Characterisation, expression and possible functions of prohibitin during spermatogenesis in the silver pomfret Pampus argenteus

Abstract

Mitochondria play an important role in spermatogenesis, and some mitochondrial proteins are specifically related to this process. In this study we investigated the cytological characteristics of spermatogenic cells, including mitochondrial dynamics, during spermatogenesis in Pampus argenteus. In addition, we characterised the mitochondria-related protein prohibitin (PHB), which has been reported to play roles in mitochondrial dynamics and animal fertility. The full-length cDNA of the P. argenteus phb gene (Pa-phb) is 1687bp, including a 102-bp 5′-untranslated region (UTR), a 772-bp 3′-UTR and an 813-bp open reading frame encoding 271 amino acids. The predicted P. argenteus PHB protein (Pa-PHB) contains three functional domains (a transmembrane domain, an SPFH domain (the conserved region of stomatins, prohibitins, flotillins and HflK/C) and a coiled-coil domain) and exhibits high similarity with its homologue in other animals. The Pa-phb gene was widely expressed in all tissues examined, especially the liver and heart. We primarily focused on Pa-phb expression during spermatogenesis after observing the cytological features of male germ cells, and found that Pa-phb transcripts were detected throughout the course of development of male germ cells. Notably, we observed colocalised signals of Pa-PHB and mitochondria, which were distributed in the cytoplasm around the nucleus in spermatogonia, spermatocytes and early spermatids, tended to move to one side of the cell in middle spermatids and, finally, were colocalised in the sperm midpiece. These observations indicate that Pa-PHB is primarily localised in mitochondria during spermatogenesis, indicating that it has a role in mitochondria. Based on the results of this and previous studies regarding the essential roles of PHB in mitochondria and spermatogenesis in animals, we propose a functional model for PHB during spermatogenesis, including possible roles in the proliferation of spermatogonia and in the regulation of mitochondrial morphology and function in spermatogenic cells.