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

今年的主要目的是比较淡水龟和陆龟体内的芘代谢。我们的选择标准也不同：在前一种情况下，我们使用远亲来获得大致的概述，在后一种情况下，我们集中在关系密切的Testudo属。有趣的是，通过对这些代谢物的分析，我们发现了两种不同的模式：所有淡水甲鱼样品中的主要代谢物是P-1-硫酸盐，而陆生物种中的主要代谢物是P-1-葡萄糖醛酸化物，变异性较大。由于所有淡水物种都是食肉动物，而龟类是严格的食草动物，这提出了两种可能的解释：第一，硫酸盐优势是由于食性(我们可以在肉食动物中看到类似的模式，功能UGT1A亚型数量较低)，或者，第二，差异是由于栖息地，即具有可访问硫酸盐的水环境负责代谢物的分布(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 1A5进行的克隆实验并不成功。(这就是我们没有建立酵母模型的原因)。同时，爬行动物的NGS数据缓慢增加创造了一个新的机会：虽然由于总体成本的原因，测序本身是不可行的，但分析各种爬行动物基因组和转录本的方法正在开发中，将在今年完成。

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.