Xu Jing, Xu Fuhua, Hennebold Jon D, Molskness Theodore A, Stouffer Richard L
Division of Neuuroscience, Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, OR 97006, USA.
Endocrinology. 2007 Nov;148(11):5385-95. doi: 10.1210/en.2007-0541. Epub 2007 Aug 9.
CRH/urocortin-receptor-binding protein (CRH/UCN-R-BP) mRNAs are dynamically expressed in the primate ovary during the menstrual cycle. Therefore, studies were designed to localize CRH/UCN-R-BP mRNAs to ovarian cell types, quantitate protein expression during the corpus luteum (CL) lifespan, and investigate the role of this system in the macaque ovary at midcycle. Monkey ovaries were removed during the preovulatory phase and through the luteal phase to localize CRH/UCN-R-BP mRNAs by in situ hybridization and determine their protein levels in CL by Western blotting. Also, vehicle or a CRH receptor antagonist (astressin) was injected into the preovulatory follicle; daily serum samples were analyzed for hormone levels, and ovaries were removed on d 9 of the luteal phase for histological analysis. There was minimal ligand mRNA staining, whereas receptor and CRHBP was detected in the granulosa and theca cells of the preovulatory follicle. However, ligand and receptor mRNA staining was appreciable in luteal cells of the CL during the early luteal phase (ECL) and diminished in the late luteal phase (LCL). CRHBP staining was low in the ECL and increased markedly in the LCL. Ligand and receptor protein expression was also highest during ECL, whereas CRHBP expression was highest at the LCL. Although astressin injection did not prevent follicle rupture, progesterone levels were significantly less by the mid-luteal phase, and estradiol levels never increased above baseline during the CL lifespan. Histological indices of cell degeneration were observed in the astressin-treated CL. Thus, CRH/UCN-R-BP components are expressed in an ovarian cell-specific manner. The expression pattern and results from antagonist injection are consistent with the hypothesis that CRH/UCN-R activation promotes luteal development and/or structure-function in monkeys during the menstrual cycle.
促肾上腺皮质激素释放激素/尿皮质素受体结合蛋白(CRH/UCN-R-BP)mRNA在月经周期中于灵长类动物卵巢中动态表达。因此,开展了多项研究,旨在将CRH/UCN-R-BP mRNA定位到卵巢细胞类型,定量黄体(CL)生命周期中的蛋白质表达,并研究该系统在猕猴卵巢周期中期的作用。在排卵前期至黄体期切除猴卵巢,通过原位杂交定位CRH/UCN-R-BP mRNA,并通过蛋白质印迹法测定CL中的蛋白质水平。此外,将溶媒或CRH受体拮抗剂(阿片促黑皮质素)注入排卵前卵泡;每天分析血清样本中的激素水平,并在黄体期第9天切除卵巢进行组织学分析。配体mRNA染色极少,而在排卵前卵泡的颗粒细胞和卵泡膜细胞中检测到受体和CRHBP。然而,在黄体早期(ECL)CL的黄体细胞中配体和受体mRNA染色明显,而在黄体晚期(LCL)减少。CRHBP染色在ECL中较低,在LCL中显著增加。配体和受体蛋白表达在ECL期间也最高,而CRHBP表达在LCL时最高。尽管注射阿片促黑皮质素并未阻止卵泡破裂,但到黄体中期孕酮水平显著降低,并且在CL生命周期中雌二醇水平从未升高至基线以上。在阿片促黑皮质素处理的CL中观察到细胞变性的组织学指标。因此,CRH/UCN-R-BP成分以卵巢细胞特异性方式表达。拮抗剂注射的表达模式和结果与以下假设一致,即CRH/UCN-R激活在月经周期中促进猕猴黄体发育和/或结构功能。