A novel superfusion chamber for the measurement of endogenous glutamate release from cerebellar slices

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.

Abstract

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.