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首页> 外文期刊>Neuroscience: An International Journal under the Editorial Direction of IBRO >Fluorescent labeling of both GABAergic and glycinergic neurons in vesicular GABA transporter (VGAT)-venus transgenic mouse.
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Fluorescent labeling of both GABAergic and glycinergic neurons in vesicular GABA transporter (VGAT)-venus transgenic mouse.

机译：囊泡GABA转运蛋白（VGAT）-金星转基因小鼠中GABA能和甘氨酸能神经元的荧光标记。

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摘要

Inhibitory neurons play important roles in a number of brain functions. They are composed of GABAergic neurons and glycinergic neurons, and vesicular GABA transporter (VGAT) is specifically expressed in these neurons. Since the inhibitory neurons are scattered around in the CNS, it is difficult to identify these cells in living brain preparations. The glutamate decarboxylase (GAD) 67-GFP knock-in mouse has been widely used for the identification of GABAergic neurons, but their GAD67 expression was decreased compared to the wild-type mice. To overcome such a problem and to highlight the function and morphology of inhibitory neurons, we generated four lines of VGAT-Venus transgenic mice (lines #04, #29, #39 and #49) expressing Venus fluorescent protein under the control of mouse VGAT promoter. We found higher expression level of Venus transcripts and proteins as well as brighter fluorescent signal in line #39 mouse brains, compared to brains of other lines examined. By Western blots and spectrofluorometric measurements of forebrain, the line #39 mouse showed stronger GFP immunoreactivity and brighter fluorescent intensity than the GAD67-GFP knock-in mouse. In addition, Venus was present not only in somata, but also in neurites in the line #39 mouse by histological studies. In situ hybridization analysis showed that the expression pattern of Venus in the line #39 mouse was similar to that of endogenous VGAT. Double immunostaining analysis in line #39 mouse showed that Venus-expressing cells are primarily immunoreactive for GABA in cerebral cortex, hippocampus and cerebellar cortex and for GABA or glycine in dorsal cochlear nucleus. These results demonstrate that the VGAT-Venus line #39 mouse should be useful for studies on function and morphology of inhibitory neurons in the CNS.
机译：抑制性神经元在许多脑功能中起重要作用。它们由GABA能神经元和甘氨酸能神经元组成，水泡GABA转运蛋白（VGAT）在这些神经元中特异性表达。由于抑制性神经元散布在CNS中，因此很难在活脑准备中鉴定这些细胞。谷氨酸脱羧酶（GAD）67-GFP敲入小鼠已广泛用于鉴定GABA能神经元，但与野生型小鼠相比，它们的GAD67表达下降。为了克服这个问题并突出抑制神经元的功能和形态，我们在小鼠VGAT的控制下生成了四行表达维纳斯荧光蛋白的VGAT-维纳斯转基因小鼠（＃04，＃29，＃39和＃49行）启动子。我们发现，与其他受检品系的大脑相比，＃39品系小鼠脑中金星转录本和蛋白质的表达水平更高，荧光信号更亮。通过Western印迹和前脑的荧光分光光度测定，与GAD67-GFP敲入小鼠相比，＃39系小鼠表现出更强的GFP免疫反应性和更强的荧光强度。另外，通过组织学研究，金星不仅存在于躯体中，而且还存在于＃39系小鼠的神经突中。原位杂交分析表明，在＃39系小鼠中金星的表达模式与内源性VGAT相似。 39号小鼠的双重免疫染色分析表明，表达维纳斯的细胞主要对大脑皮层，海马和小脑皮质的GABA以及对背侧耳蜗核的GABA或甘氨酸具有免疫反应性。这些结果表明，VGAT-维纳斯线＃39小鼠应该用于研究中枢神经系统抑制神经元的功能和形态。

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