Chen Meijing, Yin Xuejiao, Duan Chunhui, Xie Yuchun, Ji Chenghao, Wang Yong, Liu Yueqin, Zhang Yingjie
College of Animal Science and Technology, Hebei Agricultural University, Baoding 071000, China.
College of Animal Science and Technology, Hebei Normal University of Science & Technology, Qinhuangdao 066000, China.
Animals (Basel). 2025 Jun 26;15(13):1884. doi: 10.3390/ani15131884.
Prolactin is a polypeptide hormone that plays a critical role in male reproduction. However, the underlying mechanisms of prolactin-regulated testosterone secretion and the roles of lncRNAs in this process remain unclear. We performed a comprehensive analysis of the testicular tissues of cashmere goats with different prolactin levels by means of RNA-sequencing. Then, we constructed a lncRNA-miRNA-mRNA interaction network by integrating previously submitted testicular mRNA sequencing data. We identified a novel lncRNA named lncRWDD3 and investigated its effects on testosterone synthesis in the Leydig cells of cashmere goat. The primary Leydig cells were used to explore the biological function of lncRWDD3/miR-1388-5p/NPY1R in vitro. This study found that 200 ng/mL of prolactin achieved the highest testosterone secretion in Leydig cells. LncRWDD3 or NPY1R overexpression promoted cAMP levels, testosterone secretion, and related gene expression, while lncRWDD3 or NPY1R interference had the opposite effect. It was found that lncRWDD3 acts on miR-1388-5p as a ceRNA, and neuropeptide Y receptor Y1 (NPY1R) was confirmed to be a target of chi-miR-1388-5p. Our research shows that prolactin regulates the testicular function of cashmere goats via the lncRNA-miRNA-mRNA ceRNA network, and lncRWDD3 acts as a ceRNA to activate NPY1R/cAMP signaling via the sponging of miR-1388-5p in order to govern testosterone synthesis in the Leydig cells of cashmere goats. Our results provide insights for future studies on the molecular mechanism of the prolactin regulation of testicular function in goats.
催乳素是一种在雄性生殖中起关键作用的多肽激素。然而，催乳素调节睾酮分泌的潜在机制以及lncRNAs在此过程中的作用仍不清楚。我们通过RNA测序对不同催乳素水平的绒山羊睾丸组织进行了全面分析。然后，我们整合先前提交的睾丸mRNA测序数据构建了lncRNA-miRNA-mRNA相互作用网络。我们鉴定了一种名为lncRWDD3的新型lncRNA，并研究了其对绒山羊睾丸间质细胞睾酮合成的影响。利用原代睾丸间质细胞在体外探索lncRWDD3/miR-1388-5p/NPY1R的生物学功能。本研究发现，200 ng/mL的催乳素使睾丸间质细胞中的睾酮分泌达到最高水平。lncRWDD3或NPY1R的过表达促进了cAMP水平、睾酮分泌及相关基因表达，而lncRWDD3或NPY1R的干扰则产生相反的效果。研究发现lncRWDD3作为ceRNA作用于miR-1388-5p，并且神经肽Y受体Y1（NPY1R）被证实是chi-miR-1388-5p的靶标。我们的研究表明，催乳素通过lncRNA-miRNA-mRNA ceRNA网络调节绒山羊的睾丸功能，并且lncRWDD3作为ceRNA通过海绵吸附miR-1388-5p来激活NPY1R/cAMP信号，从而调控绒山羊睾丸间质细胞中的睾酮合成。我们的结果为未来研究山羊催乳素调节睾丸功能的分子机制提供了见解。
