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其他陆龟体内芘的代谢。我们的选择标准也不同：在前一种情况下，我们使用远科来获得总体概况，在后一种情况下，我们集中在密切相关的Testudo属。有趣的是，通过对代谢产物的分析，我们发现了两种截然不同的模式：在所有淡水龟样本中，芘-1-硫酸盐占主导地位，而在陆地栖息物种中，芘-1-葡萄糖醛酸盐是主要的代谢产物，且差异更大。由于所有的淡水物种都是食肉动物，而陆龟则完全是食草动物，这就提出了两种可能的解释：首先，硫酸盐的优势是由于摄食习惯（我们可以在食肉动物的情况下看到类似的模式，其中功能UGT1A同种异构体数量很低），或者，B，差异是由于栖息地，即具有可接触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，但我对细胞色素P450 1A5的克隆实验并不成功。（这就是我们没有创建酵母模型的原因）。与此同时，爬行动物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的原因是不可行的，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。

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.