Rambags B P B, van Tol H T A, van den Eng M M, Colenbrander B, Stout T A E
Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 112, 3584 CM Utrecht, The Netherlands.
Theriogenology. 2008 Feb;69(3):366-75. doi: 10.1016/j.theriogenology.2007.10.011. Epub 2007 Nov 26.
Progesterone and oestrogen play essential roles in the maintenance of pregnancy in eutherian mammals and are thought to exert their effects on the developing conceptus indirectly, via the endometrium. In some species, early embryos have themselves been shown to express steroid receptors, thereby suggesting that reproductive steroids may also influence embryonic development directly. The aim of this study was to determine whether early intrauterine equine conceptuses express either the classical intracellular progesterone (PR) and oestrogen receptors (ERalpha and ERbeta) or the more recently characterised membrane-bound progesterone receptors (PGRMC1 and mPR). Horse conceptuses recovered on days 7, 10 and 14 after ovulation (n=8 at each stage) were examined for steroid receptor mRNA expression using quantitative rtPCR. Where commercial antibodies were available (PR, ERbeta), receptor localisation was examined immunohistochemically in day 10, 12, 14, 15 and 16 conceptuses (n=2 at each stage). mRNA for PR, PGRMC1 and mPR was detected at all stages examined, but while PGRMC1 and mPR expression increased during the day 7-14 period, PR expression decreased. ERalpha mRNA was not detected at any stage examined, whereas ERbeta mRNA was detected in all day 14, some day 10 and no day 7 conceptuses. Immunoreactive ERbeta receptors were localised to the trophectoderm of day 14-16 conceptuses; PR were not detected immunohistochemically in conceptus tissue. In summary, this study demonstrates that equine conceptuses express mRNA and, in the case of ERbeta, protein for steroid hormone receptors during the period encompassing rapid conceptus growth, differentiation and maternal pregnancy recognition.
孕酮和雌激素在真兽亚纲哺乳动物维持妊娠过程中发挥着重要作用,并且被认为是通过子宫内膜间接对发育中的孕体产生影响。在一些物种中,早期胚胎已被证明自身能够表达类固醇受体,这表明生殖类固醇也可能直接影响胚胎发育。本研究的目的是确定早期子宫内的马属动物孕体是否表达经典的细胞内孕酮受体(PR)和雌激素受体(ERα和ERβ),或者最近鉴定出的膜结合孕酮受体(PGRMC1和mPR)。对排卵后第7、10和14天回收的马属动物孕体(每个阶段n = 8),使用定量逆转录聚合酶链反应(rtPCR)检测类固醇受体mRNA表达。在有商用抗体的情况下(PR、ERβ),通过免疫组织化学方法检测第10、12、14、15和16天孕体(每个阶段n = 2)中的受体定位。在所检测的所有阶段均检测到PR、PGRMC1和mPR的mRNA,但在第7 - 14天期间,PGRMC1和mPR的表达增加,而PR的表达减少。在所检测的任何阶段均未检测到ERα mRNA,而在所有第14天、部分第10天的孕体中检测到了ERβ mRNA,第7天的孕体中未检测到。免疫反应性ERβ受体定位于第14 - 16天孕体的滋养外胚层;在孕体组织中未通过免疫组织化学方法检测到PR。总之,本研究表明,在孕体快速生长、分化以及母体识别妊娠的这段时间内,马属动物孕体表达类固醇激素受体的mRNA,对于ERβ而言,还表达其蛋白。