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  • 1990-1994  (2)
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  • 1
    Electronic Resource
    Electronic Resource
    Localization of Endo-Oligopeptidase (EC 3.4.22.19) in the Rat Nervous Tissue (1990)
    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 55 (1990), S. 0 
    Details
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: The subcellular and regional distribution of endooligopeptidase (EC 3.4.22.19), an enzyme capable of generating enkephalin by single cleavage from enkephalin-containing peptides, was determined by an enzymatic assay using metorphamide and by immunochemical techniques in the CNS of the rat. The rat CNS contains a membrane-associated form of endo-oligopeptidase, an enzyme predominantly associated with the soluble fraction of brain homogenates. Sub-cellular fractionation showed that ∼17% of the total activity of the enzyme is associated with membrane fractions including synaptosomes. Synaptosomal membranes were prepared from neocortex, striatum, hypothalamus, medulla, spinal cord, and cerebellum. The amount of EC 3.4.22.19 activity solubilized by 3-([3-cholamidopropyl]dimethylammonio)-1-propanesulfonate from synaptosomal membranes was similar in neocortex, striatum, and hypothalamus, being three- to 10-fold greater than in spinal cord, cerebellum, and medulla. A polyclonal antibody exhibiting high affinity for endo-oligopeptidase was raised in rabbits against the purified rat brain enzyme and used to localize endo-oligopeptidase by Western blotting and by immunoperoxidase techniques. A strong band corresponding to the Mr of EC 3.4.22.19 was found in solubilized proteins obtained from synaptosomal membranes prepared from hypothalamus, neocortex, and striatum when subjected to Western blotting. The immunohistochemical localization of endo-oligopeptidase indicated that the immunoreactivity was confined to gray matter in regions known to be rich in peptide-containing neurons such as the striatum. In the cerebellum, a region poor in peptides, no staining could be detected. The nonuniform distribution of endo-oligopeptidase in rat brain suggests a role in neurotransmitter processing in the CNS.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1471-4159.1990.tb03113.x
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  • 2
    Electronic Resource
    Electronic Resource
    C-fos Induction in the Spinal Cord after Peripheral Nerve Lesion (1991)
    Oxford, UK : Blackwell Publishing Ltd
    European journal of neuroscience 3 (1991), S. 0 
    Details
    ISSN: 1460-9568
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Immunocytochemical localization of a product of the proto-oncogene c-fos, Fos protein, was used to map the activity of a subset of rat spinal neurons at 3 days, 3 weeks and 3 months following section of the sciatic nerve. In a well-established experimental paradigm, the gene was induced by activation of primary afferent fibres with brief noxious sensory stimulation under anaesthetic. Central sciatic projections were demonstrated with isolectin B4 counterstain and GAP43 immunocytochemistry. In Rexed's lamina II of the spinal cord, in which there is somatotopic organization of afferent terminals, Fos-positive neurons were largely restricted to the projection area of intact peripheral nerves. Three days after a sciatic nerve lesion, the number of Fos-positive neurons in a cord region innervated by the saphenous nerve was similar to control levels, but was markedly increased by 3 weeks, remaining elevated at 3 months. Three weeks after sciatic nerve section the lectin stain in the area of sciatic representation had almost completely disappeared, and conversely GAP43 staining had greatly intensified. There was no evidence of invasion by Fos-immunoreactive cells of the area of sciatic representation. After 3 months both the size and the intensity of the lectin gap, and of the corresponding area of increased GAP43 immunoreactivity, appeared reduced. Thus a peripheral nerve lesion was followed by a delayed increase in excitability of the spinal cord as assessed by c-fos expression, so that greater numbers of second-order neurons were activated by sensory stimulation of an adjacent intact nerve. These changes may be related to the sensory abnormalities which follow nerve damage.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1460-9568.1991.tb00100.x
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