Intracellular chloride activity (alpha iCl) was measured in quiescent cat papillary muscles with chloride liquid ion-exchanger microelectrodes. Muscles were superfused in bicarbonate-buffered solutions containing a control concentration of chloride (146.1 mM) and either 2.7, 4.4, or 5.4 mM potassium. The resulting mean membrane potentials were -96.2, -86.8, and -80.6 mV, respectively. In contrast, alpha iCl and the chloride equilibrium potential were unchanged by these alterations in external potassium and remained stable at 11 mM and -60 mV for up to 10 h following removal of the tissue from the animal. This value of alpha iCl is approximately 2-4 times greater than that predicted assuming chloride is passively distributed. The upper limit of intracellular interference was estimated from low external chloride superfusion and found to be no greater than 2-3 mM. Prolonged exposure to bicarbonate-free solutions buffered with N-2-hydroxyethyl piperazine-N'-2-ethanesulfonic acid (HEPES) did not reduce the apparent value of alpha iCl, suggesting that intracellular interference from intracellular bicarbonate does not significantly influence the measurement of alpha iCl. These results suggest that chloride can move into cat papillary muscle against an electrochemical gradient.
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