摘要
旨在探究Ezrin对小鼠卵母细胞成熟、受精及早期胚胎发育的影响,揭示其可能作用机制。分别向小鼠GV期卵母细胞和原核期胚胎注射Ezrin的siRNA(si-Ez)或阴性对照siRNA(si-NC),观察对小鼠卵母细胞成熟、受精和早期胚胎发育的影响,并分别通过扫描电镜和免疫荧光染色的方法观察GV期注射si-Ez对小鼠成熟卵母细胞表面微绒毛生长、纺锤体定位和皮质颗粒迁移的影响。结果显示,GV期敲低Ezrin可显著降低小鼠卵母细胞成熟率,极显著降低受精后早期胚胎的囊胚率,虽可降低成熟卵母细胞的受精率,但差异不显著;在原核期敲低Ezrin,可极显著降低小鼠囊胚细胞数,桑椹胚存活率和囊胚率虽有下降趋势,但与对照组无显著差异;扫描电镜观察发现,敲低小鼠GV期卵母细胞Ezrin后显著降低了MⅡ期卵母细胞表面微绒毛的长度和密度;免疫荧光结果显示,敲低小鼠GV期卵母细胞Ezrin影响了纺锤体和皮质颗粒的迁移。结果表明,Ezrin通过影响微绒毛形成、纺锤体定位和皮质颗粒的正常迁移阻滞了小鼠卵母细胞成熟,进而影响了受精和早期胚胎发育。
The aim of this study was to investigate the effect of Ezrin on the maturation of mouse oocytes,fertilization,and early embryo development,as well as to reveal its mechanisms.Mouse germinal vesicle(GV)stage oocytes and pronuclear-stage embryos were injected with siRNA targeting Ezrin(si-Ez)or a negative control siRNA(si-NC),then the rates of oocyte maturation,fertilization,and early embryo development were calculated,the length and density of microvilli on the surface of the MⅡstage oocytes were detected with scanning electron microscopy,the spindle localization and cortical granule migration were observed using immunofluorescence staining.The results showed that knocking down Ezrin during the GV stage significantly reduced the maturation rate of mouse oocytes and extremely significantly lowered the developmental rate of early-stage embryos after fertilization.While it decreased the fertilization rate of mature oocytes,the difference was not significant.Knocking down Ezrin during the pronuclear stage extremely significantly reduced the total cell number of mouse blastocysts.Although there was a downward trend in the rates of morula and blastocyst survival,there was no significant difference compared to the control group.Scanning electron microscopy revealed that knocking down Ezrin in GV stage mouse oocytes significantly decreased the length and density of microvilli on the surface of MⅡstage oocytes.Immunofluorescence results showed that knocking down Ezrin in GV stage mouse oocytes affected the migration of the spindle and cortical granules.These results suggest that Ezrin,through its influence on microvilli formation,spindle localization,and cortical granule migration,blocks the maturation of mouse oocytes,consequently affecting fertilization and early embryo development.
作者
谢辉辉
童彬彬
李巧丹
潘富强
张博超
廖洪艳
贾雨珂
崔剑坤
李运生
刘亚
XIE Huihui;TONG Binbin;LI Qiaodan;PAN Fuqiang;ZHANG Bochao;LIAO Hongyan;JIA Yuke;CUI Jiankun;LI Yunsheng;LIU Ya(College of Animal Science and Technology,Anhui Agricultural University,Hefei 230036,China;Feidong County Animal Health Supervision Institute,Hefei 231600,China;Laboratory Animal Center,University of Science and Technology of China,Hefei 230026,China)
出处
《华北农学报》
CSCD
北大核心
2024年第4期231-238,共8页
Acta Agriculturae Boreali-Sinica
基金
安徽省科技重大专项(202103b06020023)
安徽省重大基础研究项目(2023z04020003)。
作者简介
谢辉辉(1998-),男,安徽阜阳人,硕士,主要从事动物生殖调控研究。谢辉辉、童彬彬为同等贡献作者。;通信作者:刘亚(1972-),男,安徽亳州人,副教授,博士,主要从事动物生殖调控研究。
