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第41卷第4期 南京医科大学学报(自然科学版)
2021年4月 Journal of Nanjing Medical University(Natural Sciences) ·477 ·
·基础医学·
神经损伤诱导蛋白2在卵巢发育中的作用
尚彦星,周 蒙,鄂裘恺,张晓倩,张学森 *
南京医科大学生殖医学国家重点实验室,江苏 南京 211166
[摘 要] 目的:通过基因敲除模型研究神经损伤诱导蛋白 2(Ninjurin2,Ninj2)在小鼠卵巢发育过程的作用。方法:利用
CRISPR/Cas9技术构建Ninj2基因敲除小鼠,通过HE染色、免疫荧光染色、体外受精(in vitro fertilization,IVF)以及脂多糖(lipo⁃
polysaccharide,LPS)暴露对敲除小鼠表型进行分析,研究Ninj2在小鼠卵巢发育中的作用。结果:成功构建Ninj2基因敲除小鼠
模型;小鼠体重及相关生化指标检测结果显示,Ninj2纯合敲除鼠与野生型小鼠相比无显著差异;HE染色显示,Ninj2敲除小鼠
在新生和成年卵巢中各级卵泡发育情况与野生型小鼠相比均无明显差别;IVF结果表明,Ninj2敲除小鼠早期胚胎发育正常;
LPS对Ninj2敲除小鼠卵巢发育没有影响。结论:在正常生理或炎性条件下,敲除Ninj2不影响小鼠卵巢发育。
[关键词] CRISPR/Cas9;Ninj2;卵巢发育
[中图分类号] R321.1 [文献标志码] A [文章编号] 1007⁃4368(2021)04⁃477⁃06
doi:10.7655/NYDXBNS20210401
Effects of nerve injury inducing protein 2 during ovarian development
SHANG Yanxing,ZHOU Meng,E Qiukai,ZHANG Xiaoqian,ZHANG Xuesen *
State Key Laboratory of Reproductive Medicine,Nanjing Medical University,Nanjing 211166,China
[Abstract] Objective:To construct nerve injury⁃inducing protein 2(Ninj2)knockout mice and study its role during mouse ovarian
development. Methods:We generated Ninj2 knockout mice by CRISPR/Cas9 technology,and detected the role of Ninj2 in mouse
ovarian development by HE staining,immunofluorescence,in vitro fertilization(IVF)and lipopolysaccharide(LPS)exposure. Results:
Ninj2 knockout mouse model was successfully constructed. Body weight and detection of related biochemical indicators showed that
there was no significant difference between Ninj2 homozygous knockout mice and wild⁃type mice;HE staining showed that there was
no significant difference in follicular development between Ninj2 knockout mice and wild⁃type controls at all levels in newborn and
adult ovaries;IVF results showed that the early embryos of Ninj2 knockout mice developed normally;LPS had no effect on ovarian
development in Ninj2 knockout mice. Conclusion:Depletion of Ninj2 does not affect the mouse ovarian development in normal
physiological or inflammatory condition.
[Key words] CRISPR/Cas9;Ninj2;ovary development
[J Nanjing Med Univ,2021,41(04):477⁃482]
下丘脑⁃垂体⁃卵巢轴(hypothalamic⁃pituitary⁃ (gonadotropin⁃releasing hormone,GNRH)神经元分泌
ovarian axis,HPOA)是影响雌性生殖能力的重要调 GNRH,调节垂体中促性腺细胞分泌卵泡刺激素
控因素,在生殖功能中发挥主要的调节作用。卵巢 (follicle stimulating hormone,FSH)和促黄体生成素
作为雌性性腺器官,主要功能在于产生卵子和性激 (luteinizing hormone,LH),GNRH的脉冲式释放是促
素,接受下丘脑和垂体激素的调节,同时反馈性调 性腺激素呈周期变化的必需条件,可以确保卵泡的
[1]
[2]
节下丘脑及垂体的功能 。促性腺激素释放激素 正常发育和生殖功能 。许多文章也报道了下丘脑
和垂体激素对排卵和卵巢激素分泌等卵巢功能的
[基金项目] 国家重点研发计划(2018YFC1004203) 调节 [3-4] 。然而,尽管一些研究描述了迷走副交感神
∗ 经和交感神经对卵巢的支配 [5-6] ,但是这些自主神经
通信作者(Corresponding author),E⁃mail:zhang_xuesen@163.com