Characterization of mitochondrial prohibitin from Boleophthalmus pectinirostris and evaluation of its possible role in spermatogenesis

Abstract

Prohibitin (PHB) is an evolutionarily conserved mitochondrial membrane protein. It plays a vital role in cell proteolysis, senescence, and apoptosis and is associated with spermatogenesis and sperm quality control in mammals. To study the characteristics of the PHB gene and its potential roles during spermatogenesis in Boleophthalmus pectinirostris, we cloned a 1153-bp full-length cDNA from the testis of B. pectinirostris with an open reading frame of 816 bp, which encodes 272 amino acid residues. Real-time quantitative PCR (qPCR) analysis revealed the presence of phb mRNA in all the tissues examined, with higher expression levels found in the testis, kidney, intestine, and muscle tissues. We examined the localization of phb mRNA during spermatogenesis by in situ hybridization (ISH), showing that phb mRNA was distributed in the periphery of the nucleus in primary and secondary spermatocytes. In spermatid and mature sperm, the phb mRNA gradually moved toward one side, where the flagellum is formed. Immunofluorescence (IF) results showed co-localization of the PHB and mitochondria at different stages during spermatogenesis of B. pectinirostris. The signals obtained for PHB decreased as spermatogenesis proceeded; the strongest detection signal was found in secondary spermatocytes, with lower levels of staining in other stages. Additionally, in the mature germ cells, the PHB signals were weak and aggregate in the midpiece of the flagellum.

Prohibitin-mediated mitochondrial ubiquitination during spermiogenesis in Chinese mitten crab Eriocheir sinensis

“…In Homo sapiens , R. norvegicus , S. cerevisiae and other species, PHB was identified as a mitochondrial inner membrane protein [23–28]. PHB plays intracellular roles through the formation of similar ring-like PHB complexes embedded in the mitochondrial membrane [33, 34].…”

“…In mitochondria, PHB was also associated with mitophagy, stabilizing mitochondria genome through TFAM (Transcription Factor A, Mitochondrial), ROS (Reactive Oxygen Free Radical) reduction, mitochondrial morphology and mitochondrial apoptosis [18–22]. The function of PHB during spermatogenesis has been reported in many species, including Rattus norvegicus [23], Saccharomyces cerevisiae [24], Caenorhabditis elegans [25], Procambarus clarkii [26], Cynops orientalis [27], and Boleophthalmus pectinirostris [28]. In C. orientalis and B. pectinirostris , the distribution of phb gene or PHB protein in spermatogenesis was reported [25, 28].…”

“…The function of PHB during spermatogenesis has been reported in many species, including Rattus norvegicus [23], Saccharomyces cerevisiae [24], Caenorhabditis elegans [25], Procambarus clarkii [26], Cynops orientalis [27], and Boleophthalmus pectinirostris [28]. In C. orientalis and B. pectinirostris , the distribution of phb gene or PHB protein in spermatogenesis was reported [25, 28]. In both R. norvegicus and P. clarkia , the PHB signal was detected in spermatogonia, spermatocytes and spermatids [23, 27].…”

Characterization of Mitochondrial Prohibitin in Opsariichthys bidens and Its Potential Functions in Spermatogenesis

“…In these species, PHB is highly expressed in spermatids, where it not only regulates mitochondrial degradation but also participates in nuclear shaping and acrosome formation [30][31][32]. In addition, in some teleosts such as the mudskipper Boleophthalmus pectinirostris and silver pomfret Pampus argenteus, PHB is expressed at all stages of spermatogenesis and maintains the normal morphology and function of mitochondria [33,34]. Therefore, these results promote an understanding of the relationship between PHB and spermatogenesis.…”

Comparative Transcriptome Analyses of Schistosoma japonicum Derived From SCID Mice and BALB/c Mice: Clues to the Abnormality in Parasite Growth and Development

“…They are interdependent on the protein level, and loss of one simultaneously leads to the loss of the other. The homologous gene of prohibitin in other animals was reported to play a vital role in the mitochondria's function, cell proteolysis, senescence, and apoptosis and is essential for embryonic viability and germline function-spermatogenesis and folliculogenesis (Artal-Sanz et al, 2003;He et al, 2008;Tyc et al, 2010;Peng et al, 2015;Chowdhury et al, 2016;Jin et al, 2016;Nguyen et al, 2016;Chai et al, 2017;Xu et al, 2017;Wang D. et al, 2017).…”