Active transport and exchange diffusion of Cl across the isolated skin of Rana pipiens

Abstract

Transepithelial Cl influx and efflux were measured in pairs of frog skin (Rana pipiens) matched according to short-circuit current, tissue conductance, and transepithelial potential (TEP). The skins were bathed symmetrically in NaCl Ringer and voltage clamped at TEP values ranging from -60 to +60 mV. At 0 TEP, Cl influx and net inward Cl movement (in neq X h-1 X cm-2) were, respectively, 961 +/- 116 and 463 +/- 68 in NaCl Ringer, 509 +/- 52 and 202 +/- 53 in amiloride-treated skins, 4,168 +/- 777 and 1,444 +/- 447 in theophylline-treated skins, and 587 +/- 38 and 97 +/- 44 in Na-free Ringer. A correlation was discovered between short-circuit current and Cl fluxes corresponding to a 2:6:1 relationship between changes in active Na transport and active Cl transport. Deviations from the predicted Cl flux ratio indicate the presence of exchange diffusion in the range of spontaneously occurring TEPs, in contrast to observations on R. temporaria and R. esculenta. The experiments indicate that a substantial portion of transepithelial Cl movement proceeds transcellularly 1) via active Cl transport that is Na dependent, amiloride sensitive, stimulated by theophylline, and apparently correlated with active Na transport, and 2) by means of exchange diffusion that not only occurs under short-circuit conditions but also at positive TEPs. It is possible to explain both the exchange diffusion and the properties of active Cl transport by a Cl-HCO3 exchange system at the apical side of the transporting cell that interacts with a Na-H exchange mechanism, a notion consistent with the recent observation of an amiloride-induced decrease in intracellular pH.