Hovde Moriah J, Bolland Danielle E, Kleinsasser Corey D, Shetty Madhur, Blackwell Aaron C, Golovko Mikhail Y, Golovko Svetlana A, Brown Christopher R, Foster James D, Vaughan Roxanne A
Department of Biomedical Sciences, University of North Dakota, School of Medicine and Health Sciences, Grand Forks, ND 58202.
bioRxiv. 2025 May 8:2025.05.02.651918. doi: 10.1101/2025.05.02.651918.
The dopamine transporter (DAT) exerts temporal and spatial control over dopaminergic neurotransmission through reuptake of extracellular dopamine (DA). The functional capacity of DAT is under the control of signaling inputs and post-translational modifications that confer acute presynaptic regulation of reuptake in response to physiological needs, and dysregulation of these processes may contribute to DA imbalances in mood disorders and drug addiction. A key modification of DAT is palmitoylation, a lipid adduction that enhances transport velocity, is suppressed by protein kinase C, and opposes protein kinase C-mediated down-regulation. Here we now show in rat striatum and heterologous cells that transporter palmitoylation is also linked to methamphetamine (METH), undergoing rapid and transient reductions in response to the drug. The time course and other characteristics of palmitoylation reduction parallel those of METH-induced transport down-regulation, and a palmitoylation-deficient DAT mutant shows enhanced down-regulation to METH, supporting a mechanistic link between reduction of the modification and reduced reuptake activity. Recovery rates differed, however, with palmitoylation returning to starting levels more rapidly than reuptake, indicating that down-regulation mechanisms remain engaged with transporters that have undergone repalmitoylation. These results support palmitoylation as a rapid response mechanism that modulates DAT entry into METH-induced down-regulation states and suggest a broader role for the modification in control of reuptake in additional physiological and pathophysiological conditions.
多巴胺转运体(DAT)通过重摄取细胞外多巴胺(DA)对多巴胺能神经传递进行时空控制。DAT的功能能力受信号输入和翻译后修饰的调控,这些调控赋予了对重摄取的急性突触前调节以响应生理需求,而这些过程的失调可能导致情绪障碍和药物成瘾中的DA失衡。DAT的一个关键修饰是棕榈酰化,这是一种脂质加成作用,可提高转运速度,受蛋白激酶C抑制,并对抗蛋白激酶C介导的下调作用。我们现在在大鼠纹状体和异源细胞中发现,转运体棕榈酰化也与甲基苯丙胺(METH)有关,对该药物会发生快速且短暂的减少。棕榈酰化减少的时间进程和其他特征与METH诱导的转运下调相似,并且一个棕榈酰化缺陷的DAT突变体对METH表现出增强的下调作用,支持了修饰减少与重摄取活性降低之间的机制联系。然而,恢复速率有所不同,棕榈酰化比重摄取更快地恢复到起始水平,这表明下调机制仍与经历了再棕榈酰化的转运体相关。这些结果支持棕榈酰化作为一种快速反应机制,可调节DAT进入METH诱导的下调状态,并表明该修饰在控制其他生理和病理生理条件下的重摄取中具有更广泛的作用。