A micro-respirometry system for measuring oxygen consumption in mitochondria, cultured cells, invertebrates and small fishes

申请人和Wilfrid Laurier的三位合作者将在广泛的应用中使用微量呼吸测量系统，以确定水生生物如何在亚细胞，细胞和整个动物水平上对金属，农药，内分泌干扰化合物和纳米颗粒等新兴污染物做出反应。 该仪器最初将用于测量线粒体的耗氧量，以更好地了解如何暴露于3-三氟甲基-4-硝基苯酚（TFM），用于控制五大湖入侵的海七鳃鳗种群的杀虫剂，干扰七鳃鳗的燃料（ATP）生产，以及在TFM治疗期间经常短暂暴露于这种化学品的非目标鱼类。由于线粒体也是金属和代谢废物（如氨）的目标，线粒体呼吸研究将确定这些毒物如何干扰不同鱼类的ATP生产。 微呼吸测量系统亦会被用来研究鱼类胚胎及幼体在接触内分泌干扰物质（即在许多沃茨发现的污染物）时，其呼吸过程如何受到影响。还将使用微量呼吸测定系统测量培养细胞（例如脑、鳃、肝）的代谢活性，以评估暴露于金属、氨和纳米颗粒期间的细胞活力和生理应激。通过开发这种测试细胞活力和生理应激的非致命手段，将有可能在生态毒理学研究中更有效地跟踪细胞对环境污染物的反应。 为了更好地了解金属如何损害无脊椎动物的气体交换和代谢，还将在模式生物中测量氧气消耗，例如常见的水蚤，其通常用于监测市政和工业废水的毒性。 所要求的仪器在威尔弗里德·劳里埃公司是不可用的，它将满足HQP在四个实验室的研究需要，这些实验室目前属于劳里埃公司新兴的鱼类生理学研究小组。

A micro-respirometry system will be used by the applicant and three collaborators at Wilfrid Laurier in a wide range of applications to determine how aquatic organisms respond to metals, pesticides, endocrine disrupting compounds, and emerging contaminants such as nanoparticles, at the sub-cellular, cellular and whole animal level. The instrument will initially be used to measure oxygen consumption of mitochondria to better understand how exposure to 3-trifluoromethyl-4-nitrophenol (TFM), the pesticide used to control invasive sea lamprey populations in the Great Lakes, interferes with fuel (ATP) production in lampreys, as well as non-target fishes which are often transiently exposed to this chemical during TFM treatments. As the mitochondria are also targets of metals and metabolic wastes such as ammonia, mitochondrial respiration studies will determine how such toxicants interfere with ATP production in different fishes. The micro-respirometry system will also be used to elucidate how respiratory processes are compromised in developing fish embryos and larvae when exposed to endocrine disrupting substances, which are contaminants found in many waters. The metabolic activity of cultured cells (e.g. brain, gills, liver) will also be measured with the micro-respirometry system to assess cell viability and physiological stress during exposure to metals, ammonia and nanoparticles. By developing such a non-lethal means of testing cell viability and physiological stress, it will be possible to more effectively track how cells respond to environmental contaminants in ecotoxicology studies. To better understand how metals compromise gas exchange and metabolism in invertebrates, oxygen consumption will also be measured in model organisms such as the common water flea, which is commonly used to monitor the toxicity of municipal and industrial effluents. The requested instrument is not available at Wilfrid Laurier, and will fulfill the research needs of HQP in the four laboratories that presently belong to Laurier's emerging Fish Physiology Research Group.