Cocaine-regulated trafficking of dopamine transporters in cultured neurons revealed by a pH sensitive reporter.
Cocaine-regulated trafficking of dopamine transporters in cultured neurons revealed by a pH sensitive reporter.
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DOI：
10.1016/j.isci.2022.105782
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发表时间：
2023-01-20
期刊：
影响因子：
5.8
通讯作者：
Pan, Ping-Yue
中科院分区：
文献类型：
--
作者：
Saenz, Jacqueline;Yao, Oscar;Khezerlou, Elnaz;Aggarwal, Meha;Zhou, Xiaofeng;Barker, David J.;DiCicco-Bloom, Emanuel;Pan, Ping-Yue
可卡因通过抑制质膜多巴胺转运蛋白(DAT)功能和改变其表面表达而发挥作用。可卡因调节DAT贩运的确切方式和机制，特别是在神经元过程中，人们仍然知之甚少。在这项研究中，我们设计并验证了DAT-pHluorin用于研究DAT在培养的小鼠中脑神经元突起中的定位及其动态转运。我们证明，与神经元胞体和树突不同，神经元胞体和树突中包含大多数DAT在弱酸性的细胞内隔室，而轴突和外周的DAT主要(75%)定位于质膜，而大的静脉曲张在酸性的细胞内结构中包含丰富的细胞内DAT。我们还证明，可卡因暴露导致突触素1敏感的DAT内化过程，随后膜重新插入，持续数天。因此，我们的研究揭示了以前未知的可卡因调节神经元突起中DAT运输的动力学和分子调控。DAT-pHluorin用于研究多巴胺转运蛋白转运DAT主要定位于轴突的质膜，可卡因诱导DAT瞬间内化，然后重新插入突触素1调节可卡因诱导的DAT转运神经科学；生物技术；细胞生物学
Cocaine acts by inhibiting plasma membrane dopamine transporter (DAT) function and altering its surface expression. The precise manner and mechanism by which cocaine regulates DAT trafficking, especially at neuronal processes, are poorly understood. In this study, we engineered and validated the use of DAT-pHluorin for studying DAT localization and its dynamic trafficking at neuronal processes of cultured mouse midbrain neurons. We demonstrate that unlike neuronal soma and dendrites, which contain a majority of the DATs in weakly acidic intracellular compartments, axonal DATs at both shafts and boutons are primarily (75%) localized to the plasma membrane, whereas large varicosities contain abundant intracellular DAT within acidic intracellular structures. We also demonstrate that cocaine exposure leads to a Synaptojanin1-sensitive DAT internalization process followed by membrane reinsertion that lasts for days. Thus, our study reveals the previously unknown dynamics and molecular regulation for cocaine-regulated DAT trafficking in neuronal processes. DAT-pHluorin is engineered for studying dopamine transporter trafficking DAT is predominantly localized to the plasma membrane in the axons Cocaine induces a transient internalization of DAT followed by reinsertion Synaptojanin1 regulates the cocaine-induced DAT trafficking Neuroscience; Biotechnology; Cell biology
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GLUT4 动员支持活跃突触的能量需求。
影响因子：
16.2
作者：
Ashrafi G;Wu Z;Farrell RJ;Ryan TA
通讯作者：
Ryan TA
DOI：
10.1083/jcb.150.3.589
发表时间：
2000-08-07
期刊：
The Journal of cell biology
影响因子：
--
作者：
Harris TW;Hartwieg E;Horvitz HR;Jorgensen EM
通讯作者：
Jorgensen EM
影响因子：
4.8
作者：
deHeuvel, E;Bell, AW;McPherson, PS
通讯作者：
McPherson, PS
影响因子：
5.3
作者：
Bagalkot, Tarique R.;Block, Ethan R.;Sorkin, Alexander
通讯作者：
Sorkin, Alexander
影响因子：
4.7
作者：
Boudanova, Ekaterina;Navaroli, Deanna A.;Melikian, Haley E.
通讯作者：
Melikian, Haley E.