The efflux of organic osmolytes such as taurine is an important mechanism by which cells regulate their volume. The effects of hypotonicity and thrombin on taurine efflux were studied in BC3H1 and C2C12 cells, two mouse myoblastic cell lines that can be induced to differentiate with serum deprivation. In proliferating cultures of both cell types preloaded with [3H]taurine, exposure to 27% hypotonicity activated a 10- to 20-fold increase in [3H]taurine efflux (Jtau). This effect was blocked by the C1- channel inhibitors NPPB and flufenamic acid. Thrombin and the thrombin receptor agonist SFLLRN also activated Jtau that was abolished by NPPB and flufenamic acid. Together, hypotonicity and thrombin synergistically activated Jtau. In differentiated myocytes, the effect of thrombin was abolished, while that of hypotonicity was significantly reduced. These results suggest that (i) hypotonicity and thrombin activate taurine-permeable anion channels in BC3H1 and C2C12 cells, and (ii) these anion channels may be involved in cell proliferation.