IRTA, San Carlos de La Rápita, 43540, Tarragona, Spain.
Scottish Fish Immunology Research Centre, University of Aberdeen, Zoology Building, Tillydrone Avenue, Aberdeen AB24 2TZ, Scotland, UK.
Fish Shellfish Immunol. 2019 Nov;94:819-832. doi: 10.1016/j.fsi.2019.10.009. Epub 2019 Oct 6.
The present study is the first report of some representative innate immune genes in meagre (Argyrosomus regius) larvae. This study has specifically focused on the growth period from hatching to the juvenile stage, a critical time in marine fish development when reliance on innate immune mechanisms are required for survival. We report molecular cloning of partial open reading frames and expression patterns for some innate immune genes (c3, cox2, met, lyzc, mxp, myd88, nod2, nod3). In addition, phylogenetic analyses of some of the sequences obtained was performed where confusion among closely allied isoforms may have existed. These results show the met isoform from meagre is met II, an isoform more similar to a homolog described in Larimichthys crocea; lysozyme (lyzc) corresponds to the c-type and NOD isoforms (nod2, nod3) separate into different clades confirming their distinctness within a common evolutionary history. Gene expression profiles of innate genes were investigated, for nine developmental stages, from 8 days post-hatching (dph) to 120 dph. Present results demonstrated that c3, cox2, met II, lyzc, mxp, myd88, nod2, and nod3 were expressed in all stages of larval development and displayed distinct expression profiles in separate tissues (kidney, spleen gut and gill). Moreover, expression patterns suggested theses innate immune genes may be influenced by feeding practices, i.e. switching from live prey (rotifer and Artemia) and weaning onto an inert commercial diet. In addition to evaluating changes in gene expression during early development, this study evaluated the modulation of gene expression by means of in vivo trials in juveniles that were stimulated with PAMPs (LPS, poly I:C, β-glucan). These results revealed significant changes in mRNA levels of target genes in the kidney, spleen, gut and gills. However, expression profiles differed in magnitude depending on the stimulant and/or tissue. These results are discussed in terms of their relevance and potential application in aquaculture practices.
本研究首次报道了一些代表性的鲷(Argyrosomus regius)幼虫固有免疫基因。本研究特别关注从孵化到幼鱼阶段的生长时期,这是海洋鱼类发育的关键时期,需要依赖固有免疫机制才能生存。我们报告了一些固有免疫基因(c3、cox2、met、lyzc、mxp、myd88、nod2、nod3)部分开放阅读框的分子克隆和表达模式。此外,对获得的一些序列进行了系统发育分析,其中可能存在密切相关同工型之间的混淆。这些结果表明,来自鲷的 met 同工型是 met II,与 Larimichthys crocea 中描述的同源物更相似;溶菌酶(lyzc)对应于 c 型和 NOD 同工型(nod2、nod3)分为不同的分支,在共同的进化历史中证实了它们的独特性。为了研究固有基因的表达谱,我们从孵化后 8 天(dph)到 120 天(dph)共检测了 9 个发育阶段。目前的结果表明,c3、cox2、met II、lyzc、mxp、myd88、nod2 和 nod3 在幼虫发育的所有阶段均有表达,并在不同组织(肾脏、脾脏、肠道和鳃)中呈现出不同的表达谱。此外,表达模式表明这些固有免疫基因可能受到喂养方式的影响,例如从活饵料(轮虫和丰年虫)到断奶后转为惰性商业饲料。除了评估早期发育过程中基因表达的变化外,本研究还通过体内试验评估了在幼鱼中刺激 PAMPs(LPS、poly I:C、β-葡聚糖)后基因表达的调节。这些结果显示,肾脏、脾脏、肠道和鳃中的靶基因的 mRNA 水平发生了显著变化。然而,表达谱的幅度取决于刺激物和/或组织。这些结果从其相关性和在水产养殖实践中的潜在应用方面进行了讨论。