Preliminary Research Of Pumilio Gene In The Reproductive System Of Schistosoma Japonicum

Objective:.Understanding the reproductive regulation of schistosoma is crucial for the Schistosomiasis.Pumilio can regulate the reproductive organs development in many species,but how it works in schistosoma is unknown.This paper aims to explore the possible functions of Schistosoma japonicum Pumilio2(Pum2),especially in reproductive system.Methods: The complete coding sequence of Schistosoma japonicum Pumilio1(SjPum1)and Schistosoma japonicum Pumilio2(SjPum2)gene were cloned and characterized.The expression of SjPum1 and SjPum2 were analyzed at different lige-cycle stages and in various tissues by qPCR.We selected the Pum2 gene to explore its functions.In situ hybridization was used to observe the location of SjPum2.Then,using RNA interference(RNAi)techniques to explored the reproductive function of SjPum2.As for siRNA gene-silencing efficiency,morphological alternations and the egg production,qPCR,confocal laser scanning microscopy and light microscopy were used.Subsequently,the expression of those genes,which may interact with apoptosis,were examined using qPCR.Results: We obtained full-length SjPum1(2613 nucleotides)and SjPum2(4479 nucleotides)gene.Bioinformatics analysis showed that SjPum1 and SjPum2 belong to PUF family.Quantitative Real-time PCR analysis revealed that SjPum1 and SjPum2 mRNA were differentially expressed throughout the life-cycle and were highly expressed in reproductive organs.In this paper,we used SjPum2 as the research object.In situ hybridization showed SjPum2 were highly expressed in the ovary and testis.After knockdown SjPum2 gene,laser confocal microscope showed morphological alterations in reproductive organs.We also found the number of eggs was decreased after silencing SjPum2 gene.Moreover,SjPum2 gene silence affected the changes of these genes interacting with apoptosis,including Caspase3 and Caspase7.Conclusion: We amplified the full-length gene of SjPum1 and SjPum2.Using qPCR,we found that Pum1 and Pum2 expressed in different period of Schistosoma japonicum and mainly expressed in reproductive organs.By silencing the expression of SjPum2,obviously reproductive morphological changes can be seen.SjPum2 gene silencing also significantly reduced egg production.Therefore,we conclude that SjPum2 gene affects the development of the reproductive organs and may also be associated with the apoptosis of Schistosoma japonicum.