Hyposmolarity-induced taurine release in cerebellar granule cells is associated with diffusion and not with high-affinity transport.
The effects of hyposmotic conditions on taurine uptake and release were studied in mice cultured cerebellar granule cells. The effect of DIDS (4,4'-diisothiocyanatostilbene-2,2'-disulfonate) and of the divalent cations Mg++ and Mn++ on the hyposmolarity-induced changes in these parameters was investigated. Mg++ (20 mM) and Mn++ (5 mM) inhibited by 25% and 41%, respectively, the release of taurine observed in 30% hyposmolar media. DIDS (100 microM) inhibited this release by 46%. Taurine efflux evoked by 50% hyposmolar solutions was reduced about 40% by Mg++ and 55% by Mn++. Taurine uptake into the granule cells could be resolved into a high-affinity carrier-mediated component plus a nonsaturable diffusion component. The kinetic constants (Km and Vmax) for the high-affinity uptake were unaffected by a 50% decrease in the osmolarity. The diffusion constant for the nonsaturable taurine uptake was increased from 1.5 x 10(-4) in isosmotic media to 4.6 x 10(-4) ml x min-1 x mg-1 in hyposmotic (50%) media. This increase in the diffusional component of taurine uptake elicited by the hyposmotic condition was inhibited approximately 25% by either 100 microM DIDS or 5 mM Mn++. These results strongly suggest that the increase in taurine release induced by swelling under hyposmotic conditions is mediated by a diffusional process and not by a reversal of the high-affinity taurine carrier.
Schousboe
A
A
Department of Biological Sciences, PharmaBiotec Research Center, Royal Danish School of Pharmacy, Copenhagen.
Sánchez Olea
R
R
Morán
J
J
Pasantes-Morales
H
H
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D001667:Binding, Competitive; D001692:Biological Transport; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism*; D003470:Culture Media; D004058:Diffusion; D006098:Granulocytes / Q000378:metabolism*; D007477:Ions; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D009994:Osmolar Concentration; D013654:Taurine / Q000378:metabolism*
D003470:Culture Media; D007477:Ions; D013654:Taurine; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; D008345:Manganese; D008274:Magnesium; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid