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低渗休克对离体虹鳟肝细胞中牛磺酸转运的影响。

Effects of hyposmotic shock on taurine transport in isolated trout hepatocytes.

作者信息

Michel F, Fossat B, Porthé-Nibelle J, Lahlou B, Saint-Marc P

机构信息

Laboratoire de Physiologie Cellulaire et Comparée, Université de Nice-Sophia Antipolis, UFR Faculté des Sciences, France.

出版信息

Exp Physiol. 1994 Nov;79(6):983-95. doi: 10.1113/expphysiol.1994.sp003823.

Abstract

Isolated trout hepatocytes exposed to hypotonic medium undergo a regulatory volume decrease (RVD) that occurs via two separate routes, K(+)-Cl- cotransport and amino acid release, the ion efflux accounting for 70% of the total osmolyte loss. Taurine, glutamine and glutamic acid are the most important and represent 73% of the total amino acid content (53 mmol (l cell water)-1). The osmolarity-sensitive release of amino acids was studied using [3H]taurine. Kinetic studies indicated two components for taurine influx: a linear Na(+)-independent transport and a saturable Na(+)-dependent system with a Michaelis-Menten constant (Km) of 122 microM and a maximum velocity (Vmax) of 31.2 pmol (mg protein)-1 min-1. This second way of uptake was also chloride dependent and indicated an apparent coupling ratio Na+:Cl-:taurine of 2:1:1. The latter component and the taurine efflux were stimulated during RVD, leading to intracellular amino acid loss. Taurine efflux activation during volume recovery was transient and also dependent on the presence of both Na+ and Cl- in the extracellular medium. Furthermore, taurine release and RVD were slowed down when Ca2+ was omitted from the medium. These results suggested two distinct and complementary mechanisms for volume regulation in trout hepatocytes during hypotonic conditions.

摘要

暴露于低渗培养基中的离体虹鳟肝细胞会经历调节性体积减小(RVD),这通过两条独立途径发生,即K(+)-Cl-协同转运和氨基酸释放,离子外流占总渗透溶质损失的70%。牛磺酸、谷氨酰胺和谷氨酸是最重要的,占总氨基酸含量的73%(53 mmol(每升细胞内液)-1)。使用[3H]牛磺酸研究了氨基酸的渗透压敏感性释放。动力学研究表明牛磺酸内流有两个组分:一种线性的不依赖Na(+)的转运和一种可饱和的依赖Na(+)的系统,其米氏常数(Km)为122 μM,最大速度(Vmax)为31.2 pmol(每毫克蛋白质)-1 分钟-1。这种第二种摄取方式也依赖氯化物,表明Na+:Cl-:牛磺酸的表观偶联比为2:1:1。在RVD期间,后一组分和牛磺酸外流受到刺激,导致细胞内氨基酸损失。体积恢复期间牛磺酸外流的激活是短暂的,并且也依赖于细胞外培养基中Na+和Cl-的存在。此外,当培养基中省略Ca2+时,牛磺酸释放和RVD会减慢。这些结果表明在低渗条件下虹鳟肝细胞体积调节存在两种不同且互补的机制。

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