A new method for the measurement of endogenous transmitter release in localized regions of hippocampal slices

We describe here a method that allows measurement of the release of endogenous amino acids from localized regions of brain slices combined with conventional electrophysiological experiments. Hippocampal slices were placed in fully submerged chambers and a cannula was positioned just above the dendritic layers of CA1. The cannula was connected to a peristaltic pump and the content of amino acids in the perfusate was measured by HPLC. Extracellular field potentials were concomitantly recorded. Stable levels of aspartate and glutamate were found above the stratum radiatum of CA1. No detectable release was found when the cannula was located above the alveus, the fimbria or in the effluent of the slice. A pulse of K+ (50 mM) produced a brief 3-fold increase in glutamate, aspartate and a detectable release of GABA in CA1. Brief high frequency trains (10 Hz) also increased significantly the release. This method will be useful in determining alterations in transmitter release in the slice in relation to anoxia, epilepsy and long term potentiation.