Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, China.
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China.
FASEB J. 2023 Jun;37(6):e22989. doi: 10.1096/fj.202300187R.
Neuromedin S (NMS) is a neuroregulatory substance and has many important roles in regulating physiological functions in animal cells, while their specific functions and mechanisms in Leydig cells (LCs) of the testis remain unclear. The current study aims to investigate the role and potential mechanisms of NMS and its receptors in regulating steroidogenesis and proliferation in goat LCs. We found that NMS and its receptors were mainly expressed in LCs of goat testes at different ages (1-day-old, 3-month-old, and 9-month-old), and the highest expressions detected at age three months. NMS addition significantly enhanced the testosterone secretion, STAR, CYP11A1, 3BHSD, and CYP17A1 expressions, cell proliferation, and PCNA expression in vitro cultured goat LCs. Mechanistically, NMS addition increased G1/S cell population, the expressions of CCND1, CDK4 and CDK6, the activities of SOD2 and CAT, and enhanced the mitochondrial fusion, the production of ATP, and mitochondrial membrane potential, while inhibited cellular ROS production, and maintained a low ubiquitination level of mitochondrial proteins. Notably, these effects of NMS addition on goat LCs were suppressed by co-treatment with NMUR2 knockdown. Therefore, these data suggest that activating NMUR2 with NMS enhances testosterone production and cell proliferation in goat LCs through modulating mitochondrial morphology, function, and autophagy. These findings may provide a novel view of the regulatory mechanisms involved in male sexual maturation.
神经调节素 S（NMS）是一种神经调节物质，在调节动物细胞的生理功能方面具有许多重要作用，但其在睾丸间质细胞（LCs）中的具体功能和机制尚不清楚。本研究旨在探讨 NMS 及其受体在调节山羊 LCs 类固醇生成和增殖中的作用及其潜在机制。我们发现，NMS 及其受体主要在不同年龄（1 天、3 个月和 9 个月）的山羊睾丸 LCs 中表达，其中 3 个月时表达最高。NMS 的添加显著增强了体外培养的山羊 LCs 中睾酮的分泌、STAR、CYP11A1、3BHSD 和 CYP17A1 的表达、细胞增殖和 PCNA 的表达。在机制上，NMS 的添加增加了 G1/S 期细胞群、CCND1、CDK4 和 CDK6 的表达、SOD2 和 CAT 的活性、增强了线粒体融合、ATP 的产生和线粒体膜电位，同时抑制了细胞 ROS 的产生，并保持了低水平的线粒体蛋白泛素化。值得注意的是，NMS 对山羊 LCs 的这些作用被 NMUR2 敲低的共同处理所抑制。因此，这些数据表明，NMS 通过调节线粒体形态、功能和自噬来激活 NMUR2，从而增强山羊 LCs 中的睾酮产生和细胞增殖。这些发现可能为男性性成熟所涉及的调节机制提供了新的视角。
