College of Life and Health Science, Anhui Science and Technology University, Fengyang, China.
Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan, China.
PeerJ. 2023 Jan 16;11:e14612. doi: 10.7717/peerj.14612. eCollection 2023.
Communication between oocytes and granulosa cells ultimately dictate follicle development or atresia. Melatonin is also involved in follicle development. This study aimed to investigate the effects of melatonin and its receptor antagonists on hormone secretion, as well as gene expression related to hormone synthesis, TGF- superfamily, and follicle development in bovine granulosa cells, and assess the effects of melatonin in the presence of 4-P-PDOT and luzindole.
Bovine ovaries were collected from a local abattoir and follicular fluid (follicle diameter 5-8 mm) was collected for granulosa cell isolation and culture. Granulosa cells and culture medium were collected 48 h after treatment with melatonin at high dose concentrations (10 M) and low dose concentrations (10 M) in the absence/presence of 4-P-PDOT and luzindole (10 M or 10 M). Furthermore, the expression level of genes related to hormonal synthesis (, , and ), TGF- superfamily (, , , and ), and development (, and ) were detected in each experimental group by real-time quantitative PCR. In addition, the level of hormones in culture medium were detected using ELISA.
Both 10 M and 10 M melatonin doses promoted the secretion of inhibin A and progesterone without affecting the production of inhibin B and estradiol. In addition, both promoted the gene expression of , , , , and , and inhibited the expression of , , , and . When combined with different doses of 4-P-PDOT and luzindole, they exhibited different effects on the secretion of inhibin B, estradiol, inhibin A, and progesterone, and the expression of , , , , and induced by melatonin.
High and low dose melatonin receptor antagonists exhibited different effects in regulating hormone secretion and the expression of various genes in response to melatonin. Therefore, concentration effects must be considered when using luzindole or 4-P-PDOT.
卵母细胞与颗粒细胞之间的通讯最终决定卵泡的发育或闭锁。褪黑素也参与卵泡的发育。本研究旨在探讨褪黑素及其受体拮抗剂对牛颗粒细胞激素分泌以及与激素合成、TGF-超家族和卵泡发育相关基因表达的影响，并评估在 4-P-PDOT 和 luzindole 存在的情况下褪黑素的作用。
从当地屠宰场采集牛卵巢，采集卵泡液（卵泡直径 5-8mm）用于颗粒细胞分离和培养。在高剂量（10 μM）和低剂量（10 μM）褪黑素处理 48 小时后，在无/存在 4-P-PDOT 和 luzindole（10 μM 或 10 μM）的情况下，收集颗粒细胞和培养基。此外，通过实时定量 PCR 检测每个实验组与激素合成（、、和）、TGF-超家族（、、、和）以及发育（、和）相关的基因表达水平。此外，通过 ELISA 检测培养基中激素的水平。
10 μM 和 10 μM 褪黑素剂量均促进抑制素 A 和孕酮的分泌，而不影响抑制素 B 和雌二醇的产生。此外，两者均促进、、、、和的基因表达，并抑制、、、和的表达。当与不同剂量的 4-P-PDOT 和 luzindole 联合使用时，它们对褪黑素诱导的抑制素 B、雌二醇、抑制素 A 和孕酮的分泌以及、、、、和的表达产生不同的影响。
高剂量和低剂量褪黑素受体拮抗剂在调节激素分泌和对褪黑素的各种基因表达方面表现出不同的作用。因此，在使用 luzindole 或 4-P-PDOT 时必须考虑浓度效应。
