Tools for High Resolution Respirometry

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Our application outlines a request by a group of 4 biologists for high resolution respirometry (HRR) equipment that is essential to test hypotheses regarding effects of stressors and local anesthetics on mitochondrial function. This equipment will allow accurate, quantitative measurements in real-time of respiration, membrane potential, calcium, nitric oxide and reactive oxygen species in mitochondria and cells. It will support research programs focusing on (a) interactions of metals, hypoxia and thermal stress in fish, (b) effects of local anesthetics on mitochondrial bioenergetics in fish, (c) effects of organic pollutants on aquatic systems, and (d) effects of viral infections on fish. Our research is expected to generate important information regarding the role of cellular energy homeostasis in handing stressors of diverse origin. Unlike comparable systems in the market, the requested HRR system is constructed of materials that minimize oxygen diffusion and has advanced electronics to provide high signal stability of the oxygen sensors. The high signal stability minimizes the amount of required biological sample and provides the basis for resolution in the physiologically relevant low-oxygen tension range. Although this instrument is designed primarily to measure respiration, it includes fluorescence, potentiometric and amperometric modules that provide the capability to measure membrane potential, cations such as calcium, pH and nitric oxide. Indeed, other add-ons can be purchased as required to increase the repertoire of measurements. Overall, the equipment will enable our researchers to generate a comprehensive framework for using mitochondrial functional indices to predict organismal health that will positively impact fundamental studies on the effect of multiple stressors and anesthetics on energy homeostasis, as well as inform applied studies on management of aquatic pollution and diseases. Moreover, this equipment will be crucial for the training of highly qualified personnel in progressive research methodologies essential in the current competitive job market.