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凡纳滨对虾不同类型血细胞和淋巴器官的免疫功能研究 | |
孙明哲![]() | |
Subtype | 博士 |
Thesis Advisor | 李富花 |
2019-05 | |
Degree Grantor | 中国科学院大学 |
Place of Conferral | 中国科学院海洋研究所 |
Degree Discipline | 海洋生物学 |
Keyword | 凡纳滨对虾 血细胞 Oka Trim9 Wssv |
Abstract | 血细胞和淋巴器官(Oka)是对虾重要的免疫组织或器官,在对虾先天免疫中发挥着重要作用。尽管已有研究对血细胞在对虾先天免疫中的功能有了一定认识,也探讨了对虾淋巴器官的形态特征和免疫功能。然而,目前对不同类型血细胞在对虾先天免疫中的功能关系尚缺乏系统认识,对对虾淋巴器官的免疫学功能缺乏系统而深入的研究。本论文以凡纳滨对虾为研究对象,建立了不同类型血细胞分离的技术,并通过转录组测序和生物信息学分析,系统探讨了对虾不同类型血细胞之间的功能关系;分析了对虾淋巴器官发挥生物学功能的分子基础,并筛选获得了对虾淋巴器官特异表达的基因,研究了相关基因的功能。研究结果为系统阐释对虾的先天免疫提供了重要基础,也为发展对虾病害防治的有效策略提供了重要参考。论文的主要进展如下:
对虾E3泛素连接酶TRIM9基因的鉴定和功能分析:在凡纳滨对虾中鉴定了两个TRIM9基因——LvTRIM9-1和LvTRIM9-2。LvTRIM9-1主要在Oka中表达,而LvTRIM9-2呈现泛表达模式,在肠和神经等组织中表达量较高。在WSSV感染后,LvTRIM9-1和LvTRIM9-2的表达呈现明显不同的表达模式,说明其在WSSV感染中的作用存在差异。利用RNAi技术分别沉默两者的表达,均能抑制WSSV在对虾体内的增殖。酵母双杂交与基因表达调控分析结果显示,LvTRIM9-1能够作用于LvIMD从而正向调控NF-κB途径,而LvTRIM9-2可以作用于Lvβ-TrCP来负向调控NF-κB途径。说明LvTRIM9-1和LvTRIM9-2通过不同的作用方式来调控对虾的NF-κB信号通路,从而在对虾体液免疫中发挥作用。 |
Other Abstract | Hemocytes and lymphoid organ (named as Oka) are key immune tissues or organs in shrimp, and they play essential roles in innate imunity of shrimp. Till now, previous studies focused on function of all hemocytes in innate immunity and the morphological observation of Oka, but systematic knowledge of the immune relationship among different types of hemocytes and immunological function of Oka is very limited. Therefore, based on the separation technique of different types of hemocytes, we conducted transcriptome sequencing and bioinformatics analysis to systematically investigate the immunological function of different types of hemocytes in Pacific white shrimp Litopenaeus vannamei. Moreover, we filtered the specifically expressed genes of Oka using the differential expressed genes (DEGs) based on the transcriptome of different tissues, and further analyzed the immunological functions of these selected genes to systematically uncover the immune roles of Oka in the dissertation. This study provided important basis for systematic interpretation of innate immunity, and essential reference for effective stragy of disease prevention in shrimp.
Identification and functional study of E3 ubiquitin ligase TRIM9 genes in shrimp. Two TRIM9 genes (designated as LvTRIM9-1 and LvTRIM9-2) were identified from the transcriptome of the Pacific whiteleg shrimp. The transcript of LvTRIM9-1 was specially detected in Oka, while the transcript of LvTRIM9-2 was detected in most tissues of the shrimp, with relatively higher levels in intestines and nerves. The expression levels of LvTRIM9-1 and LvTRIM9-2 were different post white spot syndrome virus (WSSV) infection, indicating that they played distinct roles in WSSV infection. Knockdown of two LvTRIM9 genes’ expressions by double-strand RNA mediated interference could both lead to a decrease of virus copy number in WSSV-infected shrimp. Yeast two-hybrid analysis and gene regulation analysis showed that LvTRIM9-1 could positively regulate NF-κB pathway via directly interact with immune deficiency (LvIMD), and LvTRIM9-2 could negatively regulate NF-κB pathway via directly interact with beta-transducin repeat-containing protein of shrimp (Lvβ-TrCP). These data indicated that LvTRIM9-1 and LvTRIM9-2 affected humoral immunity in different ways via distinct regulation on NF-κB pathway in shrimp. |
MOST Discipline Catalogue | 理学::海洋科学 |
Language | 中文 |
Document Type | 学位论文 |
Identifier | http://ir.qdio.ac.cn/handle/337002/156781 |
Collection | 实验海洋生物学重点实验室 |
Recommended Citation GB/T 7714 | 孙明哲. 凡纳滨对虾不同类型血细胞和淋巴器官的免疫功能研究[D]. 中国科学院海洋研究所. 中国科学院大学,2019. |
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孙明哲-毕业论文.pdf(17300KB) | 学位论文 | 暂不开放 | No License | Application Full Text |
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