Department of Sciences, University of Basilicata, Viale dell'Ateneo Lucano 10, Potenza 85100, Italy.
Fish Shellfish Immunol. 2013 Sep;35(3):988-92. doi: 10.1016/j.fsi.2013.05.026. Epub 2013 Jun 10.
The innate immune system is a fundamental defense weapon of fish, especially during early stages of development when acquired immunity is still far from being completely developed. The present study aims at looking into ontogeny of innate immune system in the brown trout, Salmo trutta, using RT-PCR based approach. Total RNA extracted from unfertilized and fertilized eggs and hatchlings at 0, 1 h and 1, 2, 3, 4, 5, 6, 7 weeks post-fertilization was subjected to RT-PCR using self-designed primers to amplify some innate immune relevant genes (TNF-α, IL-1β, TGF-β and lysozyme c-type). The constitutive expression of β-actin was detected in all developmental stages. IL-1β and TNF-α transcripts were detected from 4 week post-fertilization onwards, whereas TGF-β transcript was detected only from 7 week post-fertilization onwards. Lysozyme c-type transcript was detected early from unfertilized egg stage onwards. Similarly, tissues such as muscle, ovary, heart, brain, gill, testis, liver, intestine, spleen, skin, posterior kidney, anterior kidney and blood collected from adult brown trout were subjected to detection of all selected genes by RT-PCR. TNF-α and lysozyme c-type transcripts were expressed in all tissues. IL-1β and TGF-β transcripts were expressed in all tissues except for the brain and liver, respectively. Taken together, our results show a spatial-temporal expression of some key innate immune-related genes, improving the basic knowledge of the function of innate immune system at early stage of brown trout.
先天免疫系统是鱼类的基本防御武器,尤其是在获得性免疫尚未完全发育的早期发育阶段。本研究旨在采用基于 RT-PCR 的方法研究褐鳟(Salmo trutta)先天免疫系统的个体发生。从未受精和受精的卵以及孵化后 0、1 h 和 1、2、3、4、5、6、7 周的幼鱼中提取总 RNA,使用自行设计的引物进行 RT-PCR,以扩增一些先天免疫相关基因(TNF-α、IL-1β、TGF-β和溶菌酶 c 型)。所有发育阶段均检测到β-肌动蛋白的组成型表达。IL-1β和 TNF-α转录本从受精后 4 周开始检测到,而 TGF-β转录本仅从受精后 7 周开始检测到。溶菌酶 c 型转录本从未受精的卵期开始就被早期检测到。同样,从成年褐鳟采集的肌肉、卵巢、心脏、大脑、鳃、睾丸、肝脏、肠、脾脏、皮肤、后肾、前肾和血液等组织也通过 RT-PCR 检测所有选定的基因。TNF-α和溶菌酶 c 型转录本在所有组织中表达。IL-1β和 TGF-β转录本在除大脑和肝脏之外的所有组织中表达。总之,我们的结果显示了一些关键先天免疫相关基因的时空表达,提高了褐鳟早期先天免疫系统功能的基础知识。