Rapidly inactivating and non-inactivating calcium-activated potassium currents in frog saccular hair cells

Using a semi-intact epithelial preparation we examined the Ca2+-activated K + (K-Ca) currents of frog (Rana pipiens) saccular hair cells. After blockin g voltage-dependent K+ (K-V) currents with 4-aminopyridine (4-AP) an outwar d current containing inactivating (I-transient) and noninactivating (I-stea dy) components remained. 2. The contribution of each varied greatly from cell to cell, with I-transi ent contributing from 14 to 90% of the total outward current. Inactivation of I-transient was rapid (tau approximate to 2-3 ms) and occurred within th e physiological range of membrane potentials(V-1/2 = -63 mV). Recovery from inactivation was also rapid (tau approximate to 10 ms). 3. Suppression of both I-transient and I-steady by depolarizations that app roached the Ca2+ equilibrium potential and by treatments that blocked Ca2influx (application Ca2+-free saline or Cd2+), suggest both are Ca2+ depend ent. Both were blocked by iberiotoxin, a specific blocker of large-conducta nce K-Ca channels (BK), but not by apamin, a specific blocker of small-cond uctance K-Ca channels. 4. Ensemble-variance analysis showed that I-transient and I-steady flow thr ough two distinct populations of channels, both of which have a large singl e-channel conductance (similar to 100 pS in non-symmetrical conditions). To gether, these data indicate that both I-transient and I-steady are carried through BK channels, one of which undergoes rapid inactivation while the ot her does not. 5. Inactivation of I-transient could be removed by extracellular papain and could later be restored by intracellular application of the 'ball' domain of the auxiliary subunit (beta2) thought to mediate BK channel inactivation in rat chromaffin cells. We hypothesize that I-transient results from the association of a similar beta subunit with some of the BK channels and that papain removes inactivation by cleaving extracellular sites required for t his association.