Inhibitory neuron-specific Cre-dependent red fluorescent labeling using VGAT BAC-based transgenic mouse lines with identified transgene integration sites

Ryosuke Kaneko*, Yusuke Takatsuru, Ayako Morita, Izuki Amano, Asahi Haijima, Itaru Imayoshi, Nobuaki Tamamaki, Noriyuki Koibuchi, Masahiko Watanabe, Yuchio Yanagawa

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)373-396
Number of pages24
JournalJournal of Comparative Neurology
Issue number3
Publication statusPublished - 2018 Feb 15
Externally publishedYes


  • Cre
  • RRID: AB_10000343
  • RRID: AB_10000347
  • RRID: AB_1587367
  • RRID: AB_2179313
  • RRID: AB_2255365
  • RRID: AB_2571622
  • RRID: AB_2571847
  • RRID: AB_2631185
  • RRID: AB_442102
  • RRID: AB_477652
  • RRID: AB_88410
  • X-chromosome inactivation
  • inhibitory neuron
  • reporter mouse
  • tdTomato
  • transgene integration site

ASJC Scopus subject areas

  • Neuroscience(all)


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