Hostname: page-component-8448b6f56d-wq2xx Total loading time: 0 Render date: 2024-04-25T04:51:45.869Z Has data issue: false hasContentIssue false
  • English
  • Français

Generation, characterization, and molecular cloning of the Noerg-1 mutation of rhodopsin in the mouse

Published online by Cambridge University Press:  06 December 2005

LAWRENCE H. PINTO ,
MARTHA H. VITATERNA ,
KAZUHIRO SHIMOMURA ,
SANDRA M. SIEPKA ,
ERIN L. MCDEARMON ,
DEBORAH FENNER ,
STEPHEN L. LUMAYAG ,
CHIAKI OMURA ,
ANNE W. ANDREWS  and
MATTHEW BAKER
...Show all authors
LAWRENCE H. PINTO
Affiliation:
Department of Neurobiology and Physiology and Center for Functional Genomics, Northwestern University, Evanston
MARTHA H. VITATERNA
Affiliation:
Department of Neurobiology and Physiology and Center for Functional Genomics, Northwestern University, Evanston
KAZUHIRO SHIMOMURA
Affiliation:
Department of Neurobiology and Physiology and Center for Functional Genomics, Northwestern University, Evanston
SANDRA M. SIEPKA
Affiliation:
Department of Neurobiology and Physiology and Center for Functional Genomics, Northwestern University, Evanston
ERIN L. MCDEARMON
Affiliation:
Department of Neurobiology and Physiology and Center for Functional Genomics, Northwestern University, Evanston
DEBORAH FENNER
Affiliation:
Department of Neurobiology and Physiology and Center for Functional Genomics, Northwestern University, Evanston
STEPHEN L. LUMAYAG
Affiliation:
Department of Neurobiology and Physiology and Center for Functional Genomics, Northwestern University, Evanston
CHIAKI OMURA
Affiliation:
Department of Neurobiology and Physiology and Center for Functional Genomics, Northwestern University, Evanston
ANNE W. ANDREWS
Affiliation:
Department of Neurobiology and Physiology and Center for Functional Genomics, Northwestern University, Evanston
MATTHEW BAKER
Affiliation:
Department of Neurobiology and Physiology and Center for Functional Genomics, Northwestern University, Evanston
BRANDON M. INVERGO
Affiliation:
Department of Neurobiology and Physiology and Center for Functional Genomics, Northwestern University, Evanston
MARISSA A. OLVERA
Affiliation:
Department of Ophthalmology & Visual Sciences, University of Iowa, Iowa City
EDWARD HEFFRON
Affiliation:
Department of Ophthalmology & Visual Sciences, University of Iowa, Iowa City
ROBERT F. MULLINS
Affiliation:
Department of Ophthalmology & Visual Sciences, University of Iowa, Iowa City
VAL C. SHEFFIELD
Affiliation:
Departments of Pediatrics and Genetics, University of Iowa, Iowa City Howard Hughes Medical Institute, Carver College of Medicine, Iowa City
EDWIN M. STONE
Affiliation:
Department of Ophthalmology & Visual Sciences, University of Iowa, Iowa City
JOSEPH S. TAKAHASHI
Affiliation:
Department of Neurobiology and Physiology and Center for Functional Genomics, Northwestern University, Evanston Howard Hughes Medical Institute, Carver College of Medicine, Iowa City
Get access Rights & Permissions [Opens in a new window]

