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S-opsin knockout mice with the endogenous M-opsin gene replaced by an L-opsin variant

Published online by Cambridge University Press:  17 December 2013

SCOTT H. GREENWALD ,
JAMES A. KUCHENBECKER ,
DANIEL K. ROBERSON ,
MAUREEN NEITZ  and
JAY NEITZ
SCOTT H. GREENWALD
Affiliation:
Vision Sciences, University of Washington, Graduate Program in Neurobiology and Behavior, Seattle, Washington
JAMES A. KUCHENBECKER
Affiliation:
Vision Sciences, University of Washington, Seattle, Washington
DANIEL K. ROBERSON
Affiliation:
Vision Sciences, University of Washington, Seattle, Washington
MAUREEN NEITZ*
Affiliation:
Vision Sciences, University of Washington, Seattle, Washington
JAY NEITZ*
Affiliation:
Vision Sciences, University of Washington, Seattle, Washington
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Abstract

Specific variants of human long-wavelength (L) and middle-wavelength (M) cone opsin genes have recently been associated with a variety of vision disorders caused by cone malfunction, including red-green color vision deficiency, blue cone monochromacy, myopia, and cone dystrophy. Strikingly, unlike disease-causing mutations in rhodopsin, most of the cone opsin alleles that are associated with vision disorders do not have deleterious point mutations. Instead, specific combinations of normal polymorphisms that arose by genetic recombination between the genes encoding L and M opsins appear to cause disease. Knockout/knock-in mice promise to make it possible to study how these deleterious cone opsin variants affect the structure, function, and viability of the cone photoreceptors. Ideally, we would like to evaluate different variants that cause vision disorders in humans against a control pigment that is not associated with vision disorders, and each variant should be expressed as the sole photopigment in each mouse cone, as is the case in humans. To evaluate the feasibility of this approach, we created a line of mice to serve as the control in the analysis of disease-causing mutations by replacing exon 2 through 6 of the mouse M-opsin gene with the corresponding cDNA for a human L-opsin variant that is associated with normal vision. Experiments reported here establish that the resulting pigment, which differs from the endogenous mouse M opsin at 35 amino acid positions, functions normally in mouse cones. This pigment was evaluated in mice with and without coexpression of the mouse short wavelength (S) opsin. Here, the creation and validation of two lines of genetically engineered mice that can be used to study disease-causing variants of human L/M-opsins, in vivo, are described.

Keywords

Cone photoreceptorCone photopigmentElectroretinogramImmunohistochemistryCone functionTargeted gene replacementPhotopigment mutationS-opsin knockout
Type
Research Articles
Information
Visual Neuroscience , Volume 31 , Issue 1 , January 2014 , pp. 25 - 37
Copyright
Copyright © Cambridge University Press 2013 

