Rearrangement Of Actin Cytoskeleton By Zika Virus Infection Facilitates Blood-Testis Barrier Hyperpermeability

Zika virus(ZIKV)belongs to the genus Flavivirus of the family Flaviviridae,and its infection can cause severe diseases,such as neonatal microcephaly in children,Guillain-Barré syndrome,oligospermia,haematospermia in adults.The existence of ZIKV in semen and the sexual transmission of ZIKV indicate that it is important to study the pathogenic mechanism of ZIKV causing male reproductive damage,especially the potential mechanism of destroying the blood testis barrier(BTB).Meanwhile,the cellular actin skeleton is closely related to dengue virus,west nile virus and other flavivirus infections and is an important component of BTB,suggesting that the cellular actin skeleton may be closely related to the male reproductive damage caused by ZIKV infection.Objective: This article focuses on the molecular mechanism of the cell actin filament skeleton in ZIKV infection and the destruction of the BTB,and will provide the theoretical and experimental basis for the symptoms of clinical male disease caused by ZIKV and the destruction of the BTB.Methods: Several host molecules interacting with ZIKV were identified by co-immunoprecipitation(Co-IP)and LC-MS/MS,and the interaction between the viral envelope protein and actin was verified by Co-IP.The changes of the microfilament skeleton of mouse Sertoli cells after virus infection or overexpression of viral envelope protein were observed by immunofluorescence assay(IFA).Cytochalasin D or Jasplakinolide was used to treat mouse Sertoli cells before and after infection,and the viral content was measured by real-time quantitative PCR(RT-q PCR),western blotting(WB)and viral plaque assay.In vitro BTB model was established by isolated primary mouse Sertoli cells,and transepithelial resistance was detected after ZIKV infection or envelope protein overexpression,the role of cell actin filament skeleton in the destruction of BTB caused by ZIKV infection and the underlying molecular mechanisms were explored by Co-IP and IFA.Results: LC-MS/MS identified several cytoskeleton molecules that interact with ZIKV,including actin,Tubulin beta-5 chain,etc.ZIKV envelope protein interacts with actin,and both ZIKV-E(1-133)and ZIKV-E(134-301)truncation can interact with actin.ZIKV infection or envelope protein overexpression causes the reorganization of the Sertoli cell actin filament skeleton.Cytochalasin D or Jasplakinolide promotes ZIKV infection of mouse Sertoli cells.ZIKV or envelope protein destroys the in vitro mouse BTB model,which is achieved by reducing the interaction between actin and tight junction protein ZO-1.Conclusion: In mouse Sertoli cells,ZIKV envelope protein interacts with actin,ZIKV infection or viral envelope protein overexpression can lead to the reorganization of the cell actin filament skeleton.The interruption of actin filament dynamics caused by Cytochalasin D or Jasplakinolide will be beneficial to ZIKV infection.ZIKV infection or envelope protein overexpression leads to weakened interaction between actin and tight junction protein ZO-1,which may be one of the mechanisms by which ZIKV destroys BTB.