Description of the effects of hypotonic cell swelling and ethanol on maxi Ca2+-activated K+ channel (BK channel) activity and Cl− channel activity in GH4/C1 pituitary tumour cells.Methods
Whole cell-, cell attached- and outside-out patch clamp measurements, fluorescence (fluo-3) measurements of intracellular Ca2+ concentration, cell size video monitoring.Results
GH4/C1 pituitary tumour cells respond to both hypotonicity and ethanol with cell swelling which is followed by a regulatory volume decrease (RVD). Tetraethylammonium and 4,4′-diisothiocyanatostilbene-2,2′-disulphonic acid (DIDS) induced cell swelling per se and inhibited hypotonicity induced RVD. Ethanol-induced swelling is paralleled by an increase in the intracellular Ca2+ concentration and augmented by DIDS. BK channel activation by hypotonicity and ethanol is demonstrated in patch clamp experiments both in intact cells (cell attached configuration) and a subset of excised membrane patches (outside-out configuration). Cell swelling and addition of ionomycin under isotonic conditions leads to the activation of outwardly rectifying Cl− currents with time dependent activation at positive potentials.Conclusions
In GH4/C1 cells both hypotonicity and ethanol lead to cell swelling, RVD and to activation of BK channels. The hypotonicity-induced BK channel activation can also be observed in cell free outside-out patches. Hypotonicity, but not ethanol leads to the activation of Cl− channels with features of Ca2+-activated Cl− currents.