An S-opsin knock-in mouse (F81Y) reveals a role for the native ligand 11-cis-retinal in cone opsin biosynthesis

Christine Insinna, Lauren L. Daniele, Jason A. Davis, DeLaine D. Larsen, Colleen Kuemmel, Jinhua Wang, Sergei S. Nikonov, Barry E. Knox, Edward N Pugh Jr

Research output: Contribution to journalArticle

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Abstract

In absence of their natural ligand, 11-cis-retinal, cone opsin G-protein-coupled receptors fail to traffic normally, a condition associated with photoreceptor degeneration and blindness. We created a mouse with a point mutation (F81Y) in cone S-opsin. As expected, cones with this knock-in mutation respond to light with maximal sensitivity red-shifted from 360 to 420 nm, consistent with an altered interaction between the apoprotein and ligand, 11-cis-retinal. However, cones expressing F81Y S-opsin showed an ~3-fold reduced absolute sensitivity that was associated with a corresponding reduction in S-opsin protein expression. The reduced S-opsin expression did not arise from decreased S-opsin mRNA or cone degeneration, but rather from enhanced endoplasmic reticulum (ER)-associated degradation of the nascent protein. Exogenously increased 11-cis-retinal restored F81Y S-opsin protein expression to normal levels, suggesting that ligand binding in the ER facilitates proper folding. Immunohistochemistry and electron microscopy of normal retinas showed that Mueller cells, which synthesize a precursor of 11-cis-retinal, are closely adjoined to the cone ER, so they could deliver the ligand to the site of opsin synthesis. Together, these results suggest that the binding of 11-cis-retinal in the ER is important for normal folding during cone opsin biosynthesis.

Original languageEnglish (US)
Pages (from-to)8094-8104
Number of pages11
JournalJournal of Neuroscience
Volume32
Issue number23
DOIs
StatePublished - Jun 6 2012

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Cone Opsins
Retinaldehyde
Retinal Cone Photoreceptor Cells
Opsins
Ligands
Endoplasmic Reticulum
Protein S
Endoplasmic Reticulum-Associated Degradation
Apoproteins
Blindness
G-Protein-Coupled Receptors
Point Mutation
Retina
Electron Microscopy
Immunohistochemistry
Light
Messenger RNA
Mutation

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

An S-opsin knock-in mouse (F81Y) reveals a role for the native ligand 11-cis-retinal in cone opsin biosynthesis. / Insinna, Christine; Daniele, Lauren L.; Davis, Jason A.; Larsen, DeLaine D.; Kuemmel, Colleen; Wang, Jinhua; Nikonov, Sergei S.; Knox, Barry E.; Pugh Jr, Edward N.

In: Journal of Neuroscience, Vol. 32, No. 23, 06.06.2012, p. 8094-8104.

Research output: Contribution to journalArticle

Insinna, C, Daniele, LL, Davis, JA, Larsen, DD, Kuemmel, C, Wang, J, Nikonov, SS, Knox, BE & Pugh Jr, EN 2012, 'An S-opsin knock-in mouse (F81Y) reveals a role for the native ligand 11-cis-retinal in cone opsin biosynthesis', Journal of Neuroscience, vol. 32, no. 23, pp. 8094-8104. https://doi.org/10.1523/JNEUROSCI.0131-12.2012
Insinna, Christine ; Daniele, Lauren L. ; Davis, Jason A. ; Larsen, DeLaine D. ; Kuemmel, Colleen ; Wang, Jinhua ; Nikonov, Sergei S. ; Knox, Barry E. ; Pugh Jr, Edward N. / An S-opsin knock-in mouse (F81Y) reveals a role for the native ligand 11-cis-retinal in cone opsin biosynthesis. In: Journal of Neuroscience. 2012 ; Vol. 32, No. 23. pp. 8094-8104.
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