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NT小鼠胚胎的发育异常在着床后早期就会出现。

Developmental abnormalities of NT mouse embryos appear early after implantation.

作者信息

Jouneau Alice, Zhou Qi, Camus Anne, Brochard Vincent, Maulny Linda, Collignon Jérôme, Renard Jean-Paul

机构信息

Unité de Biologie du Développement et de la Reproduction, UMR INRA-ENVA, Institut National de la Recherche Agronomique (INRA 78352, France.

出版信息

Development. 2006 Apr;133(8):1597-607. doi: 10.1242/dev.02317.

Abstract

In mammals, cloning by nuclear transfer (NT) into an enucleated oocyte is a very inefficient process, even if it can generate healthy adults. We show that blastocysts derived from embryonic stem (ES) donor cells develop at a high rate, correctly express the pluripotential marker gene Oct4 in ICM cells and display normal growth in vitro. Moreover, the majority of them implant in the uterus of recipient females. We combine embryological studies, gene expression analysis during gastrulation and generation of chimaeric embryos to identify the developmental origin (stage and tissue affected) of NT embryo mortality. The majority died before mid-gestation from defects arising early, either at peri-implantation stages or during the gastrulation period. The first type of defect is a non-cell autonomous defect of the epiblast cells and is rescued by complementation of NT blastocysts with normal ES or ICM cells. The second type of defect affects growth regulation and the shape of the embryo but does not directly impair the initial establishment of the patterning of the embryo. Only chimaeras formed by the aggregation of NT and tetraploid embryos reveal no growth abnormalities at gastrulation. These studies indicate that the trophoblast cell lineage is the primary source of these defects. These embryological studies provide a solid basis for understanding reprogramming errors in NT embryos. In addition, they unveil new aspects of growth regulation while increasing our knowledge on the role of crosstalk between the extra-embryonic and the embryonic regions of the conceptus in the control of growth and morphogenesis.

摘要

在哺乳动物中,即便通过核移植(NT)将细胞核植入去核卵母细胞能够培育出健康的成年个体,这一过程的效率却非常低。我们发现,源自胚胎干细胞(ES)供体细胞的囊胚发育率很高,在植入前胚盘细胞(ICM)中能正确表达多能性标记基因Oct4,并且在体外显示出正常生长。此外,它们中的大多数会植入受体雌性动物的子宫。我们结合胚胎学研究、原肠胚形成过程中的基因表达分析以及嵌合体胚胎的生成,以确定NT胚胎死亡的发育起源(受影响的阶段和组织)。大多数胚胎在妊娠中期之前就因早期出现的缺陷而死亡,这些缺陷要么发生在着床前后阶段,要么发生在原肠胚形成期。第一种缺陷是上胚层细胞的非细胞自主性缺陷,通过用正常的ES或ICM细胞补充NT囊胚可以挽救。第二种缺陷影响胚胎的生长调节和形状,但不会直接损害胚胎模式形成的初始建立。只有由NT胚胎和四倍体胚胎聚集形成的嵌合体在原肠胚形成时没有显示出生长异常。这些研究表明,滋养层细胞谱系是这些缺陷的主要来源。这些胚胎学研究为理解NT胚胎中的重编程错误提供了坚实的基础。此外,它们揭示了生长调节的新方面,同时增加了我们对胚胎外区域与胚胎概念的胚胎区域之间的串扰在生长和形态发生控制中的作用的认识。

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