P62 is a protein adaptor for various metabolic processes. Mice that lack p62 develop adult-onset obesity. However, investigations on p62 in reproductive dysfunction are rare. In the present study, we explored the effect of p62 on the reproductive system. P62 deficiency-induced reproductive dysfunction occurred at a young age (8 week old). Young systemic p62 knockout (p62^-/-) and pituitary-specific p62 knockout (p62^flox/flox αGSU^cre) mice both presented a normal metabolic state, whereas they displayed infertility phenotypes (attenuated breeding success rates, impaired folliculogenesis and ovulation, etc.) with decreased luteinizing hormone (LH) expression and production. Consistently, in an infertility model of polycystic ovary syndrome (PCOS), pituitary p62 mRNA was positively correlated with LH levels. Mechanistically, p62^-/- pituitary RNA sequencing showed a significant downregulation of the mitochondrial oxidative phosphorylation (OXPHOS) pathway. In vitro experiments using the pituitary gonadotroph cell line LβT2 and siRNA/shRNA/plasmid confirmed that p62 modulated LH synthesis and secretion via mitochondrial OXPHOS function, especially Ndufa2, a component molecule of mitochondrial complex I, as verified by Seahorse and rescue tests. After screening OXPHOS markers, Ndufa2 was found to positively regulate LH production in LβT2 cells. Furthermore, the gonadotropin-releasing hormone (GnRH)-stimulating test in p62^flox/flox αGSU^cre mice and LβT2 cells illustrated that p62 is a modulator of the GnRH-LH axis, which is dependent on intracellular calcium and ATP. These findings demonstrated that p62 deficiency in the pituitary impaired LH production via mitochondrial OXPHOS signaling and led to female infertility, thus providing the GnRH-p62-OXPHOS(Ndufa2)-Ca²⁺/ATP-LH pathway in gonadotropic cells as a new theoretical basis for investigating female reproductive dysfunction.

P62 是各种代谢过程的蛋白质接头。缺乏 p62 的小鼠会发展为成年期肥胖症。然而，关于生殖功能障碍中 p62 的研究很少见。在本研究中，我们探讨了 p62 对生殖系统的影响。P62 缺乏引起的生殖功能障碍发生在年轻时（8 周大）。年轻系统性 p62 敲除 (p62 ^-/- ) 和垂体特异性 p62 敲除 (p62 ^flox/flox αGSU ^cre) 小鼠都表现出正常的代谢状态，而它们表现出不育表型（繁殖成功率降低、卵泡发生和排卵受损等），黄体生成素 (LH) 的表达和产生减少。一致地，在多囊卵巢综合征 (PCOS) 的不孕模型中，垂体 p62 mRNA 与 LH 水平呈正相关。从机制上讲，p62 ^-/-垂体 RNA 测序显示线粒体氧化磷酸化 (OXPHOS) 途径显着下调。使用垂体促性腺激素细胞系 LβT2 和 siRNA/shRNA/质粒的体外实验证实 p62 通过线粒体 OXPHOS 功能调节 LH 合成和分泌，特别是 Ndufa2，线粒体复合物 I 的组成分子，已通过 Seahorse 和救援测试验证。在筛选 OXPHOS 标记后，发现 Ndufa2 正向调节 LβT2 细胞中 LH 的产生。^{此外，p62 flox/flox} αGSU ^cre中的促性腺激素释放激素 (GnRH) 刺激试验小鼠和 LβT2 细胞表明 p62 是 GnRH-LH 轴的调节剂，该轴依赖于细胞内钙和 ATP。这些发现表明，垂体中 p62 缺乏通过线粒体 OXPHOS 信号传导损害 LH 的产生并导致女性不育，从而为促性腺细胞中的 GnRH-p62-OXPHOS(Ndufa2)-Ca ²⁺ /ATP-LH 通路提供了新的理论基础用于调查女性生殖功能障碍。