Characterization and expression analysis of prohibitin in the testis of Chinese mitten crab Eriocheir sinensis

Prohibitin plays a key role in maintaining mitochondrial membrane integrity and retaining its normal function. We have initially cloned and sequenced the cDNA of prohibitin from testis of the crab Eriocheir sinensis. The 1,357 bp Prohibitin cDNA comprises a 105 bp 5' untranslated region, a 427 bp 3' untranslated region and a 825 bp open reading frame. Protein alignment substantiates that the Prohibitin has 70.2, 69.8, 70.5, 70.9, 72.4, 70.6 and 74.9% identity with its homologues in Mus musculus, Homo sapiens, Gallus gallus, Danio rerio, Xenopus tropicalis, Drosophila mojavensis and Aedes aegypti, respectively. In situ hybridization revealed that the Prohibitin mRNA was mainly localized around the proacrosomal vesicle and nucleus membrane in early-stage spermatid. In the following middle stage, Prohibitin mRNA was situated inside the invaginated region of half-moon-like nucleus and surrounded the proacrosomal vesicle. In late-stage spermatid, the mRNA was aggregated in the acrosomal tubule, the band between the acrosome and cup-like nucleus, remanent cytoplasm as well. In the mature sperm, mRNA was only found in the acrosomal tubule and the limited space between the nucleus and acrosome. Therefore, we presume that Prohibitin may fulfill critical functions in the spermiogenesis of Eriocheir sinensis.