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bioRxiv · 10.1101/2023.02.16.528856

Effects of isoflurane and urethane anesthetics on glutamate neurotransmission in rat brain using in vivo amperometry

Beitchman, J. A.·Krishna, G.·Bromberg, C. A.·Thomas, T. C.
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Abstract

Aspects of glutamate neurotransmission implicated in normal and pathological conditions are often evaluated using in vivo recording paradigms in rats anesthetized with isoflurane or urethane. Urethane and isoflurane anesthesia influence glutamate neurotransmission through different mechanisms; however real-time outcome measures of potassium chloride (KCl)-evoked glutamate overflow and glutamate clearance kinetics have not been compared within and between regions of the brain. In the following experiments, in vivo amperometric recordings of KCl-evoked glutamate overflow and glutamate clearance kinetics (uptake rate and T80) in the cortex, hippocampus and thalamus were performed using glutamate-selective microelectrode arrays (MEAs) in young adult male, Sprague-Dawley rats anesthetized with isoflurane or urethane. Potassium chloride (KCl)-evoked glutamate overflow was similar under urethane and isoflurane anesthesia in all brain regions studied. Analysis of glutamate clearance determined that the uptake rate was significantly faster (53.2%, p<0.05) within the thalamus under urethane compared to isoflurane, but no differences were measured in the cortex or hippocampus. Under urethane, glutamate clearance parameters were region dependent, with significantly faster glutamate clearance in the thalamus compared to the cortex but not the hippocampus (p<0.05). No region dependent differences were measured for glutamate overflow using isoflurane. These data support that amperometric recordings of glutamate under isoflurane and urethane anesthesia result in mostly similar and comparable data. However, certain parameters of glutamate uptake vary based on choice of anesthesia and brain region. Special considerations must be given to these areas when considering comparison to previous literature and when planning future experiments. HighlightsO_LIKCl-evoked glutamate overflow was similar under urethane and isoflurane anesthesia C_LIO_LIGlutamate clearance parameters were similar under both anesthesias in the cortex and hippocampus but not the thalamus C_LIO_LIGlutamate clearance kinetics differ between brain regions when anesthetized with urethane. C_LIO_LIExperimental design, brain region of interest, and outcome parameters of glutamate clearance should be considered when designing anesthetized amperometry recordings of glutamate. C_LI

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Beitchman, J. A., Krishna, G., Bromberg, C. A., Thomas, T. C.. 2023-02-16. Effects of isoflurane and urethane anesthetics on glutamate neurotransmission in rat brain using in vivo amperometry. https://doi.org/10.1101/2023.02.16.528856

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