Institute of Pharmacology and Experimental Toxicology, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia.
Institute of Pathophysiology, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia.
Int J Mol Sci. 2024 May 9;25(10):5135. doi: 10.3390/ijms25105135.
Astrocytes actively participate in neurotransmitter homeostasis by bidirectional communication with neuronal cells, a concept named the tripartite synapse, yet their role in dopamine (DA) homeostasis remains understudied. In the present study, we investigated the kinetic and molecular mechanisms of DA transport in cultured striatal astrocytes of adult rats. Kinetic uptake experiments were performed using radiolabeled [H]-DA, whereas mRNA expression of the dopamine, norepinephrine, organic cation and plasma membrane monoamine transporters (DAT, NET, OCTs and PMAT) and DA receptors D1 and D2 was determined by qPCR. Additionally, astrocyte cultures were subjected to a 24 h treatment with the DA receptor agonist apomorphine, the DA receptor antagonist haloperidol and the DA precursor L-DOPA. [H]-DA uptake exhibited temperature, concentration and sodium dependence, with potent inhibition by desipramine, nortriptyline and decynium-22, suggesting the involvement of multiple transporters. qPCR revealed prominent mRNA expression of the NET, the PMAT and OCT1, alongside lower levels of mRNA for OCT2, OCT3 and the DAT. Notably, apomorphine significantly altered NET, PMAT and D1 mRNA expression, while haloperidol and L-DOPA had a modest impact. Our findings demonstrate that striatal astrocytes aid in DA clearance by multiple transporters, which are influenced by dopaminergic drugs. Our study enhances the understanding of regional DA uptake, paving the way for targeted therapeutic interventions in dopaminergic disorders.
星形胶质细胞通过与神经元细胞的双向通讯积极参与神经递质的动态平衡,这一概念被称为三突触,然而它们在多巴胺(DA)动态平衡中的作用仍未得到充分研究。在本研究中,我们研究了成年大鼠纹状体星形胶质细胞中 DA 转运的动力学和分子机制。通过放射性标记 [H]-DA 进行动力学摄取实验,而多巴胺、去甲肾上腺素、有机阳离子和质膜单胺转运体(DAT、NET、OCTs 和 PMAT)和 DA 受体 D1 和 D2 的 mRNA 表达则通过 qPCR 确定。此外,星形胶质细胞培养物接受了 24 小时的多巴胺受体激动剂阿扑吗啡、多巴胺受体拮抗剂氟哌啶醇和多巴胺前体 L-DOPA 处理。[H]-DA 摄取表现出温度、浓度和钠离子依赖性,并用去甲丙咪嗪、去甲替林和去癸胺-22 进行有力抑制,表明涉及多种转运体。qPCR 显示出 NET、PMAT 和 OCT1 的显著 mRNA 表达,同时 OCT2、OCT3 和 DAT 的 mRNA 表达水平较低。值得注意的是,阿扑吗啡显著改变了 NET、PMAT 和 D1 的 mRNA 表达,而氟哌啶醇和 L-DOPA 的影响较小。我们的发现表明,纹状体星形胶质细胞通过多种转运体有助于 DA 的清除,而这些转运体受多巴胺能药物的影响。我们的研究增强了对区域 DA 摄取的理解,为多巴胺能障碍的靶向治疗干预铺平了道路。
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