Improving Brain Slice Methodology Public Deposited

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  • March 20, 2019
Creator
  • Miller, Charles
    • Affiliation: College of Arts and Sciences, Department of Chemistry
Abstract
  • Brain slices are an in vitro preparation used to simplify the complexity of the brain by isolating particular regions of interest. Dopamine neurons have been found to function differently under current brain slice protocols than in vivo. To improve the worth of the information gained from brain slice experiments, it must be determined which elements of the in vivo environment are important to normal function. Making these determinations is central to this dissertation. Carbon-fiber microelectrodes were used to monitor electrically stimulated dopamine release in brain slices with fast-scan cyclic voltammetry (FSCV). The first question asked is what role extracellular dopamine tone plays in controlling dopamine release. Basal dopamine tone in vivo stimulates D2 autoreceptors, which regulate dopamine release; however, in brain slices, there is no dopamine tone. Therefore, determining proper autoreceptor activation is important to making release biologically relevant. Second, neurons fire action potentials along their axons to cause release at their terminals. In brain slice experiments stimulation occurs at the nerve terminals. To stimulate at the axons requires visualization of the dopamine neurons. Two transgenic mouse models that make dopaminergic cells visible due to their fluorescence are evaluated. Lastly, electrochemical and immunohistochemical methods are used to account for norepinephrine and serotonin in these brain slices. D2 autoreceptor stimulation, proper electrode placement, and the appropriate electrochemical approach are found to be important for improving current brain slice methodology.
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Rights statement
  • In Copyright
Advisor
  • Papoian, Garegin A.
  • Murray, Royce W.
  • Wightman, R. Mark
  • McCarty, Greg
  • Glish, Gary
Degree
  • Doctor of Philosophy
Degree granting institution
  • University of North Carolina at Chapel Hill Graduate School
Graduation year
  • 2008
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