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体外受精或体细胞核移植产生的单个植入前牛胚胎中基因表达的比较。

Comparison of gene expression in individual preimplantation bovine embryos produced by in vitro fertilisation or somatic cell nuclear transfer.

作者信息

de A Camargo Luiz Sergio, Powell Anne M, do Vale Filho Vicente R, Wall Robert J

机构信息

Embrapa Dairy Cattle, Juiz de Fora, MG, Brazil.

出版信息

Reprod Fertil Dev. 2005;17(5):487-96. doi: 10.1071/rd04128.

DOI:10.1071/rd04128
PMID:15907273
Abstract

In vitro fertilisation (IVF) and somatic cell nuclear transfer (SCNT) have been implicated in a variety of developmental abnormalities. Aberrant gene expression is likely to account for much of the diminished viability and developmental abnormalities observed. In the present study, the expression of multiple genes in IVF and SCNT bovine blastocyst-stage embryos were evaluated and compared with in vivo-produced embryos. Eleven genes expressed at and following maternal-zygotic transcription transition were evaluated in individual blastocysts by real-time polymerase chain reaction following RNA amplification. A subset of those genes was also evaluated in individual IVF and SCNT eight-cell embryos. A fibroblast-specific gene, expressed by nuclear donor cells, was also evaluated in IVF and SCNT embryos. The observed gene expression pattern at the eight-cell stage was not different between IVF and SCNT embryos (P > 0.05). In vitro fertilisation and SCNT blastocyst expression was lower (P < 0.01) for all genes compared with their in vivo-produced counterparts, except for lactate dehydrogenase isoenzyme A (P < 0.001). The patterns of gene expression of the IVF and SCNT blastocysts were indistinguishable. Neither SCNT eight-cell nor blastocyst-stage embryos expressed the gene used as a fibroblast marker (collagen VIalpha1). For the genes evaluated, the level of expression was influenced more by the environment than by the method used to produce the embryos. These results support the notion that if developmental differences observed in IVF- and SCNT-produced fetuses and neonates are the result of aberrant gene expression during the preimplantation stage, those differences in expression are subtle.

摘要

体外受精(IVF)和体细胞核移植(SCNT)与多种发育异常有关。异常的基因表达可能是观察到的活力下降和发育异常的主要原因。在本研究中,评估了IVF和SCNT牛囊胚期胚胎中多个基因的表达,并与体内产生的胚胎进行了比较。在RNA扩增后,通过实时聚合酶链反应在单个囊胚中评估了11个在母源-合子转录转变时及之后表达的基因。还在单个IVF和SCNT八细胞胚胎中评估了其中一部分基因。还在IVF和SCNT胚胎中评估了由核供体细胞表达的成纤维细胞特异性基因。IVF和SCNT胚胎在八细胞阶段观察到的基因表达模式没有差异(P>0.05)。与体内产生的对应胚胎相比,IVF和SCNT囊胚中所有基因的表达均较低(P<0.01),乳酸脱氢酶同工酶A除外(P<0.001)。IVF和SCNT囊胚的基因表达模式无法区分。SCNT八细胞期和囊胚期胚胎均未表达用作成纤维细胞标记物的基因(胶原蛋白VIα1)。对于所评估的基因,表达水平受环境的影响大于产生胚胎的方法。这些结果支持这样一种观点,即如果在IVF和SCNT产生的胎儿和新生儿中观察到的发育差异是着床前阶段基因表达异常的结果,那么这些表达差异是细微的。

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