Rebl Alexander, Verleih Marieke, Korytář Thomáš, Kühn Carsten, Wimmers Klaus, Köllner Bernd, Goldammer Tom
Leibniz-Institut für Nutztierbiologie (FBN), Fachbereich Molekularbiologie, Wilhelm-Stahl-Allee 2, 18196 Dummerstorf, Germany.
Vet Immunol Immunopathol. 2012 Jan 15;145(1-2):305-15. doi: 10.1016/j.vetimm.2011.11.023. Epub 2011 Dec 3.
Since 1975, the rainbow trout strain BORN (Germany) has been bred in brackish water from a coastal form imported from Denmark. Accompanying phenotypic monitoring of the adapted BORN trout until now revealed that this selection strain manifested a generally elevated resistance towards high stress and pathogenic challenge including lower susceptibility towards Aeromonas salmonicida infections in comparison to other trout strains in local aqua farms. We focus on the elucidation of both, genetic background and immunological basis for the increased survivorship to infections. A first comparison of gene expression profiles in liver tissue of healthy rainbow trout from the local selection strain BORN and imported trout using a GRASP 16K cDNA microarray revealed six differentially expressed genes evoking pathogen and wounding responses, LEAP2A (encoding for liver-expressed antimicrobial peptide), SERPINA1 (alpha-1 antitrypsin), FTH1 (middle subunit of ferritin), FGL2 (fibroleukin), CLEC4E (macrophage-inducible C-type lectin), and SERPINF2 (alpha-2 antiplasmin). Since the latter gene is not described in salmonid species so far, our first aim was to characterize the respective sequence in rainbow trout. Two trout SERPINF2 genes were identified, which share only 48% identical amino acid residues and a characteristic SERPIN domain. Second, we aimed to analyse the expression of those genes after temperature challenge (8 °C and 23 °C). Only FTH1 was upregulated in BORN and import trout after increase of temperature, while SERPINA1 and FGL2 were only elevated in import trout. Third, the expression of all named genes was analyzed after pathogen challenge with A. salmonicida subsp. salmonicida. As a main finding, we detected a comparably faster regeneration of LEAP2A mRNA abundance in BORN trout following bacterial infection. Ingenuity Pathways Analysis suggested a functional interplay among the mentioned factors and the pro-inflammatory cytokine TNF, whose stronger expression was validated in liver of BORN trout. This data indicate that the examined genes contribute to an improved first barrier against invading pathogens in BORN trout.
自1975年以来,虹鳟鱼品系BORN(德国)一直利用从丹麦引进的一种沿海类型在半咸水中养殖。对适应性BORN鳟鱼进行的伴随表型监测表明,与当地水产养殖场的其他鳟鱼品系相比,这种选育品系对高应激和病原挑战表现出普遍提高的抗性,包括对杀鲑气单胞菌感染的易感性较低。我们专注于阐明感染后存活率提高的遗传背景和免疫基础。使用GRASP 16K cDNA微阵列对当地选育品系BORN的健康虹鳟鱼和引进虹鳟鱼肝脏组织中的基因表达谱进行首次比较,发现了六个差异表达基因,这些基因引发病原体和创伤反应,即LEAP2A(编码肝脏表达的抗菌肽)、SERPINA1(α-1抗胰蛋白酶)、FTH1(铁蛋白中间亚基)、FGL2(纤维白细胞介素)、CLEC4E(巨噬细胞诱导型C型凝集素)和SERPINF2(α-2抗纤溶酶)。由于到目前为止鲑科鱼类中尚未描述后一个基因,我们的首要目标是鉴定虹鳟鱼中的相应序列。鉴定出两个虹鳟鱼SERPINF2基因,它们仅共享48%相同的氨基酸残基和一个特征性的丝氨酸蛋白酶抑制剂结构域。其次,我们旨在分析温度挑战(8℃和23℃)后这些基因的表达情况。温度升高后,只有FTH1在BORN鳟鱼和引进鳟鱼中上调,而SERPINA1和FGL2仅在引进鳟鱼中升高。第三,在用杀鲑气单胞菌亚种鲑杀鲑气单胞菌进行病原体攻击后,分析所有上述基因的表达情况。作为主要发现,我们检测到细菌感染后BORN鳟鱼中LEAP2A mRNA丰度的再生速度相对较快。 Ingenuity通路分析表明,上述因素与促炎细胞因子TNF之间存在功能相互作用,TNF在BORN鳟鱼肝脏中的更强表达得到了验证。这些数据表明,所检测的基因有助于改善BORN鳟鱼对入侵病原体的第一道屏障。