Keratin 5-Cre-driven excision of nonmuscle myosin IIA in early embryo trophectoderm leads to placenta defects and embryonic lethality.
Keratin 5-Cre-driven excision of nonmuscle myosin IIA in early embryo trophectoderm leads to placenta defects and embryonic lethality.
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DOI：
10.1016/j.ydbio.2013.07.017
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发表时间：
2013-10-01
影响因子：
2.7
通讯作者：
Egelhoff, Thomas T.
中科院分区：
文献类型：
--
作者：
Crish, James;Conti, Mary Anne;Sakai, Takao;Adelstein, Robert S.;Egelhoff, Thomas T.
在最初集中于非肌肉肌球蛋白IIA（NMIIA）在小鼠表皮发育中的作用的研究中，我们发现先前描述的细胞角蛋白5（K5）-Cre基因构建体在早期胚胎发育中表达。携带非肌肉肌球蛋白II重链基因（NMHC IIAflox/flox）的floxed等位基因的小鼠与K5-Cre系杂交。新生幼崽的后代没有表现出孟德尔基因型分布，表明胚胎致死。从胚胎第9.5天到E13.5天的植入后妊娠分析显示发育不良的胚胎和有缺陷的胎盘，迷路表面积和血管血管形成显著减少。这些结果表明，牛K5启动子驱动的Cre-重组酶在产生胎盘的滋养层细胞系细胞中早期是活跃的。为了测试这种可能性，将K5-Cre转基因小鼠与mT/mG报告小鼠杂交，其中GFP表达的激活指示Cre转基因表达。我们观察到激活K5-Cre驱动的GFP表达的外胎盘锥，在胚胎外外胚层，并在滋养层巨细胞在E6.5胚胎。此外，我们观察到GFP的表达在E11.5至E13.5在迷路的胎盘和卵黄囊。在正常胚胎的这些相同类型的细胞中，以及在E13.5卵黄囊和迷路中检测到NMIIA表达。这些研究结果表明，NMHC IIA可能发挥关键作用，在早期滋养层细胞衍生的胎盘锥和胚外外外胚层，以及在卵黄囊和迷路组织，后来形成。我们的研究结果是一致的表型组成NMIIA基因敲除小鼠早期，显示迷宫和卵黄囊特异性缺陷，但我们的研究结果延伸这些观察表明可能NMIIA的作用，以及滋养层谱系。这些结果进一步证明，K5-Cre基因构建体，先前报道在形成的表皮中在大约E12.5开始被激活，也可以广泛地用作在早期胚胎胚外滋养层组织中激活基于cre/lox的基因切除的驱动器。
In studies initially focused on roles of nonmuscle myosin IIA (NMIIA) in the developing mouse epidermis, we have discovered that a previously described cytokeratin 5 (K5)-Cre gene construct is expressed in early embryo development. Mice carrying floxed alleles of the nonmuscle myosin II heavy chain gene (NMHC IIAflox/flox) were crossed with the K5-Cre line. The progeny of newborn pups did not show a Mendelian genotype distribution, suggesting embryonic lethality. Analysis of post-implantation conceptuses from embryonic day (E)9.5 to E13.5 revealed poorly developed embryos and defective placentas, with significantly reduced labyrinth surface area and blood vessel vascularization. These results suggested the novel possibility that the bovine K5 promoter-driven Cre-recombinase was active early in trophoblast-lineage cells that give rise to the placenta. To test this possibility, K5-Cre transgenic mice were crossed with the mT/mG reporter mouse in which activation of GFP expression indicates Cre transgene expression. We observed activation of K5-Cre-driven GFP expression in the ectoplacental cone, in the extraembryonic ectoderm, and in trophoblast giant cells in the E6.5 embryo. In addition, we observed GFP expression at E11.5 to E13.5 in both the labyrinth of the placenta and the yolk sac. NMIIA expression was detected in these same cell types in normal embryos, as well as in E13.5 yolk sac and labyrinth. These findings taken together suggest that NMHC IIA may play critical roles in the early trophoblast-derived ectoplacental cone and extraembryonic ectoderm, as well as in the yolk sac and labyrinth tissues that form later. Our findings are consistent with phenotypes of constitutive NMIIA knockout mice made earlier, that displayed labyrinth and yolk sac-specific defects, but our findings extend those observations by suggesting possible NMIIA roles in trophoblast lineages as well. These results furthermore demonstrate that K5-Cre gene constructs, previously reported to be activated starting at approximately E12.5 in the forming epidermis, may be widely useful as drivers for activation of cre/lox based gene excision in early embryo extraembronic trophoblast tissues as well.
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影响因子：
3.7
作者：
Betapudi V;Rai V;Beach JR;Egelhoff T
通讯作者：
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