Oct-4 expression in pluripotent cells of the rhesus monkey
Oct-4 expression in pluripotent cells of the rhesus monkey
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DOI：
10.1095/biolreprod.103.019455
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发表时间：
2003-12-01
影响因子：
3.6
通讯作者：
Wolf, DP
中科院分区：
文献类型：
--
作者：
Mitalipov, SM;Kuo, HC;Wolf, DP
POU（Pit-Oct-Unc）结构域转录因子 Oct-4 已成为小鼠多能性的有用标记。它仅存在于胚胎基因组激活后的小鼠植入前阶段胚胎中，并且是小鼠胚胎干（ES）细胞的特征，并且在敲除小鼠中它的缺失阻止了囊胚中内细胞团（ICM）的形成。 Oct-4 的表达也与灵长类细胞的多能性相关。在这里，我们对胞浆内单精子注射产生的恒河猴植入前胚胎以及暴露于体外分化条件之前和之后的 ES 细胞中的 Oct-4 表达进行了系统研究。我们还评估了 Oct-4 的表达，作为监测体细胞核移植后重编程程度的一种手段。 Oct-4 通过逆转录聚合酶链反应和单克隆抗体免疫细胞化学进行检测。猴原核阶段的受精卵和分裂至 8 细胞阶段的胚胎均未检测到 Oct-4。 Oct-4的核染色首先在16细胞阶段变得明显，并且在桑葚胚和致密桑葚胚阶段观察到强信号。猴早期囊胚的 ICM 和滋养外胚层细胞核均为 Oct-4 阳性。然而，在扩张的囊胚的滋养外胚层细胞中该信号减弱，而在 ICM 核中表达仍然很高。与小鼠相似，孵化的猴子囊胚在 ICM 中显示出强烈的 Oct-4 表达，而在滋养外胚层中没有检测到信号。来自体外产生的囊胚的 ICM 的未分化猴 ES 细胞表达 Oct-4，与其多能性质一致，而 ES 细胞分化与信号丢失相关。因此，与小鼠一样，Oct-4 在猴子中的表达为胚胎基因组激活后的多能性提供了有用的标记。最后，在猴核移植胚胎中观察到的 Oct-4 表达缺失或异常表明核重编程不充分。
The POU (Pit-Oct-Unc)-domain transcription factor, Oct-4, has become a useful marker of pluripotency in the mouse. it is found exclusively in mouse preimplantation-stage embryos after embryonic genome activation and is a characteristic of mouse embryonic stem (ES) cells, and its absence in knockout mice precludes inner cell mass (ICM) formation in blastocysts. Expression of Oct-4 has also been associated with pluripotency in primate cells. Here, we undertook a systematic study of Oct-4 expression in rhesus macaque preimplantation embryos produced by intracytoplasmic sperm injection and in ES cells before and after exposure to differentiating conditions in vitro. We also evaluated Oct-4 expression as a means of monitoring the extent of reprogramming following somatic cell nuclear transfer. Oct-4 was detected by reverse transcription-polymerase chain reaction and immunocytochemistry with a monoclonal antibody. Monkey pronuclear-stage zygotes and cleaving embryos up to the 8-cell stage showed no detectable Oct-4. Nuclear staining for Oct-4 first became obvious at the 16-cell stage, and a strong signal was observed in morula and compact morula stages. Both ICM and trophectodermal cell nuclei of monkey early blastocysts were positive for Oct-4. However, the signal was diminished in trophectodermal cells of expanded blastocysts, whereas expression remained high in ICM nuclei. Similar to the mouse, hatched monkey blastocysts showed strong Oct-4 expression in the ICM, with no detectable signal in the trophectoderm. Undifferentiated monkey ES cells derived from the ICM of in vitro-produced blastocysts expressed Oct-4, consistent with their pluripotent nature, whereas ES cell differentiation was associated with signal loss. Therefore, Oct-4 expression in the monkey, as in the mouse, provides a useful marker for pluripotency after activation of the embryonic genome. Finally, the observed lack or abnormal expression of Oct-4 in monkey nuclear transfer embryos suggests inadequate nuclear reprogramming.