Durlej M, Knapczyk-Stwora K, Duda M, Galas J, Slomczynska M
Department of Endocrinology & Tissue Culture, Institute of Zoology, Jagiellonian University, Krakow, Poland.
Reprod Domest Anim. 2011 Jun;46(3):377-84. doi: 10.1111/j.1439-0531.2010.01673.x.
This study was designed to reveal the FSHR mRNA and protein expression in the neonatal porcine ovary and to determine whether maternal administration of antiandrogen flutamide may affect FSHR expression in the ovary of newborn piglets using real-time PCR, immunohistochemistry and Western blot analysis. Pregnant sows were injected with flutamide at a dose of 50 mg/kg body weight, given five times, every second day, starting at day 20 post-coitum (p.c.) or day 80 p.c., and ovaries were obtained from neonatal pigs. The FSHR mRNA expression was significantly decreased after flutamide administration. Furthermore, higher down-regulation was observed following exposure to antiandrogen at day 20 than at day 80 p.c. Immunohistochemistry showed the positive immunostaining for FSHR in the oocytes, granulosa cells of primary follicles and the surface epithelium of the ovaries from both control and flutamide-treated pigs. However, oocytes and granulosa cells of primary follicles in the ovaries exposed in utero to flutamide were weakly immunostained when compared to those in the control ones. The presence of FSHR protein in all investigated ovaries was confirmed by Western blot analysis. Based on our findings, we suggest that FSHR may be involved in the early follicle formation in pigs, which begins during prenatal life. Furthermore, the regulation of FSHR mRNA and protein expression in neonatal porcine ovaries after maternal exposure to flutamide confirms that androgens play a crucial role in porcine folliculogenesis at the early stages.
本研究旨在揭示新生仔猪卵巢中促卵泡激素受体(FSHR)mRNA和蛋白的表达情况,并通过实时荧光定量PCR、免疫组织化学和蛋白质印迹分析,确定母体给予抗雄激素氟他胺是否会影响新生仔猪卵巢中FSHR的表达。怀孕母猪在妊娠20天或80天开始,每隔一天注射一次剂量为50mg/kg体重的氟他胺,共注射5次,然后获取新生仔猪的卵巢。给予氟他胺后,FSHR mRNA表达显著降低。此外,与妊娠80天相比,妊娠20天接触抗雄激素后FSHR mRNA表达下调更明显。免疫组织化学显示,对照组和氟他胺处理组仔猪卵巢的卵母细胞、初级卵泡的颗粒细胞以及卵巢表面上皮中FSHR免疫染色均呈阳性。然而,与对照组相比,子宫内暴露于氟他胺的仔猪卵巢中,初级卵泡的卵母细胞和颗粒细胞免疫染色较弱。蛋白质印迹分析证实了所有研究卵巢中均存在FSHR蛋白。基于我们的研究结果,我们认为FSHR可能参与了猪卵泡的早期形成过程,该过程始于产前。此外,母体暴露于氟他胺后新生仔猪卵巢中FSHR mRNA和蛋白表达的变化证实,雄激素在猪早期卵泡发生中起关键作用。