When the Na electrochemical potential difference across the skin (delta muNa) is altered by perturbing the transmembrane electrical potential difference or the external Na concentration, effects on transport and associated oxygen consumption can be described by the formalism of linear nonequilibrium thermodynamics (Vieira, Caplan & Essig, 1972, J. Gen. Physiol. 59:77; Danisi & Lacaz-Vieira, 1974, J. Gen. Physiol. 64:372; Procópio and Lacaz-Vieira, 1977, J. Membrane Biol. 35:219). We now show that with modifications of delta muNa by substitution of Li or choline for Na in the inner bathing solution, this formalism is no longer applicable. Inner Na by K substitution ((Na X K)i) causes profound alterations in short-circuit current (SCC), JinNa, K efflux (JeffK) and PD. SCC drops transiently after (Na X K)i in Cl and in SO4 media, increasing subsequently. In Cl medium, following the initial transient, there is a late decline in SCC toward a steady state. The rate of SCC decline in Cl medium is more pronounced than that observed in SO4 medium. (Na X K)i causes a transient increase in JinNa with a peak synchronous to the minimum in SCC, both in Cl and in SO4 media. This was interpreted as due to depolarization of the inner membrane. In SO4 medium, following the peak observed after (Na X K)i, JimNa drops, to increase again toward a steady state in which SCC and JinNa are not statistically different, resembling the control condition before (Na X K)i. In Cl medium, however, the JinNa steady state is approximately 100% higher than SCC. This difference is due to an important K efflux (JeffK), which builds up progressively after the substitution. The apparent K permeability [JeffK/(Ki)] is of comparable magnitude in Cl and in SO4 media before (Na X K)i and also in SO4 medium after (Na X K)i. However, in Cl medium, after (Na X K)i, the apparent K permeability increases one order of magnitude as compared to the control condition before the ionic substitution. In Cl medium, the high levels of JinNa and of Jeff(K) observed in the steady state after (Na X K)i were interpreted as being a consequence of cell swelling. SCC and PD follow very different temporal patterns after (Na X K)i which are characterized by transients in SCC and a simple fall in PD. Reasons for these differences are discussed.

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A technique for isolating the stratum corneum from the subjacent layers of the epithelium was developed which permits studying the stratum corneum as an isolated membrane mounted between half-chambers. The method basically consists of an osmotic shock induced by immersing a piece of skin in distilled water at 50 degrees C for 2 min. When the membrane is bathed on each surface by NaCl-Ringer's solution, its electrical resistance is 14.1 +/- 1.3 omega cm2 (n=10). This value is about 1/100 of the whole skin resistance in the presence of the same solution. The hydraulic filtration coefficient (Lp) measured by a hydrostatic pressure method, with identical solutions on each side of the membrane, is 8.8 X 10(-5) +/- 1.5 X 10(-5) cm sec-1 atm-1 (n=10) in distilled water and 9.2 X 10(-5) +/- 1.4 X 10(-5) cm sec-1 atm-1 (n=10) in NaCl-Ringer's solution. These values are not statistically different and are within the range of 1/80 to 1/120 of the whole skin Lp. The stratum corneum shows an amphoteric character when studied by KCl diffusion potentials at different pH'S. The membrane presents an isoelectric pH of 4.6 +/- 0.3 (n=10). Above the isoelectric pH the potassium transport number is higher than the chloride transport number; below it, the reverse situation is valid. Divalent cations (Ca++ or Cu++) reduce membrane ionic discrimination when the membrane is negatively charged and are ineffective when the membrane fixed charges are protonated at low pH.

Biological (stratum corneum) and artificial (cation-exchange resin beads, Bio-Rad AG 50W-X2) ion exchangers were impaled by glass microelectrodes filled with KCl solution. The electrical potential difference recorded in these structures in reference to the external bathing medium was shown to be dependent on the KCl concentration of both the external and the microelectrode filling solutions. The potentials were interpreted on the grounds of the fixed charge theory of membrane potentials as a consequence of two phase boundary potentials (Donnan potentials), one at the matrix-external solution interface and the other at the matrix-microelectrode solution interface. The contribution of a diffusion component for the recorded potential was considered.