(Xeno)estrogen sensitivity of smooth muscle BK channels conferred by the regulatory β1 subunit -: A study of β1 knockout mice

Estrogen and xenoestrogens (i.e. agents that are not steroids but possess estrogenic activity) increase the open probability (P-o) of large conductance Ca2+-activated K+ (BK) channels in smooth muscle. The mechanism of action may involve the regulatory beta1 subunit. We used beta1 subunit knockout (beta1(-/-)) mice to test the hypothesis that the regulatory beta1 subunit is essential for the activation of BK channels by tamoxifen, 4-OH tamoxifen (a major biologically active metabolite), and 17 beta -estradiol in native myocytes. Patch clamp recordings demonstrate BK channels from beta1(-/-) mice were similar to wild type with the exception of markedly reduced Ca2+/voltage sensitivity and faster activation kinetics. In wild type myocytes, (xeno)estrogens increased NPo (P-o x the number of channels, N), shifted the voltage of half-activation (V-1/2) to more negative potentials, and decreased unitary conductance. These effects were non-genomic and direct, because they were rapid, reversible, and observed in cell-free patches. None of the (xeno)estrogens increased the NP. of BK channels from beta1(-/-) mice, but all three agents decreased single channel conductance. Thus, (xeno)estrogens increase BK NP. through a mechanism involving the beta1 subunit. The decrease in conductance did not require the beta1 subunit and probably reflects an interaction with the pore-forming a subunit. We demonstrate regulation of smooth muscle BK channels by physiological (steroid hormones) and pharmacological (chemotherapeutic) agents and reveal the critical role of the beta1 subunit in these responses in native myocytes.