Abstract
The dopamine transporter (DAT) plays a fundamental role in dopamine (DA) neurotransmission by retrieving the transmitter from the extracellular space back into the DA nerve terminal. Several lines of evidence suggest that in addition to uptake or forward transport, DAT can also function to release DA. This process, which is referred to as DAT-mediated release or efflux, is the mechanism used by potent and highly addictive psychostimulants, such as amphetamine (AMPH) and its analogues, to increase extracellular DA levels in motivational and reward areas of the brain. Conventionally, dopaminergic neurons release DA through a calcium-dependent exocytotic vesicular process. In the past several years, our lab and others have identified signaling pathways that promote DA release through DAT. In this review chapter and step-by-step protocol, we will describe assays to study endogenous DA efflux and DAT dynamics using acute brain slices combined with high-performance liquid chromatography (HPLC) for DA detection along with a modification of a classically used radioactive uptake assay to measure levels of DA uptake. Further examining DAT functions and DA dynamics is key for the development of novel therapeutics in DA-related disorders including substance abuse. Here, we described the use of acute brain slices further dissected into precise regions of interest to examine the dual function of DAT.
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Acknowledgments
We thank the members of the Torres laboratory for their helpful discussions. We also acknowledge the support of the following agencies: Fondecyt Initiation Grant N11191049 (JAP), NIH R01 Grant DA038598 (GET).
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Awadallah, N., Pino, J.A., Koutzoumis, D.N., Torres, G.E. (2023). Characterizing Dopamine Transporter Mediated Uptake and Efflux in Brain Slices: A New Approach. In: Fuentealba-Evans, J.A., Henny, P. (eds) Dopaminergic System Function and Dysfunction: Experimental Approaches. Neuromethods, vol 193. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2799-0_4
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DOI: https://doi.org/10.1007/978-1-0716-2799-0_4
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