Spatial and temporal expression of neuronal protein NP185 in developing avian cerebellum.
Item
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Title
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Spatial and temporal expression of neuronal protein NP185 in developing avian cerebellum.
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Identifier
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AAI9119666
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identifier
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9119666
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Creator
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Perry, Douglas Gentz.
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Contributor
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Adviser: Saul Puszkin
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Date
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1991
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Language
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English
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Publisher
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City University of New York.
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Subject
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Biology, Neuroscience | Biology, Animal Physiology | Biology, General
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Abstract
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NP185 is a protein found in nervous tissue (Kohtz and Puszkin, J Biol Chem 263:7418). Biochemical results (Kohtz & Puszkin, J Neurochem 52:285; Su et al., J Neurosci Res (in press)) demonstrate that NP185 co-purifies with bovine brain clathrin-coated vesicles (CCVs), has a phosphorylation-sensitive binding affinity for tubulin, binds to sites in synaptic vesicles and nerve terminal membranes, and promotes clathrin cage assembly in vitro. These findings suggest a supportive role for NP185 in the function of neuronal CCVs. To further understand the nature of NP185, I followed its distribution in developing central nervous tissue by immunohistochemical light microscopy, compared this distribution to that of neuronal (NF68, synaptophysin) and nonneuronal (GFAP, vimentin) proteins, and demonstrated NP185 localization in the neuromuscular junction of skeletal muscle. Results: (1) Glial cells, although strongly labelled for GFAP and vimentin, are never labelled for NP185. (2) The external granular and ventricular layers of the developing cerebellum are negative for NP185. The mantle layer is labelled for NP185 from its inception. (3) Membranous labelling for NP185 is very strong in migrating neuroblasts. In the mantle layer, NP185 labelling follows a gradient consistent with the directional gradient of synaptic formation. (4) The distribution of NP185 is identical to that of synaptophysin. The labelling pattern of NF68 generates an intense cytoplasmic immunofluorescent stain in cerebellar Purkinje cells, but NP185 yields only membranous staining in these same cells. (5) In chick skeletal muscle, NP185 is co-distributed with acetylcholine receptors (AchRs). The AchR distribution pattern found in these experiments is consistent with previous studies (Buc-Caron et al., Develop Biol 95:378; Bender et al., Neurol 26:477) of the neuromuscular junction. Conclusions: (1) NP185 is strictly neuron-specific, but NP185 distribution is not limited to the central nervous system. (2) Developmental expression of NP185 follows a pattern consistent with synaptogenesis. (3) NP185 is concentrated in synaptic terminals of mature neurons. (4) NP185 is located in the neuromuscular junctions of skeletal muscle.;In view of these findings, the "docking protein" model may be appropriate for NP185 function in nerve endings.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
