Transporter maintenance of dopamine homeostasis

描述(由申请人提供)： 多巴胺转运体(DAT)是从突触间隙移除多巴胺的主要机制。一旦被DAT摄取，DA可以被酶降解，或者被囊泡单胺转运体(VMAT2)回收和重新包装成囊泡。正是这两种蛋白DAT和VMAT2的协同作用导致了多巴胺能神经元中DA的动态平衡。这两种蛋白的功能或表达不当会导致体内平衡的改变，并与许多临床病理有关，包括抑郁症和帕金森氏病。特别令人感兴趣的是由于药物滥用而产生的监管，因为DAT是可卡因和苯丙胺等精神刺激剂的主要作用场所。因此，这项建议的目标是研究在永生化细胞系中共表达的DAT和VMAT2如何相互作用以维持体内平衡，以及滥用药物如何改变它们的功能和调节。这一建议的具体目的是：1)表征这两个蛋白的共表达对细胞系统中DA动态平衡的影响，2)检测共表达对药物诱导的神经递质调节的影响，以及3)确定共表达对药物诱导的活性氧形成的影响。

DESCRIPTION (provided by applicant): The dopamine (DA) transporter (DAT) is the primary mechanism for removing dopamine from the synaptic cleft. Once taken up by the DAT, DA can be either degraded enzymatically, or recycled and repackaged into vesicles by the vesicular monoamine transporter (VMAT2). It is the synergistic interaction of these two proteins, DAT and VMAT2, that lead to DA homeostasis in dopaminergic neurons. Improper function or expression of either of these proteins results in altered homeostasis and has been associated with many clinical pathologies, including depression and Parkinson's disease. Of particular interest is regulation due to substance abuse, since the DAT is a primary site of action of psychostimulants such as cocaine and amphetamine. Therefore, the goals of this proposal are to examine how DAT and VMAT2, co-expressed in immortalized cell lines, interact to maintain homeostasis, and how their function and regulation is altered by drugs of abuse. The specific aims of this proposal are to: 1) characterize the effect of co-expression of these two proteins on DA homeostasis in a cell system, 2) examine the effect co-expression on drug-induced neurotransmitter regulation, and 3) determine the effect of co-expression on drug-induced reactive oxygen species formation.