Preliminary Study On The Interaction Proteins In Mice Reproductive Disorder Caused By Toxoplasma Gondii Chronic Infection

Toxoplasma gondii?T.gondii?is an intracellular parasitic protozoan,which widely distributes in humans and the warm-blooded animals.In general,T.gondii rarely initiates an acute infection or death,but causes chronic infection without obvious clinical symptoms.The T.gondii chronic infection can cause toxoplasma encephalopathy,adverse pregnancy,and male reproductive disorders.The disorders of male reproduction include cell and tissue damage?germ cell cycle disorders,spermatogenic retardation or cease,changes of sperm parameters,weight loss of testis and epididymal etc.?.However,the interaction mechanism between T.gondii and reproductive is unclear.To study the effect of spermatogenesis and sperm maturation in the testes and epididymides of Kunming mice after infected by T.gondii type II Prugniaud?PRU?strains,we assessed the reproductive parameters through established chronic infection model of T.gondii.The results showed that the rate of sperm deformity increased,the sperm number and survival rate of spermatozoa decreased.The offspring mice with low birth rate and 42.86%for congenital T.gondii infection.Then,we used RNA-seq technique to sequence the RNA of T.gondii infection groups and control groups.The results showed that the expression level of 250 and 418 genes respectively in the testis and epididymis were significant differences(p<0.05,|log₂^{fold change}|?1).After comparing the two sets of data,42 genes were commonly differentially expressed.Annotated to the Gene ontology?GO?database,the bioinformatics analysis found 1696 and 3806differentially expressed genes?DEGs?in the data of the testis and epididymis groups.To understand the function of these DEGs,we used KEGG to analysis.48 DEGs in the testis groups were enriched in 70 pathways;136 DEGs in the epididymis groups were enriched in 100 pathways.Among them,phagosome,cell adhesion molecules?CAMs?,antigen processing and presentation are the most enriched pathways.The dysregulated expression of key enzymes in glycolysis affects the sperm motility.The PTGDS expressed in spermatogenic cells,sertoli cells and leydig cells,which plays an important role in the formation and maintenance of the blood-testis barrier?BTB?.Co-immunoprecipitation?Co-ip?was carried out to screen T.gondii proteins which interact with PTGDS.134 proteins of T.gondii were found,including 41 unknown proteins and 25 proteins annotated to the KEGG pathway.Through various study,ribosomal proteins of T.gondii?RPL4,RPSA,RPL37,RPL6,and RPL18A?might play a crucial role in the invasion,replication and cell disintegration of T.gondii.In the metabolic pathway,SDH and GAPDH might participate in the energy supply of the T.gondii invasion,which could be further studied.To conclude,in this study,we found DEGs by the RNA-seq of testis and epididymitis tissues infected with T.gondii and screened the proteins interacted with PTGDS by Co-ip and LC-MS/MS,which provided reliable data for subsequent studies of the mechanism of reproductive damage caused by T.gondii infection.