Ola1对小鼠早期胚胎体外发育的影响  

Effects of Ola1 on Early Embryonic Development of Mouse in vitro

作　　者：罗安凤 张玉清[1] 王昕昕 杨彩侠 解迪 LUO Anfeng;ZHANG Yuqing;WANG Xinxin;YANG Caixia;XIE Di(College of Animal Science and Technology,Yangtze University,Jingzhou 434025,China;Department of Reproductive Medicine,Chinese People’s Liberation Army Central Theater Command General Hospital,Wuhan 430060,China)

机构地区：[1]长江大学动物科学技术学院,荆州434025 [2]中国人民解放军中部战区总医院生殖医学科,武汉430060

出　　处：《中国畜牧兽医》2024年第11期4871-4879,共9页China Animal Husbandry & Veterinary Medicine

基　　金：湖北省自然科学基金面上项目(2023AFB979)。

摘　　要：【目的】探究Obg样ATP酶1(Obg-like ATPase 1,Ola1)对小鼠早期胚胎发育的影响。【方法】采用体外受精技术获取小鼠合子并进行胚胎体外发育培养,利用电转siRNA干扰技术敲低Ola 1基因,检测Ola 1基因干扰效率,并统计卵裂率和囊胚率;通过免疫荧光和TUNEL法检测胚胎的DNA损伤情况和早期凋亡情况;利用实时荧光定量PCR检测胚胎多能性和凋亡相关基因的转录水平。【结果】与对照组相比,敲低Ola 1基因后,小鼠胚胎中Ola 1基因相对表达量极显著降低(P<0.01),成功实现小鼠胚胎中敲低Ola 1基因表达。敲低Ola 1基因后,早期胚胎卵裂率与对照组相比无显著差异(P>0.05),但囊胚率极显著低于对照组(P<0.01),γ-H2A.X荧光强度和囊胚凋亡率极显著高于对照组(P<0.01),表明敲低Ola 1基因引起早期胚胎体外发育受阻。实时荧光定量PCR结果显示,与对照组相比,敲低Ola 1基因后,早期胚胎的多能性相关基因性别决定区Y框蛋白2(SOX2)和多潜能性基因(Nanog)表达量极显著降低(P<0.01),促凋亡基因B淋巴细胞瘤-2相关X蛋白(Bax)、胱天蛋白酶3(Caspase3)表达量显著或极显著上调(P<0.05;P<0.01),抗凋亡基因B淋巴细胞瘤-2(Bcl-2)表达量极显著下调(P<0.05)。【结论】Ola1可能通过调控早期胚胎的多能性相关基因表达,参与DNA损伤和早期胚胎凋亡过程,从而影响早期胚胎发育潜力。【Objective】The objective of this experiment was to explore the effects of Obg-like ATPase 1(Ola1)on early embryonic development in mouse.【Method】The mouse zygotes were obtained through in vitro fertilization,followed by the culture of embryos in vitro.Ola 1 gene was knocked down using electric siRNA interference technology.The interference efficiency of Ola 1 gene was measured,and the cleavage rate and blastocyst rate were calculated.DNA damage and early apoptosis of embryos were detected by immunofluorescence and TUNEL.Real-time quantitative PCR was used to detect the transcriptional levels of genes related to pluripotency and apoptosis in embryos.【Result】Compared with control group,the relative expression of Ola 1 gene in mouse embryos was extremely significantly decreased after the knockdown of Ola 1 gene(P<0.01).Successful knockdown of Ola 1 gene in mouse embryos.After knocking down Ola 1 gene,there was no significant difference in the early embryo cleavage rate compared with control group(P>0.05),but the blastocyst rate was extremely significantly lower than that of control group(P<0.01),theγ-H2A.X fluorescence intensity and blastocyst apoptosis rate were extremely significantly higher than that of control group(P<0.01).These results indicated that knockdown of Ola 1 gene caused early embryo development to be hindered in vitro.Real-time quantitative PCR results showed that,compared with control group,the expression of pluripotency related genes,sex determining region Y box protein 2(SOX2)and Nanog in early embryos were extremely significantly decreased after Ola 1 gene knockdown(P<0.01).The expressions of pro-apoptotic genes,B-lymphocytoma-2-associated X protein(Bax)and Caspase 3 were significantly or extremely significantly up-regulated(P<0.05 or P<0.01).The expression of anti-apoptotic gene,B-lymphoblastoma-2(Bcl-2)was significantly down-regulated(P<0.05).【Conclusion】Ola1 might be involved in DNA damage and early embryo apoptosis by regulating the expression of pluripotent genes in early e

关 键 词：Ola1 小鼠 胚胎发育 DNA损伤 细胞凋亡 

分 类 号：S852.1[农业科学—基础兽医学]