Department of Anatomy, Histology, and Neuroscience, Medical School, Universidad Autónoma de Madrid (UAM), Madrid, 28029, Spain.
J Comp Neurol. 2013 Sep 1;521(13):2927-46. doi: 10.1002/cne.23323.
Muscarinic modulation of mesolimbic dopaminergic neurons in the ventral tegmental area (VTA) plays an important role in reward, potentially mediated through the M5 muscarinic acetylcholine receptor (M5R). However, the key sites for M5R-mediated control of dopamine neurons within this region are still unknown. To address this question we examined the electron microscopic immunocytochemical localization of antipeptide antisera against M5R and the plasmalemmal dopamine transporter (DAT) in single sections through the rat VTA. M5R was located mainly to VTA somatodendritic profiles (71%; n = 627), at least one-third (33.2%; n = 208) of which also contained DAT. The M5R immunoreactivity was distributed along cytoplasmic tubulovesicular endomembrane systems in somata and large dendrites, but was more often located at plasmalemmal sites in small dendrites, the majority of which did not express DAT. The M5R-immunoreactive dendrites received a balanced input from unlabeled terminals forming either asymmetric or symmetric synapses. Compared with dendrites, M5R was less often seen in axon terminals, comprising only 10.8% (n = 102) of the total M5R-labeled profiles. These terminals were usually presynaptic to unlabeled dendrites, suggesting that M5R activation can indirectly modulate non-DAT-containing dendrites through presynaptic mechanisms. Our results provide the first ultrastructural evidence that in the VTA, M5R has a subcellular location conducive to major involvement in postsynaptic signaling in many dendrites, only some of which express DAT. These findings suggest that cognitive and rewarding effects ascribed to muscarinic activation in the VTA can primarily be credited to M5R activation at postsynaptic plasma membranes distinct from dopamine transport.
毒蕈碱调节腹侧被盖区(VTA)中的中脑边缘多巴胺能神经元在奖励中起重要作用,可能通过 M5 毒蕈碱乙酰胆碱受体(M5R)介导。然而,该区域内 M5R 介导的多巴胺神经元控制的关键部位仍然未知。为了解决这个问题,我们检查了抗 M5R 肽抗血清和质膜多巴胺转运体(DAT)在大鼠 VTA 单个切片中的电子显微镜免疫细胞化学定位。M5R 主要位于 VTA 体树突突形轮廓(71%;n = 627),其中至少有三分之一(33.2%;n = 208)也含有 DAT。M5R 免疫反应性沿体细胞和大树突的细胞质管状小泡内吞膜系统分布,但在小树枝突的质膜部位更常见,其中大多数不表达 DAT。M5R 免疫反应性树突接受形成不对称或对称突触的未标记末端的平衡输入。与树突相比,M5R 在轴突末端较少见,仅占总 M5R 标记轮廓的 10.8%(n = 102)。这些末端通常是未标记树突的突触前,表明 M5R 激活可以通过突触前机制间接调节非 DAT 包含的树突。我们的研究结果首次提供了超微结构证据,表明在 VTA 中,M5R 具有亚细胞位置,有利于在许多树突中主要参与突触后信号转导,其中只有一些表达 DAT。这些发现表明,归因于 VTA 中毒蕈碱激活的认知和奖励作用主要可以归因于与多巴胺转运不同的突触后质膜上的 M5R 激活。