Smith Sadie L, Everts Robin E, Sung Li-Ying, Du Fuliang, Page Raymond L, Henderson Boyd, Rodriguez-Zas Sandra L, Nedambale Tshimangadzo L, Renard Jean-Paul, Lewin Harris A, Yang Xiangzhong, Tian X Cindy
Center for Regenerative Biology/Department of Animal Science, University of Connecticut, Storrs, Connecticut, USA.
Mol Reprod Dev. 2009 Jan;76(1):38-47. doi: 10.1002/mrd.20927.
In vitro production (IVP) has been shown to affect embryonic gene expression and often result in large offspring syndrome (LOS) in cattle and sheep. To dissect the effects of in vitro maturation, fertilization and culture on bovine embryos, we compared the expression profiles of single blastocysts generated by: (1) in vitro maturation, fertilization and culture (IVF); (2) in vivo maturation, fertilization and in vitro culture (IVD); and (3) in vivo maturation, fertilization and development (AI). To conduct expression profiling, total RNA was isolated from individual embryos, linearly amplified and hybridized to a custom bovine cDNA microarray containing approximately 6,300 unique genes. There were 306, 367, and 200 genes differentially expressed between the AI and IVD, IVF and IVD, and AI and IVF comparisons, respectively. Interestingly, 44 differentially expressed genes were identified between the AI embryos and both the IVF and IVD embryos, making these potential candidates for LOS. There were 60 genes differentially expressed between the IVF embryos and the AI and IVD embryos. The Gene Ontology category "RNA processing" was over-represented among the genes that were down-regulated in the IVF embryos, indicating an effect of in vitro oocyte maturation/fertilization on the ability to transcribe maternal RNA stores. A culture effect on the expression of genes involved in translation was also observed by the comparison of AI with IVD embryos.
体外生产（IVP）已被证明会影响胚胎基因表达，并常常导致牛和羊出现大后代综合征（LOS）。为了剖析体外成熟、受精和培养对牛胚胎的影响，我们比较了通过以下方式产生的单个囊胚的表达谱：（1）体外成熟、受精和培养（IVF）；（2）体内成熟、受精和体外培养（IVD）；以及（3）体内成熟、受精和发育（AI）。为了进行表达谱分析，从单个胚胎中分离出总RNA，进行线性扩增，并与包含约6300个独特基因的定制牛cDNA微阵列杂交。在AI与IVD、IVF与IVD以及AI与IVF的比较中，分别有306、367和200个基因差异表达。有趣的是，在AI胚胎与IVF和IVD胚胎之间鉴定出44个差异表达基因，这些基因可能是LOS的候选基因。在IVF胚胎与AI和IVD胚胎之间有60个基因差异表达。在IVF胚胎中下调的基因中，基因本体论类别“RNA加工”过度代表，这表明体外卵母细胞成熟/受精对转录母体RNA储存能力有影响。通过将AI与IVD胚胎进行比较，还观察到培养对参与翻译的基因表达有影响。
