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激活刺激DAT磷酸化和内化，并减少DA摄取。本项目的目的是研究和表征DAT磷酸化和功能调节对安非他明，可卡因和其他精神兴奋剂的反应。具体目标一通过代谢标记大鼠纹状体或培养细胞中表达的DAT来表征DAT底物或摄取阻断剂响应的DAT磷酸化。具体目标二检测PMA/O2诱导的DAT底物或摄取阻断剂响应的DAT磷酸化。具体目标3将通过[3 H]DA摄取和生物素化DAT的细胞表面表达来检查DAT功能，以响应调节DAT磷酸化的具体目标1和2的处理。这项研究计划的目的是促进DAT磷酸化和功能调节方面的滥用和神经毒性药物的理解。

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.