Ola1对小鼠早期胚胎体外发育的影响

doi:10.16431/j.cnki.1671-7236.2024.11.022

摘要/Abstract

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Ola1基因，检测Ola1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Ola1基因后，小鼠胚胎中Ola1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Ola1基因表达。敲低Ola1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Ola1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Ola1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。

关键词: Ola1; 小鼠; 胚胎发育; DNA损伤; 细胞凋亡

Abstract: 【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.Ola1 gene was knocked down using electric siRNA interference technology.The interference efficiency of Ola1 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 Ola1 gene in mouse embryos was extremely significantly decreased after the knockdown of Ola1 gene (P＜0.01).Successful knockdown of Ola1 gene in mouse embryos.After knocking down Ola1 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),the γ-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 Ola1 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 Ola1 gene knockdown (P＜0.01).The expressions of pro-apoptotic genes,B-lymphocytoma-2-associated X protein (Bax) and Caspase3 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 embryos,thus affected the development potential of early embryos.

Key words: Ola1; mice; embryonic development; DNA damage; apoptosis

中图分类号:

S852.1

罗安凤, 张玉清, 王昕昕, 杨彩侠, 解迪. Ola1对小鼠早期胚胎体外发育的影响[J]. 中国畜牧兽医, 2024, 51(11): 4871-4879.

LUO Anfeng, ZHANG Yuqing, WANG Xinxin, YANG Caixia, XIE Di. Effects of Ola1 on Early Embryonic Development of Mouse in vitro[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(11): 4871-4879.

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