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Development of medaka models for comprehensive assessment of environmental factors

环境因素综合评估青鳉模型的开发

基本信息

批准号：
10558089
负责人：
NONAKA Masaru
金额：
$ 8.13万
依托单位：
University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10558089/
关键词：
Medaka MHC Environmental assessment Genome structure DNA ligase Toxicity test Complement Innate immunity RNAi 免疫系リガーゼIV 補体化学物質

项目摘要

Nonaka : Medaka cDNA and genomic DNA clones possibly involved in adaptive and innate immunity were isolated and characterized. The medaka major histocompatibility complex (MHC) genes were dispersed on several chromosomes in contrast to the centralized mammalian MHC.However, the class IA genes and the some genes involved in class I antigen presentation were linked, possibly representing the core region of the vertebrate MHC.Several overlapping BAC clones covering the medaka MHC class I region were isolated, and the entire nucleotide sequence of one BAC clone wa determined. This BAC clone contained an about 200 kb insert, and 16 genes were identified on this clone. Comparison of the order and orientation of these genes with those of fugu and zebrafish indicated the extensive gene rearrangement even among teleost species. In addition, the complement genes, such as C3, C4 and Bf, involved in innate immunity were isolated. The medaka C3 genes were found to be amplified into about 10 copies, suggesting that innate immunity plays an essential role in body defense of telesots, whose adaptive immunity seems to be ineffective compared to mammals.Kimura : Medaka DNA ligases involved in repair DNA damages and DRP genes involved in signal transduction in neuronal cells were analyzed. The five DRP genes isolated from medaka showed one-to-one correspondence to the human genes, indicating the phylogenetic conservation of these genes throughout the vertebrate evolution. Availability of the RNAi method to medaka was also assessed using the GFP gene.Shigeoka : Efficient assay system for chemical toxicity was established. In addition, culture cell line were established from medaka fibroblasts, enabling toxicity test at the cell level.

Nonaka：青鳉cDNA和基因组DNA克隆可能参与适应性和先天性免疫分离和表征。青鳉主要组织相容性复合体（MHC）基因分散在几条染色体上，而哺乳动物MHC基因集中在几条染色体上，但Ⅰ A类基因与Ⅰ类抗原提呈相关基因连锁，可能代表脊椎动物MHC的核心区。该BAC克隆含有一个约200 kb的插入片段，在该克隆上鉴定了16个基因。这些基因的顺序和方向与河豚和斑马鱼的比较表明，广泛的基因重排，甚至在硬骨鱼物种。此外，还分离了与先天免疫有关的补体基因，如C3、C4和Bf。发现青鳉C3基因扩增到约10个拷贝，这表明先天免疫在青鳉的身体防御中起着重要作用，与哺乳动物相比，青鳉的适应性免疫似乎是无效的。木村：青鳉DNA连接酶参与修复DNA损伤和DRP基因参与神经细胞中的信号转导进行了分析。从青鳉中分离到的5个DRP基因与人类的基因一一对应，表明这些基因在脊椎动物进化过程中具有系统发育保守性。利用GFP基因对青鳉进行了RNAi方法的有效性评估。Shigeoka：建立了有效的化学毒性测定系统。此外，从青鳉成纤维细胞建立培养细胞系，使得能够在细胞水平上进行毒性测试。