Key Laboratory of Aquacultural Biotechnology, Ningbo University, Ministry of Education, Ningbo 315211, China.
Key Laboratory of Marine Biotechnology of Zhejiang Province, Ningbo University, Ningbo 315211, China.
Int J Mol Sci. 2023 Jun 12;24(12):10030. doi: 10.3390/ijms241210030.
Mitochondria are essential for spermiogenesis. Prohibitins (PHBs; prohibitin 1, PHB1 or PHB, and prohibitin 2, PHB2) are evolutionarily conserved and ubiquitously expressed mitochondrial proteins that act as scaffolds in the inner mitochondrial membrane. In this study, we analyzed the molecular structure and dynamic expression characteristics of -PHBs, observed the colocalization of -PHB1 with mitochondria and polyubiquitin, and studied the effect of knockdown on mitochondrial DNA (mtDNA) content, reactive oxygen species (ROS) levels, and apoptosis-related gene expression in spermatids. Our aim was to explore the effect of -PHBs on mitochondrial function during the spermiogenesis of (), an economically important species in China. The predicted -PHB1/PHB2 proteins contained an N-terminal transmembrane, a stomatin/prohibitin/flotillin/HflK/C (SPFH) domain (also known as the prohibitin domain), and a C-terminal coiled-coil domain. mRNA were widely expressed in the different tissues, with elevated expression in the testis. Further, PHB1 and PHB2 were highly colocalized, suggesting that they may function primarily as an -PHB compiex in . PHB1 proteins were mainly expressed and localized in mitochondria during spermiogenesis, implying that their function may be localized to the mitochondria. In addition, PHB1 was colocalized with polyubiquitin during spermiogenesis, suggesting that it may be a polyubiquitin substrate that regulates mitochondrial ubiquitination during spermiogenesis to ensure mitochondrial quality. To further investigate the effect of -PHBs on mitochondrial function, we knocked down - and observed a decrease in mtDNA content, along with increases in ROS levels and the expressions of mitochondria-induced apoptosis-related genes , , and mRNA. These findings indicate that PHBs might influence mitochondrial function by maintaining mtDNA content and stabilizing ROS levels; in addition, PHBs might affect spermatocyte survival by regulating mitochondria-induced apoptosis during spermiogenesis in .
线粒体对于精子发生是必不可少的。抑制素（PHBs；抑制素 1，PHB1 或 PHB，和抑制素 2，PHB2）是进化上保守的和普遍表达的线粒体蛋白，它们在内膜作为支架。在这项研究中，我们分析了 PHB 的分子结构和动态表达特征，观察了 PHB1 与线粒体和多聚泛素的共定位，并研究了 PHB 敲低对精子细胞中线粒体 DNA（mtDNA）含量、活性氧（ROS）水平和凋亡相关基因表达的影响。我们的目的是探讨 PHB 在精子发生过程中线粒体功能的影响（），在中国是一种经济上重要的物种。预测的 PHB1/PHB2 蛋白含有一个 N 端跨膜结构域、一个 stomatin/prohibitin/flotillin/HflK/C（SPFH）结构域（也称为抑制素结构域）和一个 C 端卷曲螺旋结构域。mRNA 在不同组织中广泛表达，在睾丸中表达升高。此外，PHB1 和 PHB2 高度共定位，表明它们可能主要作为一个 -PHB 复合物在中发挥作用。PHB1 蛋白在精子发生过程中主要表达和定位于线粒体，这意味着它们的功能可能定位于线粒体。此外，PHB1 在精子发生过程中与多聚泛素共定位，表明它可能是一种多聚泛素底物，可调节精子发生过程中线粒体的泛素化，以确保线粒体质量。为了进一步研究 PHB 对线粒体功能的影响，我们敲低了 PHB，并观察到 mtDNA 含量减少，ROS 水平升高，以及线粒体诱导的凋亡相关基因、、和 mRNA 的表达增加。这些发现表明 PHB 可能通过维持 mtDNA 含量和稳定 ROS 水平来影响线粒体功能；此外，PHB 可能通过调节精子发生过程中线粒体诱导的凋亡来影响精母细胞的存活。
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