STABLE EXPRESSION AND COUPLING OF CARDIAC L-TYPE CA2-ADRENOCEPTORS( CHANNELS WITH BETA(1))

A number of neurotransmitters modulate cardiac dihydropyridine-sensiti ve L-type Ca2+ channels through several homologous G protein-coupled r eceptors. Previous studies that have examined receptor-Ca2+ channel in teractions have suffered because of the coexpression of various recept or subtypes in native cells. To study the functional coupling of a par ticular receptor subtype to these channels, rabbit cardiac Ca2+ channe l alpha(1) and skeletal beta and (alpha(2)/delta subunits were stably expressed in baby hamster kidney cells. In this stable cell line, Ca2 channels remained at high levels (>1000 fmol/mg protein, or 2700 chan nels per cell) over extended times. The expressed recombinant Ca2+ cha nnels displayed the voltage dependence of activation and inactivation, unitary conductance, and pharmacology characteristic of native cardia c L-type Ca2+ channels. Subsequent coexpression of the beta(1)-adrenoc eptors (150 to 300 fmol/mg protein) with the Ca2+ channels resulted in cell responsiveness to the extracellular application of isoproterenol . These results indicate that heterogeneous expression in mammalian ce lls provides a useful system for studying both biophysical analysis of Ca2+ channel properties and receptor-coupled regulatory processes.