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

Abstract

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

CLC Number:

S852.1

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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Effects of Ola1 on Early Embryonic Development of Mouse in vitro

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