Effect of various agents on the cytoplasmic calcium concentration in cultured human muscle cells

1. We determined the cytoplasmic Ca2+ concentration (Ca2+]i) in cultured human muscle cells using the fluorescent indicator Quin-2. 2. The Ca2+]i was dependent on the external Ca2+ concentration. Acetylcholine in the presence of external Ca2+ caused a transient increase in Ca2+]i. Inhibition by nifedipine indicated that this response was mediated through activated voltage-operated channels. In nominally Ca2(+)-free buffer acetylcholine did not markedly increase Ca2+]i. Therefore, the increase in Ca2+]i as a response to depolarization is mainly due to influx of external Ca2+. 3. Various concentrations of caffeine did not influence the Ca2+]i. Dantrolene decreased Ca2+]i, both in the presence and absence of external Ca2+. The reduction probably resulted from an action of dantrolene on the intracellular Ca2+ stores, since dantrolene did not influence 45Ca2+ influx or efflux and caffeine partially counteracted the reduction.