爬虫類の有鱗目と亀目における生体異物代謝経路－生物学的防御システムの機序と特徴

爬行动物有鳞动物和海龟的异生代谢途径 - 生物防御系统的机制和特征

• 批准号: 13J01324
• 负责人: オロスラーニュ バラージュ (2014-2015)
• 金额: $ 1.73万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for JSPS Fellows
• 财政年份: 2013
• 资助国家: 日本
• 起止时间: 2013-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13J01324/
• 关键词: reptile; turtle; tortoise; metabolism; pyrene; PAH; freshwater; yeast; 爬虫類; カメ類; 異物代謝; 種差; ピレン; 多環芳香族炭化水素; SULT

The main aim of this year was to compare in vivo freshwater turtle pyrene metabolism with tortoise species. Our selection criteria was also different: in the former case we used distant families to get a general overview, in the later we concentrated on the closely related Testudo genus. Interestingly, we have found two distinct pattern by the analysis of the metabolites: pyrene-1-sulfate dominated all the freshwater turtle samples, while in land dwelling species pyrene-1-glucuronide was the main metabolite, with more variability. Since all the freshwater species were carnivores, while the tortoises strictly herbivorous, this suggests two possible explanation: A, the sulfate-dominance is due to feeding habits (we can see a similar pattern in the case of Carnivores, where functioning UGT1A isoform numbers are low), or, B, the difference is due to the habitat, i.e. the water environment with accessible sulfates is responsible for the metabolite distribution (this would explain why we can see a similar pattern of pyrene metabolism in amphibians).Further understanding of the genetic background of this differences would be necessary, but my cloning experiments with Cytochrome P450 1A5 was not successful. (This is the reason we did not created a yeast model).At the same time, the slowly increasing NGS data from reptiles created a new opportunity: while sequencing itself, due to the overall costs, was not feasible, methods to analyse the various reptile genomes and transcriptomes were under development, and will be finalized in this year.

本年度的主要目的是比较淡水龟和龟类的体内芘代谢。我们的选择标准也不同：在前一种情况下，我们使用远缘科来获得一般概述，在后一种情况下，我们集中在密切相关的陆龟属。有趣的是，我们发现了两个不同的模式，芘-1-硫酸盐占主导地位的所有淡水龟样品，而在陆栖物种芘-1-葡萄糖醛酸苷是主要的代谢产物，具有更大的变异性。由于所有的淡水物种都是食肉动物，而陆龟是严格的食草动物，这表明了两种可能的解释：硫酸盐占主导地位是由于摄食习惯（我们可以在食肉动物中看到类似的模式，其中功能性UGT 1A同种型数量很低），或者，B，差异是由于栖息地，也就是说，具有可获得硫酸盐的水环境是代谢物分布的原因（这可以解释为什么我们可以在两栖动物中看到类似的芘代谢模式）进一步了解这种差异的遗传背景是必要的，但我用细胞色素P450 1A 5进行的克隆实验没有成功。(This与此同时，爬行动物的NGS数据的缓慢增长创造了一个新的机会：虽然测序本身由于总体成本而不可行，但分析各种爬行动物基因组和转录组的方法正在开发中，并将在今年完成。

项目成果

The characteristics and mechanism of pyrene metabolism in freshwater turtles and the suggested genetic background

淡水龟芘代谢特征、机制及遗传背景

• 发表时间: 2013
• 作者: 衣川智弥;宮本 晃伸;神田展行;中村卓史;Oroszlany Balazs
• 通讯作者: Oroszlany Balazs

Pyrene metabolism and the role of phase I and II enyzmes in freshwater turtles

芘代谢以及淡水龟中 I 相和 II 相酶的作用

• 发表时间: 2013
• 作者: M. Morita;H. Onoe;M. Yanagisawa;H. Ito;M. Ichikawa;K. Fujiwara;H. Saito and M. Takinoue;Oroszlany Balazs
• 通讯作者: Oroszlany Balazs

Phase II. Conjugation Ability for Pyrene Metabolism in Tortoises and Comparison to Freshwater Turtle Models

第二阶段。

• 发表时间: 2015
• 作者: Daichi Mukunoki;Daisuke Takahashi;Oroszlany Balazs
• 通讯作者: Oroszlany Balazs