Li Fei-Xue, Yu Jiao-Jiao, Liu Ying, Miao Xiao-Ping, Curry Thomas E
1 Zhejiang Key Laboratory of Organ Development and Regeneration, Institute of Developmental and Regenerative Biology, College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou, China.
2 Department of Obstetrics and Gynecology, Chandler Medical Center, University of Kentucky, Lexington, KY, USA.
Reprod Sci. 2017 Jul;24(7):1033-1040. doi: 10.1177/1933719116676394. Epub 2016 Nov 20.
Ectonucleotide pyrophosphatase/phosphodiesterase 3 ( Enpp3) is involved in multiple physiological processes, such as morphological changes and inflammatory processes. The present study investigated the spatiotemporal expression pattern and regulatory mechanisms controlling expression of Enpp3 in the rat ovary during the periovulatory period. Immature female rats were injected with pregnant mare serum gonadotropin to stimulate follicular development. Ovaries, granulosa cells, or theca-interstitial cells were collected at various times after human chorionic gonadotropin (hCG) administration. Real-time polymerase chain reaction analysis revealed that messenger RNA (mRNA) for Enpp3 was highly induced in both granulosa cells and theca-interstitial cells by hCG. In situ hybridization analysis demonstrated that Enpp3 mRNA expression was induced in theca cells at 4 hours after hCG, and the expression remained elevated until 12 hours after hCG. The expression of Enpp3 mRNA was stimulated in granulosa cells at 8 hours and reached the highest expression at 12 hours. Localization of Enpp3 mRNA was observed in newly forming corpora lutea by in situ hybridization. The hCG-stimulated expression of Enpp3 mRNA was blocked by a protein kinase C inhibitor (GF109203) instead of the protein kinase A inhibitor (H89). Furthermore, Enpp3 induction is dependent on new protein synthesis. Inhibition of progesterone action did not alter Enpp3 mRNA expression, whereas inhibition of prostaglandin synthesis or the epidermal growth factor pathway diminished Enpp3 mRNA levels. In conclusion, our findings suggest that the induction of the Enpp3 mRNA may be important for the morphological changes and inflammatory response during ovulation and luteinization.
胞外核苷酸焦磷酸酶/磷酸二酯酶3(Enpp3)参与多种生理过程,如形态变化和炎症过程。本研究调查了围排卵期大鼠卵巢中Enpp3的时空表达模式及其表达的调控机制。对未成熟雌性大鼠注射孕马血清促性腺激素以刺激卵泡发育。在人绒毛膜促性腺激素(hCG)给药后的不同时间收集卵巢、颗粒细胞或卵泡膜-间质细胞。实时聚合酶链反应分析显示,hCG可在颗粒细胞和卵泡膜-间质细胞中高度诱导Enpp3的信使核糖核酸(mRNA)表达。原位杂交分析表明,hCG注射后4小时,卵泡膜细胞中诱导了Enpp3 mRNA表达,且该表达在hCG注射后12小时一直保持升高。Enpp3 mRNA表达在颗粒细胞中于8小时被刺激,并在12小时达到最高表达。通过原位杂交在新形成的黄体中观察到Enpp3 mRNA的定位。hCG刺激的Enpp3 mRNA表达被蛋白激酶C抑制剂(GF109203)阻断,而不是被蛋白激酶A抑制剂(H89)阻断。此外,Enpp3的诱导依赖于新的蛋白质合成。抑制孕酮作用并未改变Enpp3 mRNA表达,而抑制前列腺素合成或表皮生长因子途径则降低了Enpp3 mRNA水平。总之,我们的研究结果表明,Enpp3 mRNA的诱导可能对排卵和黄体化过程中的形态变化和炎症反应很重要。