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RAG-1和IgM基因,是鳕鱼科黑线鳕(Melanogrammus aeglefinus,L.)免疫系统早期发育的标志物。

RAG-1 and IgM genes, markers for early development of the immune system in the gadoid haddock, Melanogrammus aeglefinus, L.

作者信息

Corripio-Miyar Y, Bird S, Treasurer J W, Secombes C J

机构信息

Scottish Fish Immunology Research Centre, University of Aberdeen, Aberdeen AB24 2TZ, Scotland, UK.

出版信息

Fish Shellfish Immunol. 2007 Jul;23(1):71-85. doi: 10.1016/j.fsi.2006.09.006. Epub 2006 Oct 4.

Abstract

The full sequence of the heavy chain of Immunoglobulin M (IgM) and a partial fragment of the recombination activating gene-1 (RAG-1) gene were sequenced in haddock, Melanogrammus aeglefinus, L. The complete transcript of haddock IgM consisted of 1865 bp and translated into a 572-aa peptide. The RAG-1 fragment was 1776 bp and was identified as the core region of RAG-1. These two immune genes were used in expression studies as markers of early development in haddock larvae. A DIG labelled oligoprobe of the RAG-1 gene was used in whole-mount in situ hybridisation (WISH). A hybridisation signal for RAG-1 was first detected in larvae at 25 days post-hatching (dph) in two bilateral symmetric regions of the head identified as the thymus. Further expression studies were carried out by RT-PCR analysis of RAG-1 and IgM on larval samples obtained during early development, i.e. from fertilisation to weaning. Haddock RAG-1 expression was detected after 21 days post-fertilisation (dpf) whilst IgM transcripts were not detected until 40 dpf, equivalent to day 29 post-hatching. These results suggest that the immune system in haddock starts to develop in larvae of 6-7 mm in length (25-29 dph).

摘要

对黑线鳕(Melanogrammus aeglefinus, L.)的免疫球蛋白M(IgM)重链的完整序列和重组激活基因1(RAG-1)基因的部分片段进行了测序。黑线鳕IgM的完整转录本由1865个碱基对组成,翻译成一个572个氨基酸的肽段。RAG-1片段为1776个碱基对,被鉴定为RAG-1的核心区域。这两个免疫基因被用于表达研究,作为黑线鳕幼体早期发育的标志物。使用地高辛标记的RAG-1基因寡核苷酸探针进行全胚胎原位杂交(WISH)。在孵化后25天(dph)的幼体头部两个双侧对称区域(被确定为胸腺)首次检测到RAG-1的杂交信号。通过对早期发育阶段(即从受精到断奶)获得的幼体样本进行RAG-1和IgM的逆转录聚合酶链反应(RT-PCR)分析,进一步开展表达研究。受精后21天(dpf)检测到黑线鳕RAG-1的表达,而直到40 dpf(相当于孵化后第29天)才检测到IgM转录本。这些结果表明,黑线鳕的免疫系统在体长6-7毫米(25-29 dph)的幼体中开始发育。

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