Vallée Maud, Beaudry Daniele, Roberge Claude, Matte J Jacques, Blouin Richard, Palin Marie-France
Département de Biologie, Université de Sherbrooke, Sherbrooke, Québec, Canada J1K 2R1.
Biol Reprod. 2003 Nov;69(5):1697-706. doi: 10.1095/biolreprod.103.019307. Epub 2003 Jul 30.
The implantation period is a critical time for embryonic survival in pigs. During this period, numerous growth factors are secreted by the conceptuses and the uterine endometrium in order to establish pregnancy and to provide a proper environment for embryonic development. It is well known that the Chinese Meishan sows have a larger litter size when compared with occidental sows mainly because of a superior embryonic survival rate. As a further step toward understanding the mechanisms involved in embryonic survival, we used a suppression subtractive hybridization technique to identify genes that were differentially expressed in Meishan-Landrace conceptuses and endometrial tissue at Day 15 of gestation when compared with conventional Landrace sows. Of the 1000 subtractive clones isolated from each library, 137 endometrial and 166 conceptus-enriched cDNAs were single-pass sequenced and examined by BLAST analysis for identification. Sixty-two percent of the clones found in the endometrial library and 78% of the clones found in the conceptus library showed homology with known genes. Among these genes, the 20 most relevant to embryonic survival based on the available literature were validated through real-time reverse-transcriptase polymerase chain reaction (RT-PCR) analysis. Our results show that suppression subtractive hybridization is a powerful method applicable in identifying putative candidate genes that might be used for selection of high litter-size breeds.
着床期对猪胚胎的存活至关重要。在此期间,孕体和子宫内膜会分泌多种生长因子,以建立妊娠并为胚胎发育提供适宜环境。众所周知,与西方母猪相比,中国梅山母猪产仔数更多,这主要是因为其胚胎存活率更高。为了进一步了解胚胎存活所涉及的机制,我们采用抑制性消减杂交技术,来鉴定与传统长白母猪相比,妊娠第15天梅山-长白杂交孕体和子宫内膜组织中差异表达的基因。从每个文库中分离出的1000个消减克隆中,对137个子宫内膜和166个孕体富集的cDNA进行了单通道测序,并通过BLAST分析进行鉴定。在内膜文库中发现的克隆有62%,在孕体文库中发现的克隆有78%与已知基因具有同源性。在这些基因中,根据现有文献,选取了与胚胎存活最相关的20个基因,通过实时逆转录聚合酶链反应(RT-PCR)分析进行验证。我们的结果表明,抑制性消减杂交是一种强大的方法,可用于鉴定可能用于选择高产仔数品种的假定候选基因。