Institute of Marine Research (IIM), National Research Council (CSIC), Eduardo Cabello, 6, 36208, Vigo, Spain.
Laboratory of Biotechnology and Aquatic Genomics, Interdisciplinary Center for Aquaculture Research (INCAR), University of Concepción, P.O. Box 160-C, Concepción, Chile.
Sci Rep. 2019 Mar 5;9(1):3569. doi: 10.1038/s41598-019-39870-3.
Mediterranean mussels (Mytilus galloprovincialis) are sessile filter feeders that live in close contact with numerous marine microorganisms. As all invertebrates, they lack an adaptive immune response and how these animals are able to respond to a bacterial infection and discriminate it from their normal microbiome is difficult to understand. In this work, we conducted Illumina sequencing of the transcriptome of individual mussels before and after being infected with Vibrio splendidus. The control mussels were injected with filtered seawater. We demonstrate that a great variability exists among individual transcriptomes and that each animal showed an exclusive repertoire of genes not shared with other individuals. The regulated genes in both the control and infected mussels were also analyzed and, unexpectedly, the sampling before the injection was considered a stress stimulus strong enough to trigger and modulate the response in hemocytes, promoting cell migration and proliferation. We found a clear response against the injection of filtered seawater, suggesting a reaction against a tissue injury in which the myticins, the most expressed antimicrobial peptides in mussel, appeared significantly up regulated. Functional experiments with flow cytometry confirmed the transcriptomic results since a significant alteration of hemocyte structures and a decrease in the number of hemocytes positive for myticin C were found only after a Vibrio infection and not observed when mussels were bled before, generating a tissue injury. Therefore, we report the involvement of myticins in the response to a danger signal such as a simple injection in the adductor muscle.
地中海贻贝(Mytilus galloprovincialis)是一种固着滤食动物,与许多海洋微生物密切接触。与所有无脊椎动物一样,它们缺乏适应性免疫反应,因此很难理解这些动物如何能够对细菌感染做出反应,并将其与正常微生物组区分开来。在这项工作中,我们对感染灿烂弧菌前后的单个贻贝进行了 Illumina 转录组测序。对照组贻贝注射过滤海水。我们证明个体转录组之间存在很大的可变性,并且每个动物都表现出与其他个体不共享的独特基因库。还分析了对照组和感染组的调节基因,出乎意料的是,注射前的采样被认为是一种足够强的应激刺激,可以触发并调节血细胞中的反应,促进细胞迁移和增殖。我们发现对过滤海水的注射有明显的反应,这表明对组织损伤的反应,其中贻贝类抗菌肽中表达最丰富的肌肽 C 明显上调。流式细胞术的功能实验证实了转录组结果,因为只有在感染灿烂弧菌后才发现血细胞结构发生明显改变,并且对肌肽 C 呈阳性的血细胞数量减少,而在贻贝采血前不会发生这种情况,从而导致组织损伤。因此,我们报告了肌肽 C 参与对危险信号的反应,例如在闭壳肌中简单注射。