Abstract：
Neuromedin S (NMS) plays key roles in reproductive regulation, while its function and mechanism in follicular development remain unclear. The current study aims to investigate the specific role and mechanisms of NMS and its receptors in regulating the proliferation and steroidogenesis of ovarian granulosa cells (GCs). Phenotypically, a certain concentration of NMS addition promoted the proliferation and estrogen production of goat GCs, accompanied by an increase in the G1/S cell population and upregulation of the expression levels of cyclin D1, cyclin dependent kinase 6, steroidogenic acute regulatory protein, cytochrome P450, family 11, subfamily A, polypeptide 1, 3beta-hydroxysteroid dehydrogenase, and cytochrome P450, family 11, subfamily A, polypeptide 1, while the effects of NMS treatment were effectively hindered by knockdown of neuromedin U receptor type 2 (NMUR2). Mechanistically, activation of NMUR2 with NMS maintained endoplasmic reticulum (ER) calcium (Ca2+) homeostasis by triggering the PLCG1-IP3R pathway, which helped preserve ER morphology, sustained an appropriate level of endoplasmic reticulum unfolded protein response (UPRer), and suppressed the nuclear translocation of activating transcription factor 4. Moreover, NMS maintained intracellular Ca2+ homeostasis to activate the calmodulin 1-large tumor suppressor kinase 1 pathway, ultimately orchestrating the regulation of goat GC proliferation and estrogen production through the Yes1 associated transcriptional regulator-ATF4-c-Jun pathway. Crucially, the effects of NMS were mitigated by concurrent knockdown of the NMUR2 gene. Collectively, these data suggest that activation of NMUR2 by NMS enhances cell proliferation and estrogen production in goat GCs through modulating the ER and intracellular Ca2+ homeostasis, leading to activation of the YAP1-ATF4-c-Jun pathway. These findings offer valuable insights into the regulatory mechanisms involved in follicular growth and development, providing a novel perspective for future research.
摘要：
NeuromedinS(NMS)在生殖调节中起关键作用，而其在卵泡发育中的作用和机制尚不清楚。本研究旨在探讨NMS及其受体在调节卵巢颗粒细胞（GCs）增殖和类固醇生成中的具体作用和机制。表型，添加一定浓度的NMS可促进山羊GCs的增殖和雌激素的产生，伴随着G1/S细胞群体的增加和细胞周期蛋白D1，细胞周期蛋白依赖性激酶6，类固醇生成急性调节蛋白的表达水平的上调，细胞色素P450家族11亚家族A,多肽1，3β-羟基类固醇脱氢酶，细胞色素P450家族11亚家族A,多肽1，而NMS治疗的作用被神经介肽U受体2型（NMUR2）的敲低有效地阻碍。机械上,用NMS激活NMUR2通过触发PLCG1-IP3R途径维持内质网（ER）钙（Ca2）稳态，有助于保持ER形态，维持适当水平的内质网未折叠蛋白反应（UPRer），并抑制激活转录因子4的核易位。此外，NMS维持细胞内Ca2稳态以激活钙调蛋白1-大肿瘤抑制激酶1途径，最终通过Yes1相关的转录调节因子-ATF4-c-Jun途径协调山羊GC增殖和雌激素产生的调节。至关重要的是，NMUR2基因的同时敲除减轻了NMS的影响.总的来说，这些数据表明，NMS激活NMUR2通过调节内质网和细胞内Ca2+稳态增强山羊GCs的细胞增殖和雌激素产生，导致激活YAP1-ATF4-c-Jun途径。这些发现为卵泡生长和发育的调节机制提供了有价值的见解，为未来的研究提供了新的视角。
