Ms4a6d 基因敲除对雌性小鼠生育力的影响  

Effects of Ms4a6d knockout on fertility in female mice

作　　者：亢恩川 王莹 曾继涛 何畏 KANG Enchuan;WANG Ying;ZENG Jitao;HE Wei(Reproductive Medicine Center,First Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing,400038,China)

机构地区：[1]陆军军医大学(第三军医大学)第一附属医院生殖医学中心,重庆400038

出　　处：《陆军军医大学学报》2024年第10期1115-1122,共8页Journal of Army Medical University

基　　金：重庆市自然科学基金面上项目(CSTB2022NSCQ-MSX0155);国家自然科学基金面上项目(81873860)。

摘　　要：目的探究巨噬细胞表面特异分子跨膜4域亚家族A成员6D(membrane-spanning 4-domains subfamily A member 6D,Ms4a6d)基因敲除对雌性小鼠生育力的影响。方法构建Ms4a6d基因敲除小鼠,以各周龄纯合Ms4a6d基因敲除(Ms4a6d^(-/-))小鼠为实验组;采用qPCR、琼脂糖凝胶法鉴定小鼠基因型;运用HE染色、免疫荧光、ELISA等方法检测血清抗缪勒管激素(anti-müllerian hormone,AMH)和雌性Ms4a6d^(-/-)小鼠卵巢中巨噬细胞数量及各级卵泡构成变化;通过生育力实验比较成年Ms4a6d^(-/-)雌鼠妊娠率及平均产仔数变化。结果与同周龄野生型(Ms4a6d+/+)雌鼠比较,2周龄和4周龄的Ms4a6d^(-/-)雌鼠卵巢组织中巨噬细胞显著减少(P<0.01);8周龄时2组卵巢巨噬细胞差异无统计学意义;8周龄Ms4a6d^(-/-)雌鼠卵巢系数显著降低(P<0.01);8周龄时卵巢组织中原始卵泡、初级卵泡、次级卵泡和窦卵泡数量显著减少(P<0.05),各年龄段Ms4a6d^(-/-)雌鼠血清中AMH显著降低(P<0.05)。成年Ms4a6d^(-/-)雌鼠的平均妊娠率为56%,每胎产仔数为(4.1±1.1)只,均显著低于同龄野生型雌鼠的妊娠率(89%)和每胎产仔数[(6.3±1.2)只]。结论Ms4a6d基因敲除导致青春期前雌鼠卵巢组织中巨噬细胞数量减少,抑制其生长卵泡发育,最终表现为成年雌鼠卵巢储备减少,生育力降低。Objective To investigate the effect of membrane-spanning 4-domains subfamily member 6D(MS4A6D)gene knockout on the fertility of female mice.Methods On the basis of the successful construction of Ms4a6d gene knockout model mice,the homozygous Ms4a6d gene knockout(Ms4a6d^(-/-))mice at an age of different weeks were subjected.Mouse genotype was identified with RT-qPCR and agarose gel electrophoresis.HE staining,immunofluorescence assay and ELISA were used to detect serum level of anti-müllerian hormone(AMH),composition of macrophages and follicles at all levels in the ovaries of Ms4a6d^(-/-)mice,respectively.The fertility changes in pregnant rate and average litter size of female Ms4a6d^(-/-)mice were observed by fertility experiments.Results Compared with the wild-type(Ms4a6d+/+)female mice at the same age,there were less macrophages in the ovarian tissues of the 2-and 4-week-old Ms4a6d^(-/-)female mice(P<0.01),but no significant difference was observed between the 2 types of mice at 8 weeks of age.For the adult Ms4a6d^(-/-)female mice at an age of 8 weeks,the ovarian coefficient was obviously lower(P<0.01),and the numbers of primordial,primary,secondary and antral follicles in the ovarian tissues was statistically lower(P<0.05)when compared with the adult wild-type female mice at the same age.The serum AMH level was notably lower in the Ms4a6d^(-/-)female mice of all ages than the wild-type ones at corresponding ages(P<0.05).The average pregnancy rate was 56%and the litter size was 4.1±1.1 in adult Ms4a6d^(-/-)female mice,which were significantly lower than those of wild-type mice of the same age(89%and 6.3±1.2).Conclusion Ms4a6d gene knockout decreases the number of macrophages in the ovarian tissues of adolescent females,interferes with the development of follicles in the ovarian tissues of adolescent females,and leads to a decrease in ovarian reserve in adult females and a decrease in fertility in adult females.

关 键 词：Ms4a6d 巨噬细胞 卵巢 生育力 

分 类 号：R339.22[医药卫生—人体生理学] R394-33[医药卫生—基础医学] R394.1