Dynamic coupling between TRPV4 and Ca²⁺-activated SK1/3 and IK1 K⁺ channels plays a critical role in regulating the K⁺-secretory BK channel in kidney collecting duct cells.

The large-conductance Ca²⁺-activated K⁺ channel, BK (KCNMA1), is expressed along the connecting tubule (CNT) and cortical collecting duct (CCD) where it underlies flow- and Ca²⁺-dependent K⁺ secretion. Its activity is partially under the control of the mechanosensitive transient receptor potential vanilloid type 4 (TRPV4) Ca²⁺-permeable channel. Recently, we identified three small-/intermediate-conductance Ca²⁺-activated K⁺ channels, SK1 (KCNN1), SK3 (KCNN3), and IK1 (KCNN4), with notably high Ca²⁺-binding affinities, that are expressed in CNT/CCD and may be regulated by TRPV4-mediated Ca²⁺ influx. The K⁺-secreting CCD mCCDcl1 cells, which express these channels, were used to determine whether SK1/3 and IK1 are activated on TRPV4 stimulation and whether they contribute to Ca²⁺ influx and activation of BK. Activation of TRPV4 (GSK1016790A) modestly depolarized the membrane potential and robustly increased intracellular Ca²⁺, [Ca²⁺]_i Inhibition of both SK1/3 and IK1 by application of apamin and 1-[(2-chlorophenyl)diphenylmethyl]-1H-pyrazole (TRAM-34), respectively, further depolarized the membrane potential and markedly suppressed the TRPV4-mediated rise in [Ca²⁺]_i Application of BK inhibitor iberiotoxin after activation of TRPV4 without apamin/TRAM-34 also reduced [Ca²⁺]_i and further intensified membrane depolarization, demonstrating BK involvement. However, the BK-dependent effects on [Ca²⁺]_i and membrane potential were largely abolished by pretreatment with apamin and TRAM-34, identical to that observed by separately suppressing TRPV4-mediated Ca²⁺ influx, demonstrating that SK1/3-IK1 channels potently contribute to TRPV4-mediated BK activation. Our data indicate a direct correlation between TRPV4-mediated Ca²⁺ signal and BK activation but where early activation of SK1/3 and IK1 channels are critical to sufficiently enhanced Ca²⁺ entry and [Ca²⁺]_i levels required for activation of BK.

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