Tohoku University Graduate School of Agricultural Science, Sendai, Miyagi, Japan.
Division of Behavioral Neuroscience and Psychiatric Disorders, Emory University, Atlanta, GA, USA.
Methods Mol Biol. 2022;2384:139-152. doi: 10.1007/978-1-0716-1759-5_9.
Social and prosocial behaviors, including communication, social bonding, and affiliation, parental behaviors, and empathy are key features of a highly social mammalian species. However, the neuronal mechanism in the brain underlying these behaviors remains unclear because of limited information on the social and prosocial behavioral levels in rodent models generally used in behavioral neuroscience studies.The rodent species, prairie vole (Microtus ochrogaster), is one of the nontraditional animal models with several advantages in experimental science over other rodent models, such as mice or rats. Additionally, it demonstrates characteristics advantageous in the study of social and prosocial behaviors, such as monogamous pair bonding behavior, biparental care, and consoling behavior toward partners stressed by aversive foot shock stimulus. Recent studies of prairie voles have highlighted the importance of oxytocin (OXT) and oxytocin receptor (OXTR)-mediated mechanisms in the regulation of these behaviors.Recently, we established assisted reproductive technologies for prairie voles, and successfully and efficiently generated an OXTR gene knockout (KO) prairie vole using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9 ), a powerful genome editing tool with artificially developed single-strand guide RNAs (sgRNA) and Cas9 endonucleases.Herein, we describe the method for CRISPR /Cas9-mediated generation of OXTR KO prairie vole. This OXTR KO prairie vole can be a valuable tool to understand their unique social and prosocial behaviors and elucidate how the oxytocin system influences or modulates these behaviors in the brain.
社交和亲社会行为，包括交流、社交联系和依附、亲代行为和同理心，是高度社会性哺乳动物物种的关键特征。然而，由于在行为神经科学研究中通常使用的啮齿动物模型的社交和亲社会行为水平的信息有限，大脑中这些行为的神经机制仍不清楚。草原田鼠（Microtus ochrogaster）是一种非传统的动物模型，与其他啮齿动物模型（如老鼠或大鼠）相比，它在实验科学中有几个优势。此外，它表现出有利于研究社交和亲社会行为的特征，例如一夫一妻制的配对结合行为、双亲照顾和对受到厌恶的足部电击刺激的伴侣的安慰行为。最近对草原田鼠的研究强调了催产素（OXT）和催产素受体（OXTR）介导的机制在调节这些行为中的重要性。最近，我们为草原田鼠建立了辅助生殖技术，并成功高效地使用 Clustered Regularly Interspaced Short Palindromic Repeats（CRISPR）/CRISPR-associated protein 9（Cas9），一种具有人工开发的单链引导 RNA（sgRNA）和 Cas9 内切酶的强大基因组编辑工具，生成了 OXTR 基因敲除（KO）草原田鼠。在此，我们描述了使用 CRISPR/Cas9 介导生成 OXTR KO 草原田鼠的方法。这种 OXTR KO 草原田鼠可以成为一种有价值的工具，用于了解它们独特的社交和亲社会行为，并阐明催产素系统如何在大脑中影响或调节这些行为。
