The present study demonstrates that B-type Ca2+ channels observed in rat ventricular myocytes markedly reacted to agents known to affect the ion-motive plasma membrane Ca2+-ATPase (PMCA) pump. Chlorpromazine (CPZ)-activated B-type Ca2+ channels were completely blocked by internal application of PMCA pump inhibitors, namely La3+ (100 microm), eosin (10 microm) and AIF(3) (100 microm). Calmodulin (50 U/ml), the main endogenous positive regulator of PMCA, was unable to activate but significantly reduced CPZ-activated B-type channel activity. In the same manner, ATP (1 and 4 mm), the main energizing substrate of PMCA, was able to reversibly and significantly reduce this activity in a dose-dependent manner. Interestingly, anti-PMCA antibody 5F10, but not anti-Na/K ATPase antibody (used as a negative control) induced a marked Ba2+-conducting channel activity that shared the same characteristics with that of CPZ-activated B-type channels. 5F10-Activated channels were mostly selective towards Ba2+ , mainly had three observed conductance levels (23, 47 and 85 pS), were observed with a frequency of about 1 out of 5 membrane patches and were completely blocked by 10 microm eosin. These results suggest that B-type Ca2+ channels are some form of the PMCA pump.