红螯螯虾CHH2基因的表达特征及其在卵巢发育中的功能  

Expression profiles of CHH2 gene in redclaw crayfish Cherax quadricarinatus and its role in ovarian development

作　　者：陈乐然 郑建波 贾永义[2] 迟美丽[2] 李飞[2] 程顺[2] 刘士力[2] 刘一诺 蒋文枰[2] 顾志敏 CHEN Leran;ZHENG Jianbo;JIA Yongyi;CHI Meili;LI Fei;CHENG Shun;LIU Shili;LIU Yinuo;JIANG Wenping;GU Zhimin(Key Laboratory of Freshwater Aquatic Genetic Resources,Ministry of Agriculture and Rural Affairs,College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306,China;Key Laboratory of Healthy Freshwater Aquaculture,Ministry of Agriculture and Rural Affairs,Key Laboratory of Freshwater Aquaculture Genetic and Breeding of Zhejiang Province,Zhejiang Institute of Freshwater Fisheries,Huzhou 313001,China)

机构地区：[1]上海海洋大学水产与生命学院,农业农村部淡水水产种质资源重点实验室,上海201306 [2]浙江省淡水水产研究所农业农村部淡水渔业健康养殖重点实验室,浙江省淡水水产遗传育种重点实验室,浙江湖州313001

出　　处：《浙江农业学报》2023年第1期33-40,共8页Acta Agriculturae Zhejiangensis

基　　金：湖州市乡村振兴专项(2019ZD2028);浙江省农业(水产)新品种选育重大科技专项(2021C02069-4)。

摘　　要：为研究高血糖激素基因(CHH)在红螯螯虾卵巢发育中的作用,基于转录组学数据鉴定到红螯螯虾CHH2基因序列,并对其基本特性进行生物信息学分析。采用实时荧光定量PCR(qRT-PCR)检测了CHH2在红螯螯虾不同组织和卵巢发育阶段的表达情况。结果表明,CHH蛋白在甲壳类物种中高度保守,同源性高达66.84%。qRT-PCR结果表明,在红螯螯虾的肝胰腺中几乎检测不到CHH2表达,在其他组织均有表达,包括肌肉、鳃、肠、心,且在卵巢中的表达水平最高;另外,CHH2在卵巢发育各个时期(未发育、发育期、快速发育期、成熟期)均有表达,表达水平先升高后降低,在卵巢发育成熟期表达水平最低。此外,在切除单侧眼柄和雌二醇活体注射的红螯螯虾卵巢中,CHH2表达水平显著降低。通过注射体外合成CHH2-dsRNA对红螯螯虾进行RNAi干扰试验,结果显示,卵巢发育重要标记卵黄蛋白原基因(VTG)表达水平显著上升。综上所述,CHH2可能负调控红螯螯虾卵巢发育,研究结果为进一步开发基于RNAi的促性腺成熟技术奠定理论基础。To study the role of crustacean hyperglycemic hormone gene(CHH) in the ovarian development of Cherax quadricarinatus, sequence of CHH2 gene was identified based on transcriptomic data, and the basic characteristics were analyzed by bioinformatic. qRT-PCR was used to detect the expression level of CHH2 in different tissues and ovarian development stages. The results showed that CHH proteins were highly conserved among crustacean species, and they had high homologies with 66.84%. The tissue distribution at transcriptional level displayed that CHH2 was hardly to be detected in hepatopancreas, and was detected at low level in muscle, gills, heart and intestine, while highly expressed in ovary. In addition, CHH2 was expressed in all stages of ovarian development and exhibited an abnormal expression pattern with the lowest expression level at mature stage. On the other hand, CHH2 expression was significantly reduced in ovarian tissues after ESA treatment and E2 injection. Furthermore, VTG was regarded as an important marker for ovarian development, VTG-mRNA appeared to be increased through RNAi-induced gene silencing. In conclusion, CHH2 might negatively regulate ovarian development, and this study would provide new insights for further exploring the mechanism of crustacean ovarian development.

关 键 词：红螯螯虾 高血糖激素基因2(CHH2) RNAI 卵巢发育 卵黄蛋白原 

分 类 号：S917.4[农业科学—水产科学]