Morphological Structure Observation And Differential Expressed MiRNAs Analysis Of Normal Developed And Underdevelop Female Schitosoma Japonicum

Female worms from single sex infections(SSIF) can’t develop to mature worm, and also can’t produce normal eggs. Eggs produced by normal female worms are the main factor that cause pathological damage and transmission of schistosomiasis, this made the study of female reproductive system development important. In order to further understand the mechanism of female reproductive system development, we isolate female worms from single-sex or double-sex cercariae-infected mice. Those parasites’ morphological structure observed and differential expressed mi RNA were analysed.1. Morphological structure observation and apoptosis detection of normal developed and underdevelop female worms.CLMS were adopted to observe the morphological structure. TEM were adopted to observe the vitelline sub-cellular structure. TUNEL were used to detect apoptosis of vitelline cells. The signficant different of worm size, organ size and cell status were found. TEM results show there are no droplet in the underdeveloped female worms which was a frequent structure in the developed female worms. Base of the above result, we detect the apoptopsis in the vitelline there are no droplet in the underdeveloped female worm’s vitelline cells, which we found both sides show mildly apoptopsis. During our experiment, we found an interesting phenomenon that adult female worms contain specific fluorescence phenomenon, which failure to found in male worms and immature(Infectious after 14 days) female worms, but its substance basis need further exploration.2. Analysis of differential expressed miRNA between normal developed and underdeveled female wormsHiseq deep sequencing were adopted to reveal the differential expressed mi RNAs. After date clean and bioinformatic analysis, 9358376 and 9314275 clear resds were got respectively. Among them 5237397 and 5244859 tags were matched to genome respectively, 2301243 and 3955847 reads were matched to precursor/mature mi RNA(Align to mi RBase19). After compare the known mi RNA expression between two samples, 38 differentially expressed mi RNA were found(21 were up regulated, 17 were down regulated in underdevelop female). mi RNA which was abundant expressed such as: mi R-1, mi R-let-7, mi R-7-5P, mi R-124-3p, mi R-36-3p were found higher level in underdeveloped female. Bantam,one of highly expressed mi RNAs, was found lower level in underdeveloped female worms. After compare to the known function of similar mi RNA involved in other species, we hypothesis that those abundant expressed mi RNAs may inhibit underdeveloped female reproduction system’s cells differentiation, but it still need more detail work to verified.3. Preliminary explore of let-7 functionBased on the Hiseq sequence result, let-7 was found higher level in immature female, thus made us try to explore its function via injection mi RNA mimics in the host. Firstly we test the potential target mi RNAs by RT-PCR, and observe the tegument of female to value the effection. The results show let-7 mimics can down regulated many predicted target genes. Secondly, female worms 42 d after infection were counted, individual female worm’s egg produce amount and egg hatching rates were statistics, the results show egg hatching rates significantly rised and egg granuloma reduced obviously. SEM results show there are no significant difference between groups.