Ultrarapid delayed rectifier K(+) current in H9c2 rat ventricular cell line: biophysical property and molecular identity.

Ultrarapid delayed rectifier K(+) currents (I(Kur)s) contribute importantly to cardiac repolarization. However, understanding of I(Kur)s has been hampered by the difficulty of dissecting them from overlapping currents in primary cells. We found with whole-cell patch-clamp recordings that H9c2 cells, a rat ventricular cell line, under 50% confluence (single myoblasts) express exclusively I(Kur)-like current which activates rapidly upon depolarization and partially inactivates during 150-ms pulses. The H9c2 I(Kur) activates within the same voltage range as native cardiac I(Kur)s, with a half-activation voltage of -13 mV. The H9c2 I(Kur) can be completely blocked by tetraethylammonium and 4-aminopyridine. Reversal potential (-79 mV) and envelope-of tail analyses indicate that the H9c2 I(Kur) is carried by a single population of K(+) channels. H9c2 I(Kur) was increased by beta-adrenoceptor-PKA and decreased by alpha(1)-adrenoceptor-PKC activation by isoproterenol and phenylephrine, respectively. Immunocytochemistry was performed with antibodies against 11 different K(+) channels. Positive immuno-stain-ing of the cytoplasmic membrane of H9c2 cells was seen only with Kv3.1b antibody. Antisense oligodeoxynucleotides directed against Kv3.1b subunit sequence significantly inhibited the H9c2 I(Kur). We conclude that the H9c2 cells at the myoblast stage express mainly I(Kur) and Kv3.1b may be a molecular component of the H9c2 I(Kur), and H9c2 cells provide a suitable system for studying I(Kur).