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Title: Characterization of the Retinal Proteome During Rod Photoreceptor Genesis

Author
item Barnhill, Alison
item HECKER, LAURA - Mayo Clinic
item KOHUTYUK, OKSANA - Iowa State University
item BUSS, JANICE - Iowa State University
item HONAVAR, VASANT - Iowa State University
item WEST-GREENLEE, HEATHER - Iowa State University

Submitted to: BMC Research Notes
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/27/2010
Publication Date: 1/27/2010
Citation: Barnhill, A.E., Hecker, L.A., Kohutyuk, O., Buss, J.E., Honavar, V., West-Greenlee, H.M. 2010. Characterization of the Retinal Proteome During Rod Photoreceptor Genesis. BMC Research Notes 2010. 3:25. Available: http://www.biomedcentral.com/1756-0500/3/25.

Interpretive Summary: The structure and function of the retina is well understood but the way that a particular cell makes a decision to become that cell type is not. Rod photoreceptors are cells in the retina that are responsible for sensing low light levels. There are diseases that affect these rod photoreceptor cells causing varying levels of blindness. If the cell fate decision can be understood then ways to manipulate them to replace photoreceptors becomes a possibility. One way that we addressed this question was to look at the proteome of the retina at different developmental time points. The proteome is the sum total of all proteins being expressed in a cell or tissue at any given point in time. We looked for proteins that were peaking at ages important to rod photoreceptor development. We identified 26 dynamically expressed proteins that had not been associated with retinal development before.

Technical Abstract: Background: The process of rod photoreceptor genesis, cell fate determination and differentiation is complex and multi-factorial. Previous studies have defined a model of photoreceptor differentiation that relies on intrinsic changes within the presumptive photoreceptor cells as well as changes in surrounding tissue that are extrinsic to the cell. We have used a proteomics approach to identify proteins that are dynamically expressed in the mouse retina during rod genesis and differentiation. Findings: A series of six developmental ages from E13 to P5 were used to define changes in retinal protein expression during rod photoreceptor genesis and early differentiation. Retinal proteins were separated by isoelectric focus point and molecular weight. Gels were analyzed for changes in protein spot intensity across developmental time. Protein spots that peaked in expression at E17, P0 and P5 were picked from gels for identification. There were 239 spots that were picked for identification based on their dynamic expression during the developmental period of maximal rod photoreceptor genesis and differentiation. Of the 239 spots, 60 of them were reliably identified and represented a single protein. Ten proteins were represented by multiple spots, suggesting they were post-translationally modified. Of the 42 unique dynamically expressed proteins identified, 16 had been previously reported to be associated with the developing retina. Conclusions: Our results represent the first proteomics study of the developing mouse retina that includes prenatal development. We identified 26 dynamically expressed proteins in the developing mouse retina whose expression had not been previously associated with retinal development.