Neuronal protein NP185 is developmentally regulated, initially expressed during synaptogenesis, and localized in synaptic terminals.
Evidence is presented here that demonstrates the presence of NP185 (AP3) in neuronal cells, specifically within syn-aptic terminals of the central nervous system and in the peripheral nervous system, particularly in the neuro-muscular junction of adult chicken muscle. Biochemical results obtained in our laboratories indicate that NP185 is associated with brain synaptic vesicles, with clathrin-coated vesicles, and with the synaptosomal plasma membrane. Also, NP185 binds to tubulin and clathrin light chains and the binding is regulated by phosphorylation (Su et al., 1991). Based on these properties and the data reported here, we advance the postulate that NP185 fulfills multiple functions in synaptic terminals. One function is that of a plasma membrane docking or channel protein, another of a signaling molecule for brain vesicles to reach the synaptic terminal region, and a third is that of a recycling molecule by binding to protein components on the lipid bilayer of the synaptic plasma membrane during the process of endocytosis. In support of these premises, a thorough study of NP185 using the developing chick brain, adult mouse brain, and chicken straited muscle was begun by temporally and spatially mapping the expression and localization of NP185 in evolving and mature nerve endings. To achieve these objectives, monoclonal antibodies to NP185 were used for immunocytochemistry in tissue sections of chicken and mouse cerebella. The distribution of NP185 was compared with those of other cytoskeletal and cytoplasmic proteins of axons and synapses, namely synaptophysin, vimentin, neurofilament NF68, and the intermediate filaments of glial cells (GFAP). The data indicate that expression of NP185 temporally coincides with synaptogenesis, and that the distribution of this protein is specific for synaptic terminal buttons of the CNS and the PNS.
Puszkin
S
S
Department of Pathology, Mount Sinai School of Medicine, CUNY, NY 10029.
Perry
D
D
Li
S
S
Hanson
V
V
D033963:Adaptor Protein Complex 3; D033942:Adaptor Proteins, Vesicular Transport; D000375:Aging / Q000502:physiology*; D000818:Animals; D001921:Brain / Q000196:embryology / Q000254:growth & development / Q000502:physiology*; D002531:Cerebellum / Q000502:physiology*; D002642:Chick Embryo; D002645:Chickens; D034041:Monomeric Clathrin Assembly Proteins; D009411:Nerve Endings / Q000378:metabolism / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009474:Neurons / Q000502:physiology*; D013569:Synapses / Q000378:metabolism / Q000502:physiology*
D033963:Adaptor Protein Complex 3; D033942:Adaptor Proteins, Vesicular Transport; D034041:Monomeric Clathrin Assembly Proteins; D009419:Nerve Tissue Proteins; C439023:clathrin assembly protein AP180