渔业研究2024，Vol.46Issue(6)：563-571,9.DOI:10.14012/j.jfr.2024087
AC3介导草鱼GCRV感染的功能机制研究
Study on functional mechanisms of AC3-mediated GCRV infection in Ctenopharyngodon idella
摘要
Abstract
[Background]Grass carp reovirus(GCRV)infection can cause leakage bleeding by increasing vascu-lar endothelial permeability.[Objective]To investigate the functional mechanism of adenylate cyclase 3(AC3)mediated GCRV infection,a series of studies were conducted in Ctenopharyngodon idella kidney(CIK)cells.[Methods]AC3 overexpression was successfully achieved in CIK cells.Real-time fluorescent quantitative polymerase chain reaction(qPCR)was used to detect the relative expression of ac3,claudin c,claudin 15,claudin 18,irf3,irf7,ifn1 and ifn2 at different time points(0 h,12 h,and 24 h)of GCRV infection.The protein level of GCRV VP7 was detected by Western blot(WB).The transendothelial electrical resistance(TEER)of the CIK cell layer under various conditions was measured using a cell resistance meter,while the connections between CIK cells were examined using a transmission electron microscope.[Results]The results showed that AC3 overexpression significantly enhanced the expression of tight junction molecules claudin 15 and claudin 18 in CIK cells,but had no significant effect on the expression levels of interfering semaphorin pathway genes irf3,irf7,ifn1 and ifn2.However,the protein amount of GCRV VP7 was inhibited.The transepithelial electrical res-istance assay demonstrated that AC3 overexpression significantly increased the resistance of the CIK cell layer.Transmission electron microscopy results indicated that AC3 overexpression reduced the extent of damage to CIK cell-to-cell junctions caused by GCRV infection.[Conclusion]In summary,AC3 can influence the relat-ive expression of claudin molecules and the barrier function of cells,inhibiting GCRV proliferation within these cells.[Significance]This study introduces a novel approach for discovering GCRV resistance-related mo-lecules from the perspective of cell permeability.关键词
草鱼/腺苷酸环化酶3(AC3)/草鱼呼肠孤病毒(GCRV)/细胞连接/细胞层通透性Key words
Ctenopharyngodon idella/adenylate cyclase 3(AC3)/grass carp reovirus(GCRV)/cell junction/cell permeability分类
农业科技引用本文复制引用
段伟,唐浩,孙明雪,廖伊健,肖调义,李耀国..AC3介导草鱼GCRV感染的功能机制研究[J].渔业研究,2024,46(6):563-571,9.基金项目
湖南省自然科学基金项目(2024JJ5199) （2024JJ5199）
