Use of gene regulatory sensors to monitor environmental pollutant levels in small mammals and birds

由于人类活动，环境中大量的杀虫剂和二恶英类污染物大量增加。其中一些化学品具有持久性，并在食物链中积累，这引起了人们对人类甚至野生动物健康中长期接触的重大关切。测量环境中这类污染物的经典方法是通过生物化学手段对水和土壤来源进行采样，以评估其丰度。 一旦确定了这些测量值，就会评估它们是否在被认为对生物“安全”的范围内。 然而，这种方法的主要缺陷是，人们如何知道需要多少这些化学物质才能影响单个物种的健康？ 细胞将至少部分地通过芳烃受体（AhR）的作用对污染物和农药进行解毒，所述芳烃受体（AhR）调节属于细胞色素P450 1A家族（包括CYP 1A 1和CYP 1B 1）的异生物质代谢酶。 从理论上讲，这些基因表达得越多，特定物种就越需要处理大量特定污染物。 我的实验室已经开发出分子工具，通过qRT-PCR测量细胞中的hepatocyte基因表达。 因此，我的研究计划将是测量CYP 1A 1和CYP 1B 1基因在生活在密集农业实践地区附近的动物中的表达，以及在农药使用最少的地区。 这些物种将包括（但不限于）树燕、蒙面鼩、浣熊和小棕蝠。我们还将监测这些动物中雌激素信号传导是否受损，并直接检测遗传毒性雌激素代谢产物的存在。这个多学科的研究计划旨在解决几个重要的生物/环境问题。 我坚信，我们的遗传方法（结合HPLC/质谱法）比传统的生物化学/化学方法更敏感和准确，可以测量环境中的污染。 因此，我们的遗传工具在评估密集农业对野生动物和人类健康的潜在影响方面可能是非常有用和具有成本效益的。

The abundance of pesticides and dioxin-like pollutants has massively increased in the environment due to human activity. Some of these chemicals are persistent and accumulate in the food chain, which raises major concerns regarding mid- and long-term exposure on human and even wildlife health. The classical way of measuring such pollutants in the environment is to sample water and soil sources by biochemical means to assess their abundance. Once these measurements are determined, it is assessed whether they are within a range that is deemed "safe" for living creatures. However the major flaw with this is that how does one know how much of these chemicals are required to affect the health of individual species? Cells will be detoxified of pollutants and pesticides, at least in part, by the action of the aryl hydrocarbon receptor (AhR), which regulates xenobiotic metabolism enzymes belonging to the cytochrome P450 1A family (including CYP1A1 and CYP1B1). In theory, the more these CYP genes are expressed, the more a particular species is dealing with a significant dose of a particular pollutant. My laboratory has developed molecular tools to measure CYP gene expression in cells by qRT-PCR. Thus, my research plan will be to measure CYP1A1 and CYP1B1 gene expression in animals that live close to regions of intense agricultural practices, and also in regions where the use of pesticides is minimal. These species will include (but not limited to) the tree swallow, the masked shrew, the raccoon, and the little brown bat. We will also monitor if estrogen signaling is compromised in these animals and directly test for the presence of genotoxic estrogen metabolites. This multidisciplinary research program aims at addressing several important biological/environmental problems. I firmly believe that our genetic approach (coupled to HPLC/mass spectrometry) is significantly more sensitive and accurate than conventional biochemical/chemical methods to measure contamination in the environment. Hence our genetic tools could prove to be extremely useful and cost effective in assessing the potential effects of intense agriculture on wildlife and human health.