title: Cerebellar granule cells express a specific isoform of agrin that lacks the acetylcholine receptor aggregating activity.
abstract: Agrin is a synapse-organizing molecule that mediates nerve-induced aggregation of acetylcholine receptors and other postsynaptic components at the developing and regenerating vertebrate neuromuscular junctions. Several lines of evidence indicate that agrin might play a similar role in directing the organization of postsynaptic specifications of neuron-neuron synapse formation. Here we used immunological methods and polymerase chain reaction to identify the expression of agrin protein and alternatively spliced mRNA isoforms in the culture of rat granule cells. Anti-agrin polyclonal antibody labeled the cultured granule cells and it detected a protein of over 200 kDa in size from the lysate of the cultured cells. Analysis by polymerase chain reaction showed that the granule cells in culture expressed predominantly the B0 isoform of agrin mRNA. When granule cells were co-cultured with primary chick myotubes, there was no detectable effect on the aggregation of acetylcholine receptors on the surface of the myotubes. These results show that the cerebellar granule cells, similar to motor neurons in vitro, express and secrete agrin but it lacks the acetylcholine receptor aggregating activity.
authors.affiliation: Department of Biology, Hong Kong University of Science and Technology.
authors.forename: K L
authors.initials: KL
authors.lastname: So
authors.forename: S
authors.initials: S
authors.lastname: Pun
authors.forename: D C
authors.initials: DC
authors.lastname: Wan
authors.forename: K W
authors.initials: KW
authors.lastname: Tsim
mesh_terms: D018171:Agrin / Q000235:genetics / Q000378:metabolism*; D017398:Alternative Splicing; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism*; D002642:Chick Embryo; D017931:DNA Primers / Q000235:genetics; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011950:Receptors, Cholinergic / Q000378:metabolism*
chemical_list: D018171:Agrin; D017931:DNA Primers; D012333:RNA, Messenger; D011950:Receptors, Cholinergic