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Cumulus-specific genes are transcriptionally silent following somatic cell nuclear transfer in a mouse model

机译:在小鼠模型中体细胞核转移后积云特异性基因转录沉默

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摘要

This study investigated whether four cumulus-specific genes: follicular stimulating hormone receptor (FSHr), hyaluronan synthase 2 (Has2), prostaglandin synthase 2 (Ptgs2) and steroidogenic acute regulator protein (Star), were correctly reprogrammed to be transcriptionally silent following somatic cell nuclear transfer (SCNT) in a murine model. Cumulus cells of C57×CBA F1 female mouse were injected into enucleated oocytes, followed by activation in 10 µmol/L strontium chloride for 5 h and subsequent in vitro culture up to the blastocyst stage. Expression of cumulus-specific genes in SCNT-derived embryos at 2-cell, 4-cell and day 4.5 blastocyst stages was compared with corresponding in vivo fertilized embryos by real-time PCR. It was demonstrated that immediately after the first cell cycle, SCNT-derived 2-cell stage embryos did not express all four cumulus-specific genes, which continually remained silent at the 4-cell and blastocyst stages. It is therefore concluded that all four cumulus-specific genes were correctly reprogrammed to be silent following nuclear transfer with cumulus donor cells in the mouse model. This would imply that the poor preimplantation developmental competence of SCNT embryos derived from cumulus cells is due to incomplete reprogramming of other embryonic genes, rather than cumulus-specific genes.
机译:这项研究调查了四个卵丘特异性基因:卵泡刺激素受体(FSHr),透明质酸合酶2(Has2),前列腺素合酶2(Ptgs2)和类固醇生成的急性调节蛋白(Star)是否被正确地重新编程为在体细胞后转录沉默鼠模型中的核转移(SCNT)。将C57×CBA F1雌性小鼠的卵细胞注入去核卵母细胞,然后在10 µmol / L氯化锶中活化5 h,然后进行体外培养直至胚泡期。通过实时PCR,比较了2细胞,4细胞和4.5天囊胚阶段SCNT衍生胚胎中的卵丘特异性基因的表达与相应的体内受精胚胎的表达。已经证明,在第一个细胞周期后,SCNT衍生的2细胞期胚胎并不表达所有四个积云特异性基因,这些基因在4细胞和胚泡期一直保持沉默。因此得出的结论是,在小鼠模型中,用积云供体细胞进行核转移后,所有四个积云特异性基因均正确地重新编程为沉默。这意味着来自积云细胞的SCNT胚胎植入前发育能力差是由于其他胚胎基因而不是积云特异性基因的不完全重编程。

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