Abstract

We performed genome-wide mutagenesis of C57BL/6J mice using the mutagen N-ethyl-N-nitrosourea (ENU) and screened the third generation (G3) offspring for visual system alterations using electroretinography and fundus photography. Several mice in one pedigree showed characteristics of retinal degeneration when tested at 12–14 weeks of age: no recordable electroretinogram (ERG), attenuation of retinal vessels, and speckled pigmentation of the fundus. Histological studies showed that the retinas undergo a photoreceptor degeneration with apoptotic loss of outer nuclear layer nuclei but visual acuity measured using the optomotor response under photopic conditions persists in spite of considerable photoreceptor loss. The Noerg-1 mutation showed an autosomal dominant pattern of inheritance in progeny. Studies in early postnatal mice showed degeneration to occur after formation of partially functional rods. The Noerg-1 mutation was mapped genetically to chromosome 6 by crossing C57BL/6J mutants with DBA/2J or BALB/cJ mice to produce an N2 generation and then determining the ERG phenotypes and the genotypes of the N2 offspring at multiple loci using SSLP and SNP markers. Fine mapping was accomplished with a set of closely spaced markers. A nonrecombinant region from 112.8 Mb to 115.1 Mb was identified, encompassing the rhodopsin (Rho) coding region. A single nucleotide transition from G to A was found in the Rho gene that is predicted to result in a substitution of Tyr for Cys at position 110, in an intradiscal loop. This mutation has been found in patients with autosomal dominant retinitis pigmentosa (RP) and results in misfolding of rhodopsin expressed in vitro. Thus, ENU mutagenesis is capable of replicating mutations that occur in human patients and is useful for generating de novo models of human inherited eye disease. Furthermore, the availability of the mouse genomic sequence and extensive DNA polymorphisms made the rapid identification of this gene possible, demonstrating that the use of ENU-induced mutations for functional gene identification is now practical for individual laboratories.

