New and Improved Sensor Platforms and Quantification of Nitric Oxide for In Vitro and In Vivo Systems

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PROJECT SUMMARY/ABSTRACT Nitric oxide (NO) is a key signaling molecule in biological systems, but it is poorly understood due to a lack of temporal and spatial sensors that measure NO. A direct, fast and accurate sensor for measuring NO would be of immense value to ongoing and future research. Single walled carbon nanotubes (SWNT) have excellent potential for use as NO sensors both in vitro and in vivo, but they are not widely used by the research community because they are not readily available in an easy-to-use platform. The goals of this project are to 1) develop new SWNT sensor platforms and modify the current systems to improve sensor specificity and handling, and 2) determine nitric oxide concentration and dynamics with a highly sensitive and accurate sensor. The goals will be accomplished through the completion of four specific projects. In the first project we will develop a ratiometric sensor to quantify NO over long time periods, the second project will involve the development of an easy-to-use platform for the in vivo delivery and stabilization of SWNT sensors that are capable of both spatial and temporal analyte quantification. Projects 3 and 4 will use the SWNT sensors to investigate NO concentrations, specifically for project 3 we will determine the intracellular dynamics of NO in relation to organelles and membranes and project 4 will involve the investigation of extracellular NO concentration values and gradients associated with healthy and diseased cells. This work will leverage the expertise of the Iverson Laboratory to engineer new and improved sensor delivery platforms and use the sensors to investigate intracellular and extracellular NO signaling. By developing these platforms a template for the development of other SWNT sensor systems will also be provided, allowing researchers to learn about reactive oxygen species, small molecules and proteins in a spatial and temporal fashion at the sub-cellular level. The investigation of the NO signaling for the cells investigated in this project will provide researchers with a basic understanding of NO’s role in cell signaling and provide a template for investigation of other cell types.