Lee Kai-Fai, Yao Yuan-Qing, Kwok Ka-Leung, Xu Jia-Sen, Yeung William S B
Department of Obstetrics and Gynaecology, University of Hong Kong, Hong Kong, People's Republic of China.
Biochem Biophys Res Commun. 2002 Mar 29;292(2):564-70. doi: 10.1006/bbrc.2002.6676.
Fertilization and development of mouse embryos occur in the ampullae of oviduct. We hypothesize that fetal-maternal communication exists in the preimplantation period, allowing optimal development of embryos. It is known that embryotrophic factors from oviduct affect the development of embryos. Although embryos affect their own transport in the oviduct, the mechanism of action is unknown. As a step toward understanding the action of embryos on oviductal physiology, we adopted suppression subtractive hybridization (SSH) to compare the gene expression in the mouse oviduct containing early embryos with that of oviduct containing oocytes. Ten to twelve 1-cell mouse embryos were transferred to one oviduct of a foster mother and similar number of oocytes were transferred to the contralateral oviduct. The animals were sacrificed after 48 h and their oviducts were excised for mRNA study. Using SSH, we screened out 250 putative positive clones from the subtracted embryo-containing oviduct library and 97 of them were screened positive by reverse dot-blot analysis. DNA sequence analysis identified genes that shared high homology with sequences in GenBank/EMBL database with unknown functions. Overall, 13 of the 90 high-quality sequences (14%) were homologous to 6 different genes previously described. Reverse Northern analysis confirmed that the expression of these genes were higher in the embryo-containing oviduct than in the oocyte-containing oviduct. About 12% of these clones (11/90) were novel. This article is the first to report identification of genes in the oviduct that are upregulated in the presence of embryos during the preimplantation period.
小鼠胚胎的受精和发育发生在输卵管壶腹部。我们推测在植入前阶段存在胎儿与母体之间的交流,以促进胚胎的最佳发育。已知来自输卵管的胚胎营养因子会影响胚胎的发育。虽然胚胎会影响其自身在输卵管中的运输,但其作用机制尚不清楚。作为理解胚胎对输卵管生理作用的第一步,我们采用抑制性消减杂交(SSH)技术,比较含有早期胚胎的小鼠输卵管与含有卵母细胞的输卵管中的基因表达。将10至12个1细胞期的小鼠胚胎移植到代孕母鼠的一侧输卵管中,将数量相近的卵母细胞移植到对侧输卵管中。48小时后处死动物,切除其输卵管用于mRNA研究。通过SSH技术,我们从消减后的含胚胎输卵管文库中筛选出250个推定的阳性克隆,其中97个经反向点杂交分析呈阳性。DNA序列分析鉴定出与GenBank/EMBL数据库中功能未知的序列具有高度同源性的基因。总体而言,90条高质量序列中的13条(14%)与先前描述的6个不同基因同源。反向Northern分析证实,这些基因在含胚胎的输卵管中的表达高于含卵母细胞的输卵管。这些克隆中约12%(11/90)是新发现的。本文首次报道了在植入前阶段,胚胎存在时输卵管中上调基因的鉴定。