Skip to main content Accessibility help
×
Home

Generation, identification and functional characterization of the nob4 mutation of Grm6 in the mouse

  • LAWRENCE H. PINTO (a1), MARTHA H. VITATERNA (a1), KAZUHIRO SHIMOMURA (a1), SANDRA M. SIEPKA (a1), VICTORIA BALANNIK (a1), ERIN L. MCDEARMON (a1) (a2), CHIAKI OMURA (a1), STEPHEN LUMAYAG (a1), BRANDON M. INVERGO (a1), BRETT GLAWE (a1), DONALD R. CANTRELL (a3), SAMSOON INAYAT (a3) (a4), MARISSA A. OLVERA (a5), KIRSTAN A. VESSEY (a6), MAUREEN A. McCALL (a6) (a7), DENNIS MADDOX (a8), CATHERINE W. MORGANS (a9), BRANDON YOUNG (a10), MATHEW T. PLETCHER (a10), ROBERT F. MULLINS (a5), JOHN B. TROY (a3) and JOSEPH S. TAKAHASHI (a1) (a2)...

Abstract

We performed genome-wide chemical mutagenesis of C57BL/6J mice using N-ethyl-N-nitrosourea (ENU). Electroretinographic screening of the third generation offspring revealed two G3 individuals from one G1 family with a normal a-wave but lacking the b-wave that we named nob4. The mutation was transmitted with a recessive mode of inheritance and mapped to chromosome 11 in a region containing the Grm6 gene, which encodes a metabotropic glutamate receptor protein, mGluR6. Sequencing confirmed a single nucleotide substitution from T to C in the Grm6 gene. The mutation is predicted to result in substitution of Pro for Ser at position 185 within the extracellular, ligand-binding domain and oocytes expressing the homologous mutation in mGluR6 did not display robust glutamate-induced currents. Retinal mRNA levels for Grm6 were not significantly reduced, but no immunoreactivity for mGluR6 protein was found. Histological and fundus evaluations of nob4 showed normal retinal morphology. In contrast, the mutation has severe consequences for visual function. In nob4 mice, fewer retinal ganglion cells (RGCs) responded to the onset (ON) of a bright full field stimulus. When ON responses could be evoked, their onset was significantly delayed. Visual acuity and contrast sensitivity, measured with optomotor responses, were reduced under both photopic and scotopic conditions. This mutant will be useful because its phenotype is similar to that of human patients with congenital stationary night blindness and will provide a tool for understanding retinal circuitry and the role of ganglion cell encoding of visual information.

Copyright

Corresponding author

Address correspondence and reprint requests to: Lawrence H. Pinto, Northwestern University, Dept. Neurobiology & Physiology, 2205 Tech Dr., Northwestern University, Evanston, IL 60208, USA. E-mail: larry-pinto@northwestern.edu

