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Intra-retinal visual cycle required for rapid and complete cone dark adaptation


Daytime vision is mediated by retinal cones, which, unlike rods, remain functional even in bright light and dark-adapt rapidly. These cone properties are enabled by rapid regeneration of their pigment. This in turn requires rapid chromophore recycling that may not be achieved by the canonical retinal pigment epithelium visual cycle. Recent biochemical studies have suggested the presence of a second, cone-specific visual cycle, although its physiological function remains to be established. We found that the Müller cells in the salamander neural retina promote cone-specific pigment regeneration and dark adaptation that are independent of the pigment epithelium. Without this pathway, dark adaptation of cones was slow and incomplete. Notably, the rates of cone pigment regeneration by the retina and pigment epithelium visual cycles were essentially identical, suggesting a possible common rate-limiting step. Finally, we also observed cone dark adaptation in the isolated mouse retina.

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Figure 1: Effect of bleach on pigment content in salamander photoreceptors in dissociated and intact retina.
Figure 2: Effect of bleach on sensitivity in salamander photoreceptors in dissociated and intact retina.
Figure 3: Effect of the Müller cell inhibitor L-α-AAA on the recovery of cone sensitivity following a bleach.
Figure 4: Rod and cone responses from salamander whole-retina ERG recordings.
Figure 5: Kinetics of cone dark adaptation from whole-retina ERG recordings.
Figure 6: Rod and cone responses from mouse whole-retina ERG recordings.


  1. 1

    Ebrey, T. & Koutalos, Y. Vertebrate photoreceptors. Prog. Retin. Eye Res. 20, 49–94 (2001).

    CAS  Article  Google Scholar 

  2. 2

    Wald, G., Brown, P.K. & Smith, P.H. Iodopsin. J. Gen. Physiol. 38, 623–681 (1955).

    CAS  Article  Google Scholar 

  3. 3

    Palczewski, K. et al. Rod outer segment retinol dehydrogenase: substrate specificity and role in phototransduction. Biochemistry 33, 13741–13750 (1994).

    CAS  Article  Google Scholar 

  4. 4

    Jones, G.J., Crouch, R.K., Wiggert, B., Cornwall, M.C. & Chader, G.J. Retinoid requirements for recovery of sensitivity after visual-pigment bleaching in isolated photoreceptors. Proc. Natl. Acad. Sci. USA 86, 9606–9610 (1989).

    CAS  Article  Google Scholar 

  5. 5

    Pepperberg, D.R. et al. Interphotoreceptor retinoid-binding protein (IRBP). Molecular biology and physiological role in the visual cycle of rhodopsin. Mol. Neurobiol. 7, 61–85 (1993).

    CAS  Article  Google Scholar 

  6. 6

    Hecht, S., Haig, C. & Chase, A.M. The influence of light adaptation on subsequent dark adaptation of the eye. J. Gen. Physiol. 20, 831–850 (1937).

    CAS  Article  Google Scholar 

  7. 7

    Rushton, W.A. Visual Adaptation. Proc. R. Soc. Lond. B 162, 20–46 (1965).

    CAS  Article  Google Scholar 

  8. 8

    Saari, J.C. Biochemistry of visual pigment regeneration: the Friedenwald lecture. Invest. Ophthalmol. Vis. Sci. 41, 337–348 (2000).

    CAS  PubMed  Google Scholar 

  9. 9

    Kefalov, V.J. et al. Breaking the covalent bond: a pigment property that contributes to desensitization in cones. Neuron 46, 879–890 (2005).

    CAS  Article  Google Scholar 

  10. 10

    Matsumoto, H., Tokunaga, F. & Yoshizawa, T. Accessibility of the iodopsin chromophore. Biochim. Biophys. Acta 404, 300–308 (1975).

