Identification And Functional Study Of Key Molecules Of Schistosoma Japonicum SjSpenito Regulating Growth And Development Through Spoc Domain

Schistosomiasis is a highly neglected zoonotic parasitic disease that is endemic in 78 countries and territories in Africa,the Americas and Asia,with approximately240 million people infected worldwide and nearly 800 million more threatened.The epidemic of this parasitic disease is a serious threat to the health of people in infected areas,but the current treatment of schistosomiasis relies only on a single drug Praziquantel,and the long-term large-scale use of this drug has led to resistance of schistosomes in some infected areas,so the search for new drug target point and therapeutic means is urgent.Schistosoma eggs are the key to schistosome transmission and pathogenesis,so inhibiting schistosome egg production is essential to interrupting schistosomiasis transmission and reducing the pathological damage caused by eggs to the host organism.Based on the purpose of this study,the laboratory has clarified the differential genes in the early stages of worm pairing through the dynamic expression profiles of transcripts in different developmental stages of male and female worms,and identified key molecules that may be involved in the growth and development of schistosomes and the initiation of sexual maturation in females through analysis and finding that one of those genes knock-down significantly inhibited the growth and development of Schistosoma japonicum.Structural analysis of the protein sequence of this gene revealed that it contains a conserved SPOC domain at the C-terminus and two RRMs domains at the N-terminus,which are consistent with the structural features of the Split end protein family.To further investigate the specific function of this gene in Schistosoma japonicum,we inhibited the expression of the SjSpenito gene in vitro cultures of Schistosoma japonicum and showed that inhibition of the gene resulted in a significant increase in the activity frequency of mature worms,wore pairing of male and female worms,specific bamboo-like movements,and ultimately death of the worms,confirming that this gene is essential for the survival of adult Schistosoma japonicum.Based on the functional characteristics of the SPOC domain,which recruits multiple co-transcription repressors to regulate downstream signal pathway,we performed RNA-seq and ATAC-seq sequencing analysis on interfering worms to investigate more deeply the molecular mechanisms involved in the effects of SjSpenito on worm viability.RNA-seq results show that inhibition of SjSpenito expression can cause significant changes in the expression of a large number of genes in the worm;Inhibiting the expression of can lead to a significant decrease in the vitality of the larvae;In particular,knockdown of SjSpenito caused a decrease in the expression of the co-localized gene,and inhibition of expression also significantly reduced the viability of the worm,suggesting that SjSpenito may affect the viability of Schistosoma japonicum by regulating the expression of downstream genes;In addition,the cellular localization results showed that the significantly differentially expressed genes after knockdown of SjSpenito were mainly located in the neural cell population of the worm,suggesting that SjSpenito may affect the viability of the worm by regulating the normal function of the nervous system.ATAC-seq results show that significant changes in chromatin accessibility that inhibit SjSpinito expression of some transcription factors of the worm;Inhibition of the expression of its reciprocal transcription factor leads to increased excitability and signature bamboo-like movements of the worm;Inhibition of expression leads to a reduction in worm viability;Among them,co-localization of transcription factors with SjSpenito was observed,suggesting that SjSpenito may the movement and vitality of worms by affecting the chromatin accessibility of.In summary,inhibition of SjSpenito expression in vitro cultured worms can lead to increased excitability and ultimately to death of the worms;It was also confirmed that SjSpenito could influence the motility and excitability of the nervous system of the worm by regulating the expression of the downstream gene;Influencing worm viability by regulating chromatin accessibility of reciprocal transcription factors such as.This study not only reveals the molecular mechanism of SjSpenito’s regulation of worm viability,but also provides a theoretical basis for the development of new anti-schistosome vaccines and drug target points.