Schousboe A, Sánchez Olea R, Morán J, Pasantes-Morales H
Department of Biological Sciences, PharmaBiotec Research Center, Royal Danish School of Pharmacy, Copenhagen.
J Neurosci Res. 1991 Dec;30(4):661-5. doi: 10.1002/jnr.490300409.
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.
在培养的小鼠小脑颗粒细胞中研究了低渗条件对牛磺酸摄取和释放的影响。研究了4,4'-二异硫氰基芪-2,2'-二磺酸盐(DIDS)以及二价阳离子Mg++和Mn++对低渗诱导的这些参数变化的影响。Mg++(20 mM)和Mn++(5 mM)分别抑制了在30%低渗培养基中观察到的牛磺酸释放的25%和41%。DIDS(100 μM)抑制这种释放达46%。50%低渗溶液引起的牛磺酸外流被Mg++降低了约40%,被Mn++降低了55%。颗粒细胞对牛磺酸的摄取可分为高亲和力载体介导成分和非饱和扩散成分。高亲和力摄取的动力学常数(Km和Vmax)不受渗透压降低50%的影响。非饱和牛磺酸摄取的扩散常数从等渗培养基中的1.5×10(-4)增加到低渗(50%)培养基中的4.6×10(-4) ml·min-1·mg-1。低渗条件引起的牛磺酸摄取扩散成分的这种增加被100 μM DIDS或5 mM Mn++抑制了约25%。这些结果有力地表明,低渗条件下肿胀诱导的牛磺酸释放增加是由扩散过程介导的,而不是由高亲和力牛磺酸载体的逆转介导的。