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Kinetics of Rhodopsin’s Chromophore Monitored in a Single Photoreceptor

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Rhodopsin

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1271))

Abstract

Absorption of light isomerizes the retinyl chromophore of the photoreceptor pigment rhodopsin from 11-cis to all-trans, generating the photoactivated rhodopsin form. The photoisomerization of the chromophore however destroys rhodopsin, and its regeneration requires the removal of the all-trans and the supply of fresh 11-cis chromophore. The all-trans chromophore is removed through a series of steps beginning with its release from photoactivated rhodopsin in the form of all-trans-retinal, leaving behind the apoprotein opsin. All-trans-retinal is then reduced to all-trans-retinol, which is transported out of the photoreceptor. Rhodopsin is regenerated from opsin and fresh 11-cis-retinal arriving to the photoreceptor from the retinal pigment epithelium. Both all-trans and 11-cis-retinal can form precursors of lipofuscin, a pigment that accumulates with age in the lysosomal compartment of the retinal pigment epithelium. All-trans-retinal, all-trans-retinol, and lipofuscin precursors all emit significant and distinct fluorescence signals, allowing their monitoring in single photoreceptor cells with fluorescence imaging. Here we describe the procedures for measuring these fluorophores in single mouse rod photoreceptors.

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Authors and Affiliations

  1. Departments of Ophthalmology and Neurosciences, Medical University of South Carolina, 167 Ashley Ave., Charleston, SC, 29425, USA

    Leopold Adler IV, Nicholas P. Boyer, Chunhe Chen & Yiannis Koutalos Ph.D.

Authors
  1. Leopold Adler IV
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  2. Nicholas P. Boyer
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  3. Chunhe Chen
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  4. Yiannis Koutalos Ph.D.
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Corresponding author

Correspondence to Yiannis Koutalos Ph.D. .

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Editors and Affiliations

  1. Department of Pharmacology, School of Medicine, Case Western Reserve University, Cleveland, Ohio, USA

    Beata Jastrzebska

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Adler, L., Boyer, N.P., Chen, C., Koutalos, Y. (2015). Kinetics of Rhodopsin’s Chromophore Monitored in a Single Photoreceptor. In: Jastrzebska, B. (eds) Rhodopsin. Methods in Molecular Biology, vol 1271. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2330-4_21

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  • DOI: https://doi.org/10.1007/978-1-4939-2330-4_21

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2329-8

  • Online ISBN: 978-1-4939-2330-4

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