.e. yohimbine and idazoxan. Yohimbine was found to increase the CSF levels of the noradrenergic metabolites MHPG and VMA and of the dopaminergic metabolite HVA and to have serotonergic effects, whereas idazoxan was shown to exhibit noradrenergic effect only.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1076-5174
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Physics
    Notes: An N-terminal proteolytic processing product of chromogranin A was obtained from bovine chromaffin granules using two steps of C18 solid-phase extraction and reversed-phase high-performance liquid chromatography. Electrospray mass spectrometry revealed a protein with a molecular mass of 8632.0 for the fraction showing immunoreactivity against the N-terminus of chromogranin A, which differed by 48 u from that of the N-terminal processing product, vasostatin I (CGA1-76). Derivatization with mercaptoethanol showed that the peptide had an intact S—S bridge, which is a key structural feature of vasostatin I. Peptide mapping experiments involving reduction/alkylation with vinylpyridine and trypsin digestion were consistent with oxidation of the three methionine residues of vasostatin I to their sulphoxide forms. The oxidation of the methionine residues was found to occur during the C18 solid-phase extraction procedure. The use of freshly prepared Tris-HCl buffer and eluents and flushing the buffer and eluents with nitrogen were shown to result in the isolation of the non-oxidized form of vasostatin I.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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