The Journal of General Physiology
Cell MicroControls
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Published online Sep 27 2004. doi:10.1085/jgp.200409078
The Rockefeller University Press, 0022-1295 $8.00
JGP, Volume 124, Number 4, 429-443
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Physiological and Microfluorometric Studies of Reduction and Clearance of Retinal in Bleached Rod Photoreceptors

Efthymia Tsina1, Chunhe Chen2, Yiannis Koutalos2, Petri Ala-Laurila1, Marco Tsacopoulos3, Barbara Wiggert4, Rosalie K. Crouch5, and M. Carter Cornwall1

1 Department of Physiology and Biophysics, Boston University School of Medicine, Boston, MA 02118
2 Department of Physiology and Biophysics, University of Colorado School of Medicine, Denver, CO 80262
3 Department of Experimental Physiology, University of Athens, Athens, Greece
4 National Eye Institute, National Institutes of Health, Bethesda, MD 20892
5 Department of Ophthalmology, Medical University of South Carolina, Charleston, SC 29425

Address correspondence to M. Carter Cornwall, Department of Physiology and Biophysics, Boston University School of Medicine, 715 Albany St., Boston, MA 02118. Fax: (617) 638-4273; email: cornwall{at}bu.edu

The visual cycle comprises a sequence of reactions that regenerate the visual pigment in photoreceptors during dark adaptation, starting with the reduction of all-trans retinal to all-trans retinol and its clearance from photoreceptors. We have followed the reduction of retinal and clearance of retinol within bleached outer segments of red rods isolated from salamander retina by measuring its intrinsic fluorescence. Following exposure to a bright light (bleach), increasing fluorescence intensity was observed to propagate along the outer segments in a direction from the proximal region adjacent to the inner segment toward the distal tip. Peak retinol fluorescence was achieved after ~30 min, after which it declined very slowly. Clearance of retinol fluorescence is considerably accelerated by the presence of the exogenous lipophilic substances IRBP (interphotoreceptor retinoid binding protein) and serum albumin. We have used simultaneous fluorometric and electrophysiological measurements to compare the rate of reduction of all-trans retinal to all-trans retinol to the rate of recovery of flash response amplitude in these cells in the presence and absence of IRBP. We find that flash response recovery in rods is modestly accelerated in the presence of extracellular IRBP. These results suggest such substances may participate in the clearance of retinoids from rod photoreceptors, and that this clearance, at least in rods, may facilitate dark adaptation by accelerating the clearance of photoproducts of bleaching.

Key Words: retinol • vitamin A • photoreceptor • visual cycle • IRBP


Abbreviations used in this paper: IRBP, interphotoreceptor retinoid binding protein; Meta II, metarhodopsin II; NADPH, nicotine adenine dinucleotide phosphate; RDH, retinol dehydrogenase; RPE, retinal pigment epithelium.


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