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Inhibitory neuron‐specific Cre‐dependent red fluorescent labeling using VGAT BAC‐based transgenic mouse lines with identified transgene integration sites
Author(s) -
Kaneko Ryosuke,
Takatsuru Yusuke,
Morita Ayako,
Amano Izuki,
Haijima Asahi,
Imayoshi Itaru,
Tamamaki Nobuaki,
Koibuchi Noriyuki,
Watanabe Masahiko,
Yanagawa Yuchio
Publication year - 2018
Publication title -
journal of comparative neurology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.855
H-Index - 209
eISSN - 1096-9861
pISSN - 0021-9967
DOI - 10.1002/cne.24343
Subject(s) - biology , transgene , inhibitory postsynaptic potential , genetically modified mouse , fate mapping , microbiology and biotechnology , neuron , neuroscience , genetics , gene , embryonic stem cell
Inhibitory neurons are crucial for shaping and regulating the dynamics of the entire network, and disturbances in these neurons contribute to brain disorders. Despite the recent progress in genetic labeling techniques, the heterogeneity of inhibitory neurons requires the development of highly characterized tools that allow accurate, convenient, and versatile visualization of inhibitory neurons in the mouse brain. Here, we report a novel genetic technique to visualize the vast majority and/or sparse subsets of inhibitory neurons in the mouse brain without using techniques that require advanced skills. We developed several lines of Cre‐dependent tdTomato reporter mice based on the vesicular GABA transporter (VGAT)‐BAC, named VGAT‐stop‐tdTomato mice. The most useful line (line #54) was selected for further analysis based on two characteristics: the inhibitory neuron‐specificity of tdTomato expression and the transgene integration site, which confers efficient breeding and fewer adverse effects resulting from transgene integration‐related genomic disruption. Robust and inhibitory neuron‐specific expression of tdTomato was observed in a wide range of developmental and cellular contexts. By breeding the VGAT‐stop‐tdTomato mouse (line #54) with a novel Cre driver mouse line, Galntl4‐CreER, sparse labeling of inhibitory neurons was achieved following tamoxifen administration. Furthermore, another interesting line (line #58) was generated through the unexpected integration of the transgene into the X‐chromosome and will be used to map X‐chromosome inactivation of inhibitory neurons. Taken together, our studies provide new, well‐characterized tools with which multiple aspects of inhibitory neurons can be studied in the mouse.