Monte A P O, Barros V R P, Santos J M, Menezes V G, Cavalcante A Y P, Gouveia B B, Bezerra M E S, Macedo T J S, Matos M H T
Nucleus of Biotechnology Applied to Ovarian Follicle Development, Federal University of San Francisco Valley, 56300-990, Petrolina, PE, Brazil.
Nucleus of Biotechnology Applied to Ovarian Follicle Development, Federal University of San Francisco Valley, 56300-990, Petrolina, PE, Brazil.
Theriogenology. 2019 Apr 15;129:61-69. doi: 10.1016/j.theriogenology.2019.02.005. Epub 2019 Feb 8.
This study analyzed IGF-1 protein immunostaining in sheep ovaries, the effect of IGF-1 alone or associated with FSH on the culture of secondary follicles, and the immunostaining of LHR protein in antral follicles before and after culture. Ovaries were collected for IGF-1 protein analysis. In experiment 1, secondary follicles were cultured in α-MEM (control) or α-MEM supplemented with IGF-1 (10, 50 or 100 ng/mL). In experiment 2, follicles were cultured in the same media of experiment 1 plus 750 ng/mL FSH. Moreover, LHR immunostaining was analyzed in fresh antral follicles and after culture in 50 ng/mL IGF-1 + FSH. The IGF-1 protein was immunolocalized in oocytes from all stages of follicle development and in the granulosa cells from secondary and antral follicles. IGF-1 did not influence (P > 0.05) follicular viability and growth (experiment 1). However, in experiment 2, 50 ng/mL IGF-1 + FSH stimulated oocyte growth (P < 0.05) and LHR immunostaining in antral follicles. Control medium, 10 or 50 ng/mL IGF-1 + FSH showed similar levels of reactive oxygen species, glutathione and active mitochondria (P > 0.05). In conclusion, the IGF-1 protein is present in all ovarian follicle stages in sheep. Moreover, the association between 50 ng/mL IGF-1 and FSH has a synergistic effect in vitro, increasing the percentage of fully grown oocytes and the intensity of immunostaining of LHR protein in oocytes and granulosa cells of cultured antral follicles.
本研究分析了绵羊卵巢中胰岛素样生长因子-1（IGF-1）蛋白的免疫染色情况、IGF-1单独作用或与促卵泡激素（FSH）联合作用对次级卵泡培养的影响，以及培养前后窦状卵泡中促黄体生成素受体（LHR）蛋白的免疫染色情况。采集卵巢用于IGF-1蛋白分析。在实验1中，将次级卵泡培养于α-改良 Eagle 培养基（对照）或添加了IGF-1（10、50或100 ng/mL）的α-改良 Eagle 培养基中。在实验2中，卵泡培养于实验1的相同培养基中并添加750 ng/mL FSH。此外，分析了新鲜窦状卵泡以及在50 ng/mL IGF-1 + FSH中培养后的LHR免疫染色情况。IGF-1蛋白在卵泡发育各阶段的卵母细胞以及次级和窦状卵泡的颗粒细胞中均有免疫定位。IGF-1对卵泡活力和生长无影响（P > 0.05，实验1）。然而在实验2中，50 ng/mL IGF-1 + FSH刺激了卵母细胞生长（P < 0.05）以及窦状卵泡中的LHR免疫染色。对照培养基、10或50 ng/mL IGF-1 + FSH显示出相似水平的活性氧、谷胱甘肽和活性线粒体（P > 0.05）。总之，IGF-1蛋白存在于绵羊所有卵巢卵泡阶段。此外，50 ng/mL IGF-1与FSH联合在体外具有协同作用，增加了完全成熟卵母细胞的百分比以及培养的窦状卵泡卵母细胞和颗粒细胞中LHR蛋白的免疫染色强度。
