Studies On The Different Proteome Between Male And Female Worm And The Functions Of Gynecophoral Canal Protein Gene In Schistosoma Japonicum

Schistosomiasis caused by Schistosoma japonicum is an important zoonosis in China and southeast Asia. Pairing between male and female worm of schistosomes is the key process for successful development of female maturation. Adult female worm can produce a large number of eggs that are retained in the liver of final host where they elicit inflammatory immune responses and also are responsible for the dissemination of parasite. Base on above characters of schistosomes, comparative proteome between male and female worm and the function of Gynecophoral canal protein gene of Schistosoma japonicum (SjGCP) were investigated by two dimensional electrophoresis in combination with mass spectrometry and RNA interference in order to reveal the mechanism of development after pairing.1. Proteomic analysis of differentially expressed proteins between male and female worm of Schistosoma japonicumSoluble proteins, hydrophobic proteins and membrane proteins that were extracted from eggs, schistosomulum, adult male and female worm of Schistosoma japonicum were separated by two dimensional electrophoresis. There were 1016 ± 67, 1808 ± 89, 1142 ± 45 and 1288 ± 32 spots for soluble proteins, 1425±108, 952 ± 59, 847 ± 75 and 965 ± 69 spots for hydrophobic proteins and 78 ± 3, 67 ± 3, 108±4 and 122 ± 4 spots for membrane proteins from egg, schistosomulum, female and male worms of adult stage, respectively. The differentially and uniquely expressed proteins for male worm and female worm after pairing (42 days) include 41±4 and 23 ± 2 unique spots for soluble proteins, 11 ± 1 and 26 ± 3 unique spots for hydrophobic proteins and 4 and 5 unique spots for membrane proteins. MALDI-TOF MS and ESI-MS/MS were employed to analyze 12 unique spots from female worm and 16 unique spots from male worm for peptide mass fingerprinting and sequencing. The results showed that a series of intracellular signal molecules, molecules involved in cellular metabolism and transcriptional regulation were expressed differentially between male and female worm in Schistosoma japonicum. Comparing the proteome of schistosomes between male and female worm after pairing may permit the identification of protein candidates for the development of vaccine or new targets for drug development against schistosomiasis.2. Analysis the functions of gynecophoral canal protein gene in Schistosoma japonicum Gynecophoral canal protein gene of Schistosoma japonicum (SjGCP) is specifically expressed in theadult male worm and its protein shows wide distribution in the adult female worm after pairing. Three dsRN As were designed according to the sequence of gynecophoral canal protein gene. The SjGCP of RNAi parasites was successfully inhibited using RT-PCR, Western blotting and immunofluorescence analysis. Parasites made an 84% reduction in the transcript of SjGCP. Interestingly, pairing between male and female worm was found totally abolished in two groups (sl,s2) that were treated with high concentration of different dsRNA. Electron scanning microscope analysis of the ultrastructure on the surface of gynecophoral tegument in treated parasites shows irregular ridges and short spine. The...