CRISPR/Cas9-mediated in vivo gene editing reveals that neuronal 5-HT1A receptors in the dorsal raphe nucleus contribute to body temperature regulation in mice

Nishitani, Naoya; Ohmura, Yu; Nagayasu, Kazuki; Shibui, Norihiro; Kaneko, Shuji; Ohashi, Akiko; Yoshida, Takayuki; Yamanaka, Akihiro; Yoshioka, Mitsuhiro

CRISPR/Cas9-mediated in vivo gene editing reveals that neuronal 5-HT1A receptors in the dorsal raphe nucleus contribute to body temperature regulation in mice

Nishitani, Naoya; Ohmura, Yu; Nagayasu, Kazuki et al.

Brain research, 2019, 1719, 243-252

Permalink : https://hdl.handle.net/2115/79239

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論文情報

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アクセス権	open access
URI	https://hdl.handle.net/2115/79239
タイトル	CRISPR/Cas9-mediated in vivo gene editing reveals that neuronal 5-HT1A receptors in the dorsal raphe nucleus contribute to body temperature regulation in mice
著者	Nishitani, Naoya ; Ohmura, Yu ; Nagayasu, Kazuki ; Shibui, Norihiro ; Kaneko, Shuji ; Ohashi, Akiko ; Yoshida, Takayuki ; Yamanaka, Akihiro ; Yoshioka, Mitsuhiro
著者名 Nishitani, Naoya
e-Rad_Researcher 80597659 著者名 Ohmura, Yu
e-Rad_Researcher 00717902 著者名 Nagayasu, Kazuki
著者名 Shibui, Norihiro
e-Rad_Researcher 60177516 著者名 Kaneko, Shuji
e-Rad_Researcher 00571019 著者名 Ohashi, Akiko
e-Rad_Researcher 60374229 著者名 Yoshida, Takayuki
e-Rad_Researcher 60323292 著者名 Yamanaka, Akihiro
e-Rad_Researcher 40182729 著者名 Yoshioka, Mitsuhiro
言語	英語
キーワード	Genome editing
	Serotonergic
	Rectal temperature
発行日	2019-09-15
出版者	Elsevier
収録物名	Brain research
巻(号)	1719
ページ	243-252
抄録	Serotonin (5-HT) in the central nervous system regulates a variety of biological functions, from the basic homeostatic control to higher brain functions, by acting on fourteen known receptor subtypes. However, it is still usually unclear which receptor subtype is responsible for a specific function due to the lack of highly selective ligands for most of these receptors. Although 5-HT receptor knockout mice are useful, the brain-wide distribution of various receptors makes it difficult to dissect receptor functions in specific and brain regions and cell types. Recent advances in CRISPR/Cas9-mediated in vivo genome editing technology may overcome this problem. In this study, we constructed a viral vector expressing a single guide (sg)RNA targeting Htr1a (sgHtr1a) and Cre recombinase under the control of a neuron-specific promoter. Injection of the viral vector into the dorsal raphe nucleus (DRN) of Cre-dependent Cas9 knock-in mice induced Cre-dependent Cas9 expression mainly in DRN serotonin and GABA neurons. Mismatch cleavage assay and Sanger sequencing showed insertion or deletion formation at the target site. 5-HT1A receptor agonist-induced hypothermia was attenuated and antidepressant effect of a selective serotonin reuptake inhibitor (SSRI) was enhanced by microinjection of the viral vector expressing sgHtr1a into the DRN of Cre-dependent Cas9 knock-in mice. These results suggest that this in vivo CRISPR/Cas9-mediated 5-HT receptor gene knockout strategy provides a reliable and low-cost method for elucidating 5-HT receptor functions in specific cell types and brain regions. Further, we demonstrate that the neuronal 5-HT1A receptor in the DRN regulates body temperature and antidepressant effect of SSRI.
資源タイプ	学術雑誌論文
出版タイプ	Accepted Manuscript
権利情報	Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
権利情報	© 2019. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/
PISSN	0006-8993
関連情報	PMID 31194947
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1016/j.brainres.2019.06.009