Thomas Rich
University of South Alabama, USA
Scientific Tracks Abstracts: Neurochem Neuropharm
Cyclic nucleotides are ubiquitous second messengers known to differentially regulate many cellular functions over a wide range of timescales. Several lines of evidence suggest that intracellular distributions of cAMP and cGMP are not uniform, and that compartmentalization is largely responsible for signaling specificity within these signaling pathways. However, real time measurements of cAMP and cGMP signals have been hampered by the low signal-to-noise ratio of fluorescence and FRET probes, as well as the inability to simultaneously measure multiple signals at discrete subcellular locations within the same cell. In addition, measurements are typically performed in two spatial dimensions (2D), further limiting insight into second messenger signaling systems. Here, we present novel hyperspectral imaging approaches that increase signal-to-noise ratios of fluorescence and FRET probes and allow real time, multiplexed measurements of intracellular signaling pathways in three spatial dimensions (3D). We have used hyperspectral approaches to visualize cyclic nucleotide gradients in several cell types, including pulmonary endothelial cells. Interestingly, in endothelial cells gradients primarily form along the apical to basolateral axis. These gradients would not be discerned by traditional imaging approaches. These data suggest that 2D imaging studies of cyclic nucleotide compartmentalization may lead to erroneous conclusions about the existence of second messenger gradients, and that 3D studies are required to assess mechanisms of signaling specificity. Results also demonstrated that novel hyperspectral imaging technologies are powerful tools for measuring biochemical processes in discrete subcellular domains.
Thomas Rich received his BAE and MSc in Aerospace Engineering from the Georgia Institute of Technology in 1988 and 1990 respectively. He has done his PhD in Biomedical Engineering from Vanderbilt University in 1996. He is currently a Professor in the Center for Lung Biology and Department of Pharmacology at the University of South Alabama. His research interests include the development of novel approaches to measure second messenger signals in real time, understanding mechanisms underlying signal specificity, and the roles of intracellular signaling pathways in Pulmonary Physiology and Pathophysiology.
Email: trich@southalabama.edu
