BK-beta1 subunit: immunolocalization in the mammalian connecting tubule and its role in the kaliuretic response to volume expansion

Large, Ca2+-activated K+ channels (BK), comprised of alpha- and beta-subunits, mediate K+ secretion during high flow rates in distal nephron segments. Because the BK-beta1 subunit enhances Ca2+ sensitivity of BK in a variety of cells, we determined its role in flow-induced K+ secretion and its localization in the mammalian nephron. To determine the role of BK-beta1 in the kaliuretic response to volume expansion, the rate of K+ excretion (UKV) vs. varied urinary flow rates were determined in wild-type and BK-1 knockout mice (BK-beta 1-/-). When flow rate was varied by volume expansion (2 ml x h-1 x 25 g body wt-1) for 30 to 60 min in wild-type mice, we found that the UKV increased significantly with increasing urine flow rates (r2 = 0.50, P < 0.00001, n = 31), as demonstrated previously in distal nephron of rats and rabbits. However, in BK-beta1-/- mice, UKV did not vary with changing flow rates (r2 = 0.15, P = 0.08, n = 20). Using immunohistochemical techniques, we found that BK-beta1 was strongly expressed in the apical membrane of the murine distal nephron and that 98% of BK-beta1 protein detected by histochemistry colocalized with NCX, a marker of connecting tubules (CNT). Both BK-beta1 and NCX colocalized with BK-beta in separate experiments. Furthermore, we confirmed BK-beta1 protein expression in the apical membrane of connecting tubules in rabbits. BK-beta1 RNA from rabbit CNT was sequenced and was identical to previously published rabbit muscle sequences. These data show that the BK-beta1 accessory subunit is present in the CNT segment of the mammalian distal nephron and has a significant role in the kaliuretic response to increased urinary flow induced by volume expansion. [PUBLICATION ABSTRACT]