Hascup ER, Hascup KN, Stephens M, Pomerleau F, Huettl P, Gratton A et al. Rapid microelectrode measurements and the origin and regulation of extracellular glutamate in rat prefrontal cortex. J Neurochem 115: 1608-1620

Department of Psychiatry, Douglas Mental Health University Institute, McGill University, Montreal, Canada.
Journal of Neurochemistry (Impact Factor: 4.28). 10/2010; 115(6):1608-20. DOI: 10.1111/j.1471-4159.2010.07066.x
Source: PubMed


Glutamate in the prefrontal cortex (PFC) plays a significant role in several mental illnesses, including schizophrenia, addiction and anxiety. Previous studies on PFC glutamate-mediated function have used techniques that raise questions on the neuronal versus astrocytic origin of glutamate. The present studies used enzyme-based microelectrode arrays to monitor second-by-second resting glutamate levels in the PFC of awake rats. Locally applied drugs were employed in an attempt to discriminate between the neuronal or glial components of the resting glutamate signal. Local application of tetrodotoxin (sodium channel blocker), produced a significant (∼ 40%) decline in resting glutamate levels. In addition significant reductions in extracellular glutamate were seen with locally applied ω-conotoxin (MVIIC; ∼ 50%; calcium channel blocker), and the mGluR(2/3) agonist, LY379268 (∼ 20%), and a significant increase with the mGluR(2/3) antagonist LY341495 (∼ 40%), effects all consistent with a large neuronal contribution to the resting glutamate levels. Local administration of D,L-threo-β-benzyloxyaspartate (glutamate transporter inhibitor) produced an ∼ 120% increase in extracellular glutamate levels, supporting that excitatory amino acid transporters, which are largely located on glia, modulate clearance of extracellular glutamate. Interestingly, local application of (S)-4-carboxyphenylglycine (cystine/glutamate antiporter inhibitor), produced small, non-significant bi-phasic changes in extracellular glutamate versus vehicle control. Finally, pre-administration of tetrodotoxin completely blocked the glutamate response to tail pinch stress. Taken together, these results support that PFC resting glutamate levels in rats as measured by the microelectrode array technology are at least 40-50% derived from neurons. Furthermore, these data support that the impulse flow-dependent glutamate release from a physiologically -evoked event is entirely neuronally derived.

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    • "Before the in vivo experiment, each MEA assembly was calibrated in vitro at constant applied potential of +0.7 V versus an Ag/AgCl reference electrode for parameters including slope (electrode sensitivity), linearity (R2) and selectivity (Figure 1b) as previously described.16,19 During calibration, the MEA was placed into a beaker containing a continuously stirred phosphate-buffered saline (pH 7.4 at 37 °C) and challenged to final concentrations of 250 μM ascorbic acid (Sigma-Aldrich), 20, 40 and 60 μM Glu (Sigma-Aldrich), 2 μM dopamine (Sigma-Aldrich) and 8.8 μM H2O2 (Sigma-Aldrich). "
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