Yu Jun, Huang Qiuru, Fu Yangbo, He Lei, Shen Cong, Chen Xia, Li Zhiran, Li Jiaxin, Wang Chenyu, Wang Xinda, Yang Binbin, Lin Ziwen, Qiao Chen, Tan Xiaofang, Yang Xiaoqing, Chen Hao, Zheng Ying, Zheng Bo, Sun Fei
Institute of Reproductive Medicine, Medical School, Nantong University, Nantong, 226001, China.
State Key Laboratory of Reproductive Medicine and Offspring Health, Center for Reproduction and Genetics, Suzhou Municipal Hospital, Gusu School, The Affiliated Suzhou Hospital of Nanjing Medical University, Nanjing Medical University, Suzhou, 215002, China.
Cell Mol Biol Lett. 2025 May 9;30(1):58. doi: 10.1186/s11658-025-00737-3.
Testicular aging has profound effects on spermatogenesis, sperm function, and the spermatogenic microenvironment, contributing to reduced male fertility. However, the precise molecular mechanisms by which mitochondria influence spermiogenesis during aging still remain largely unclear.
Vha68-3 KO flies were generated using the CRISPR/Cas9 technique. Testicular phenotypes and functions were mainly observed through immunofluorescence staining and transmission electron microscopy. Multi-omics study was mainly conducted through single-cell RNA sequencing and transcriptome-metabolomics association analysis. Vha68-3 binding proteins were identified via liquid chromatography-tandem mass spectrometry. The therapeutic potential of modulating mitochondrial metabolism for testicular aging mainly relied on the dietary intake of related compounds in fruit flies.
In this study, we identified Vha68-3, a testis-specific subunit of the V-type adenosine triphosphate (ATP) synthase, predominantly localized in the tails of elongated spermatids, as a key age-related regulator of male fertility and spermatid elongation in Drosophila testes. Crucially, Vha68-3 deficiency impaired mitochondrial homeostasis in elongated spermatids during testicular aging. Through a multi-omics approach, including single-cell transcriptomics, protein interaction mapping of Vha68-3, and transcriptome-metabolome integration, we identified pyruvate metabolism as a critical pathway disrupted by Vha68-3 deficiency. Moreover, dietary supplementation with pyruvate (PA), S-lactoylglutathione (SLG), and phosphoenolpyruvate (PEP) effectively alleviated mitochondrial dysfunction and testicular aging linked to Vha68-3 deficiency.
Our findings uncover novel mechanisms by which mitochondrial metabolism regulates spermatid elongation and propose potential therapeutic strategies to combat mitochondrial metabolic disorders in aging testes.
睾丸衰老对精子发生、精子功能和生精微环境有深远影响，导致男性生育能力下降。然而，线粒体在衰老过程中影响精子形成的精确分子机制仍 largely不清楚。
使用CRISPR/Cas9技术构建Vha68-3基因敲除果蝇。主要通过免疫荧光染色和透射电子显微镜观察睾丸表型和功能。多组学研究主要通过单细胞RNA测序和转录组-代谢组关联分析进行。通过液相色谱-串联质谱鉴定Vha68-3结合蛋白。调节线粒体代谢对睾丸衰老的治疗潜力主要依赖于果蝇饮食中相关化合物的摄入。
在本研究中，我们鉴定出Vha68-3，一种V型三磷酸腺苷（ATP）合酶的睾丸特异性亚基，主要定位于延长型精子细胞的尾部，是果蝇睾丸中与年龄相关的男性生育力和精子细胞延长的关键调节因子。至关重要的是，Vha68-3缺陷在睾丸衰老过程中损害了延长型精子细胞中的线粒体稳态。通过多组学方法，包括单细胞转录组学、Vha68-3的蛋白质相互作用图谱绘制以及转录组-代谢组整合，我们确定丙酮酸代谢是被Vha68-3缺陷破坏的关键途径。此外，饮食中补充丙酮酸（PA）、S-乳酰谷胱甘肽（SLG）和磷酸烯醇丙酮酸（PEP）可有效减轻与Vha68-3缺陷相关的线粒体功能障碍和睾丸衰老。
我们的研究结果揭示了线粒体代谢调节精子细胞延长的新机制，并提出了对抗衰老睾丸中线粒体代谢紊乱的潜在治疗策略。
