Taurocholate depolarizes rat hepatocytes in primary culture by increasing cell membrane Na+ conductance

Abstract

Rat hepatocytes in primary culture were impaled with conventional microelectrodes. Addition of 5-100 mumol/l taurocholate led to a slowly developing depolarization that was maximal at 50 mumol/l (10.5 +/- 1.5 mV, n = 15) and not reversible. The effect was Na+ dependent and decreased in cells preincubated with 1 mumol/l taurocholate. Increasing external K+ tenfold depolarized the cells by 12.3 +/- 2.3 mV under control conditions and by 6.3 +/- 1.2 mV with 50 mumol/l taurocholate present (n = 7). Depolarization by 1 mmol/l Ba2+ was 7.6 +/- 0.8 mV and 6.0 +/- 0.7 mV (n = 9) before and after addition of taurocholate, respectively. Cable analysis and Na+ substitution experiments reveal that this apparent decrease in K+ conductance reflects an actual increase in Na+ conductance: in the presence of taurocholate, specific cell membrane resistance decreased from 2.8 to 2.3 k omega x cm2 x Na+ substitution by 95% depolarized cell membranes by 8.9 +/- 2.9 mV (n = 9), probably due to indirect effects on K+ conductance via changes in cell pH. With taurocholate present, the same manoeuvre changed membrane voltages by -0.8 +/- 2.6 mV. When Na+ concentration was restored to 100% from solutions containing 5% Na+, cells hyperpolarized by 3.5 +/- 3.6 mV (n = 7) under control conditions and depolarized by 4.4 +/- 2.9 mV in the presence of taurocholate, respectively. In Cl- substitution experiments, there was no evidence for changes in Cl- conductance by taurocholate. These results show that taurocholate-induced membrane depolarization is due to an increase in Na+ conductance probably via uptake of the bile acid.