title: Highly basic 30- and 32-kilodalton proteins associated with synapse formation on polylysine-coated beads in enriched neuronal cell cultures.
abstract: Neuronal proteins involved in axonal outgrowth and synapse formation were examined in an enriched neuronal cell culture system of the cerebellum. In rat cerebellar cell cultures, 98.9% of the cells are neurons and the remaining 1.1% of the cells are flat nonneuronal cells. These enriched neuronal cultures, examined with two-dimensional gel electrophoresis, showed protein patterns similar to those of neonatal cerebellum, but very different patterns from glial enriched cultures. High levels of a neuronal membrane acidic 29-kilodalton (kD) protein were found. It has been shown previously that neuronal cultures incubated with polylysine-coated beads will develop numerous presynaptic elements on the bead surface. We report here that isolation of the beads from enriched neuronal cell cultures incubated with [35S]methionine showed, with two-dimensional nonequilibrium pH gradient gel electrophoresis (2D-NEPHGE), levels of a basic 32-kD protein (pI 8) note detected in cultures alone, and increased levels of a 30-kD protein (pI 10). When culture medium was examined with 2D-NEPHGE, three acidic proteins were identified that were secreted by the cultured neurons. In summary, a neuronal enriched cell culture system was used with isolated polylysine-coated beads to identify basic 30-kD and 32-kD proteins that may be involved in synapse formation.
authors.affiliation: Department of Anatomy, College of Medicine, Ohio State University, Columbus 43210-1239.
authors.forename: R W
authors.initials: RW
authors.lastname: Burry
authors.forename: D M
authors.initials: DM
authors.lastname: Hayes
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002531:Cerebellum / Q000032:analysis* / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007150:Immunohistochemistry; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D008715:Methionine / Q000378:metabolism; D008863:Microspheres; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000378:metabolism; D009474:Neurons / Q000032:analysis* / Q000378:metabolism; D011107:Polylysine; D051381:Rats; D013569:Synapses / Q000378:metabolism*
chemical_list: D009419:Nerve Tissue Proteins; D011107:Polylysine; D008715:Methionine