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首页> 外文期刊>IBRO Reports >VGAT and VGLUT2 expression in MCH and orexin neurons in double transgenic reporter mice
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VGAT and VGLUT2 expression in MCH and orexin neurons in double transgenic reporter mice

机译：VGAT和VGLUT2在MCH和orexin神经元的双重转基因记者小鼠中的表达

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The neuropeptides orexin and melanin-concentrating hormone (MCH), as well as the neurotransmitters GABA (γ-Aminobutyric acid) and glutamate are chief modulators of the sleep-wake states in the posterior hypothalamus. To investigate co-expression of vesicular GABA transporter (VGAT, a marker of GABA neurons) and the vesicular glutamate transporter-2 (VGLUT2, a marker of glutamate neurons) in orexin and MCH neurons, we generated two transgenic mouse lines. One line selectively expressed the reporter gene EYFP in VGAT+ neurons, whereas the other line expressed reporter gene tdTomato in VGLUT2+ neurons. Co-localization between these genetic reporters and orexin or MCH immunofluorescent tags was determined using 3D computer reconstructions of Z stacks that were acquired using a multiphoton laser confocal microscope. Our results demonstrated that MCH neurons expressed neither VGAT nor VGLUT2, suggesting MCH neurons are a separate cluster of cells from VGAT+ GABAergic neurons and VGLUT2+ glutamatergic neurons. Moreover, most orexin neurons expressed VGLUT2, indicating these neurons are glutamatergic. Our data suggested that in the posterior hypothalamus there are four major distinct groups of neurons: VGAT+, orexin+/VGLUT2+, orexin-/VGLUT2+, and MCH neurons. This study facilitated our understanding of the role of these neurotransmitters and neuropeptides in relation to sleep/wake regulation.
机译：神经肽食欲素和黑色素浓缩激素（MCH）以及神经递质GABA（γ-氨基丁酸）和谷氨酸是下丘脑后部睡眠觉醒状态的主要调节剂。为了研究水泡GABA转运蛋白（VGAT，GABA神经元的标志物）和水泡谷氨酸转运蛋白2（VGLUT2，谷氨酸神经元的标志物）在orexin和MCH神经元中的共表达，我们产生了两个转基因小鼠品系。一系在VGAT +神经元中选择性表达报告基因EYFP，而另一系在VGLUT2 +神经元中表达报告基因tdTomato。这些遗传报告基因与orexin或MCH免疫荧光标签之间的共定位是通过使用多光子激光共聚焦显微镜获得的Z堆栈的3D计算机重建来确定的。我们的结果表明，MCH神经元既不表达VGAT也不表达VGLUT2，表明MCH神经元是来自VGAT + GABA能神经元和VGLUT2 +谷氨酸能神经元的单独细胞簇。此外，大多数食欲素神经元表达VGLUT2，表明这些神经元具有谷氨酸能。我们的数据表明，在下丘脑后部，有四个主要的不同神经元组：VGAT +，orexin + / VGLUT2 +，orexin- / VGLUT2 +和MCH神经元。这项研究促进了我们对这些神经递质和神经肽在睡眠/唤醒调节方面的作用的理解。

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