    CAS  Article  Google Scholar 

  11. 11

    Mata, N.L., Radu, R.A., Clemmons, R.C. & Travis, G.H. Isomerization and oxidation of vitamin A in cone-dominant retinas: a novel pathway for visual-pigment regeneration in daylight. Neuron 36, 69–80 (2002).

    CAS  Article  Google Scholar 

  12. 12

    Das, S.R., Bhardwaj, N., Kjeldbye, H. & Gouras, P. Muller cells of chicken retina synthesize 11-cis-retinol. Biochem. J. 285, 907–913 (1992).

    CAS  Article  Google Scholar 

  13. 13

    Bustamante, J.J., Ziari, S., Ramirez, R.D. & Tsin, A.T. Retinyl ester hydrolase and the visual cycle in the chicken eye. Am. J. Physiol. 269, R1346–R1350 (1995).

    CAS  PubMed  Google Scholar 

  14. 14

    Trevino, S.G., Villazana-Espinoza, E.T., Muniz, A. & Tsin, A.T. Retinoid cycles in the cone-dominated chicken retina. J. Exp. Biol. 208, 4151–4157 (2005).

    Article  Google Scholar 

  15. 15

    Villazana-Espinoza, E.T., Hatch, A.L. & Tsin, A.T. Effect of light exposure on the accumulation and depletion of retinyl ester in the chicken retina. Exp. Eye Res. 83, 871–876 (2006).

    CAS  Article  Google Scholar 

  16. 16

    Feathers, K.L. et al. Nrl-knockout mice deficient in Rpe65 fail to synthesize 11-cis retinal and cone outer segments. Invest. Ophthalmol. Vis. Sci. 49, 1126–1135 (2008).

    Article  Google Scholar 

  17. 17

    Wenzel, A. et al. RPE65 is essential for the function of cone photoreceptors in NRL-deficient mice. Invest. Ophthalmol. Vis. Sci. 48, 534–542 (2007).

    Article  Google Scholar 

  18. 18

    Mariani, A.P. Photoreceptors of the larval tiger salamander retina. Proc. R. Soc. Lond. B 227, 483–492 (1986).

    CAS  Article  Google Scholar 

  19. 19

    Jablonski, M.M. & Iannaccone, A. Targeted disruption of Muller cell metabolism induces photoreceptor dysmorphogenesis. Glia 32, 192–204 (2000).

    CAS  Article  Google Scholar 

  20. 20

    Kato, S., Ishita, S., Sugawara, K. & Mawatari, K. Cystine/glutamate antiporter expression in retinal Muller glial cells: implications for dl-alpha-aminoadipate toxicity. Neuroscience 57, 473–482 (1993).

    CAS  Article  Google Scholar 

  21. 21

    Pedersen, O.O. & Karlsen, R.L. Destruction of Muller cells in the adult rat by intravitreal injection of d,l-alpha-aminoadipic acid. An electron microscopic study. Exp. Eye Res. 28, 569–575 (1979).

    CAS  Article  Google Scholar 

  22. 22

    Bonaventure, N., Roussel, G. & Wioland, N. Effects of dl-alpha-amino adipic acid on Muller cells in frog and chicken retinae in vivo: relation to ERG b wave, ganglion cell discharge and tectal evoked potentials. Neurosci. Lett. 27, 81–87 (1981).

    CAS  Article  Google Scholar 

  23. 23

    Pow, D.V. Visualising the activity of the cystine-glutamate antiporter in glial cells using antibodies to aminoadipic acid, a selectively transported substrate. Glia 34, 27–38 (2001).

    CAS  Article  Google Scholar 

  24. 24

    Goldstein, E.B. Cone pigment regeneration in the isolated frog retina. Vision Res. 10, 1065–1068 (1970).

    CAS  Article  Google Scholar 

  25. 25

    Jin, J., Jones, G.J. & Cornwall, M.C. Movement of retinal along cone and rod photoreceptors. Vis. Neurosci. 11, 389–399 (1994).

