Ulbrich S E, Kettler A, Einspanier R
Institute of Physiology, Research Centre for Milk and Food Weihenstephan, Technical University of Munich, Weihenstephaner Berg 3, Freising D-85350, Germany.
J Steroid Biochem Mol Biol. 2003 Feb;84(2-3):279-89. doi: 10.1016/s0960-0760(03)00039-6.
This study examined the regulation and localization of estrogen receptors alpha and beta (ERalpha, ERbeta) and progesterone receptor (PR) in the bovine oviduct. Oviduct epithelial cells from cycling cows (in vivo) were investigated. In addition, the reactivity of a cell suspension culture stimulated with physiological doses of estradiol-17beta (E2) or progesterone (P4) was tested (in vitro). The specific steroid receptor expression of oviductal cells was quantified for mRNA using real-time RT-PCR. Furthermore, steroid receptor proteins were analyzed by Western blotting and localized by immunohistochemistry in situ. Obvious cyclic changes of receptor expression in vivo were observed and concurrent expression patterns were detected in vitro. PR and ERalpha mRNA transcripts were elevated in vivo during the follicular phase. The highest PR and ERalpha protein expression was detected subsequently during the early-luteal phase. In vitro, E2-supplementation resulted in an upregulation of PR and ERalpha. Both ERbeta mRNA and protein expression were highest during the luteal phase in vivo and elevated ERbeta expression levels were observed in vitro after P4 treatment. Evidence is provided for a varying expression of ERalpha, ERbeta and PR in bovine oviducts at different cycle stages in vivo, respectively under steroid supplementation in vitro. The region specific and cycle dependent expression differences point towards a functional importance of the three steroid receptors in the bovine oviduct, the site of fertilization and early embryonic development.
本研究检测了雌激素受体α和β（ERα、ERβ）以及孕激素受体（PR）在牛输卵管中的调控及定位。对处于发情周期的奶牛（体内）的输卵管上皮细胞进行了研究。此外，还检测了用生理剂量的雌二醇-17β（E2）或孕酮（P4）刺激的细胞悬浮培养物的反应性（体外）。使用实时逆转录聚合酶链反应（RT-PCR）对输卵管细胞的特异性类固醇受体表达的mRNA进行定量。此外，通过蛋白质免疫印迹法分析类固醇受体蛋白，并通过原位免疫组织化学进行定位。观察到体内受体表达明显的周期性变化，并且在体外检测到了同时存在的表达模式。在卵泡期，体内PR和ERα的mRNA转录物升高。随后在黄体早期检测到PR和ERα的最高蛋白表达。在体外，补充E导致PR和ERα上调。ERβ的mRNA和蛋白表达在体内黄体期最高，并且在体外经P4处理后观察到ERβ表达水平升高。本研究为体内不同周期阶段以及体外类固醇补充条件下，牛输卵管中ERα、ERβ和PR的不同表达提供了证据。区域特异性和周期依赖性的表达差异表明这三种类固醇受体在牛输卵管（受精和早期胚胎发育的部位）中具有重要功能。
