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PMID: 15626704 Published · ppublish English Comparative Study Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Reduction of all-trans retinal to all-trans retinol in the outer segments of frog and mouse rod photoreceptors.

Biophysical journal ·Vol. 88 ·No. 3 ·2005-03-00 ·Pages 2278-87

Chen C, Tsina E, Cornwall MC, Crouch RK, Vijayaraghavan S, Koutalos Y

Abstract

The first step in the Visual Cycle, the series of reactions that regenerate the vertebrate visual pigment rhodopsin, is the reduction of all-trans retinal to all-trans retinol, a reaction that requires NADPH. We have used the fluorescence of all-trans retinol to study this reduction in living rod photoreceptors. After the bleaching of rhodopsin, fluorescence (excitation, 360 nm; emission, 457 or 540 nm) appears in frog and wild-type mouse rod outer segments reaching a maximum in 30-60 min at room temperature. With this excitation and emission, the mitochondrial-rich ellipsoid region of the cells shows strong fluorescence as well. Fluorescence measurements at different emission wavelengths establish that the outer segment and ellipsoid signals originate from all-trans retinol and reduced pyridine nucleotides, respectively. Using outer segment fluorescence as a measure of all-trans retinol formation, we find that in frog rod photoreceptors the NADPH necessary for the reduction of all-trans retinal can be supplied by both cytoplasmic and mitochondrial metabolic pathways. Inhibition of the reduction reaction, either by retinoic acid or through suppression of metabolic activity, reduced the formation of retinol. Finally, there are no significant fluorescence changes after bleaching in the rod outer segments of Rpe65(-/-) mice, which lack 11-cis retinal.

MeSH Terms
Animals Cells, Cultured Female Fluorescence Recovery After Photobleaching/methods Isomerism Kinetics Light Male Metabolic Clearance Rate Mice Mice, Inbred C57BL Microscopy, Fluorescence/methods NADP/metabolism Oxidation-Reduction/radiation effects Photoreceptor Cells Rana pipiens Retinaldehyde/metabolism Rod Cell Outer Segment/metabolism,radiation effects Species Specificity Vitamin A/metabolism
Chemicals
Vitamin A NADP Retinaldehyde
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Chen Chunhe
Department of Physiology and Biophysics, University of Colorado School of Medicine, Denver, Colorado, USA.
Tsina Efthymia
Cornwall M Carter
Crouch Rosalie K
Vijayaraghavan Sukumar
Koutalos Yiannis
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Article Info
Journal
Biophysical journal
Abbr.
Biophys J
ISSN
0006-3495
DOI
10.1529/biophysj.104.054254
Published
2005-03-00
Epub
2004-00-30
Pages
2278-87
Language
English
Region
United States
NLM ID
0370626
PMCID
PMC1305277
Subset
IM
Grants
NIDA NIH HHS · R01 DA010266 · United States
NEI NIH HHS · R24 EY014793 · United States
NEI NIH HHS · R01 EY014850 · United States
NIDA NIH HHS · DA10266 · United States
NEI NIH HHS · EY014793 · United States
NEI NIH HHS · R01 EY001157 · United States
NEI NIH HHS · R01 EY004939 · United States
NEI NIH HHS · EY04939 · United States
NEI NIH HHS · EY01157 · United States
NEI NIH HHS · EY014850 · United States
External Links
PubMed source DOI: 10.1529/biophysj.104.054254 PMC Full Text View journal details
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