Magnadóttir Bergljót, Lange Sigrun, Steinarsson Agnar, Gudmundsdóttir Sigridur
Institute for Experimental Pathology, University of Iceland, Keldur v. Vesturlandsveg, IS-112 Reykjavík, Iceland.
Comp Biochem Physiol B Biochem Mol Biol. 2004 Oct;139(2):217-24. doi: 10.1016/j.cbpc.2004.07.009.
The aim of this study was to monitor the ontogenic development of innate immune parameters of cod (Gadus morhua L.) and to determine the presence of maternal IgM. The general protein composition and enzyme activity was also studied. At intervals, samples were collected of fertilized cod eggs and larvae from 3 days after fertilization until 57 days after hatching. Cell lysates were prepared and analysed by Western blotting using antibodies prepared against cod IgM, the complement component C3 and C-reactive protein (CRP) as well as against cod serum proteins and haemoglobin. Antibodies against salmon cathepsins and against several mammalian proteins of immunological significance were also used. Maternal IgM was not detected but C3 and the closely associated apolipoprotein A-I were present from the time of embryo organogenesis. C-reactive protein was not detected and none of the antibodies against mammalian immune parameters cross-reacted with the cod material. Protein and proteomic analysis showed that the major proteins of the egg samples were vitellogenin derived maternal proteins. Other non-vitellogenin maternal proteins, not yet identified, were also detected in the fertilized eggs. Cathepsin was present in all samples, but other enzyme activity was restricted to larval samples from 4 days after hatching when feeding had commenced. Haemoglobin was not detected until 10 days after hatching.
本研究的目的是监测鳕鱼（Gadus morhua L.）先天免疫参数的个体发育，并确定母体免疫球蛋白M（IgM）的存在情况。同时还研究了其总体蛋白质组成和酶活性。在受精后3天至孵化后57天的时间段内，每隔一段时间采集受精鳕鱼卵和幼体样本。制备细胞裂解物，并通过蛋白质印迹法进行分析，所用抗体包括针对鳕鱼IgM、补体成分C3和C反应蛋白（CRP）制备的抗体，以及针对鳕鱼血清蛋白和血红蛋白制备的抗体。还使用了针对鲑鱼组织蛋白酶以及几种具有免疫学意义的哺乳动物蛋白质的抗体。未检测到母体IgM，但从胚胎器官发生时起就存在C3和与之密切相关的载脂蛋白A-I。未检测到C反应蛋白，且针对哺乳动物免疫参数的抗体均未与鳕鱼样本发生交叉反应。蛋白质和蛋白质组学分析表明，卵样本中的主要蛋白质是源自卵黄蛋白原的母体蛋白质。在受精卵中还检测到了其他尚未鉴定的非卵黄蛋白原母体蛋白质。组织蛋白酶存在于所有样本中，但其他酶活性仅限于孵化后4天开始摄食时的幼体样本。直到孵化后10天才检测到血红蛋白。
