Regulatory volume decrease and associated osmolyte fluxes in cerebellar granule neurons are calcium independent.

Abstract

To investigate a possible role for Ca as a transduction signal for regulatory volume decrease (RVD), the effects of external Ca removal, Ca channel blockers (Cd, Co, La, Gd, verapamil, diltiazem, dihydropyridines) and inhibitors of endoplasmic reticulum Ca release (dantrolene, ryanodine, TMB-8) were examined on RVD and on the swelling-activated efflux of two main osmolytes: Cl (traced by 125I) and [3H]taurine. Omission of Ca plus EGTA did not affect RVD or osmolyte release but when BAPTA was the chelator, RVD decreased 20%, 125I fluxes were unaffected and taurine stimulated efflux decreased (20%) while the basal efflux slightly increased (<10%). Verapamil, diltiazem, Co, Cd, La and Gd did not affect RVD or osmolyte fluxes. Nimodipine and nitrendipine (25-50 microM) markedly decreased RVD and osmolyte fluxes (>90%) through a mechanism independent of extracellular Ca. Swelling elicited an increase in cytosolic Ca measured by fura-2, which was notably variable ranging 50-350 nM. However, RVD and osmolyte fluxes were not affected by the blockers of endogenous Ca release dantrolene, ryanodine and TMB-8 or by the permeable Ca chelator BAPTA-AM, even when the cytosolic Ca increase was abolished by the chelator. These results indicate that 1) RVD and osmolyte fluxes are independent of extracellular Ca 2) RVD, osmolyte release and cytosolic Ca raise are only coincident events. Consequently, Ca is unlikely to be a transducing signal for RVD in neurons.