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Amperometric glucose biosensors based on the immobilization of glucose oxidase (GOx) on Pt electrodes with electropolymerized o-phenylenediamine (PPD) were implanted in the right striatum of freely-moving rats. Carbon paste electrodes for the simultaneous monitoring of ascorbic acid (AA) and/or tissue O2 were implanted in the left striatum. A detailed in vivo characterization of the Pt/PPD/GOx signal was carried out using various pharmacological manipulations. Confirmation that the biosensor responded to changing glucose levels in brain extracellular fluid (ECF) was obtained by intraperitoneal (i.p.) injection of insulin that caused a decrease in the Pt/PPD/GOx current, and local administration of glucose (1 mM) via an adjacent microdialysis probe that resulted in an increase in the biosensor current. An insulin induced increase in tissue O2 in the brain was also observed. Interference studies involved administering AA and subanaesthetic doses of ketamine i.p. Both resulted in increased extracellular AA levels with ketamine also causing an increase in O2. No significant change in the Pt/PPD/GOx current was observed in either case indicating that changes in O2 and AA, the principal endogenous interferents, have minimal effect on the response of these first generation biosensors. Stability tests over a successive 5-day period revealed no significant change in sensitivity. These in vivo results suggest reliable glucose monitoring in brain ECF.

Simultaneous monitoring of tissue O(2) and regional cerebral blood flow (rCBF) was performed in the striatum of freely-moving rats. Differential pulse amperometry and constant potential amperometry were used to monitor O(2) levels at a carbon paste electrode (CPE), while rCBF values were obtained using the H2 clearance technique. Two forms of behavioural activation were studied and the resultant changes in tissue O(2) and blood flow compared. Both tail pinch and induced grooming produced immediate and parallel increases in O(2) and blood flow which returned to baseline on cessation of activity. These findings indicate that under conditions of physiological stimulation the direct voltammetric measurement of O(2) in brain tissue with a CPE can be used as a reliable index of increases in rCBF, resulting in an improvement in time resolution from 5 min (H2 clearance) to <1 s (amperometry). Because tissue O(2) is a balance between supply by the blood stream and utilisation by the cells, increases in O(2) current are an index of increased blood flow only when supply significantly exceeds utilisation.

Tail pinch in the rat gives rise to a well characterised pattern of behaviour which includes gnawing, licking and eating. We have used both in vivo voltammetry and microdialysis to monitor neurochemical changes which accompany the behavioural response to a 5-min tail pinch. Tail pinch resulted in a increase of extracellular 5-hydroxytryptamine and a smaller and more delayed increase of 5-hydroxyindole acetic acid in the hippocampus. In the striatum there was a rise of both extracellular dopamine and ascorbate. With a recently developed constant potential voltammetric technique we can continuously monitor changes in extracellular ascorbate. Using this technique we found a very rapid rise in ascorbate current during a 5-min tail pinch; the current began to decline as soon as the clip was removed. The high time resolution of the technique also allowed us to record similar ascorbate changes during a 0.5-s tail pinch.