Association of phosphotyrosine with rapsyn expression in Xenopus embryonic cells

The postsynaptic membrane of the neuromuscular junction is highly enriched in rapsyn, which is thought to interact directly with nicotinic acetylcholine receptors (AChR) and anchor them at the synapse. We expressed rapsyn with or without AChRs in Xenopus embryos by mRNA injection. Co-expression of AChR and rapsyn caused the clustering of these two proteins in cultured cells isolated from the injected embryos. When rapsyn was expressed alone, it also became clustered at the substratum-facing membrane in cultured cells and at cell-cell contacts in whole mount embryos. No clusters were observed in cells that expressed AChRs alone. In rapsyn-expressing cells, proteins that are tyrosine phosphorylated as shown by anti-phosphotyrosine antibody labeling were concentrated at rapsyn clusters. Rapsyn itself does not appear to be a substrate for tyrosine kinase. This suggests that other phosphotyrosine-containing proteins are co-clustered with rapsyn in these cells.