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We have characterized the membrane currents in the H9c2 clonal muscle cell line derived from embryonic rat ventricle. These cells, established by selective serial passage and clonal proliferation, have been found by Hescheler and coworkers to express dihydropyridine-sensitive calcium channels that respond to beta-adrenergic stimulation. We have investigated the macroscopic and elementary currents in these cells by using standard patch-clamp methods. In cells that are kept confluent for 3-4 weeks, we have confirmed the expression of L-type calcium channels and additionally establish that the unitary conductance of many, but not all, of these channels (25 pS in 70 mM barium) is equal to that of cardiac rather than skeletal muscle. When the cells are proliferating rapidly, calcium channels are sparse or absent, but at least two distinct potassium channels and a nonspecific cation channel are observed. The nonspecific channel exhibits a conductance of 30 pS in physiological saline and conducts sodium, potassium, and calcium with nearly equal efficacy. Several unusual properties distinguish this nonspecific channel from others described previously. Gating is voltage dependent, with slow activation and marked increases in open probability at positive potentials. Unlike voltage, activation and marked increases in open probability at positive potentials. Unlike voltage, changes in [Ca2+] or in membrane stretch do not noticeably influence activity. In conclusion, our work and that of Hescheler et al indicate that H9c2 cells are potentially valuable surrogates for the investigation of ion channel regulation and muscular gene expression.

L-type calcium channels, potassium channels, and novel nonspecific cation channels in a clonal muscle cell line derived from embryonic rat ventricle.