Hypotonic swelling of liver cells is followed by regulatory volume decrease (RVD), which has been shown to involve facilitated release of K+. In this study, the role of C1- in RVD was examined by videoplanimetric analysis of cell volume and measurement of membrane potential (Vm) and resistance (Rm) in single isolated rat hepatocytes, and by measurement of 36Cl efflux in the isolated perfused liver preloaded with the isotope. Liver cells subjected to hypotonic stress by removal of 50 mM external NaCl (70% of control osmolality) swelled from an initial volume of 6.68 +/- 0.77 to 8.27 +/- 0.88 pl (24.3 +/- 3.4% increase) within 1 min and exhibited RVD at an initial rate of 0.26 +/- 0.01 pl/min. A step decrease in external Cl- accelerated the initial rate of RVD to 0.53 +/- 0.08 pl/min. RVD was abolished in cells that had been depleted of Cl-. Vm and Rm displayed biphasic responses to hypotonic stress. An initial (less than 15 s) hyperpolarization of Vm from -35.4 +/- 2.2 to -38.8 +/- 2.6 mV was followed by a gradual depolarization reaching -30.2 +/- 2.0 mV by 1 min. In parallel, Rm initially (less than 15 s) increased from 101 +/- 13 to 121 +/- 17 M omega (19 +/- 3% increase) and then declined to 55 +/- 4 M omega (59 +/- 4% of initial Rm) within 1 min. These changes were reversible upon return to isotonicity.(ABSTRACT TRUNCATED AT 250 WORDS)