Premium
Correlation of clustering peak density and total amount of acetylcholine receptor on cultured mouse myotubes
Author(s) -
Shikada K.,
Kimura I.,
Kimura M.
Publication year - 1987
Publication title -
international journal of developmental neuroscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.761
H-Index - 88
eISSN - 1873-474X
pISSN - 0736-5748
DOI - 10.1016/0736-5748(87)90048-7
Subject(s) - myogenesis , acetylcholine receptor , fluorescence , chemistry , microgram , receptor , microbiology and biotechnology , alpha (finance) , biophysics , biochemistry , biology , in vitro , medicine , physics , construct validity , nursing , quantum mechanics , patient satisfaction
Abstract The development of acetylcholine receptor (AChR) and its clustering were studied quantitatively on mouse myotubes in nerve‐muscle co‐cultures. AChR was visualized by fluorescence‐labeled antibodies (F‐Ab) against crude AChR or fluorescence‐labeled α‐bungarotoxin (F‐α‐BuTX). The F‐Ab stain was observed throughout the entire surface of the myotube at day 8 and appeared clustered at day 13. Both peak density and total amount of fluorescence in F‐Ab stained myotubes were plotted against days in culture. Both fluorescence indices markedly increased from days 8 to 13 of culture were greater in extent in myotubes incubated in the presence of spinal cord explant, as compared with its absence. Similar results were observed in myotubes stained with F‐α‐BuTX. d ‐Tubocurarine ( d ‐TC, 0.1 mg/ml) and native α‐BuTX (1 μg/ml) clearly inhibited both the total amount of fluorescence and the development of peak fluorescence density in the F‐Ab stained myotubes. But the inhibition by d ‐TC appeared at the later day in culture than α‐BuTX did. Low temperature (28°c) and cholesterol (1 μg/ml) treatment inhibited peak fluorescence density without affecting total amount of fluorescence. These results show that the development of ACh can be characterized both by clustering peak density (indicating the lateral mobility of AChR) and by total amount of fluorescence.