This proposal requests to purchase and operate a Bio-Rad Confocal Microscope in conjunction with a Nikon rE 2000U inverted microscope to enhance the rapid developing research programs in Kosair Children Hospital Research Institute (KCHRI). Since the establishment of KCHRI in 2000, this group has recruited 8 faculty members and a large number of research scientists, postdoctoral research fellows, and students with a total of 40 researchers. Many of them have thus far successfully acquired NIH grants (7 R01,3 COBRE, 2 R21,2 postdoctoral fellowships). In addition, an additional 4 R01 proposals have been recently reviewed and received favorable percentiles (13%, 15%, 16.2%, and 22%), hence they are very likely to be funded. All these newly funded projects require an extensive use of the confocal microscope. The microscope system will be employed in several different ways. Drs. D. Gozal, R. Liu, and Cheng, will use the confocal microscope to analyze chronic intermittent hypoxia-mediated and aging-induced brain neuronal cell death, reactive oxygen species (ROS) production, protein aggregation, remodeling of cardiac axons and terminals in the heart, and receptor expressions/colocalization, labeled with different dyes or immunostained. Drs Epstein and Y. Liu will examine diabetes-related proteins and ROS production in beta cells and myocardium. Dr. E. Gozal examine protein-protein interactions that regulate Akt activation by hypoxia and to identify markers of apoptosis such as Annexin V on the outer membrane of apoptotic cells in response to intermittent hypoxia. Dr. Wang will image intracellular Ca2+ distribution, measure mitochondrial membrane potentials, and study the translocation of apoptotic proteins among subcellular structures induced by hypoxia. Dr. Lassiter will study the presence of complement membrane attack complexes (MAC) deposited in vivo onto post-ischemic cerebral cells and determine the fate of MAC following deposition. The KCHRI has a strong commitment to the acquisition and maintenance of this microscope system. In support of this application, KCHRI will renovate a 100 sq ft room to house the system and will underwrite the long-term service contract for the instrument, if it is purchased.
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