Dai Yanzhenzi, Ivell Richard, Liu Xuan, Janowski Dana, Anand-Ivell Ravinder
School of Biosciences, University of NottinghamNottingham, United Kingdom.
Leibniz Institute for Farm Animal BiologyDummerstorf, Germany.
Front Physiol. 2017 Jun 6;8:359. doi: 10.3389/fphys.2017.00359. eCollection 2017.
In most mammals the peptide hormone relaxin is a key physiological component regulating early pregnancy and birth. However, synteny analysis shows that the gene encoding ovarian relaxin-2 is deleted in cows and sheep. While, these ruminants appear to exhibit a relaxin-like physiology, as in other mammals, until now a molecular understanding of this has been lacking. Cloning and expression analysis of the cognate bovine receptor for relaxin, RXFP1, as well as of the structurally related receptor, RXFP2, in female tissues, shows that these are expressed in a similar way to other mammals. RXFP1 transcripts are found in ovarian theca cells, endometrium, and myometrium, whereas RXFP2 transcripts are expressed in ovarian theca cells, oocytes, as well as in myometrium. Transfection of receptor-expressing gene constructs into HEK293 cells indicates that bovine RXFP1 has a greater EC50 at 10-50 nM for porcine or human relaxin, compared to human RXFP1. For bovine RXFP2, in contrast, the EC50 is <1 nM for its cognate ligand, bovine INSL3, but also 10-30 nM for porcine or human relaxin. Functional analysis shows that bovine myometrial cells are able to respond to exogenous relaxin and INSL3 with a significant increase in cAMP. Although expressing mRNA for both RXFP1 and RXFP2, bovine follicular theca cells only respond to INSL3 with a dose-dependent increase in cAMP. Altogether the results suggest that the cow is able to compensate for the missing hormone, and moreover imply that relaxin analogs could offer an important therapeutic option in treating female ruminant infertility.
在大多数哺乳动物中,肽类激素松弛素是调节早期妊娠和分娩的关键生理成分。然而,同线性分析表明,编码卵巢松弛素-2的基因在牛和羊中缺失。虽然这些反刍动物似乎表现出与其他哺乳动物类似的松弛素样生理功能,但迄今为止,对此缺乏分子层面的理解。对雌性组织中松弛素的同源牛受体RXFP1以及结构相关受体RXFP2进行克隆和表达分析,结果表明它们的表达方式与其他哺乳动物相似。RXFP1转录本在卵巢膜细胞、子宫内膜和子宫肌层中被发现,而RXFP2转录本则在卵巢膜细胞、卵母细胞以及子宫肌层中表达。将表达受体的基因构建体转染到HEK293细胞中表明,与人类RXFP1相比,牛RXFP1对猪或人类松弛素在10 - 50 nM时具有更高的半数有效浓度(EC50)。相比之下,对于牛RXFP2,其同源配体牛胰岛素样肽3(INSL3)的EC50 <1 nM,但对猪或人类松弛素的EC50也为10 - 30 nM。功能分析表明,牛子宫肌层细胞能够对外源松弛素和INSL3作出反应,使环磷酸腺苷(cAMP)显著增加。尽管牛卵泡膜细胞同时表达RXFP and INSL3的mRNA,但仅对INSL3作出反应,cAMP呈剂量依赖性增加。总体而言,结果表明牛能够补偿缺失的激素,此外还意味着松弛素类似物可能为治疗雌性反刍动物不孕症提供重要的治疗选择。