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体外模拟缺血期间人大脑皮质氨基酸的释放

Amino-acid release from human cerebral cortex during simulated ischaemia in vitro.

作者信息

Hegstad E, Berg-Johnsen J, Haugstad T S, Hauglie-Hanssen E, Langmoen I A

机构信息

Institute for Surgical Research, Rikshospitalet, University of Oslo, Norway.

出版信息

Acta Neurochir (Wien). 1996;138(2):234-41. doi: 10.1007/BF01411367.

Abstract

The aim of the present study was to investigate the release of amino-acids in human cerebral cortex during membrane depolarization and simulated ischaemia (energy deprivation). Superfluous tissue from temporal Iobe resections for epilepsy was cut into 500 microns thick slices and incubated in vitro. Membrane depolarization with 50 mM K+ caused a release of glutamate, aspartate, GABA and glycine, but not glutamine or leucine. The release of glutamate and GABA was Ca(++)-dependent. Slices were exposed to simulated ischaemia (energy deprivation; ED) by combined glucose/oxygen deprivation. This caused a Ca(++)-independent release of glutamate, aspartate, GABA, glycine, and taurine which started after 8 min, peaked at the end or shortly after the 27 min period of ED, and returned to control levels within 11 min following termination of ED. Preloaded D-[3H]aspartate was released both during K(+)-stimulation and ED. Release of D-[3H]aspartate during ED was delayed compared to glutamate supporting an initial phase of synaptic glutamate release. Uptake of L-[3H]glutamate was increased during the period of glutamate release, suggesting passive diffusion across the cell membrane or enhanced transport efficacy in cellular elements with functioning uptake mechanisms.

摘要

本研究的目的是调查在膜去极化和模拟缺血(能量剥夺)过程中人类大脑皮层中氨基酸的释放情况。将用于癫痫治疗的颞叶切除术中多余的组织切成500微米厚的切片并进行体外培养。用50 mM K+进行膜去极化会导致谷氨酸、天冬氨酸、γ-氨基丁酸(GABA)和甘氨酸的释放,但不会导致谷氨酰胺或亮氨酸的释放。谷氨酸和GABA的释放依赖于Ca(++)。通过联合葡萄糖/氧剥夺使切片暴露于模拟缺血(能量剥夺;ED)状态。这会导致谷氨酸、天冬氨酸、GABA、甘氨酸和牛磺酸的Ca(++)非依赖性释放,该释放在8分钟后开始,在ED的27分钟末期或末期后不久达到峰值,并在ED终止后的11分钟内恢复到对照水平。预加载的D-[3H]天冬氨酸在K(+)刺激和ED期间均会释放。与谷氨酸相比,ED期间D-[3H]天冬氨酸的释放有所延迟,这支持了突触谷氨酸释放的初始阶段。在谷氨酸释放期间,L-[3H]谷氨酸的摄取增加,这表明其通过细胞膜的被动扩散或在具有有效摄取机制的细胞成分中转运效率提高。

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