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首页> 外文期刊>Zoological Science >Inhibition of MEK and GSK3 Supports ES Cell-like Domed Colony Formation from Avian and Reptile Embryos
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Inhibition of MEK and GSK3 Supports ES Cell-like Domed Colony Formation from Avian and Reptile Embryos

机译：MEK和GSK3的抑制支持从禽类和爬行类胚胎中形成ES细胞样圆顶集落形成

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As amniotes diversified, mammals may have modified mechanisms of cellular pluripotency along with the acquisition of a placenta. What then defined pluripotent states in the ancestral amniotes? To study the evolutionary background of pluripotency in amniotes, we tested the effects of extracellular effectors on primary culture cells from avian and reptile embryos in serum-free medium. When treated with a combination of a MEK inhibitor and a GSK3 inhibitor (2i condition), chicken early embryos formed domed colonies (DCs), which were morphologically indistinguishable from the colonies formed by mouse and rat naive embryonic stem cells. However, no DCs formed when cells from further-developed embryos were cultured in the 2i condition, indicating that there is a clear boundary of DC-forming ability at around the stage of primitive streak elongation. Quail embryos at the blastoderm and cleavage stages also formed DCs in the 2i condition, which is consistent with the notion that the appearance of DCs corresponds with the presence of pluripotent cells in embryos. Gecko blastoderms also formed DCs in the 2i condition, but gastrulas did not. ERK activation by bFGF caused an effect opposite to that of the 2i condition, namely, it dispersed colonies of cells even from early embryos in all species examined. These results suggest that the regulation of pluripotency by FGF/ERK signaling may date back at least to the common ancestor of mammals, birds, and reptiles. However, gene expression analysis indicated the possibility that mammalian pluripotency transcription factors function differently in non-mammalian amniotes.
机译：随着羊膜动物的多样化，哺乳动物可能具有改变的细胞多能性机制以及获得胎盘。那么在祖先的羊膜动物中定义了什么多能态呢？为了研究羊膜细胞多能性的进化背景，我们测试了细胞外效应物对无血清培养基中禽类和爬行类胚胎原代培养细胞的影响。当用MEK抑制剂和GSK3抑制剂（2i条件）联合治疗时，鸡早期胚胎形成圆顶菌落（DCs），在形态上与小鼠和大鼠幼稚胚胎干细胞形成的菌落没有区别。然而，当在2i条件下培养来自进一步发育的胚胎的细胞时，没有形成DC，这表明在原始条纹延伸阶段附近，DC形成能力存在明确的边界。在胚盘和卵裂期的鹌鹑胚胎也在2i条件下形成DC，这与DC的出现与胚胎中多能细胞的存在相对应的观点是一致的。壁虎胚盘在2i条件下也形成了DC，但长颈鹿却没有。 bFGF激活ERK所产生的作用与2i条件相反，即，它甚至分散了所有受检物种中早期胚胎的细胞集落。这些结果表明，FGF / ERK信号传导对多能性的调节至少可以追溯到哺乳动物，鸟类和爬行动物的共同祖先。但是，基因表达分析表明，哺乳动物多能性转录因子在非哺乳动物羊膜中的功能可能不同。

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