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References

Applebury, M.L., Antoch, M.P., Baxter, L.C., Chun, L.L.Y., Kalk, J.D., Farhangfar, F., Kage, K., Krzystolik, M.G., Lyass, L.A. & Robbins, J.T. (2000). The murine cone photoreceptor: A single cone type expresses both S and M opsins with retinal spatial patterning. Neuron 27, 513–523.CrossRefGoogle Scholar
Asenjo, A.B., Rim, J. & Oprian, D.D. (1994). Molecular determinants of human red/green color discrimination. Neuron 12, 1131–1138.Google Scholar
Bush, R.A. & Sieving, P.A. (1996). Inner retinal contributions to the primate photopic fast flicker electroretinogram. Journal of the Optical Society of America. A, Optics, Image Science, and Vision 13, 557–565.Google Scholar
Carroll, J., Dubra, A., Gardner, J.C., Mizrahi-Meissonnier, L., Cooper, R.F., Dubis, A.M., Nordgren, R., Genead, M., Connor, T.B. Jr., Stepien, K.E., Sharon, D., Hunt, D.M., Banin, E., Hardcastle, A.J., Moore, A.T., Williams, D.R., Fishman, G., Neitz, J., Neitz, M. & Michaelides, M. (2012). The effect of cone opsin mutations on retinal structure and the integrity of the photoreceptor mosaic. Investigative Ophthalmology & Visual Science 53, 8006–8015.Google Scholar
Carroll, J., McMahon, C., Neitz, M. & Neitz, J. (2000). Flicker-photometric electroretinogram estimates of L:M cone photoreceptor ratio in men with photopigment spectra derived from genetics. Journal of the Optical Society of America. A, Optics, Image Science, and Vision 17, 499–509.Google Scholar
Carroll, J., Neitz, M., Hofer, H., Neitz, J. & Williams, D.R. (2004). Functional photoreceptor loss revealed with adaptive optics: An alternate cause of color blindness. Proceedings of the National Academy of Sciences of the United States of America 101, 8461–8466.Google Scholar
Carroll, J., Neitz, M. & Neitz, J. (2002). Estimates of L:M cone ratio from ERG flicker photometry and genetics. Journal of Vision 2, 531–542.Google Scholar
Crognale, M.A., Fry, M., Highsmith, J., Haegerstrom-Portnoy, G., Neitz, J., Neitz, M. & Webster, M.A. (2004). Characterization of a novel form of X-linked incomplete achromatopsia. Visual Neuroscience 21, 197–204.Google Scholar
Daniele, L.L., Insinna, C., Chance, R., Wang, J., Nikonov, S.S. & Pugh, E.N. Jr. (2011). A mouse M-opsin monochromat: Retinal cone photoreceptors have increased M-opsin expression when S-opsin is knocked out. Vision Research 51, 447–458.Google Scholar
Fliesler, S.J. & Basinger, S.F. (1985). Tunicamycin blocks the incorporation of opsin into retinal rod outer segment membranes. Proceedings of the National Academy of Sciences of the United States of America 82, 1116–1120.CrossRefGoogle ScholarPubMed
Gresh, J., Goletz, P.W., Crouch, R.K. & Rohrer, B. (2003). Structure-function analysis of rods and cones in juvenile, adult, and aged C57bl/6 and Balb/c mice. Visual Neuroscience 20, 211–220.CrossRefGoogle ScholarPubMed
Haverkamp, S., Wassle, H., Duebel, J., Kuner, T., Augustine, G.J., Feng, G. & Euler, T. (2005). The primordial, blue-cone color system of the mouse retina. The Journal of Neuroscience 25, 5438–5445.Google Scholar
Jacobs, G.H., Deegan, J.F. II & Moran, J.L. (1996). ERG measurements of the spectral sensitivity of common chimpanzee (Pan troglodytes). Vision Research 36, 2587–2594.CrossRefGoogle ScholarPubMed
Jacobs, G.H., Neitz, J. & Deegan, J.F. II. (1991). Retinal receptors in rodents maximally sensitive to ultraviolet light. Nature 353, 655–656.Google Scholar
Lem, J., Krasnoperova, N.V., Calvert, P.D., Kosaras, B., Cameron, D.A., Nicolo, M., Makino, C.L. & Sidman, R.L. (1999). Morphological, physiological, and biochemical changes in rhodopsin knockout mice. Proceedings of the National Academy of Sciences of the United States of America 96, 736–741.Google Scholar
McClements, M., Davies, W.I., Michaelides, M., Young, T., Neitz, M., MacLaren, R.E., Moore, A.T. & Hunt, D.M. (2013). Variations in opsin coding sequences cause x-linked cone dysfunction syndrome with myopia and dichromacy. Investigative Ophthalmology & Visual Science 54, 1361–1369.CrossRefGoogle ScholarPubMed
Merbs, S.L. & Nathans, J. (1993). Role of hydroxyl-bearing amino acids in differentially tuning the absorption spectra of the human red and green cone pigments. Photochemistry and Photobiology 58, 706–710.Google Scholar
Mizrahi-Meissonnier, L., Merin, S., Banin, E. & Sharon, D. (2010). Variable retinal phenotypes caused by mutations in the X-linked photopigment gene array. Investigative Ophthalmology & Visual Science 51, 3884–3892.Google Scholar
Nathans, J., Thomas, D. & Hogness, D.S. (1986). Molecular genetics of human color vision: The genes encoding blue, green, and red pigments. Science 232, 193–202.Google Scholar
Neitz, J. & Neitz, M. (2011). The genetics of normal and defective color vision. Vision Research 51, 633–651.CrossRefGoogle ScholarPubMed
Neitz, J., Neitz, M. & Jacobs, G.H. (1989). Analysis of fusion gene and encoded photopigment of colour-blind humans. Nature 342, 679–682.Google Scholar
Neitz, M., Carroll, J., Renner, A., Knau, H., Werner, J.S. & Neitz, J. (2004). Variety of genotypes in males diagnosed as dichromatic on a conventional clinical anomaloscope. Visual Neuroscience 21, 205–216.CrossRefGoogle ScholarPubMed
Neitz, M., Neitz, J. & Jacobs, G.H. (1991). Spectral tuning of pigments underlying red-green color vision. Science 252, 971–974.Google Scholar
Papermaster, D.S. & Dreyer, W.J. (1974). Rhodopsin content in the outer segment membranes of bovine and frog retinal rods. Biochemistry 13, 2438–2444.Google Scholar
Röhlich, P., van Veen, T. & Szél, Á. (1994). Two different visual pigments in one retinal cone cell. Neuron 13, 1159–1166.Google Scholar
Smallwood, P.M., Olveczky, B.P., Williams, G.L., Jacobs, G.H., Reese, B.E., Meister, M. & Nathans, J. (2003). Genetically engineered mice with an additional class of cone photoreceptors: Implications for the evolution of color vision. Proceedings of the National Academy of Sciences of the United States of America 100, 11706–11711.Google Scholar
Sun, H., Macke, J.P. & Nathans, J. (1997). Mechanisms of spectral tuning in the mouse green cone pigment. Proceedings of the National Academy of Sciences of the United States of America 94, 8860–8865.CrossRefGoogle ScholarPubMed
Szél, Á., Röhlich, P., Mieziewska, K., Aguirre, G. & van Veen, T. (1993). Spatial and temporal differences between the expression of short- and middle-wave sensitive cone pigments in the mouse retina: A developmental study. The Journal of Comparative Neurology 331, 564–577.Google Scholar
Wald, G., Brown, P.K. & Gibbons, I.R. (1963). The problem of visual excitation. Journal of the Optical Society of America. A, Optics, Image Science, and Vision 53, 20–35.Google Scholar
Wang, Z., Asenjo, A.B. & Oprian, D.D. (1993). Identification of the Cl- binding site in the human red and green color vision pigments. Biochemistry 32, 2125–2130.Google Scholar
Williams, G.A. & Jacobs, G.H. (2007). Cone-based vision in the aging mouse. Vision Research 47, 2037–2046.CrossRefGoogle ScholarPubMed