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Journal of Neuroscience, Vol 13, 586-595, Copyright © 1993 by Society for Neuroscience
A muscle cell variant defective in glycosaminoglycan biosynthesis forms nerve-induced but not spontaneous clusters of the acetylcholine receptor and the 43 kDa protein
H Gordon, M Lupa, D Bowen and Z Hall
Department of Physiology, School of Medicine, University of California- San Francisco 94143-0444.
Myotubes of the C2 mouse muscle cell line form clusters of ACh receptors
(AChRs) at apparently random sites along their length when cultured alone,
and near sites of nerve-muscle contact when cocultured with neurons. We
find in aneural cultures that myotubes of a C2 variant, S27, which is
defective in glycosaminoglycan synthesis, express the AChR on their
surface, but do not form clusters. S27 cells in aneural cultures also
express the 43 kDa protein but do not cluster it. The altered distribution
of laminin and collagen IV on the surface of S27 myotubes suggests that the
basal lamina is abnormal. Neither the addition of exogenous proteoglycans
or conditioned medium from wild- type C2 cells, nor the growth of S27 cells
on substrates rich in basal lamina elements caused clusters to appear on
S27 myotubes in aneural cultures. When cultured with primary neurons,
however, S27 myotubes formed large clusters of the AChR near sites of
neurite contact. The clusters were coincident with patches of the 43 kDa
protein. Prelabeling experiments indicate that at least some AChRs in the
clusters arise through aggregation. Although Torpedo agrin induces AChR
clusters on C2 myotubes, it does not do so on S27 cells. Our experiments
suggest that the spontaneous formation of clusters of AChRs and the 43 kDa
protein in aneural cultures of myotubes depends upon the normal synthesis
of muscle proteoglycans, and that nerve-induced clustering does not. Thus,
there appear to be multiple mechanisms for the formation of AChR clusters.
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