Ka Hakhyun, Seo Heewon, Kim Mingoo, Moon Sunjin, Kim Heebal, Lee Chang-Kyu
Department of Biological Resources and Technology, Yonsei University, Wonju 220-710, Republic of Korea.
Anim Reprod Sci. 2008 Oct;108(1-2):79-91. doi: 10.1016/j.anireprosci.2007.07.008. Epub 2007 Aug 1.
Cloning by somatic cell nuclear transfer (SCNT) in pigs has great value for research and biomedical applications. However, cloning pigs is inefficient, and cloning procedures often lead to the birth of abnormal offspring because of the inadequate nuclear remodeling of donor cells as well as inadequate subsequent development. To understand the problems of the cloning process, it is necessary to understand how the uterus interacts with cloned embryo during pregnancy and supports placentation and fetal development. In this study, we compared gene expression profiles of the uterus with SCNT embryos to those of the uterus with normal embryos by natural mating. We obtained the uterine endometrial tissues on day 30 of pregnancy and conducted gene expression profiling using the Platinum Pig 13K oligonucleotide microarrays. Of the 13,610 genes analyzed, expression of 351 genes significantly increased or decreased in the uterine tissues with SCNT embryos compared to those with normal embryos. The differentially regulated genes included enzymes involved in steroidogenesis and extracellular matrix remodeling and uterine secretory proteins. Analyses of real-time reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization of selected genes confirmed the validity of the gene expression patterns observed in the microarray analysis. Results of this study showed that the transcriptional profile of the genes in the uterus with SCNT embryos was regulated differently indicating that the maternal responsiveness to the SCNT embryos was impaired, resulting in the altered gene expression in the uterus and, in turn, abnormal placental and fetal development and increased embryonic loss.
通过体细胞核移植(SCNT)克隆猪在研究和生物医学应用方面具有重要价值。然而,克隆猪的效率较低,且由于供体细胞的核重塑不足以及随后发育不充分,克隆过程常常导致异常后代的出生。为了了解克隆过程中的问题,有必要了解子宫在妊娠期间如何与克隆胚胎相互作用,并支持胎盘形成和胎儿发育。在本研究中,我们将怀有SCNT胚胎的子宫与通过自然交配怀有正常胚胎的子宫的基因表达谱进行了比较。我们在妊娠第30天获取了子宫内膜组织,并使用Platinum Pig 13K寡核苷酸微阵列进行基因表达谱分析。在分析的13610个基因中,与怀有正常胚胎的子宫组织相比,怀有SCNT胚胎的子宫组织中有351个基因的表达显著增加或减少。差异调节的基因包括参与类固醇生成和细胞外基质重塑的酶以及子宫分泌蛋白。对选定基因的实时逆转录聚合酶链反应(RT-PCR)和原位杂交分析证实了微阵列分析中观察到的基因表达模式的有效性。本研究结果表明,怀有SCNT胚胎的子宫中基因的转录谱受到不同调节,这表明母体对SCNT胚胎的反应性受损,导致子宫中基因表达改变,进而导致胎盘和胎儿发育异常以及胚胎损失增加。