Rahbek-Clemmensen Troels, Lycas Matthew D, Erlendsson Simon, Eriksen Jacob, Apuschkin Mia, Vilhardt Frederik, Jørgensen Trine N, Hansen Freja H, Gether Ulrik
Molecular Neuropharmacology and Genetics Laboratory, Department of Neuroscience Faculty of Health and Medical Sciences, University of Copenhagen, DK-2200, Copenhagen, Denmark.
Lundbeck Foundation Center for Biomembranes in Nanomedicine, Department of Neuroscience, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2200, Copenhagen, Denmark.
Nat Commun. 2017 Sep 29;8(1):740. doi: 10.1038/s41467-017-00790-3.
Dopamine regulates reward, cognition, and locomotor functions. By mediating rapid reuptake of extracellular dopamine, the dopamine transporter is critical for spatiotemporal control of dopaminergic neurotransmission. Here, we use super-resolution imaging to show that the dopamine transporter is dynamically sequestrated into cholesterol-dependent nanodomains in the plasma membrane of presynaptic varicosities and neuronal projections of dopaminergic neurons. Stochastic optical reconstruction microscopy reveals irregular dopamine transporter nanodomains (∼70 nm mean diameter) that were highly sensitive to cholesterol depletion. Live photoactivated localization microscopy shows a similar dopamine transporter membrane organization in live heterologous cells. In neurons, dual-color dSTORM shows that tyrosine hydroxylase and vesicular monoamine transporter-2 are distinctively localized adjacent to, but not overlapping with, the dopamine transporter nanodomains. The molecular organization of the dopamine transporter in nanodomains is reversibly reduced by short-term activation of NMDA-type ionotropic glutamate receptors, implicating dopamine transporter nanodomain distribution as a potential mechanism to modulate dopaminergic neurotransmission in response to excitatory input.The dopamine transporter (DAT) has a crucial role in the regulation of neurotransmission. Here, the authors use super-resolution imaging to show that DAT clusters into cholesterol-dependent membrane regions that are reversibly regulated by ionotropic glutamate receptors activation.
多巴胺调节奖赏、认知和运动功能。多巴胺转运体通过介导细胞外多巴胺的快速重摄取，对多巴胺能神经传递的时空控制至关重要。在此，我们使用超分辨率成像显示，多巴胺转运体在突触前膨体和多巴胺能神经元的神经突的质膜中动态隔离到依赖胆固醇的纳米结构域中。随机光学重建显微镜揭示了不规则的多巴胺转运体纳米结构域（平均直径约70纳米），其对胆固醇耗竭高度敏感。实时光激活定位显微镜显示在活的异源细胞中有类似的多巴胺转运体膜组织。在神经元中，双色直接随机光学重建显微镜显示酪氨酸羟化酶和囊泡单胺转运体2独特地定位在多巴胺转运体纳米结构域附近，但不与之重叠。纳米结构域中多巴胺转运体的分子组织通过NMDA型离子型谷氨酸受体的短期激活而可逆地减少，这表明多巴胺转运体纳米结构域分布作为一种潜在机制，可响应兴奋性输入来调节多巴胺能神经传递。多巴胺转运体（DAT）在神经传递调节中起关键作用。在此，作者使用超分辨率成像显示DAT聚集到依赖胆固醇的膜区域中，这些区域受离子型谷氨酸受体激活的可逆调节。
