Association of cytoskeletal proteins with newly formed acetylcholine receptor aggregates induced by embryonic brain extract

Experimental Cell Research
M P DanielsR J Bloch

Abstract

Aggregates of acetylcholine receptors (AChR) in muscle cell membranes are associated with accumulations of certain cytoskeletal and peripheral membrane proteins. We treated cultured rat myotubes briefly with embryonic brain extract (EBX) to promote AChR aggregation and determined the distribution of several of these proteins at early stages of aggregation. EBX-treated and control cultures were stained with tetramethylrhodamine-alpha-bungarotoxin to identify AChR aggregates and were then frozen and sectioned on a cryostat. These sections were stained with primary antibodies and fluoresceinated secondary antibodies to localize cytoskeletal proteins. The distributions of AChRs and cytoskeletal proteins was examined qualitatively and analyzed by a semiquantitative assay. Qualitatively, the 43K protein had a distribution that was virtually identical to that of AChR in both control and EBX-treated cultures, and it always colocalized with early AChR aggregates. The 58K protein similarly colocalized with early AChR aggregates, but it was also in aggregate-free areas of muscle membrane. The association of vinculin with the aggregates was quantitatively similar to that of the 43K and 58K proteins, but, qualitatively, its distribution did...Continue Reading

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Citations

Oct 1, 1992·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·W D Phillips, J P Merlie
Jul 1, 1991·The Journal of Cell Biology·S C Froehner
Oct 1, 1993·Neuropathology and Applied Neurobiology·S KelićK Kristensson

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