Athanassakis I, Farmakiotis V, Aifantis I, Gravanis A, Vassiliadis S
Department of Biology, University of Crete, Heraklion, Crete, Greece.
J Endocrinol. 1999 Nov;163(2):221-7. doi: 10.1677/joe.0.1630221.
The detection of corticotropin-releasing hormone (CRH) in the pregnant and non-pregnant uterus has driven research to determine the role of this 41 amino acid neuropeptide in the female reproductive system. As concentrations of CRH mRNA and its peptide product are greater in the implantation sites of the early pregnant uterus compared with the regions between implantation sites, CRH has been hypothesised to participate in blastocyst implantation. Using the mouse system as an experimental model, we studied the distribution of CRH in the uterus during the oestrus cycle and early gestational period, and now provide evidence for its involvement in embryo implantation using cell culture techniques. The percentage of CRH-positive uterine cells and the amount of CRH released during anoestrus, pro-oestrus and oestrus were determined by immunofluorescence and ELISA experiments respectively. The highest number of intracellularly CRH-positive cells was obtained during pro-oestrus, whereas the highest CRH concentration in uterine cell culture supernatants was detected during anoestrus. At early stages of gestation, CRH was detected in the endometrium on days 2, 3 and 4 of pregnancy and in the myometrium on days 3 and 4, whereas it was undetectable on day 5. The functional role of CRH during early gestation was evaluated by administering anti-CRH antibody to mice from day 3 to day 8 of pregnancy. This treatment resulted in implantation failure in 60% of the cases, in which implantation sites, although clearly present in the uterus, had failed to host an embryo. These results provide direct evidence about the involvement of CRH in murine embryo implantation and are in agreement with hypotheses postulated in humans.
对妊娠和非妊娠子宫中促肾上腺皮质激素释放激素（CRH）的检测推动了相关研究，以确定这种由41个氨基酸组成的神经肽在女性生殖系统中的作用。由于与着床部位之间的区域相比，妊娠早期子宫着床部位的CRH mRNA及其肽产物浓度更高，因此有人推测CRH参与了胚泡着床。我们以小鼠系统作为实验模型，研究了发情周期和妊娠早期子宫中CRH的分布情况，现在利用细胞培养技术为其参与胚胎着床提供了证据。分别通过免疫荧光和ELISA实验测定了发情间期、发情前期和发情期CRH阳性子宫细胞的百分比以及释放的CRH量。发情前期细胞内CRH阳性细胞数量最多，而发情间期在子宫细胞培养上清液中检测到的CRH浓度最高。在妊娠早期，妊娠第2、3和4天在子宫内膜中检测到CRH，第3和4天在子宫肌层中检测到CRH，而在第5天未检测到。通过在妊娠第3天至第8天给小鼠注射抗CRH抗体，评估了CRH在妊娠早期的功能作用。这种处理导致60%的病例着床失败，其中着床部位虽在子宫中明显存在，但未能容纳胚胎。这些结果为CRH参与小鼠胚胎着床提供了直接证据，与人类提出的假说一致。
