Photoreceptors have several important physiological functions. They detect light but they also adapt to constant illumination and darkness. Phosphorylation of rhodopsin is a critical biochemical reaction that modulates the phototransduction cascade. In this proposal we outline methods to precisely characterize the contributions of rhodopsin phosphorylation to light and dark adaptation and to the kinetics of the photoresponse. Rhodopsin phosphorylation will be measured both in isolated intact mouse retinas and in vivo using high-resolution rapid quench and mass spectrometry methodology. The time course of rhodopsin phosphorylation will be established under a variety of physiologically important conditions. Physiological responses of rod photoreceptors will be established by quantitative electroretinography under conditions identical to those used for phosphorylation measurements. A variety of mouse strains will be examined in which genes encoding proteins that influence rhodopsin phosphorylation and regeneration are mutated. Retinoid metabolism will be monitored and it will also be compared with the kinetics of physiological responses to light and darkness. These studies will precisely define the contributions of phosphorylation and visual cycle reactions to light and dark adaptation and photoresponse kinetics in mammalian retinas.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
2R01EY006641-16
Application #
6333093
Study Section
Visual Sciences C Study Section (VISC)
Program Officer
Hunter, Chyren
Project Start
1986-07-01
Project End
2006-06-30
Budget Start
2001-07-01
Budget End
2002-06-30
Support Year
16
Fiscal Year
2001
Total Cost
$331,521
Indirect Cost
Name
University of Washington
Department
Biochemistry
Type
Schools of Medicine
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195
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Hurley, James B (2017) Warburg's vision. Elife 6:
Du, Jianhai; Rountree, Austin; Cleghorn, Whitney M et al. (2016) Phototransduction Influences Metabolic Flux and Nucleotide Metabolism in Mouse Retina. J Biol Chem 291:4698-710
Du, Jianhai; Yanagida, Aya; Knight, Kaitlen et al. (2016) Reductive carboxylation is a major metabolic pathway in the retinal pigment epithelium. Proc Natl Acad Sci U S A 113:14710-14715
Zhang, Lijuan; Du, Jianhai; Justus, Sally et al. (2016) Reprogramming metabolism by targeting sirtuin 6 attenuates retinal degeneration. J Clin Invest 126:4659-4673
Contreras, Laura; Ramirez, Laura; Du, Jianhai et al. (2016) Deficient glucose and glutamine metabolism in Aralar/AGC1/Slc25a12 knockout mice contributes to altered visual function. Mol Vis 22:1198-1212

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