Embryonic chicken skeletal muscle cells fail to develop normal excitation-contraction coupling in the absence of the alpha ryanodine receptor. Implications for a two-ryanodine receptor system.

Two ryanodine receptor (RyR), sarcoplasmic reticulum Ca2+ release channels, alpha and beta, co-exist in chicken skeletal muscles. To investigate a two-RyR Ca2+ release system, we compared electrically evoked Ca2+ transients in Crooked Neck Dwarf (cn/cn) cultured muscle cells, which do not make alpha RyR, and normal (+/?) cells. At day 3 in culture, Ca2+ release in +/? cells required extracellular Ca2+ (Ca2+o), and Ca2+ transients had slow kinetics. At day 5, Ca2+ release was Ca2+o-independent in 40% of the cells, and transients were more rapid. By day 7, all +/? cells had Ca2+o-independent Ca2+ release. Contractions were observed in +/? cells on all days. Ca2+ transients were observed in cn/cn cells on days 3, 5, and 7, but in each case they were Ca2+o-dependent and exhibited slow kinetics. Localized vesiculations, not contractions, occurred in cn/cn cells. By day 10, Ca2+ transients were no longer observed in cn/cn cells even in Ca2+o. Sarcoplasmic reticulum Ca2+ was not depleted, as caffeine induced Ca2+ transients. Thus, in the absence of alpha RyR there is a failure to develop Ca2+o-independent Ca2+ release and contractions and to sustain Ca2+o-dependent release. Moreover, contributions by the alpha RyR cannot be duplicated by the beta RyR alone.