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Identification of uterine genes induced by embryo during mice implantation.

小鼠植入过程中胚胎诱导的子宫基因的鉴定。

基本信息

批准号：
09671676
负责人：
HIGUCHI Toshihiro
金额：
$ 1.98万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671676/
关键词：
Implanation Embryo Endometrium cDNA subtraction Infertility

项目摘要

In this project, we sought to identify the genes involved in the implantation process by subtracting cDNA constructed from the uteri of non-pregnant mice from cDNA from those of pregnancy day 5 mice. One of the identified cDNA encoded the calcium binding protein D-9k (Calbindin). Northern blot analysis showed that the size of the transcript was the same as rat calbindin mRNA, and that it was expressed in the uteri of pregnancy day 5 mice. We found that the expression of calbindin mRNA increased on day 3 of pregnancy when the serum progesterone concentration is high. In fact, progesterone induced a marked increase in the expression of calbindin mRNA.In situ hybridization showed that the uterine expression of calbindin mRNA disappeared in the glandular epithelium on day 5 of pregnancy. In the pseudopregnant mice, the expression of calbindin mRNA was observed in the glandular epithelium on day 5, suggesting that this molecule is regulated not only by the sex steroid hormones, but also by the embryonal signal. To examine this possibility, we performed the embryo transfer experiments. While calbindin mRNA was detected in the glandular epithelium of the control uterine horn, it was not detected in the embryo-transferred horn 36 h after embryo transfer. These results indicated that the expression of calbindin mRNA in the glandular epithelium was suppressed by the presence of blastocysts on day 5 of pregnancy.In conclusion we could found that embryonic signals modulate the endometrium during implantation. We are now analyzing other cDNA clones cloned from subtracted cDNA library. This approach will contribute towards the clarification of reproductive physiology.

在这个项目中，我们试图通过从怀孕第 5 天小鼠的 cDNA 中减去从非怀孕小鼠子宫构建的 cDNA 来鉴定参与植入过程的基因。已鉴定的 cDNA 之一编码钙结合蛋白 D-9k (Calbindin)。 Northern印迹分析表明该转录本大小与大鼠钙结合蛋白mRNA相同，并且在妊娠第5天小鼠的子宫中表达。我们发现，当妊娠第3天血清孕酮浓度较高时，钙结合蛋白mRNA的表达增加。事实上，黄体酮诱导钙结合蛋白mRNA的表达显着增加。原位杂交显示，妊娠第5天，腺上皮中钙结合蛋白mRNA的子宫表达消失。在假孕小鼠中，第5天在腺上皮中观察到钙结合蛋白mRNA的表达，表明该分子不仅受到性类固醇激素的调节，而且还受到胚胎信号的调节。为了检验这种可能性，我们进行了胚胎移植实验。虽然在对照子宫角的腺上皮中检测到钙结合蛋白 mRNA，但在胚胎移植后 36 小时，在胚胎移植角中未检测到钙结合蛋白 mRNA。这些结果表明，腺上皮中钙结合蛋白 mRNA 的表达因妊娠第 5 天囊胚的存在而受到抑制。总之，我们可以发现胚胎信号在着床过程中调节子宫内膜。我们现在正在分析从消减 cDNA 文库中克隆的其他 cDNA 克隆。这种方法将有助于阐明生殖生理学。