The uptake and release of sulfobromophthalein (BSP) and ouabain were studied in hepatocytes isolated from rats which received thioacetamide (TA) (1, 2 or 4 mmol/kg) 6, 12, 24, 48 or 72 h before sacrifice. The effects of TA (1 mmol/kg) on hepatocyte BSP transport varied with time. Total BSP hepatocyte content was decreased 6 h and 24 h after TA treatment and increased 48 h after TA treatment due to decreased and increased uptake, respectively. Hepatocyte BSP conjugation was increased 6 h and 24 h after TA treatment and decreased at 48 h. At the higher doses of TA, 24 h after treatment total BSP hepatocyte content was increased and concomitantly BSP release was decreased at the 4 mmol/kg dose of TA. Ouabain uptake was decreased at all doses of TA and at all treatment times. However, release of ouabain was not altered by TA treatment. These results show TA depresses hepatocyte total BSP uptake (depending on dose and time factors) and depresses hepatocyte ouabain uptake.