This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 1) The John D. Ash (ECI) Project: The primary project was to characterize the proteome of mouse retinas in development and in transgenic mice expressing leukemia inhibitory factor. A secondary project was to characterize the retina proteome of the retina in a mouse model of oxygen induced retinopathy. A plan to use this facility to identify proteins modified by oxidation including 4-HNE, and nitro-tyrosine is due next year. The goal of the study is to determine whether LIF protects photoreceptors by reducing global or specific protein oxidation or promoting protection despite protein oxidation. 2) The Raju V. Rajala (ECI) Project: This project aims to investigate proteins which interact with the retinal insulin receptor. As a result of our effort we have successfully identified actin as one of the retinal insulin receptor interacting proteins. This finding opens a new area of research to study the actin-insulin receptor interactions in diseased states such as diabetic retinopathy and stress-induced retinal degeneration. Another project within this context is to identify novel proteins which bind to p85 subunit of PI3K. We also aim to identify substrates to PTP1B protein tyrosine phosphatase. The results will significantly contribute to the understanding of the retinal insulin receptor signaling. 3) The Xi-Qing Ding (PJI) Project: Photoreceptor cyclic nucleotide-gated (CNG) channel complex assembly and interaction with adjacent proteins are crucial for the channel function. We plan to identify the modulatory molecules of cone CNG channel using a proteomics approach. Affinity purified channel complexes are analyzed by the proteomics/analytical biochemical approaches. This includes one- and two- dimensional gel electrophoresis, in-gel trypsin digestion, mass spectrometry and MS/MS analysis, and database searching. 4) The Michael H. Elliott (PJI) Project: We have made moderate/heavy use of the Proteomics/Analytical Biochemistry Module in the past funding period, which will continue next year. We have applied a mass spectrometric analysis to examine the protein composition of detergent-resistant membranes from photoreceptor rod outer segments using the Proteomics Module and this work resulted in a 2008 publication in the Journal of Neurochemistry. In addition, the Li-Cor Odyssey IR Imaging system was essential for quantitative immunoblot analysis in this paper and in another study. In the upcoming grant period, we plan to use the Proteomics/Analytical Biochemistry Module to identify protein targets that are present in caveolae isolated from retinal pigment epithelium. The results will significantly contribute to a better understanding of the role of pigment epithelium for keeping the functional integrity of retinal photoreceptor cells. 5) Anne Kasus-Jacobi (PJI) Project: We plan to use a proteomic approach 1) to Identify oxidative modification of the retinol dehydrogenase RDH12 in mouse retina, during light-induced oxidative stress, and 2) to identify the proteins forming Michael's adducts with 4-Hydroxynonenal in wild-type and Rdh12 knockout mouse retinas, during light-induced oxidative stress. The results to be obtained from these specific aims will reveal the pathological process during the light-induced oxidative stress, which will shed light to the understanding of photoreceptor degeneration.
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