Short-range differential pulse voltammetry for fast, selective analysis of basal levels of cerebral compounds in vivo

Abstract

Differential pulse voltammetry (DPV) with pretreated biosensors (carbon fibre microelectrodes (mCFE), 10-30 microns diameter) allows selective in vivo measurement of basal endogenous levels of dopamine (DA), serotonin (5-HT), their metabolites (dihydroxyphenylacetic acid, DOPAC; 5-hydroxyindoleacetic acid, 5-HIAA), and neuropeptides. We have now modified DPV in order to reduce the time of analysis from tens of seconds to 1-2 s without losing selectivity. We call this newly reported method short-range differential pulse voltammetry (SRDPV). Simply, while in DPV the complete oxidation peak is recorded, SRDPV measures only the top of each oxidation peak. For example, to monitor peak 2 which corresponds to the in vivo oxidation of extracellular DOPAC and occurs at approximately +85 +/- 10 mV, the initial (Ei) and final (Ef) potentials applied with DPV were -100 mV and +200 mV, respectively, while they were +75 mV (Ei) and +95 mV (Ef) with SRDPV. At the typical scan range of 10 mV.s-1, the effective time of measurement was 30 s for DPV and 2 s for SRDPV. A similar procedure was performed to analyze peak 3 (5-HIAA, occurring at +230 +/- 11 mV) with Ei + 50 mV and Ef + 350 mV for DPV, or +220 mV and +240 mV for SRDPV. DPV and SRDPV were compared in vitro by quantitating DOPAC and 5-HIAA in solutions of increasing concentrations (chosen on the basis of the suggested in vivo content of these two compounds). Data indicated that similar sensitivity and selectivity were obtained with both methods at all concentrations, supporting the applicability of SRDPV for in vitro studies. In vivo experiments were performed in anesthetized adult male rats prepared for voltammetry by inserting the electrically pretreated biosensor (mCFE) into the striatum. DPV measurements were performed automatically every 3-5 min and were alternated every 10-20 min with a sequence of 5-10 SRDPV scans performed every 10-30 s. Subsequent pharmacological or electrical manipulations of the two biogenic amine systems studied were monitored by alternate use of DPV and SRDPV. The data presented support the capability of SRDPV with pretreated biosensors to measure in vivo electroactive compounds with selectivity and sensitivity comparable to that of DPV, but with improved time resolution.