Combined electrochemical and electrophysiological studies of monoamine overflow in rat hippocampal slices

In vivo electrochemical measurements of chronoamperometric recordings from Nafion-coated electrodes were used to investigate monoamine overflow from selected regions of the rat hippocampal slice. Concurrent electrophysiological measurements of evoked CA1 pyramidal cell population spike responses were used to characterize changes in the electrical activity in the slices that occur during potassium-induced neurotransmitter overflow. Superfusion with elevated K⁺ (10-50 mM, 5 min) elicited consistent concentration-dependent increases in the electrochemical responses recorded from the dentate gyrus. At the onset of K⁺ perfusion, there was an initial increase in the population spike response, followed by electrical silence, which usually lasted 5-10 min following the return to normal medium, and required 20-30 min for complete recovery of the response. The potassium-induced electrochemical signal always increased following the decline in the electrophysiological response. Although the electrochemical signal usually returned to baseline much before the electrophysiological response (usually within 5 min), both signals remained refractory for some time. Cocaine pretreatment (10-50 μM) caused a dose-dependent augmentation of the electrochemical responses. Local pressure ejection of K⁺ via a micropipette elicited dose-dependent increases in the electrochemical signals that were of relativity brief duration as compared to superfusion with K⁺. Such potassium-evoked responses were highly localized, and were attenuated in amplitude in animals that had been previously treated with the selective noradrenergic neurotoxin, DSP-4. In addition to K⁺, local applications of methyl-amphetamine, tyramine and veratridine also elicited electrochemical signals, and the time courses of these responses were specific to the releasing agent that was used. Preliminary data obtained using high-speed electrochemical 3ecordings of both oxidation and reduction current suggested that tyramine ejections evoked pimarily norepinephrine overflow, while K⁺ evoked the overflow of both norepinephrine and serotonin. The present experiments demonstrate that simultaneous electrophysiological and electrochemical experiments can be used in an isolated preparation of brain such as the hippocampal slice to characterize the electrophysiological events that occur during stimulated transmitter release.