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首页> 外文期刊>Comparative Medicine >Decreased growth factor expression through RNA interference inhibits development of mouse preimplantation embryos
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Decreased growth factor expression through RNA interference inhibits development of mouse preimplantation embryos

机译：通过RNA干扰减少的生长因子表达抑制小鼠植入前胚胎的发育

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Mitogenic growth factors play an important role in cellular development and differentiation. The purpose of this study was to assess the extent to which epidermal growth factor (EGF) and transforming growth factor alpha (TGFalpha) and their cognate receptor (EGFR) are crucial for normal preimplantation embryo development. We used RNA interference to decrease expression of growth factors in preimplantation mouse embryos. We microinjected 1-cell mouse embryos individually with short interfering RNA (siRNA) specific to EGF, TGFalpha, and EGFR and then analyzed temporal and spatial gene expression patterns at different stages of development before implantation. Transfection with siRNA significantly reduced growth factor expression in 1-cell, 2-cell, morula, early-blastocyst, and late-blastocyst embryos to levels similar to those in untreated 'cloned' embryos derived through intracytoplasmic nuclear injection. In addition, siRNA effectively decreased expression of maternally inherited genes between 24 and 72 h after transfection, with recovery of gene expression during late-blastocyst stage at 96 h after transfection. Furthermore, siRNA significantly decreased blastocyst formation, increased the number of apoptotic cells, and reduced the total number of differentiated cells in blastocysts; these changes were greatest after decreasing EGFR and of both EGF and TGFalpha simultaneously. These results support our hypothesis that EGF and TGFalpha are crucial for embryo survival and development. Further, dysregulated expression of growth factors is associated with poor development of cloned mouse embryos.
机译：有丝分裂生长因子在细胞发育和分化中起重要作用。这项研究的目的是评估表皮生长因子（EGF）和转化生长因子α（TGFalpha）及其同源受体（EGFR）在正常植入前胚胎发育中至关重要的程度。我们使用RNA干扰来减少植入前小鼠胚胎中生长因子的表达。我们分别对EGF，TGFalpha和EGFR短干扰RNA（siRNA）分别微注射了1细胞小鼠胚胎，然后在植入前分析了不同发育阶段的时空基因表达模式。 siRNA转染可将1细胞，2细胞，桑ula，胚泡早期和胚泡晚期胚中的生长因子表达显着降低，其水平与通过胞浆内核注射获得的未经处理的“克隆”胚胎的水平相似。此外，siRNA有效地降低了转染后24至72 h之间母本遗传基因的表达，并在转染后96 h的胚泡晚期阶段恢复了基因表达。此外，siRNA显着减少了胚泡的形成，增加了凋亡细胞的数量，并减少了胚泡中分化细胞的总数。在同时降低EGFR以及同时降低EGF和TGFalpha时，这些变化最大。这些结果支持我们的假设，即EGF和TGFalpha对于胚胎存活和发育至关重要。此外，生长因子的表达失调与克隆的小鼠胚胎的发育不良有关。

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