燕山羊绒是季节性繁殖动物,是重要的国家遗传资源。本研究旨在探讨催乳素(PRL)在雄鹿附睾功能中的作用。将21个月大的羊绒雄鹿随机分为对照组(CON)和溴隐亭(BCR,催乳素抑制剂,0.06mg/kg体重(BW))医治组。该实验于2020年9月至10月在秦皇岛市进行,中国,持续了30天.在BCR处理前的最后一天(第0天)和BCR处理后的第15天和第30天(第15天和第30天)收集血液。在第30天,所有的钱都被运到了当地的屠宰场,屠宰后立即收集附睾样本。左附睾保存在4%多聚甲醛中进行组织学观察,右附睾立即保存在液氮中进行RNA测序(RNA-seq)。结果表明,到第30天,PRL抑制剂降低了血清PRL和雌二醇(E2)浓度(p<0.05),并趋于降低黄体生成素(LH)浓度(p=0.052)。但在第0天或第15天没有差异(p>0.05)。在卵泡刺激素(FSH)中没有观察到差异(p>0.05),睾酮(T),和二氢睾酮(DHT)浓度在两组之间。PRL受体(PRLR)蛋白主要位于附睾上皮细胞的细胞质和细胞间物质中。PRL抑制剂降低(p<0.05)附睾中PRLR蛋白的表达。在BCR组中,附睾上皮的高度增加,附睾管的直径也是如此(p<0.05)。然而,附睾管的直径减小(p<0.05)。此后,在附睾组织中鉴定出358个差异表达基因(DEGs),其中191人上调,167人下调。基因本体论和京都百科全书的基因和基因组分析显示,ESR2,MAPK10,JUN,ACTL7A,CALML4主要富集在雌激素信号通路,类固醇结合,钙离子结合,GnRH信号通路,cAMP信号通路,和化学致癌作用-活性氧途径,与附睾功能有关。总之,抑制PRL可能通过减少PRLR蛋白的表达和E2的分泌而影响附睾的结构。ESR2,MAPK10,JUN,ACTL7A,CALML4可能是PRL调节附睾生殖功能的关键基因。
Yanshan Cashmere bucks are seasonal breeding animals and an important national genetic resource. This study aimed to investigate the involvement of prolactin (PRL) in the epididymal function of bucks. Twenty eleven-month-old Cashmere bucks were randomly divided into a control (CON) group and a bromocriptine (BCR, a prolactin inhibitor, 0.06 mg/kg body weight (BW)) treatment group. The experiment was conducted from September to October 2020 in Qinhuangdao City,
China, and lasted for 30 days. Blood was collected on the last day before the BCR treatment (day 0) and on the 15th and 30th days after the BCR treatment (days 15 and 30). On the 30th day, all bucks were transported to the local slaughterhouse, where epididymal samples were collected immediately after slaughter. The left
epididymis was preserved in 4% paraformaldehyde for histological observation, and the right
epididymis was immediately preserved in liquid nitrogen for RNA sequencing (RNA-seq). The results show that the PRL inhibitor reduced the serum PRL and estradiol (E2) concentrations (p < 0.05) and tended to decrease luteinizing hormone (LH) concentrations (p = 0.052) by the 30th day, but no differences (p > 0.05) occurred by either day 0 or 15. There were no differences (p > 0.05) observed in the follicle-stimulating hormone (FSH), testosterone (T), and dihydrotestosterone (DHT) concentrations between the two groups. The PRL receptor (PRLR) protein was mainly located in the cytoplasm and intercellular substance of the epididymal epithelial cells. The PRL inhibitor decreased (p < 0.05) the expression of the PRLR protein in the
epididymis. In the BCR group, the height of the epididymal epithelium in the caput and cauda increased, as did the diameter of the epididymal duct in the caput (p < 0.05). However, the diameter of the cauda epididymal duct decreased (p < 0.05). Thereafter, a total of 358 differentially expressed genes (DEGs) were identified in the epididymal tissues, among which 191 were upregulated and 167 were downregulated. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed that ESR2, MAPK10, JUN, ACTL7A, and CALML4 were mainly enriched in the estrogen signaling pathway, steroid binding, calcium ion binding, the GnRH signaling pathway, the cAMP signaling pathway, and the chemical carcinogenesis-reactive oxygen species pathway, which are related to epididymal function. In conclusion, the inhibition of PRL may affect the structure of the
epididymis by reducing the expression of the PRLR protein and the secretion of E2. ESR2, MAPK10, JUN, ACTL7A, and CALML4 could be the key genes of PRL in its regulation of epididymal reproductive function.