This request is for a Bio-Rad Confocal Scanning Laser Microscope system in conjunction with a Zeiss microscope equipped with episcopic fluorescence optics. The system will significantly enhance the research programs of four major users who are conducting N.I.H. funded R01 research projects and three users, two of whom are R15 awardees. The microscope system will be employed in several distinctly different ways. Drs. Dun, Ardell and Hoover will employ the microscope system to reconstruct brainstem, spinal neurons and intracardiac ganglion cells filled with lucifer yellow or biotin-dextran in vitro preparations and to image the spatial relationship between nerve terminals containing a specific neurotransmitter and dye-filled neuron. Dr. Ganote will determine the site(s) of translocation of heat shock proteins and several other cytoskeletal proteins in myocytes. Dr. Defore will examine the role of integrin adhesion receptors in attachment-dependent survival of retinal pigment epithelial cells in vitro, using confocal microscopy to image both ligand-activated and clustered receptors. Dr. Harrison will examine gustatory axon terminals in relation to hypothalamic neurons containing specific neuropeptides that influence food intake and feeding behavior. Dr. Ecay will address the issue of trafficking of myosin-coated trans- Golgi network vesicles to apical and/or basolateral membrane domains. The Bio-Rad system is the most optimal instrument available for the applications outlined here. The investigators whose projects comprise this application have evaluated the systems developed by Bio-Rad, Zeiss and Olympus. The preliminary data included in the application were obtained using the Bio-Rad microscope. The East Tennessee State University College of Medicine has a strong institutional commitment to the acquisition and maintenance of the microscope system. A highly experienced and full-time research assistant, Mr. David Neely, will oversee the day-to-day operation of the microscope and a room about 100 ft/2 in the Department of Pharmacology will be renovated to house the system. The College of Medicine has also agreed to underwrite the service contract for the instrument, if it is purchased.
