Psychostimulant Regulated DAT Phosphorylation

描述(由申请人提供)： DAT是一种神经递质转运体，在神经去极化和突触释放后清除突触中的多巴胺(DA)。蛋白激酶C的激活刺激DAT的磷酸化和内化，减少DA的摄取。本项目的目的是检测和表征DAT的磷酸化和功能调节对苯丙胺、可卡因和其他精神刺激剂的反应。特定目的1通过代谢标记在大鼠纹状体或培养细胞中表达的DAT来表征DAT对底物或摄取阻滞剂的反应。特定目的2检测PMA/O诱导的DAT对底物或摄取阻滞剂的反应。特定目标三将通过[~3H]DA摄取和细胞表面生物素化DAT表达来检测DAT的功能，以响应特定目标一和特定目标二调节DAT磷酸化的处理。这项研究计划的目的是促进对滥用和神经毒性药物的DAT磷酸化和功能调节的理解。

DESCRIPTION (provided by applicant): DAT is a neurotransmitter transporter that clears the synapse of dopamine (DA) following neural depolarization and synaptic release. Protein kinase C activation stimulates DAT phosphorylation and internalization and reduces DA uptake. The purpose of this project is to examine and characterize DAT phosphorylation and functional regulation in response to amphetamines, cocaine and other psychostimulants. Specific Aim One characterizes DAT phosphorylation in response to DAT substrates or uptake blockers by metabolic labeling of DATs expressed in rat striatum or cultured cells Specific Aim Two examines PMA/Oainduced DAT phosphorylation in response to DAT substrates or uptake blockers. Specific Aim Three will examine DAT function by [3H]DA uptake and cell surface expression of DAT by biotinylation, in response to treatments from Specific Aims One and Two that modulate DAT phosphorylation. The purpose of this research proposal is to advance the understanding of DAT phosphorylation and functional regulation with respect to abused and neurotoxic drugs.