Fine visualization of oxytocin receptor expressing neurons via the novel labeling system

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K06467
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 42040:Laboratory animal science-related
• Research Institution: National Center of Neurology and Psychiatry
• Principal Investigator: Inoue U. Yukiko 国立研究開発法人国立精神・神経医療研究センター, 神経研究所 疾病研究第六部, リサーチフェロー (30611777)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: オキシトシン受容体 / ゲノム編集 / ノックインマウス

Outline of Final Research Achievements

Oxytocin, which acts as a neurotransmitter in the brain, has attracted widespread interest because it modulates the function of neurons that express its receptor to facilitatively regulate social behaviors such as nursing behavior and family bonding. Hence, research to better understand the oxytocin neural network is very active now. However, existing antibodies and mouse models are insufficient to accurately visualize the expression of this receptor. In this study, we newly generated multiple knock-in mouse lines by genome editing using fertilized eggs, to detect the sub-cellular localization of oxytocin receptors with unprecedented precision. In addition, we provided domestic and foreign researchers with mouse tools expressing fluorescent reporters and Cre recombinase, which are essential for manipulating oxytocin neural circuits in the brain. By developing these resources, we have contributed to the promotion of oxytocin research on a worldwide level.

Free Research Field

神経科学、ゲノム編集、遺伝子改変マウス

Academic Significance and Societal Importance of the Research Achievements

オキシトシン受容体は高脂溶性の膜タンパク質のため、これを認識する抗体を作製することは難しく、マウス脳組織上で研究に使用できる良好な抗体が存在しなかった。そこで、受容体タンパク質の末端にタグを付加したノックインマウスをゲノム編集により作出し、タグに対する抗体を用いて超解像度顕微鏡解析を行い、受容体の神経細胞内局在を初めて観察することができた。また、オキシトシン神経回路の操作による機能解析に必須の蛍光レポーターやCre組換え酵素発現マウスも作出し、国際的なバイオリソースに寄託することにより、国内外のオキシトシン研究者への提供を容易にし、社会性行動を制御する神経回路網の理解を深めることに貢献した。