Mutant rhodopsin transgene expression on a null background

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Publication Type Journal Article
School or College College of Science; School of Medicine
Department Ophthalmology; Neurobiology & Anatomy; Biology
Creator Baehr, Wolfgang; Frederick, Jeanne M.; Church-Kopish, Jill; Howes, Kimberly
Other Author Krasnoperova, Nataliia V.; Ruther, Klaus.; Lem, Janice; Hoffmann, Kirstin
Title Mutant rhodopsin transgene expression on a null background
Date 2001-03
Description PURPOSE. To study mechanisms leading to photoreceptor degeneration in mouse models for autosomal dominant retinitis pigmentosa (adRP) based on the rhodopsin P23H mutation. METHODS. Mice of a transgenic line expressing a rhodopsin triple mutant, V20G, P23H, and P27L (GHL), were mated with rhodopsin (rho) knockout mice. Littermates of various ages and genotypes (GHL+rho+/+, GHL+rho+/-, and GHL+rho-/-) were examined for outer nuclear layer thickness and outer segment formation (histology), fate of mutant rhodopsin (immunocytochemistry), and photoreceptor function (electroretinogram; ERG). RESULTS. Mice expressing GHL-rhodopsin in the absence of wild-type rhodopsin had severe retinopathy, which was nearly complete by postnatal day (P)30. GHL-rhodopsin formed homodimers nearly exclusively on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels, whereas wild-type rhodopsin predominantly formed monomers. Expression level of mutant rhodopsin in predegenerate (P10) GHL+rho-/- retinas was low, approximately 10% to 25% of normal levels. No elaboration of disc membrane or outer segment formation was observed at any time point examined. The mutant rhodopsin was found mostly in perinuclear locales (endoplasmic reticulum; ER) as evidenced by colocalization using the antibodies Rho1D4 and calnexin-NT. CONCLUSIONS. GHL-rhodopsin dimerizes, localizes to the ER, and fails to transport and support outer segment formation. Additionally, the mutant protein does not support a scotopic ERG a-wave and accelerates photoreceptor degeneration over that occurring with the rhodopsin knockout alone. These findings indicate a cytotoxic effect of the mutant protein, probably elicited by an unfolded protein response.
Type Text
Publisher Association for Research in Vision and Ophthalmology
Volume 42
Issue 3
First Page 826
Last Page 833
Subject Mutant Rhodopsin; Photoreceptor Degeneration; Autosomal Dominant Retinitis Pigmentosa; Rhodopsin
Subject MESH Electroretinography; Gene Expression; Retinal Degeneration;Rhodopsin; Transgenes
Language eng
Bibliographic Citation Frederick JM, Krasnoperova NV, Hoffmann K, Church-Kopish J, Ruther K, Howes K, Lem J, Baehr W. (2001). Mutant rhodopsin transgene expression on a null background. Invest Ophthalmol Vis Sci, 42(3), 826-33
Rights Management (c) Association for Research in Vision and Ophthalmology
Format Medium application/pdf
Identifier ir-main,1705
ARK ark:/87278/s6mg872r
Setname ir_uspace
Date Created 2012-06-13
Date Modified 2012-06-13
ID 706338
Reference URL https://collections.lib.utah.edu/ark:/87278/s6mg872r
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