Nie G Y, Li Y, Batten L, Griffiths B, Wang J, Findlay J K, Salamonsen L A
Prince Henry's Institute of Medical Research, P.O. Box 5152, Clayton, Victoria 3168, Australia.
Mol Hum Reprod. 2000 Dec;6(12):1131-9. doi: 10.1093/molehr/6.12.1131.
RNA differential display was applied to identify genes critical for the establishment of pregnancy in the mouse. One of the gene fragments identified was homologous to human SC35 splicing factor; the mouse counterpart had not then been cloned. To obtain the full cDNA sequence of the mouse gene, a cDNA library was screened and four positive clones were fully analysed. Sequencing analysis indicated that we had cloned alternatively spliced mRNA species of mouse SC35 splicing factor. A map of splicing structure for this gene's pre-mRNA was then proposed and region-specific mRNA species were tested on Northern blots. This analysis indicated that the overall expression level of SC35 mRNA was much higher in implantation sites than in inter-implantation sites in the mouse uterus during early pregnancy. The expression of alternatively spliced mRNAs for SC35 was differently regulated both during early pregnancy and by steroid hormones. Embryo-derived factors were also implicated in the up-regulation of SC35 mRNA at implantation sites. These results demonstrate, for the first time, that an essential splicing factor is regulated in a complex manner during implantation in the mouse uterus. Hence, its correct regulation could be important for the success of pregnancy.
运用RNA差异显示技术来鉴定对小鼠妊娠建立至关重要的基因。所鉴定出的一个基因片段与人类SC35剪接因子同源;当时小鼠的对应基因尚未克隆出来。为了获得小鼠基因的完整cDNA序列,对一个cDNA文库进行了筛选,并对四个阳性克隆进行了全面分析。序列分析表明,我们克隆到了小鼠SC35剪接因子的可变剪接mRNA种类。随后提出了该基因前体mRNA的剪接结构图,并在Northern印迹上检测了区域特异性mRNA种类。该分析表明,在妊娠早期,小鼠子宫中植入部位的SC35 mRNA总体表达水平远高于非植入部位。SC35可变剪接mRNA的表达在妊娠早期和类固醇激素作用下受到不同的调控。胚胎来源的因子也与植入部位SC35 mRNA的上调有关。这些结果首次证明,在小鼠子宫植入过程中,一种重要的剪接因子受到复杂的调控。因此,其正确调控对于妊娠成功可能很重要。