Rapid acid extrusion response triggered by alpha(1) adrenoceptor in CHO cells

1. Using a microphysiometer with synchronized valve switching, we investigated real-time acid extrusion from Chinese hamster ovary (CHO) cells in which human alpha(1) adrenoceptor (AR) is stably expressed, in response to noradrenaline (NA). 2. In the cells expressing alpha(1a) AR, the time course of extracellular acidification after stimulation had two phases; in the first phase it transiently reached a rate several times greater than the base rate with a peak at around 10 s, and in the second it increased to 2 times the base rate and reached a plateau in 2 min. Both phases showed a concentration-dependent increase of acidification rate in response to NA, but had distinct pEC(50) values; 5.6 for the transient phase and 7.2 for the steady phase. 3. In the cells expressing alpha(1b) AR, the transient phase was not detected but the steady phase was observed. The pEC(50) value was 7.1, although the magnitude of the response was much smaller than that with alpha(1a) AR. 4. Both 5-(N-ethyl-N-isopropyl)amiloride (EIPA) and HOE642 inhibited the acid extrusion response by either AR in a concentration-dependent manner. EIPA and HOE642 had high pIC(50) values (7.4 and 7.3, respectively) for inhibition of the transient phase response via alpha(1a) AR. In the inhibition of the steady phase response via alpha(1a) AR, both drugs revealed the presence of two components in the response; one had high pIC(50) values (8.1 and 8.2 for EIPA and HOE642, respectively) and the other had low pIC(50) values (5.6 and 6.0, respectively). In contrast, the steady phase response via alpha(1b) AR was inhibited by EIPA and HOE642 with low pIC(50) values (5.3 and 5.9, respectively). 5. As Ca2+ was depleted, the alpha(1a) AR-induced transient phase disappeared, while the steady phase was not affected. 6. These results suggest that alpha(1a) AR drives two acid extrusion systems in CHO cells upon stimulation; one elicits the transient response, which is largely mediated by an EIPA/HOE642-sensitive and Ca(2+)-dependent Na(+)-H(+) exchanger (NHE), presumably NHE1, and the other induces the steady acid extrusion that is mediated by NHE1 and another NHE which has low sensitivity to both EIPA and HOE642. alpha(1b) AR drives only the steady phase acid extrusion response, which is mainly mediated by NHEs other than NHE1.