On-line, continuous measurement of extracellular striatal glucose using microdialysis sampling and electrochemical...

A sensitive, enzymatic glucose electrode was coupled with the microdialysis sampling technique to enable the continuous, on-line measurement of dialysate glucose. The glucose sensitive electrode was fabricated by immobilizing glucose oxidase onto the surface of an osmium-polyvinylpyrridine horse radish peroxidase gel (Os-gel-HRP) which had been cast coated onto a glassy carbon electrode. This 'bilayer' electrode generated a reductive current to glucose at a potential of 0 mV thereby minimizing faradic oxidative interferences. The system utilized the continuous mixing of two fluids immediately prior to the 'bilayer' electrode. One fluid was the dialysate. The other was an oxygenated, low pH phosphate buffer which minimized oxidative interference, buffered the electrode from variations of pH and maximized enzyme efficiency. In practical terms, the 'bilayer' electrode was simple to manufacture, quick to reach stable basal currents (less than 60 min), sensitive (2.5 microM glucose could be detected in the dialysate) and durable (usable for up to 3 days). In vivo experiments, used the smallest commercially available microdialysis probes to demonstrate that on-line, continuous measurements of EC striatal glucose in the dialysate were receptive to pharmacological (local perfusion with veratridine (50 microM), systemic hyperglycemia (1.5 ml of 0.55 M glucose intraperitoneal (i.p.)) and anesthesia (Nembutal 40 mg/kg i.p.)) and behavioral (restraint) manipulations. This technique allows for greater temporal resolution than conventional HPLC procedures whilst requiring significantly less technical outlay or analytical expertise. The high sensitivity of the analytical technique could facilitate the study of EC glucose levels in very localized regions of the brain if coupled to microdialysis probes of small dimensions.