Genetic reprogramming of transcription factor ap-2gamma in bovine somatic cell nuclear transfer preimplantation embryos and placentomes.

Kenneth I Aston, Gugan-Peng Li, Brady A Hicks, Quinton A Winger, Kenneth L White
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引用次数: 7

Abstract

Bovine somatic cell nuclear transfer (SCNT) efficiency remains very low despite a tremendous amount of research devoted to its improvement over the past decade. Frequent early and mid-gestational losses are commonly accompanied by placental abnormalities. A transcription factor, activating protein AP-2gamma, has been shown to be necessary for proper placental development in the mouse. We first evaluated the expression of the gene coding for AP-2gamma (Tfap2c) in several bovine fibroblast donor cell lines and found it was not expressed. Subsequently we determined the expression profile of Tfap2c in oocytes and various stages of preimplantation in vitro fertilized (IVF) embryos. Tfap2c was undetectable in oocytes and early embryos, and was detectable at relatively high levels in morula and blastocyst IVF embryos. The lack of expression in oocytes and donor cells means Tfap2c must be induced in the zygote at the morula stage in properly reprogrammed embryos. SCNT embryos expressed Tfap2c at the eight-cell stage, 2 days earlier than control embryos. Control embryos first expressed Tfap2c at the morula stage, and at this stage Tfap2c was significantly lower in the SCNT embryos. No differences in expression were detected at the blastocyst stage. To determine whether Tfap2c was properly reprogrammed in the placenta of SCNT pregnancies, we evaluated its expression in cotyledons and caruncles of SCNT and control pregnancies between days 55 and 90 gestation. Expression of Tfap2c in caruncles significantly increased between days 55 and 90, while expression in cotyledons was relatively consistent over that same period. Expression levels in SCNT tissues were not different from controls. This data indicates Tfap2c expression is altered in early preimplantation SCNT embryos, which may have developmental consequences resulting from genes influenced by Tfap2c, but expression was not different at the blastocyst stage and in placentomes.

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转录因子ap-2 γ在牛体细胞核移植胚胎和植入胚胎中的遗传重编程。
牛体细胞核移植(SCNT)的效率仍然很低,尽管大量的研究致力于其改进在过去的十年。频繁的早期和中期妊娠损失通常伴随着胎盘异常。一种转录因子,激活蛋白ap -2 γ,已被证明是小鼠胎盘正常发育所必需的。我们首先在几种牛成纤维细胞供体细胞系中评估了AP-2gamma (Tfap2c)编码基因的表达,发现它不表达。随后,我们测定了Tfap2c在卵母细胞和体外受精(IVF)胚胎着床前不同阶段的表达谱。在卵母细胞和早期胚胎中检测不到Tfap2c,而在桑葚胚和囊胚IVF胚胎中检测到相对较高的水平。在卵母细胞和供体细胞中缺乏表达意味着Tfap2c必须在经过适当重编程的胚胎的桑葚胚期的受精卵中被诱导。SCNT胚胎在8细胞期表达Tfap2c,比对照胚胎早2天。对照胚胎在桑葚胚期首次表达Tfap2c, SCNT胚胎在这一阶段的Tfap2c水平明显较低。在囊胚期未检测到表达差异。为了确定Tfap2c是否在SCNT妊娠胎盘中被正确重编程,我们评估了其在SCNT子叶和结节中的表达,并在妊娠55天至90天的对照妊娠中进行了评估。第55 ~ 90天,Tfap2c在结节中的表达显著增加,而在子叶中的表达在同一时期相对稳定。SCNT组织中的表达水平与对照组没有差异。这些数据表明,Tfap2c表达在早期植入前SCNT胚胎中发生改变,这可能与受Tfap2c影响的基因的发育后果有关,但在囊胚期和胎盘期的表达没有差异。
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