Department of Veterinary Biochemistry, College of Veterinary Science, Sri Venkateswara Veterinary University, Tirupati, Andhra Pradesh, India.
Department of Veterinary Biochemistry, College of Veterinary Science, Sri Venkateswara Veterinary University, Proddatur, Andhra Pradesh, India.
Reprod Domest Anim. 2023 Sep;58(9):1270-1278. doi: 10.1111/rda.14430. Epub 2023 Jul 13.
The present study was conducted to ascertain whether the role of kisspeptin in promoting in vitro development of preantral follicles was through the regulation of P450 aromatase gene expression and steroidogenesis in sheep. Accordingly, the cumulus cells and oocytes were collected from different development stages of preantral follicles grown in vivo and cultured in vitro in TCM199B (Group I), TCM199B + KP (10 μg/mL) (Group II) and Standard medium + KP (10 μg/mL). To measure the steroid (Estradiol-17β; E and Progesterone; P ) synthesis through ELISA, spent culture medium was collected separately from the same in vitro groups. E synthesis in the spent medium collected from all the three groups showed an increasing trend from PFs' exposed to respective culture media for 3 min to 2-day culture stage but decreased thereafter till 6-day culture stage. This is followed by a sharp increase in E concentration in the spent medium collected after in vitro maturation. However, P synthesis in group III followed increased pattern as the development progressed from PFs' exposed to culture medium for 3 min to in vitro maturation stage. The steroid production was observed at all stages of in vitro development in altered supplemented conditions. The steroid synthesis in the spent medium was highest in the 6 day cultured PFs' in Standard medium + KP matured in vitro for 24 h. Therefore, supplementation of kisspeptin along with other growth factors promoted steroid production in cultured preantral follicles far better than in other media.
本研究旨在确定 kisspeptin 是否通过调节 P450 芳香化酶基因表达和类固醇生成来促进绵羊前腔卵泡的体外发育。因此,从体内生长和体外培养的不同发育阶段的前腔卵泡中收集卵丘细胞和卵母细胞,分别在 TCM199B(第 I 组)、TCM199B+KP(10μg/mL)(第 II 组)和标准培养基+KP(10μg/mL)(第 III 组)中培养。为了通过 ELISA 测量类固醇(雌二醇-17β;E 和孕酮;P)的合成,分别从相同的体外组中收集用过的培养物。从所有三组中收集的用过的培养基中 E 的合成显示出从 PF 暴露于各自的培养基 3 分钟到 2 天培养阶段逐渐增加的趋势,但此后直到 6 天培养阶段减少。此后,在体外成熟后收集的用过的培养基中 E 浓度急剧增加。然而,第 III 组中 P 的合成在从 PF 暴露于培养基 3 分钟到体外成熟阶段的发育过程中遵循增加的模式。在改变的补充条件下,观察到体外发育的所有阶段的类固醇生成。在标准培养基+KP 中培养 6 天的 PF 在体外成熟 24 小时后,用过的培养基中的类固醇合成最高。因此,与其他生长因子一起补充 kisspeptin 可促进培养的前腔卵泡中的类固醇生成,效果远优于其他培养基。