A glycine transporter 2-Cre knock-in mouse line for glycinergic neuron-specific gene manipulation.
A glycine transporter 2-Cre knock-in mouse line for glycinergic neuron-specific gene manipulation.
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DOI：
10.1016/j.ibror.2017.07.002
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发表时间：
2017-12
期刊：
影响因子：
2.6
通讯作者：
Yanagawa Y
中科院分区：
文献类型：
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作者：
Kakizaki T;Sakagami H;Sakimura K;Yanagawa Y
甘氨酸是脑干和脊髓中的一种抑制性神经递质。甘氨酸转运蛋白2 （GLYT2）负责细胞外甘氨酸的摄取。GLYT2在甘氨酸能神经元中特异性表达，因此被用作甘氨酸能神经元的标记物。在这里，我们产生了GLYT2启动子驱动的Cre重组酶（Cre）表达小鼠（GLYT2-Cre敲入小鼠），以开发一种操纵甘氨酸能神经元中基因表达的工具。通过将GLYT2-Cre敲入小鼠与以Cre依赖方式表达β-半乳糖苷酶（β-gal）的Cre报告小鼠系R26R杂交来检测Cre活性。对GLYT2-Cre/R26R双转基因小鼠脑和脊髓进行X-gal染色发现，Cre活性主要分布在脑干、小脑和脊髓。这些区域富含甘氨酸能神经元。此外，我们在GLYT2-Cre/R26R双转基因小鼠大脑中对β-gal进行免疫组织化学结合GLYT2的原位杂交，以确定Cre活性是否特异性定位于甘氨酸能神经元。β-gal蛋白和GLYT2 mrna共定位于小脑高尔基细胞、耳蜗背核、巨细胞网状核、脊髓三叉核、梯形体核和外侧小网膜。这些脑区中超过98%的GLYT2 mrna表达细胞也表达β-gal，而90-98%的β-gal阳性细胞表达GLYT2 mrna。因此，在GLYT2-Cre敲入小鼠中，Cre活性特异性地高保真地定位于甘氨酸能神经元。GLYT2-Cre敲入小鼠系将为研究甘氨酸能神经元和神经传递提供一个有用的工具。
Glycine is an inhibitory neurotransmitter in the brainstem and spinal cord. Glycine transporter 2 (GLYT2) is responsible for the uptake of extracellular glycine. GLYT2 is specifically expressed in glycinergic neurons and thus has been used as a marker of glycinergic neurons. Here, we generated GLYT2 promotor-driven Cre recombinase (Cre)-expressing mice (GLYT2-Cre knock-in mice) to develop a tool for manipulating gene expression in glycinergic neurons. Cre activity was examined by crossing the GLYT2-Cre knock-in mice with a Cre reporter mouse line, R26R, which express β-galactosidase (β-gal) in a Cre-dependent manner. X-gal staining of GLYT2-Cre/R26R double transgenic mouse brains and spinal cords revealed that the Cre activity was primarily distributed in the brainstem, cerebellum, and spinal cord. These areas are rich in glycinergic neurons. Furthermore, we performed immunohistochemistry for β-gal combined with in situ hybridization for GLYT2 in the GLYT2-Cre/R26R double transgenic mouse brains to determine whether Cre activity is specifically localized to glycinergic neurons. The β-gal protein and GLYT2 mRNAs were colocalized in the cerebellar Golgi cells, dorsal cochlear nucleus, gigantocellular reticular nucleus, spinal trigeminal nucleus, nucleus of the trapezoid body, and lateral lemniscus. More than 98% of the GLYT2 mRNA-expressing cells in these brain regions also expressed β-gal, whereas 90–98% of the β-gal-positive cells expressed the GLYT2 mRNAs. Thus, Cre activity is specifically localized to glycinergic neurons with high fidelity in the GLYT2-Cre knock-in mice. The GLYT2-Cre knock-in mouse line will be a useful tool for studying glycinergic neurons and neurotransmission.
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影响因子：
3.6
作者：
Saito K;Kakizaki T;Hayashi R;Nishimaru H;Furukawa T;Nakazato Y;Takamori S;Ebihara S;Uematsu M;Mishina M;Miyazaki J;Yokoyama M;Konishi S;Inoue K;Fukuda A;Fukumoto M;Nakamura K;Obata K;Yanagawa Y
通讯作者：
Yanagawa Y
影响因子：
16.2
作者：
Foster, Edmund;Wildner, Hendrik;Tudeau, Laetitia;Haueter, Sabine;Ralvenius, William T.;Jegen, Monika;Johannssen, Helge;Hoesli, Ladina;Haenraets, Karen;Ghanem, Alexander;Conzelmann, Karl-Klaus;Boesl, Michael;Zeilhofer, Hanns Ulrich
通讯作者：
Zeilhofer, Hanns Ulrich
影响因子：
3.1
作者：
Rahman, Jamilur;Besser, Stefanie;Huelsmann, Swen
通讯作者：
Huelsmann, Swen
DOI：
10.1016/j.bbrc.2004.03.125
发表时间：
2004-05-07
影响因子：
3.1
作者：
Ebihara, S;Yamamoto, T;Yanagawa, Y
通讯作者：
Yanagawa, Y
影响因子：
30.8
作者：
通讯作者：
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