Faculty of Biosciences and Aquaculture, Bodø University College, 8049 Bodø, Norway.
Fish Shellfish Immunol. 2010 May-Jun;28(5-6):879-86. doi: 10.1016/j.fsi.2010.02.006. Epub 2010 Feb 11.
Antimicrobial peptides (AMP) are important components of the innate immune system in metazoans. They have been studied widely in several fishes, but little is known about these defence factors in Atlantic cod, which is thought to have a less sophisticated adaptive immune system compared to other teleosts. The aim of the present study was to screen for potential AMPs in various tissues of Atlantic cod and to examine their spectra of activity. Acidic crude extracts were prepared from thirteen tissues (i.e. mucus, gills, skin, intestine, rectum, head kidney, spleen, blood, gall bladder, liver, ovary, muscle and peritoneal wall). Following partial purification by solid-phase extraction, 78 fractions were obtained and these were assayed for antimicrobial activity using a two-layer radial diffusion assay. Some of the fractions prepared from several tissues examined had potent activity against the test bacteria. In general, acetonitrile rich fractions displayed higher antibacterial activity than the aqueous ones. The most potent fractions were obtained from the gall bladder and they exhibited potent antimicrobial activity against 8 of the 9 test bacteria, including the cod pathogen Vibrio anguillarum. Antibacterial activity was completely eliminated or reduced upon treatment with proteinase K in most fractions. Protein profiles obtained by SDS-PAGE and two-dimensional gel electrophoresis showed that antimicrobial activity of the partially purified tissue extracts might be due to cationic, low molecular weight peptides.
抗菌肽 (AMP) 是后生动物先天免疫系统的重要组成部分。它们在几种鱼类中得到了广泛的研究,但在大西洋鳕鱼中,这些防御因子的了解甚少,与其他硬骨鱼相比,人们认为大西洋鳕鱼的适应性免疫系统不太复杂。本研究的目的是筛选大西洋鳕鱼各种组织中的潜在 AMP,并研究它们的活性谱。从 13 种组织(即粘液、鳃、皮肤、肠、直肠、头肾、脾、血、胆囊、肝、卵巢、肌肉和腹膜壁)中制备酸性粗提取物。通过固相萃取进行部分纯化后,获得 78 个馏分,并使用双层放射状扩散测定法测定这些馏分的抗菌活性。从几种检查的组织中制备的一些馏分对测试细菌具有很强的活性。一般来说,富含乙腈的馏分比水相的馏分具有更高的抗菌活性。最有效的馏分来自胆囊,它们对 9 种测试细菌中的 8 种具有很强的抗菌活性,包括鳕鱼病原体鳗弧菌。大多数馏分经蛋白酶 K 处理后,抗菌活性完全消除或降低。SDS-PAGE 和二维凝胶电泳获得的蛋白质图谱表明,部分纯化的组织提取物的抗菌活性可能归因于阳离子、低分子量肽。