Effect of melittin on the apical membrane Na+ and C1- conductances of frog cornea epithelium

Abstract

The transepithelial conductance increased with 10(-6) M melittin on the tear side of the frog cornea. This effect was attributed to the opening of an apical membrane Na+ conductance. However, the potency of the venom at this concentration (apical membrane fractional resistance (fRo) decreased to near zero) masked this and other effects. With 3 x 10(-7) M on the tear side, the effects were finite (fRo > 0) and reversible. With fRo > 0, the effects of melittin could be readily studied on the transport parameters of Na+, Cl-, and K+. Epithelial cells of intact bullfrog corneas were impaled with microelectrodes using an in vitro preparation. Under short-circuit current (Isc) conditions, 20 min after melittin Isc increased by 3.1 from 8.2 microA/cm2., fRo decreased by 18 from 51%; the intracellular potential, Vo, depolarized by 19.4 from -56.5 mV; and the transepithelial conductance, gv increased by 0.57 from 0.29 mS/cm2. Tenfold decreases in tear Na+ or Cl- concentrations changed the transport parameters consistent with the formation of a Na+ conductance and an increase in the apical membrane Cl- conductance by the venom. These conclusions were further supported by the minimal effect of melittin in Cl(-)-fre and, particularly, in Na(+)-free solutions. Changes in K+ concentration had no effect on transport parameters. These findings indicate that the effect of melittin at this low concentration is upon a Na+ channel protein and not due to nonspecific conductances.