    CAS  Article  Google Scholar 

  26. 26

    Sarantis, M. & Mobbs, P. The spatial relationship between Muller cell processes and the photoreceptor output synapse. Brain Res. 584, 299–304 (1992).

    CAS  Article  Google Scholar 

  27. 27

    Lamb, T.D. & Pugh, E.N. Jr. Dark adaptation and the retinoid cycle of vision. Prog. Retin. Eye Res. 23, 307–380 (2004).

    CAS  Article  Google Scholar 

  28. 28

    Redmond, T.M. et al. Rpe65 is necessary for production of 11-cis-vitamin A in the retinal visual cycle. Nat. Genet. 20, 344–351 (1998).

    CAS  Article  Google Scholar 

  29. 29

    Ahmad, K.M., Klug, K., Herr, S., Sterling, P. & Schein, S. Cell density ratios in a foveal patch in macaque retina. Vis. Neurosci. 20, 189–209 (2003).

    Article  Google Scholar 

  30. 30

    Green, D.G., Guo, H. & Pillers, D.A. Normal photoresponses and altered b-wave responses to APB in the mdx(Cv3) mouse isolated retina ERG supports role for dystrophin in synaptic transmission. Vis. Neurosci. 21, 739–747 (2004).

    Article  Google Scholar 

  31. 31

    Thoreson, W.B. & Ulphani, J.S. Pharmacology of selective and non-selective metabotropic glutamate receptor agonists at l-AP4 receptors in retinal ON bipolar cells. Brain Res. 676, 93–102 (1995).

    CAS  Article  Google Scholar 

  32. 32

    Yu, W. & Miller, R.F. NBQX, an improved non-NMDA antagonist studied in retinal ganglion cells. Brain Res. 692, 190–194 (1995).

    CAS  Article  Google Scholar 

  33. 33

    Coleman, P.A. & Do Miller, R.F. N-methyl-D-aspartate receptors mediate synaptic responses in the mudpuppy retina? J. Neurosci. 8, 4728–4733 (1988).

    CAS  Article  Google Scholar 

  34. 34

    Green, D.G. & Kapousta-Bruneau, N.V. A dissection of the electroretinogram from the isolated rat retina with microelectrodes and drugs. Vis. Neurosci. 16, 727–741 (1999).

    CAS  Article  Google Scholar 

  35. 35

    Levine, J.S. & MacNichol, E.F. Jr. Microspectrophotometry of primate photoreceptors: art, artifact, and analysis. in The Visual System (eds A. Fein & J.S. Levine) 73–78 (Liss, New York, 1985).

    Google Scholar 

  36. 36

    MacNichol, E.F. Jr. A photon-counting microspectrophotometer for the study of single vertebrate photoreceptor cells. in Frontiers of Visual Science (eds S.J. Cool & E.L. Smith) 194–208 (Springer, Berlin, 1978).

    Chapter  Google Scholar 

  37. 37

    Govardovskii, V.I., Fyhrquist, N., Reuter, T., Kuzmin, D.G. & Donner, K. In search of the visual pigment template. Vis. Neurosci. 17, 509–528 (2000).

    CAS  Article  Google Scholar 

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We thank R. Crouch for the generous gift of 11-cis retinal and 11-cis retinol, P. Ala-Laurila for his expert assistance with the analysis of microspectrophotometric data and K.-W. Yau, P. Lukasiewicz, J. Corbo, R. Crouch and P. Ala-Laurila for comments on the manuscript. This work was supported by a Career Development Award from Research to Prevent Blindness, the Karl Kirchgessner Foundation and NIH grants EY 019312 (V.J.K.), EY 01157 (M.C.C.) and EY 02687 (Department of Ophthalmology and Visual Sciences at Washington University).

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Correspondence to Vladimir J Kefalov.

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Wang, JS., Estevez, M., Cornwall, M. et al. Intra-retinal visual cycle required for rapid and complete cone dark adaptation. Nat Neurosci 12, 295–302 (2009).

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