Exploration of Molecular Mechanisms of Immunity in the Pacific Oyster (Crassostrea gigas) in Response to Vibrio alginolyticus Invasion

E. Zhang, Zan Li, Luyao Dong, Yanwei Feng, Guohua Sun, Xiaohui Xu, Zhongping Wang, Cuiju Cui, Weijun Wang, Jianmin Yang
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Abstract

Simple Summary As a filter-feeding and sessile invertebrate living in estuaries and intertidal areas, Crassostrea gigas must cope with and adapt to a dynamic and changeable environment. Vibrio alginolyticus is a Gram-negative bacterium that is widespread in oceans and estuaries, and is one of the main Vibrio species that cause oyster disease. In this study, we used transcriptome sequencing to help us better understand how the giant oyster adapts to pathogen-rich environments. By focusing on the gills, which play a crucial role in the immune response, we aimed to shed light on the molecular processes underlying the interaction between the oyster and the pathogen. Abstract Over the years, oysters have faced recurring mass mortality issues during the summer breeding season, with Vibrio infection emerging as a significant contributing factor. Tubules of gill filaments were confirmed to be in the hematopoietic position in Crassostrea gigas, which produce hemocytes with immune defense capabilities. Additionally, the epithelial cells of oyster gills produce immune effectors to defend against pathogens. In light of this, we performed a transcriptome analysis of gill tissues obtained from C. gigas infected with Vibrio alginolyticus for 12 h and 48 h. Through this analysis, we identified 1024 differentially expressed genes (DEGs) at 12 h post-injection and 1079 DEGs at 48 h post-injection. Enrichment analysis of these DEGs revealed a significant association with immune-related Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. To further investigate the immune response, we constructed a protein–protein interaction (PPI) network using the DEGs enriched in immune-associated KEGG pathways. This network provided insights into the interactions and relationships among these genes, shedding light on the underlying mechanisms of the innate immune defense mechanism in oyster gills. To ensure the accuracy of our findings, we validated 16 key genes using quantitative RT-PCR. Overall, this study represents the first exploration of the innate immune defense mechanism in oyster gills using a PPI network approach. The findings provide valuable insights for future research on oyster pathogen control and the development of oysters with enhanced antimicrobial resistance.
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探索太平洋牡蛎(Crassostrea gigas)应对溶藻弧菌入侵的免疫分子机制
简单摘要 作为一种生活在河口和潮间带的滤食性无脊椎动物,巨牡蛎必须应对和适应动态多变的环境。溶藻弧菌是一种革兰氏阴性细菌,广泛存在于海洋和河口地区,是导致牡蛎疾病的主要弧菌之一。在这项研究中,我们利用转录组测序帮助我们更好地了解巨牡蛎如何适应病原体丰富的环境。通过重点研究在免疫反应中起关键作用的鳃,我们旨在揭示牡蛎与病原体之间相互作用的分子过程。摘要 多年来,牡蛎在夏季繁殖季节经常面临大量死亡的问题,弧菌感染是一个重要因素。经证实,巨头鲈的鳃丝小管处于造血位置,可产生具有免疫防御能力的血细胞。此外,牡蛎鳃的上皮细胞会产生免疫效应物质来抵御病原体。有鉴于此,我们对感染藻溶性弧菌 12 小时和 48 小时的巨牡蛎鳃组织进行了转录组分析。这些 DEG 的富集分析表明,它们与免疫相关的基因本体(Gene Ontology,GO)术语和京都基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路有显著关联。为了进一步研究免疫反应,我们利用富集在免疫相关 KEGG 通路中的 DEGs 构建了一个蛋白质-蛋白质相互作用(PPI)网络。该网络揭示了这些基因之间的相互作用和关系,揭示了牡蛎鳃先天免疫防御机制的内在机制。为确保研究结果的准确性,我们利用定量 RT-PCR 验证了 16 个关键基因。总之,本研究是首次利用 PPI 网络方法探索牡蛎鳃的先天免疫防御机制。这些发现为今后牡蛎病原体控制研究和开发具有更强抗菌能力的牡蛎提供了宝贵的见解。
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