Evaluation of stearate-graphite paste electrodes for chronic measurement of extracellular dopamine concentrations in the mammalian brain

Abstract

Chronoamperometric procedures, in combination with pharmacological treatments, were used to verify whether stearate-modified graphite paste recording electrodes (SGEs) could measure basal extracellular dopamine (DA) concentrations in the striatum of awake rats over a 3-week period of implantation. Baseline Chronoamperometric signals were unaffected by systemic injections of ascorbate (AA) or the monoamine oxidase inhibitor pargyline, or by intraventricular infusions of the AA degrading enzyme AA-oxidase. In contrast, systemic injections of d-amphetamine or nomifensine increased, and γ-butyrolactone decreased, the signal in a reproducible fashion over a similar test period. In addition, 6-hydroxydopamine lesions of the nigrostriatal DA pathway attenuated the ability of d-amphetamine to increase, and γ-butyrolactone to decrease, the Chronoamperometric signal. In separate studies, reverse microdialysis, performed with dialysis probes implanted directly adjacent to SGEs in the striatum, permitted the assessment of electrode selectivity, sensitivity, response linearity, and detection limits to DA. Perfusion of the probe with normal Ringer solution (5 μl/min) decreased the baseline Chronoamperometric signal by 10 nA. Comparable decreases in the baseline signal were observed after systemic injections of γ-butyrolactone or medial forebrain bundle infusions of tetrodotoxin, suggesting these decreases reflected depletion of extracellular DA to levels below the electrode's detection limit. Reverse dialysis with high concentrations of AA, DOPAC, 5-HT, or 5-HIAA, failed to reverse the decrease in the Chronoamperometric signal induced by dialysis. In contrast, reverse dialysis with a physiologically relevant range of DA concentrations, in rats pretreated with the DA uptake blocker nomifensine, increased the Chronoamperometric signal in a linear fashion with a detection threshold of < 20 nM. Combined, these results indicate that the baseline Chronoamperometric signals recorded at +0.20 V in the striatum with SGEs do not reflect changes in extracellular concentrations of AA, DA metabolites, or indoles, but rather represent neuronally mediated nanomolar changes in extracellular DA concentrations, even after extended periods of implantation in brain tissue.