Barnes S, Leighton G E, Davies J A
Department of Pharmacology and Therapeutics, University of Wales College of Medicine, Cardiff, U.K.
J Neurosci Methods. 1988 Feb;23(1):57-61. doi: 10.1016/0165-0270(88)90022-2.
Methods are described for use in the study of potassium-stimulated release of endogenous glutamate and aspartate from perfused cerebellar slices using high performance liquid chromatography (HPLC) with fluorometric detection. Slices were cut at 250 micron in one direction only, and were thus similar to those used for electrophysiological techniques. Details of an easily constructed, simple superfusion system are given. HPLC isocratic separation was used to assay aspartate and glutamate release, glutamate alone showing calcium-dependent release. Repetitive pulses of high potassium-containing artificial cerebrospinal fluid induced release of glutamate (six pulses), demonstrating that the tissue was viable and capable of repeated release.
描述了用于研究钾刺激灌注小脑切片释放内源性谷氨酸和天冬氨酸的方法，该方法采用带荧光检测的高效液相色谱（HPLC）。切片仅在一个方向切成250微米厚，因此与用于电生理技术的切片相似。给出了一个易于构建的简单灌流系统的详细信息。采用HPLC等度分离法测定天冬氨酸和谷氨酸的释放，仅谷氨酸显示出钙依赖性释放。含高钾的人工脑脊液重复脉冲诱导谷氨酸释放（六个脉冲），表明组织是活的且能够重复释放。
