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孕酮受体在小鼠和人类生殖器官中的表达与定位

Expression and localization of the progesterone receptor in mouse and human reproductive organs.

作者信息

Teilmann Stefan Cuoni, Clement Christian Alexandro, Thorup Jørgen, Byskov Anne Grete, Christensen Søren Tvorup

机构信息

Laboratory of Reproductive Biology, Juliane Marie Center, Rigshospitalet, Copenhagen, Denmark.

出版信息

J Endocrinol. 2006 Dec;191(3):525-35. doi: 10.1677/joe.1.06565.

Abstract

The effects of gonadotropins on progesterone receptor (PR) expression and localization in the mouse oviduct, uterus, and ovary was examined. In the oviduct ciliated epithelial cells of adult mice and human revealed a unique PR localization to the lower half of the motile cilia whereas the nuclei were unstained or faintly stained. Pubertal female mice were further studied by confocal laser scanning microscopy and western blotting before and after injection with FSH and LH followed by human chorionic gonadotropin (hCG) injection after a 48-h period. PR immunolocalization to the oviduct cilia was greatly increased in pubertal mice upon hCG stimulation. In neighboring goblet cells, the PR staining was confined to the nuclei. Nuclear PR localization was evident in epithelial cells of the uterus as well as in a fraction of stromal and muscle cells. Staining intensity and number of stained cells was not affected by hormone stimulation. In the ovary, weak PR immunolocalization was observed in unprimed animals but increased significantly after hCG stimulation. In granulosa cells of preovulatory follicles PR was exclusively observed in mural cells, whereas cumulus cells remained negative. At all stages examined, primary granulosa cell cilia lacked PR staining. SDS-PAGE and western blotting analysis of tissues from oviduct, uterus, and ovary confirmed antibody specificity, and identified two bands corresponding to the PR isoforms PR-A and PR-B. Upon hCG stimulation, a new band cross-reacting with anti-PR emerged above the PR-A form in oviduct fractions, suggesting LH-induced phosphorylation of PR-A. We suggest that ciliary PR in the oviduct plays a role in progesterone signaling after ovulation, possibly via non-genomic events. These novel findings warrant further studies of oviduct and postovulatory signaling events and suggest a sensory role for oviduct cilia in the process of oocyte transport/fertilization.

摘要

研究了促性腺激素对小鼠输卵管、子宫和卵巢中孕激素受体(PR)表达及定位的影响。在成年小鼠和人类的输卵管纤毛上皮细胞中,PR独特地定位于活动纤毛的下半部分,而细胞核未染色或染色很浅。对青春期雌性小鼠在注射促卵泡激素(FSH)和促黄体生成素(LH)前后,以及48小时后注射人绒毛膜促性腺激素(hCG)后,通过共聚焦激光扫描显微镜和蛋白质免疫印迹法进行了进一步研究。hCG刺激后,青春期小鼠输卵管纤毛上的PR免疫定位显著增加。在相邻的杯状细胞中,PR染色局限于细胞核。子宫上皮细胞以及部分基质细胞和肌肉细胞中可见细胞核PR定位。激素刺激不影响染色强度和染色细胞数量。在卵巢中,未预处理的动物中观察到PR免疫定位较弱,但hCG刺激后显著增加。在排卵前卵泡的颗粒细胞中,PR仅在壁层细胞中观察到,而卵丘细胞呈阴性。在所检查的所有阶段,初级颗粒细胞纤毛均缺乏PR染色。对输卵管、子宫和卵巢组织进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质免疫印迹分析,证实了抗体的特异性,并鉴定出两条对应于PR异构体PR-A和PR-B的条带。hCG刺激后,输卵管部分中出现一条与抗PR交叉反应的新条带,位于PR-A条带上方,提示LH诱导了PR-A的磷酸化。我们认为,输卵管中的纤毛PR可能通过非基因组事件在排卵后孕酮信号传导中发挥作用。这些新发现值得对输卵管和排卵后信号传导事件进行进一步研究,并提示输卵管纤毛在卵母细胞运输/受精过程中具有传感作用。

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