Development and maturation of the immune system in zebrafish, Danio rerio:: a gene expression profiling, in situ hybridization and immunological study
Development and maturation of the immune system in zebrafish, Danio rerio:: a gene expression profiling, in situ hybridization and immunological study
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DOI：
10.1016/s0145-305x(03)00103-4
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发表时间：
2004-01-01
影响因子：
2.9
通讯作者：
Sin, YM
中科院分区：
文献类型：
--
作者：
Lam, SH;Chua, HL;Sin, YM
通过实时定量聚合酶链反应、原位杂交（ISH）、免疫亲和纯化和蛋白质印迹法检测免疫球蛋白（Ig）以及浸没免疫实验来研究免疫相关基因表达谱，研究斑马鱼免疫系统的发育和成熟。 Ikaros 表达首次在受精后 1 天 (dpf) 检测到，此后在 28 至 42 dpf 之间逐渐增加至两倍以上，然后在成年鱼（105 dpf 龄）中降至低于最初 1 dpf 的表达水平。重组激活基因 1 (Rag-1) 的表达水平在 3 至 17 dpf 之间迅速增加（10 倍），在 21 至 28 dpf 之间达到最大值，然后逐渐下降。然而，在105 dpf的成鱼中，Rag-1的表达水平明显下降，与3 dpf时的表达水平相当。 T 细胞受体 α 恒定区和免疫球蛋白轻链恒定区 (IgLC) 同型 1、2 和 3 mRNA 在 3 dpf 时检测到低水平，并且其表达水平在受精后 4 至 6 周 (wpf) 期间稳定增加至成人范围。使用组织切片 ISH，2 周后在头肾中检测到 Rag-1 表达，而 3 周后在头肾和胸腺中检测到 IgLC-1、2 和 3。分泌的 Ig 只能在 4 周后使用免疫亲和纯化和蛋白质印迹法检测到。分别在 4 周和 6 周免疫的斑马鱼中观察到对 T 独立抗原（福尔马林杀死的嗜水气单胞菌）和 T 依赖性抗原（人丙种球蛋白）的体液反应，表明获得了免疫能力。研究结果表明，斑马鱼免疫系统的形态和功能在 4-6 周后就已经成熟。 (C) 2003 年，爱思唯尔有限公司出版。
The development and maturation of the immune system in zebrafish was investigated using immune-related gene expression profiling by quantitative real-time polymerase chain reaction, in situ hybridization (ISH), inummoglobulin (Ig) detection by immuno-affinity purification and Western blotting as well as immersion immunization experiments. Ikaros expression was first detected at 1 day post-fertilization (dpf) and thereafter increased gradually to more than two-fold between 28 and 42 dpf before decreasing to less than the initial 1 dpf expression level in adult fish (aged 105 dpf). Recombination activating gene-1 (Rag-1) expression levels increased rapidly (by 10-fold) between 3 and 17 dpf, reaching a maximum between 21 and 28 dpf before decreasing gradually. However, in adult fish aged 105 dpf, the expression level of Rag-1 had dropped markedly, and was equivalent to the expression level at 3 dpf. T-cell receptor alpha constant region and immunoglobulin light chain constant region (IgLC) isotype-1, 2 and 3 mRNAs were detected at low levels by 3 dpf and their expression levels increased steadily to the adult range between 4 and 6 weeks post-fertilization (wpf). Using tissue-section ISH, Rag-1 expression was detected in head kidney by 2 wpf while IgLC- 1, 2 and 3 were detected in the head kidney and the thymus by 3 wpf onwards. Secreted Ig was only detectable using immuno-affinity purification and Western blotting by 4 wpf. Humoral response to T-independent antigen (formalin-killed Aeromonas hydrophila) and T-dependent antigen (human gamma globulin) was observed in zebrafish immunized at 4 and 6 wpf, respectively, indicating that immunocompetence was achieved. The findings reveal that the zebrafish immune system is morphologically and functionally mature by 4-6 wpf. (C) 2003 Published by Elsevier Ltd.