Striatal Dopamine D2-Muscarinic Acetylcholine M1 Receptor-Receptor Interaction in a Model of Movement Disorders
Striatal Dopamine D2-Muscarinic Acetylcholine M1 Receptor-Receptor Interaction in a Model of Movement Disorders
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DOI：
10.3389/fphar.2020.00194
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发表时间：
2020-03-13
影响因子：
5.6
通讯作者：
Ciruela, Francisco
中科院分区：
文献类型：
--
作者：
Crans, Rene A. J.;Wouters, Elise;Ciruela, Francisco
帕金森病（PD）是一种以运动控制缺陷为特征的神经退行性疾病，与黑质纹状体多巴胺能神经元的丧失有关。在多巴胺能失神经支配的同时，纹状体内乙酰胆碱增加，导致纹状体多巴胺能-胆碱能神经传递失衡。目前，可用的PD药物治疗（例如，原多巴胺能药物）并不能恢复改变的多巴胺能-胆碱能平衡。此外，它最终会引起与胆碱能相关的不良反应。在这里，我们通过评估多巴胺D-2和毒毒碱乙酰胆碱M-1受体（分别为D2R和M1R）的物理和功能相互作用来研究多巴胺能和胆碱能系统之间的相互作用，这两种受体均表达于纹状体中棘神经元。首先，我们通过生化（即共免疫沉淀）和生物物理（即BRET1和NanoBiT (R)）测定，在瞬时转染的HEK293T细胞中提供了D2R-M1R复合物存在的证据。随后，通过双免疫荧光染色和AlphaLISA (R)免疫分析，观察D2R-M1R在小鼠纹状体中的共分布。最后，我们通过行为学研究评估了两种受体之间的功能相互作用，通过实施经典的急性利血平药理动物模型实验性帕金森病。利血平化小鼠给予d2r选择性激动剂（sumanirole）和/或m1r选择性拮抗剂（VU0255035），并评估pd相关行为任务（即运动活动）的变化。重要的是，VU0255035 （10 mg/kg）增强了无效剂量（3 mg/kg）的抗帕金森样作用（即增加运动活动和减少嗜睡）。总之，我们的数据表明存在假定的纹状体D2R/M1R异构体，这可能是治疗PD运动障碍的相关靶点，副作用更少。
Parkinson's disease (PD) is a neurodegenerative disorder characterized by motor control deficits, which is associated with the loss of striatal dopaminergic neurons from the substantia nigra. In parallel to dopaminergic denervation, there is an increase of acetylcholine within the striatum, resulting in a striatal dopaminergic-cholinergic neurotransmission imbalance. Currently, available PD pharmacotherapy (e.g., prodopaminergic drugs) does not reinstate the altered dopaminergic-cholinergic balance. In addition, it can eventually elicit cholinergic-related adverse effects. Here, we investigated the interplay between dopaminergic and cholinergic systems by assessing the physical and functional interaction of dopamine D-2 and muscarinic acetylcholine M-1 receptors (D2R and M1R, respectively), both expressed at striatopallidal medium spiny neurons. First, we provided evidence for the existence of D2R-M1R complexes via biochemical (i.e., co-immunoprecipitation) and biophysical (i.e., BRET1 and NanoBiT (R)) assays, performed in transiently transfected HEK293T cells. Subsequently, a D2R-M1R co-distribution in the mouse striatum was observed through double-immunofluorescence staining and AlphaLISA (R) immunoassay. Finally, we evaluated the functional interplay between both receptors via behavioral studies, by implementing the classical acute reserpine pharmacological animal model of experimental parkinsonism. Reserpinized mice were administered with a D2R-selective agonist (sumanirole) and/or an M1R-selective antagonist (VU0255035), and alterations in PD-related behavioral tasks (i.e., locomotor activity) were evaluated. Importantly, VU0255035 (10 mg/kg) potentiated the antiparkinsonian-like effects (i.e., increased locomotor activity and decreased catalepsy) of an ineffective sumanirole dose (3 mg/kg). Altogether, our data suggest the existence of putative striatal D2R/M1R heteromers, which might be a relevant target to manage PD motor impairments with fewer adverse effects.