Year of Publication

2006

Document Type

Dissertation

College

Medicine

Department

Anatomy and Neurobiology

First Advisor

Greg A. Gerhardt

Abstract

L-glutamate is the predominant excitatory amino acid neurotransmitter inthe mammalian central nervous system. Prior aging studies have focusedprimarily on dopaminergic circuitry of the striatum, and data obtained studyingglutamate regulation in the striatum have been largely equivocal. Thesediscrepancies are due in large part to the limitations of microdialysis; while it isextremely sensitive to minute concentrations of analyte, it is lacking in terms ofthe temporal resolution necessary to study a neurotransmitter with rapid releaseand clearance kinetics such as glutamate. In order to address this matter, ourlaboratory has designed a ceramic-based multisite microelectrode with thecapability to detect and analyze fluctuations in extracellular glutamateconcentrations on a sub-second basis. These microelectrodes were utilized tostudy the phasic release and uptake dynamics of potassium-evoked glutamate inthe striatum of young (6 month), late-middle aged (18 month) and aged (24month) Fischer 344 rats. Our results showed a reduced glutamate clearancerate and an attenuated response to potassium depolarization in the corticostriatalprojections of aged animals in comparison to other age groups. In addition,average maximal glutamate release amplitudes were decreased in the striatumof aged animals. Pressure ejection of exogenous glutamate solution furtherconfirmed the decreased glutamate clearance ability of the aged striatum. Thesepotassium and exogenous glutamate data also highlighted a markeddorsoventral gradient in the striatum in terms of glutamate release and clearanceability. We further explored this phenomenon of age-related decreased glutamateuptake by coupling our in vivo technology with classical immunoblotting andbiotinylation techniques in order to investigate glutamate transporter regulation.Decreased glutamate clearance in the aged rats cannot be attributed to areduction in steady-state total transporter protein levels. Rather, our resultsindicate that reduced plasma membrane surface trafficking of GLAST in the agedstriatum may be partially responsible for this effect. Finally, we modified ourmicroelectrodes to study basal glutamate levels in the striatum of the aging,freely moving rat. This approach allowed us to study extracellular glutamateregulation free from the potential confounding variable of anesthesia. Our resultsdemonstrate that there is no significant alteration in basal glutamate levels inaging in the brain regions investigated. More importantly, this study validated theefficacy of the utilization of ceramic-based multisite microelectrodes for the studyof alterations in glutamate neurotransmission in the aging, freely moving rat, andit lays the foundation for future work correlating such changes with age associatedimpairments in motor function.

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