Neuronal fast activating and meningeal silent modulatory BK channel splice variants cloned from rat

The big conductance calcium-activated K(+) channel (BK) is involved in regulating neuron and smooth muscle cell excitability. Functional diversity of BK is generated by alpha-subunit splice variation and co-expression with beta subunits. Here, we present six different splice combinations cloned from rat brain or cerebral vascular/meningeal tissues, of which at least three variants were previously uncharacterized (X1, X2(92), and X2(188)). An additional variant was identified by polymerase chain reaction but not cloned. Expression in Xenopus oocytes showed that the brain-specific X1 variant displays reduced current, faster activation, and less voltage sensitivity than the insert-less Zero variant. Other cloned variants Strex and Slo27,3 showed slower activation than Zero. The X1 variant contains sequence inserts in the S1-S2 extracellular loop (8 aa), between intracellular domains RCK1 and RCK2 (4 aa at SS1) and upstream of the calcium "bowl" (27 aa at SS4). Two other truncated variants, X2(92) and X2(188), lacking the intracellular C-terminal (stop downstream of S6), were cloned from cerebral vascular/meningeal tissue. They appear non-functional as no current expression was observed, but the X2(92) appeared to slow the activation of the Zero variant when co-expressed. Positive protein expression of X2(92) was observed in oocytes by immunoblotting and fluorescence using a yellow fluorescent protein-tagged construct. The functional characteristics of the X1 variant may be important for a subpopulation of BK channels in the brain, while the "silent" truncated variants X2(92) and X2(188) may play a role as modulators of other BK channel variants in a way similar to well known beta subunits.