References

Hide All

REFERENCES

Chang, B., Heckenlively, J.R., Bayley, P.R., Brecha, N.C., Davisson, M.T., Hawes, N.L., Hirano, A.A., Hurd, R.E., Ikeda, A., Johnson, B.A., McCall, M.A., Morgans, C.W., Nusinowitz, S., Peachey, N.S., Rice, D.S., Vessey, K.A. & Gregg, R.G. (2006). The nob2 mouse, a null mutation in Cacna1f: Anatomical and functional abnormalities in the outer retina and their consequences on ganglion cell visual responses. Visual Neuroscience 23, 1124.
Dawson, W.W., Trick, G.L. & Litzkow, C.A. (1979). Improved electrode for electroretinography. Investigative Ophthalmology & Visual Science 18, 988991.
Dryja, T.P., McGee, T.L., Berson, E.L., Fishman, G.A., Sandberg, M.A., Alexander, K.R., Derlacki, D.J. & Rajagopalan, A.S. (2005). Night blindness and abnormal cone electroretinogram ON responses in patients with mutations in the GRM6 gene encoding mGluR6. Proceedings of the National Academy of Science USA 102, 48844889.
Gregg, R.G., Mukhopadhyay, S., Candille, S.I., Ball, S.L., Pardue, M.T., McCall, M.A. & Peachey, N.S. (2003). Identification of the gene and the mutation responsible for the mouse nob phenotype. Investigative Ophthalmology & Visual Science 44, 378384.
Kuffler, S.W. (1953). Discharge patterns and functional organization of mammalian retina. Journal of Neurophysiology 16, 3768.
Kunishima, N., Shimada, Y., Tsuji, Y., Sato, T., Yamamoto, M., Kumasaka, T., Nakanishi, S., Jingami, H. & Morikawa, K. (2000). Structural basis of glutamate recognition by a dimeric metabotropic glutamate receptor. Nature 407, 971977.
Masu, M., Iwakabe, H., Tagawa, Y., Miyoshi, T., Yamashita, M., Fukuda, Y., Sasaki, H., Hiroi, K., Nakamura, Y., Shigemoto, R., Takada, M., Nakamura, K., Nakao, K., Katsuki, M. & Nakanishi, S. (1995). Specific deficit of the ON response in visual transmission by targeted disruption of the mGluR6 gene. Cell 80, 757765.
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 & Visual Science 45, 932936.
Morgans, C.W., Bayley, P.R., Oesch, N.W., Ren, G., Akileswaran, L. & Taylor, W.R. (2005). Photoreceptor calcium channels: Insight from night blindness. Visual Neuroscience 22, 561568.
Morgans, C.W., Ren, G. & Akileswaran, L. (2006). Localization of nyctalopin in the mammalian retina. European Journal of Neuroscience 23, 11631171.
Nirenberg, S., Carcieri, S.M., Jacobs, A.L. & Latham, P.E. (2001). Retinal ganglion cells act largely as independent encoders. Nature 411, 698701.
Nirenberg, S. & Meister, M. (1997). The light response of retinal ganglion cells is truncated by a displaced amacrine circuit. Neuron 18, 637650.
O'Connor, E., Allen, L.E., Bradshaw, K., Boylan, J., Moore, A.T. & Trump, D. (2006). Congenital stationary night blindness associated with mutations in GRM6 encoding glutamate receptor MGluR6. British Journal of Ophthalmology 90, 653654.
Pardue, M.T., McCall, M.A., LaVail, M.M., Gregg, R.G. & Peachey, N.S. (1998). A naturally occurring mouse model of X-linked congenital stationary night blindness. Investigative Ophthalmology & Visual Science 39, 24432449.
Pinto, L.H., Vitaterna, M.H., Shimomura, K., Siepka, S.M., McDearmon, E.L., Fenner, D., Lumayag, S.L., Omura, C., Andrews, A.W., Baker, M., Invergo, B.M., Olvera, M.A., Heffron, E., Mullins, R.F., Sheffield, V.C., Stone, E.M. & Takahashi, J.S. (2005). Generation, characterization, and molecular cloning of the Noerg-1 mutation of rhodopsin in the mouse. Visual Neuroscience 22, 619629.
Pinto, L.H., Vitaterna, M.H., Siepka, S.M., Shimomura, K., Lumayag, S., Baker, M., Fenner, D., Mullins, R.F., Sheffield, V.C., Stone, E.M., Heffron, E. & Takahashi, J.S. (2004). Results from screening over 9000 mutation-bearing mice for defects in the electroretinogram and appearance of the fundus. Vision Research 44, 33353345.
Renteria, R.C., Tian, N., Cang, J., Nakanishi, S., Stryker, M. & Copenhagen, M. (2006). Intrinsic ON responses of the retinal OFF pathway are suppressed by the ON pathway. Journal of Neuroscience 26, 1185711869
Rosemond, E., Wang, M., Yao, Y., Storjohann, L., Stormann, T., Johnson, E.C. & Hampson, D.R. (2004). Molecular basis for the differential agonist affinities of group III metabotropic glutamate receptors. Molecular Pharmacology 66, 834842.
Sagdullaev, B.T. & McCall, M.A. (2005). Stimulus size and intensity alter fundamental receptive-field properties of mouse retinal ganglion cells in vivo. Visual Neuroscience 22, 649659.
Saszik, S.M., Robson, J.G. & Frishman, L.J. (2002). The scotopic threshold response of the dark-adapted electroretinogram of the mouse. The Journal of Physiology 543, 899916.
Sharon, D., Vorobiov, D. & Dascal, N. (1997). Positive and negative coupling of the metabotropic glutamate receptors to a G protein-activated K+ channel, GIRK, in Xenopus oocytes. The Journal of General Physiology 109, 477490.
Shimbo, K., Brassard, D.L., Lamb, R.A. & Pinto, L.H. (1996). Ion selectivity and activation of the M2 ion channel of influenza virus. Biophysical Journal 70, 13351346.
Siepka, S.M. & Takahashi, J.S. (2005). Forward genetic screen to identify circadian rhythm mutants in mice. Methods in Enzymology 393, 217228.
Slaughter, M.M. & Miller, R.F. (1981). 2-amino-4-phosphonobutyric acid: a new pharmacological tool for retina research. Science 211, 182185.
Stern-Bach, Y., Bettler, B., Hartley, M., Sheppard, P.O., O'Hara, P.J. & Heinemann, S.F. (1994). Agonist selectivity of glutamate receptors is specified by two domains structurally related to bacterial amino acid-binding proteins. Neuron 13, 13451357.
Vitaterna, M.H., Pinto, L.H. & Takahashi, J.S. (2006). Large-scale mutagenesis and phenotypic screens for the nervous system and behavior in mice. Trends in Neuroscience 29, 233240.
Zeitz, C., van Genderen, M., Neidhardt, J., Luhmann, U.F., Hoeben, F., Forster, U., Wycisk, K., Matyas, G., Hoyng, C.B., Riemslag, F., Meire, F., Cremers, F.P. & Berger, W. (2005). Mutations in GRM6 cause autosomal recessive congenital stationary night blindness with a distinctive scotopic 15-Hz flicker electroretinogram. Investigative Ophthalmology & Visual Science 46, 43284335.

Keywords

Related content

Powered by UNSILO

Generation, identification and functional characterization of the nob4 mutation of Grm6 in the mouse

  • LAWRENCE H. PINTO (a1), MARTHA H. VITATERNA (a1), KAZUHIRO SHIMOMURA (a1), SANDRA M. SIEPKA (a1), VICTORIA BALANNIK (a1), ERIN L. MCDEARMON (a1) (a2), CHIAKI OMURA (a1), STEPHEN LUMAYAG (a1), BRANDON M. INVERGO (a1), BRETT GLAWE (a1), DONALD R. CANTRELL (a3), SAMSOON INAYAT (a3) (a4), MARISSA A. OLVERA (a5), KIRSTAN A. VESSEY (a6), MAUREEN A. McCALL (a6) (a7), DENNIS MADDOX (a8), CATHERINE W. MORGANS (a9), BRANDON YOUNG (a10), MATHEW T. PLETCHER (a10), ROBERT F. MULLINS (a5), JOHN B. TROY (a3) and JOSEPH S. TAKAHASHI (a1) (a2)...

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed.