Characterization of noradrenaline-induced increases in intracellular Ca2+ levels in Chinese hamster ovary cells stably expressing human α1A-adrenoceptor

The mechanism for noradrenaline (NA)-induced increases in intracellular Ca2+ concentration ([Ca2+];) and physiological significance of Na+ influx through receptor-operated channels (ROCs) and store-operated channels (SOCs) were studied in Chinese hamster ovary (CHO) cells stably expressing human a,A-adrenoceptor (alpha(1A)-AR). [Ca2+]; was measured using the Ca2+ indicator fura-2. NA (1 mu M) elicited transient and subsequent sustained [Ca2+]; increases, which were inhibited by YM-254890 (G(alpha q/11) inhibitor), U-73122 (phospholipase C (PLC) inhibitor), and bisindolylmaleimide I (protein kinase C (PKC) inhibitor), suggesting their dependence on G(alpha q/11)/PLC/PKC. Both phases were suppressed. by extracellular Ca2+ removal, SK&F 96365 (inhibitor of SOC and nonselective cation channel type-2 (NSCC-2)), LOE 908 (inhibitor of NSCC-1 and NSCC-2), and La3+ (inhibitor of transient receptor potential canonical (TRPC) channel). Reduction of extracellular Na+ and pretreatment with KB-87943, a Na+/Ca2+ exchanger (NCX) inhibitor, inhibited both phases of [Ca2+], increases. These results suggest that 1) stimulation of alpha(1A)-AR with NA elicits the transient and sustained increases in [Ca2+]; mediated through NSCC-2 that belongs to a TRPC family; 2) Na+ influx through these channels drives NCX in the reverse mode, causing Ca2+ influx in, exchange for Na+ efflux; and 3) the G(alpha q/11)/PLC/PKC-dependent pathway plays an important role in the increases in [Ca2+];.