Chronic second‐by‐second measures of l‐glutamate in the central nervous system of freely moving rats

l‐glutamate (Glu) is the main excitatory neurotransmitter in the central nervous system (CNS) and is associated with motor behavior and sensory perception. While microdialysis methods have been used to record tonic levels of Glu, little is known about the more rapid changes in Glu signals that may be observed in awake rats. We have reported acute recording methods using enzyme‐based microelectrode arrays (MEA) with fast response time and low detection levels of Glu in anesthetized animals with minimal interference. The current paper concerns modification of the MEA design to allow for reliable measures in the brain of conscious rats. In this study, we characterized the effects of chronic implantation of the MEA into the brains of rats. We were capable of measuring Glu levels for 7 days without loss of sensitivity. We performed studies of tail‐pinch induced stress, which caused a robust biphasic increase in Glu. Histological data show chronic implantation of the MEAs caused minimal injury to the CNS. Taken together, our data show that chronic recordings of tonic and phasic Glu can be carried out in awake rats for up to 17 days in vivo allowing longer term studies of Glu regulation in behaving rats.

[1]  L. Gorton,et al.  Redox hydrogel based bienzyme electrode for L-glutamate monitoring. , 1999, Journal of pharmaceutical and biomedical analysis.

[2]  S. Rossell,et al.  One-second time resolution brain microdialysis in fully awake rats. Protocol for the collection, separation and sorting of nanoliter dialysate volumes. , 2003, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[3]  P. Rada,et al.  Glutamate measured by 6-s resolution brain microdialysis: capillary electrophoretic and laser-induced fluorescence detection application. , 1997, Journal of chromatography. B, Biomedical sciences and applications.

[4]  G. Akopian,et al.  Short-term plasticity at inhibitory synapses in rat striatum and its effects on striatal output. , 2001, Journal of neurophysiology.

[5]  F. Mora,et al.  Glutamate-Glutamine Cycle and Aging in Striatum of the Awake Rat: Effects of a Glutamate Transporter Blocker , 2004, Neurochemical Research.

[6]  N. Dusticier,et al.  Serotonin depletion produces long lasting increase in striatal glutamatergic transmission , 2001, Journal of neurochemistry.

[7]  N. Mahy,et al.  Time‐Related Cortical Amino Acid Changes After Basal Forebrain Lesion: A Microdialysis Study , 1995, Journal of neurochemistry.

[8]  F. Pomerleau,et al.  Age-related changes in the dynamics of potassium-evoked L-glutamate release in the striatum of Fischer 344 rats , 2004, Journal of Neural Transmission.

[9]  Ralph N. Adams,et al.  Nafion-coated electrodes with high selectivity for CNS electrochemistry , 1984, Brain Research.

[10]  L. Swanson The Rat Brain in Stereotaxic Coordinates, George Paxinos, Charles Watson (Eds.). Academic Press, San Diego, CA (1982), vii + 153, $35.00, ISBN: 0 125 47620 5 , 1984 .

[11]  G. Gerhardt,et al.  Self-referencing ceramic-based multisite microelectrodes for the detection and elimination of interferences from the measurement of L-glutamate and other analytes. , 2001, Analytical chemistry.

[12]  G. Gerhardt,et al.  Microelectrode array studies of basal and potassium‐evoked release of l‐glutamate in the anesthetized rat brain , 2006, Journal of neurochemistry.

[13]  Robert T Kennedy,et al.  In vivo neurochemical monitoring by microdialysis and capillary separations. , 2002, Current opinion in chemical biology.

[14]  F. Pomerleau,et al.  Real Time in Vivo Measures of l‐Glutamate in the Rat Central Nervous System Using Ceramic‐Based Multisite Microelectrode Arrays , 2003, Annals of the New York Academy of Sciences.

[15]  F Moussy,et al.  In vitro and in vivo degradation of glucose oxidase enzyme used for an implantable glucose biosensor. , 2000, Diabetes technology & therapeutics.

[16]  Hua Yang,et al.  Voltammetric study of extracellular dopamine near microdialysis probes acutely implanted in the striatum of the anesthetized rat , 2005, Journal of Neuroscience Methods.

[17]  J. Engel,et al.  Pentylenetetrazol-induced kindling: early involvement of excitatory and inhibitory systems , 1996, Epilepsy Research.

[18]  Francois Pomerleau,et al.  Improved ceramic-based multisite microelectrode for rapid measurements of l-glutamate in the CNS , 2002, Journal of Neuroscience Methods.

[19]  B. Moghaddam,et al.  Temporal dynamics of glutamate efflux in the prefrontal cortex and in the hippocampus following repeated stress: effects of pretreatment with saline or diazepam , 1997, Neuroscience.

[20]  Zheng-Xiong Xi,et al.  The Origin and Neuronal Function of In Vivo Nonsynaptic Glutamate , 2002, The Journal of Neuroscience.

[21]  Lawrence K Duffy,et al.  An ultrastructural analysis of tissue surrounding a microdialysis probe , 1999, Journal of Neuroscience Methods.

[22]  Bita Moghaddam,et al.  Stress activation of glutamate neurotransmission in the prefrontal cortex: implications for dopamine-associated psychiatric disorders , 2002, Biological Psychiatry.

[23]  B. Westerink,et al.  Brain microdialysis of GABA and glutamate: What does it signify? , 1997, Synapse.

[24]  K. Moxon,et al.  Ceramic-based multisite microelectrodes for electrochemical recordings. , 2000, Analytical chemistry.

[25]  B. Westerink,et al.  In vivo monitoring of extracellular glutamate in the brain with a microsensor , 2006, Brain Research.

[26]  P. Kalivas,et al.  Regulation of Extracellular Glutamate in the Prefrontal Cortex: Focus on the Cystine Glutamate Exchanger and Group I Metabotropic Glutamate Receptors , 2005, Journal of Pharmacology and Experimental Therapeutics.

[27]  Robert T. Kennedy,et al.  In vivo monitoring of amino acids by microdialysis sampling with on-line derivatization by naphthalene-2,3-dicarboxyaldehyde and rapid micellar electrokinetic capillary chromatography , 2004, Journal of Neuroscience Methods.

[28]  B. Westerink,et al.  Evaluation of hydrogel-coated glutamate microsensors. , 2006, Analytical chemistry.

[29]  Nicholas A. Cellar,et al.  Microfluidic chip for low-flow push-pull perfusion sampling in vivo with on-line analysis of amino acids. , 2005, Analytical chemistry.