Keywords

Chemical mutagenesisENUForward geneticsRetinaMouse modelsRetinitis pigmentosaMolecular cloningPositional cloningGene discoveryRhodopsinProtein foldingDisulfide bond formationRodsRod photoreceptorRho-C110Y mutationOptomotor responseVisual acuity
Type
Research Article
Information
Visual Neuroscience , Volume 22 , Issue 5 , September 2005 , pp. 619 - 629
Copyright
© 2005 Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Blackshaw, S., Fraioli, R.E., Furukawa, T., & Cepko, C.L. (2001). Comprehensive analysis of photoreceptor gene expression and the identification of candidate retinal disease genes. Cell 107, 579–589.CrossRefGoogle Scholar
Blanks, J.C., Adinolfi, A.M., & Lolley, R.N. (1974). Photoreceptor degeneration and synaptogenesis in retinal-degenerative (rd) mice. Journal of Comparative Neurology 156, 95–106.CrossRefGoogle Scholar
Brand, M., Heisenberg, C.P., Warga, R.M., Pelegri, F., Karlstrom, R.O., Beuchle, D., Picker, A., Jiang, Y.J., Furutani-Seiki, M., van Eeden, F.J., Granato, M., Haffter, P., Hammerschmidt, M., Kane, D.A., Kelsh, R.N., Mullins, M.C., Odenthal, J., & Nusslein-Volhard, C. (1996). Mutations affecting development of the midline and general body shape during zebrafish embryogenesis. Development 123, 129–142.Google Scholar
Dalke, C., Loster, J., Fuchs, H., Gailus-Durner, V., Soewarto, D., Favor, J., Neuhauser-Klaus, A., Pretsch, W., Gekeler, F., Shinoda, K., Zrenner, E., Meitinger, T., de Angelis, M.H., & Graw, J. (2004). Electroretinography as a screening method for mutations causing retinal dysfunction in mice. Investigative Ophthalmology and Visual Science 45, 601–609.CrossRefGoogle Scholar
Dawson, W.W., Trick, G.L., & Litzkow, C.A. (1979). Improved electrode for electroretinography. Investigative Ophthalmology and Visual Science 18, 988–991.Google Scholar
Doyle, D.A., Cabral, J.M., Pfuetzner, R.A., Kuo, A., Gulbis, J.M., Cohen, S.L., Chait, B.T., & MacKinnon, R. (1998). The structure of the potassium channel: Molecular basis of K+ conduction and selectivity. Science 280, 69–77.CrossRefGoogle Scholar
Dryja, T.P., Hahn, L.B., Cowley, G.S., McGee, T.L., & Berson, E.L. (1991). Mutation spectrum of the rhodopsin gene among patients with autosomal dominant retinitis pigmentosa. Proceedings of the National Academy of Sciences of the U.S.A. 88, 9370–9374.CrossRefGoogle Scholar
Frederick, J.M., Krasnoperova, N.V., Hoffmann, K., Church-Kopish, J., Ruther, K., Howes, K., Lem, J., & Baehr, W. (2001). Mutant rhodopsin transgene expression on a null background. Investigative Ophthalmology and Visual Science 42, 826–833.Google Scholar
Germer, S., Holland, M.J., & Higuchi, R. (2000). High-throughput SNP allele-frequency determination in pooled DNA samples by kinetic PCR. Genome Research 10, 258–266.CrossRefGoogle Scholar
Hwa, J., Klein-Seetharaman, J., & Khorana, H.G. (2001). Structure and function in rhodopsin: Mass spectrometric identification of the abnormal intradiscal disulfide bond in misfolded retinitis pigmentosa mutants. Proceedings of the National Academy of Sciences of the U.S.A. 98, 4872–4876.CrossRefGoogle Scholar
Hwa, J., Reeves, P.J., Klein-Seetharaman, J., Davidson, F., & Khorana, H.G. (1999). Structure and function in rhodopsin: Further elucidation of the role of the intradiscal cysteines, Cys-110, -185, and -187, in rhodopsin folding and function. Proceedings of the National Academy of Sciences of the U.S.A. 96, 1932–1935.CrossRefGoogle Scholar
Illing, M.E., Rajan, R.S., Bence, N.F., & Kopito, R.R. (2002). A rhodopsin mutant linked to autosomal dominant retinitis pigmentosa is prone to aggregate and interacts with the ubiquitin proteasome system. Journal of Biological Chemistry 277, 34150–34160.CrossRefGoogle Scholar
Johnson, L.V. & Blanks, J.C. (1984). Application of acrylamide as an embedding medium in studies of lectin and antibody binding in the vertebrate retina. Current Eye Research 3, 969–974.CrossRefGoogle Scholar
Justice, M.J., Noveroske, J.K., Weber, J.S., Zheng, B., & Bradley, A. (1999). Mouse ENU mutagenesis. In Human Molecular Genetics 8, 1955–1963.CrossRefGoogle Scholar
Karnik, S.S. & Khorana, H.G. (1990). Assembly of functional rhodopsin requires a disulfide bond between cysteine residues 110 and 187. Journal of Biological Chemistry 265, 17520–17524.Google Scholar
Keverne, E.B. (1997). An evaluation of what the mouse knockout experiments are telling us about mammalian behaviour. Bioessays 19, 1091–1098.CrossRefGoogle Scholar
Kono, M., Yu, H., & Oprian, D.D. (1998). Disulfide bond exchange in rhodopsin. Biochemistry 37, 1302–1305.CrossRefGoogle Scholar
