中国水产科学2024，Vol.31Issue(7)：766-779,14.DOI:10.12264、JFSC2024-0115
驱动蛋白基因调节克氏原螯虾血淋巴细胞的吞噬作用
Kinesin-related genes regulate phagocytosis in Procambarus clarkii hemocytes
摘要
Abstract
Procambarus clarkii is an economically important aquaculture species in China,and pathogens,especially Vibrio parahaemolyticus,have caused significant economic losses in large-scale farming.However,the mechanism of the immune system response to pathogen invasion remains unclear.To delve into the molecular mechanisms behind the immune regulation of the Procambarus clarkii,this study analyzed the transcriptome data of its hemocytes that had been co-cultured with 3-hydroxybutyrate.By analyzing the transcriptome data of hemocytes from P.clarkii co-cultured with 3-hydroxybutyrate,significant differential expression of cytoskeleton genes was observed,suggesting a potential association with immune regulation in P.clarkii.This study annotated and classified cytoskeleton genes and validated their impact on cellular immune regulation.The annotation results showed that out of five cytoskeleton genes,four belonged to the kinesin family(KIF).All contained KISc domains,and they encoded amino acids primarily composed of glutamate and leucine.The secondary protein structure was mainly composed of α-helices.The genes encoding KIFs were generally similar across species,except for variations in the number,type of conserved motifs,and gene structure among kinesin-like-protein genes in different species.Quantitative reverse transcription-polymerase chain reaction results showed that the expression levels of KIFs were higher in the hemopoietic tissues and gills of P.clarkii than in tissues such as eyestalks and muscles(P<0.05).RNA interference results demonstrated that the optimal knockdown effect was achieved with a dose of 0.10 μg/g body weight of dsKIF11,reducing the expression level of KIF11 to 8.01%of its normal level(P<0.05).Moreover,compared to the control group,after the knockdown of the KIF11 gene,the phagocytic rates of hemocytes towards green-fluorescence-labeled V.parahaemolyticus and red fluorescent microspheres decreased by 36.26%(P<0.01)and 41.94%(P<0.01),respectively.The abovementioned results indicate that KIFs exhibit high conservation and are involved in the phagocytic activity of hemocytes.This finding provides a basis for further investigation of the function and mechanism of KIFs in innate immunity.关键词
克氏原螯虾/血淋巴细胞/驱动蛋白/吞噬作用/免疫Key words
Procambarus clarkii/hemocyte/kinesin/phagocytosis/immune分类
农业科技引用本文复制引用
杜国儒,解伟,左佳俊,刘伟,王雪惠,隋丽英..驱动蛋白基因调节克氏原螯虾血淋巴细胞的吞噬作用[J].中国水产科学,2024,31(7):766-779,14.基金项目
天津市科技计划项目(22YFZCSN00200) （22YFZCSN00200）
天津市海洋资源与化学重点实验室(天津科技大学)开放基金资助项目(202303). （天津科技大学）