Konopka, R.J. & Benzer, S. (1971). Clock mutants of Drosophila melanogaster. Proceedings of the National Academy of Sciences of the United States of America 68, 2112–2116.CrossRefGoogle Scholar
McGill, T.J., Douglas, R.M., Lund, R.D., & Prusky, G.T. (2004). Quantification of spatial vision in the Royal College of Surgeons rat. Investigative Ophthalmology and Visual Science 45, 932–936.CrossRefGoogle Scholar
Moldin, S.O., Farmer, M.E., Chin, H.R., & Battey, J.F., Jr. (2001). Trans-NIH neuroscience initiatives on mouse phenotyping and mutagenesis. In Mammalian Genome 12, 575–581.CrossRefGoogle Scholar
Nishimura, D.Y., Fath, M., Mullins, R.F., Searby, C., Andrews, M., Davis, R., Andorf, J.L., Mykytyn, K., Swiderski, R.E., Yang, B., Carmi, R., Stone, E.M., & Sheffield, V.C. (2004). Bbs2-null mice have neurosensory deficits, a defect in social dominance, and retinopathy associated with mislocalization of rhodopsin. Proceedings of the National Academy of Sciences of the U.S.A. 101, 16588–16593.CrossRefGoogle Scholar
Nusslein-Volhard, C. & Wieschaus, E. (1980). Mutations affecting segment number and polarity in Drosophila. Nature 287, 795–801.CrossRefGoogle Scholar
Pahl, H.L. (1999). Signal transduction from the endoplasmic reticulum to the cell nucleus. Physiological Reviews 79, 683–701.Google Scholar
Papazian, D.M., Schwarz, T.L., Tempel, B.L., Jan, Y.N., & Jan, L.Y. (1987). Cloning of genomic and complementary DNA from Shaker, a putative potassium channel gene from Drosophila. Science 237, 749–753.CrossRefGoogle Scholar
Peachey, N.S. & Ball, S.L. (2003). Electrophysiological analysis of visual function in mutant mice. Documenta Ophthalmologica 107, 13–36.CrossRefGoogle Scholar
Pearn, M.T., Randall, L.L., Shortridge, R.D., Burg, M.G., & Pak, W.L. (1996). Molecular, biochemical, and electrophysiological characterization of Drosophila norpA mutants. Journal of Biological Chemistry 271, 4937–4945.Google Scholar
Pinto, L.H. & Enroth-Cugell, C. (2000). Tests of the mouse visual system. Mammalian Genome 11, 531–536.CrossRefGoogle Scholar
Prusky, G.T., Alam, N.M., Beekman, S., & Douglas, R.M. (2004). Rapid quantification of adult and developing mouse spatial vision using a virtual optomotor system. Investigative Ophthalmology and Visual Science 45, 4611–4616.CrossRefGoogle Scholar
Rajan, R.S. & Kopito, R.R. (2005). Suppression of wild-type rhodopsin maturation by mutants linked to autosomal dominant retinitis pigmentosa. Journal of Biological Chemistry 280, 1284–1291.CrossRefGoogle Scholar
Rozen, S. & Skaletsky, H. (2000). Primer3 on the WWW for general users and for biologist programmers. Methods of Molecular Biology 132, 365–386.Google Scholar
Sanyal, S. & Bal, A.K. (1973). Comparative light and electron microscopic study of retinal histogenesis in normal and rd mutant mice. Zeitschrift Anatomische Entwicklungsgeschichte 142, 219–238.CrossRefGoogle Scholar
Siepka, S.M. & Takahashi, J.S. (2005). Forward genetic screen to identify circadian rhythm mutants in mice. Methods in Enzymology 393, 217–228.Google Scholar
Sung, C.H., Davenport, C.M., Hennessey, J.C., Maumenee, I.H., Jacobson, S.G., Heckenlively, J.R., Nowakowski, R., Fishman, G., Gouras, P., & Nathans, J. (1991). Rhodopsin mutations in autosomal dominant retinitis pigmentosa. Proceedings of the National Academy of Sciences of the U.S.A. 88, 6481–6485.CrossRefGoogle Scholar
Takahashi, J.S., Pinto, L.H., & Vitaterna, M.H. (1994). Forward and reverse genetic approaches to behavior in the mouse [see comments]. Science 264, 1724–1733.CrossRefGoogle Scholar
Tempel, B.L., Papazian, D.M., Schwarz, T.L., Jan, Y.N., & Jan, L.Y. (1987). Sequence of a probable potassium channel component encoded at Shaker locus of Drosophila. Science 237, 770–775.CrossRefGoogle Scholar
Vaithinathan, R., Berson, E.L., & Dryja, T.P. (1994). Further screening of the rhodopsin gene in patients with autosomal dominant retinitis pigmentosa. Genomics 21, 461–463.CrossRefGoogle Scholar
Vitaterna, M.H., Pinto, L.H., & Turek, F.W. (2005). Molecular genetic basis for mammalian circadian rhythms. In Principles and Practice of Sleep Medicine, ed. Kryger, M.H., pp. 363–374. Philadelphia: Saulders.CrossRef
Wu, C.-F., Ganetzky, B., Haugland, F., & Liu, A.-X. (1983). Potassium currents in drosophila: Different components affected by mutations of two genes. Science 220, 1076–1078.CrossRefGoogle Scholar
Zhou, G., Kamahori, M., Okano, K., Chuan, G., Harada, K., & Kambara, H. (2001). Quantitative detection of single nucleotide polymorphisms for a pooled sample by a bioluminometric assay coupled with modified primer extension reactions (BAMPER). Nucleic Acids Research 29, E93.CrossRefGoogle Scholar