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Cloning and molecular characterization of cGMP-gated ion channels from rod and cone photoreceptors of striped bass (M. saxatilis) retina

Published online by Cambridge University Press:  09 March 2006

CHRISTOPHE PAILLART ,
KAI ZHANG ,
TATIANA I. REBRIK ,
WOLFGANG BAEHR  and
JUAN I. KORENBROT
CHRISTOPHE PAILLART
Affiliation:
Department of Physiology, School of Medicine, University of California at San Francisco, San Francisco, California
KAI ZHANG
Affiliation:
Department of Ophthalmology and Visual Sciences, Moran Eye Center, University of Utah Health Science Center, Salt Lake City, Utah
TATIANA I. REBRIK
Affiliation:
Department of Physiology, School of Medicine, University of California at San Francisco, San Francisco, California
WOLFGANG BAEHR
Affiliation:
Department of Ophthalmology and Visual Sciences, Moran Eye Center, University of Utah Health Science Center, Salt Lake City, Utah
JUAN I. KORENBROT
Affiliation:
Department of Physiology, School of Medicine, University of California at San Francisco, San Francisco, California
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Abstract

Vertebrate photoreceptors respond to light with changes in membrane conductance that reflect the activity of cyclic-nucleotide gated channels (CNG channels). The functional features of these channels differ in rods and cones; to understand the basis of these differences we cloned CNG channels from the retina of striped bass, a fish from which photoreceptors can be isolated and studied electrophysiologically. Through a combination of experimental approaches, we recovered and sequenced three full-length cDNA clones. We made unambiguous assignments of the cellular origin of the clones through single photoreceptor RT-PCR. Synthetic peptides derived from the sequence were used to generate monospecific antibodies which labeled intact, unfixed photoreceptors and confirmed the cellular assignment of the various clones. In rods, we identified the channel α subunit gene product as 2040 bp in length, transcribed into two mRNA 1.8 kb and 2.9 kb in length and translated into a single 96-kDa protein. In cones we identified both α (CNGA3) and β (CNGB3) channel subunits. For α, the gene product is 1956 bp long, the mRNA 3.4 kb, and the protein 74 kDa. For β, the gene product is 2265 bp long and the mRNA 3.3 kb. Based on deduced amino acid sequence, we developed a phylogenetic map of the evolution of vertebrate rod and cone CNG channels. Sequence comparison revealed channels in striped bass, unlike those in mammals, are likely not N-linked-glycosylated as they are transported within the photoreceptor. Also bass cone channels lack certain residues that, in mammals, can be phosphorylated and, thus, affect the cGMP sensitivity of gating. On the other hand, functionally critical residues, such as positively charged amino acids within the fourth transmembrane helix (S4) and the Ca2+-binding glutamate in the pore loop are absolutely the same in mammalian and nonmammalian species.

Keywords

cGMP-gated ion channelsRetinaPhotoreceptorsRodCone
Type
Research Article
Information
Visual Neuroscience , Volume 23 , Issue 1 , January 2006 , pp. 99 - 113
Copyright
2006 Cambridge University Press

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References

REFERENCES

Ahmad, I., Leinders-Zufall, T., Kocsis, J.D., Shepherd, G.M., Zufall, F., & Barnstable, C.J. (1994). Retinal ganglion cells express a cGMP-gated cation conductance activatable by nitric oxide donors. Neuron 12, 155–165.Google Scholar
Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A., & Struhl, K. (2005). Current Protocols in Molecular Biology. New York: Greene Publishing Associates and Wiley-Interscience.
Barry, P.H. (2003). The relative contributions of cAMP and InsP3 pathways to olfactory responses in vertebrate olfactory receptor neurons and the specificity of odorants for both pathways. Journal of General Physiology 122, 247–250.Google Scholar
Bezanilla, F. (2000). The voltage sensor in voltage-dependent ion channels. Physiological Reviews 80, 555–92.Google Scholar
Biel, M., Seeliger, M., Pfeifer, A., Kohler, K., Gerstner, A., Ludwig, A., Jaissle, G., Fauser, S., Zrenner, E., & Hofmann, F. (1999). Selective loss of cone function in mice lacking the cyclic nucleotide-gated channel CNG3. Proceedings of the National Academy of Sciences of the U.S.A. 96, 7553–7557.Google Scholar
Chen, T.Y., Peng, Y.W., Dhallan, R.S., Ahamed, B., Reed, R.R., & Yau, K.W. (1993). A new subunit of the cyclic nucleotide-gated cation channel in retinal rods. Nature 362, 764–767.Google Scholar
Chomczynski, P. & Sacchi, N. (1987). Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Analytical Biochemistry 162, 156–159.Google Scholar
Colamartino, G., Menini, A., & Torre, V. (1991). Blockage and permeation of divalent cations through the cyclic GMP-activated channel from tiger salamander retinal rods. Journal of Physiology 440, 189–206.Google Scholar
Collin, S.P. & Trezise, A.E. (2004). The origins of colour vision in vertebrates. Clinical and Experimental Optometry 87, 217–223.Google Scholar
Cook, N.J., Hanke, W., & Kaupp, U.B. (1987). Identification, purification, and functional reconstitution of the cyclic GMP-dependent channel from rod photoreceptors. Proceedings of the National Academy of Sciences of the U.S.A. 84, 585–589.Google Scholar
Cook, N.J., Molday, L.L., Reid, D., Kaupp, U.B., & Molday, R.S. (1989). The cGMP-gated channel of bovine rod photoreceptors is localized exclusively in the plasma membrane. Journal of Biological Chemistry 264, 6996–6999.Google Scholar
Deutsch, C. (2003). The birth of a channel. Neuron 40, 265–276.Google Scholar
Ebrey, T. & Koutalos, Y. (2001). Vertebrate photoreceptors. Progress in Retinal Eye Research 20, 49–94.Google Scholar
Eismann, E., Muller, F., Heinemann, S.H., & Kaupp, U.B. (1994). A single negative charge within the pore region of a cGMP-gated channel controls rectification, Ca2+ blockage, and ionic selectivity. Proceedings of the National Academy of Sciences of the U.S.A. 91, 1109–1113.Google Scholar
Faillace, M.P., Bernabeu, R.O., & Korenbrot, J.I. (2004). Cellular processing of cone photoreceptor cyclic GMP-gated ion channels: A role for the S4 structural motif. Journal of Biological Chemistry 279, 22643–22653.Google Scholar
Felsenstein, J. (2004). Inferring Phylogenetics. Sunderland, Massachusetts: Sinauer Associates.
Frings, S., Seifert, R., Godde, M., & Kaupp, U.B. (1995). Profoundly different calcium permeation and blockage determine the specific function of distinct cyclic nucleotide-gated channels. Neuron 15, 169–179.Google Scholar
Gerstner, A., Zong, X., Hofmann, F., & Biel, M. (2000). Molecular cloning and functional characterization of a new modulatory cyclic nucleotide-gated channel subunit from mouse retina. Journal of Neuroscience 20, 1324–1332.Google Scholar
Hackos, D.H. & Korenbrot, J.I. (1997). Calcium modulation of ligand affinity in the cyclic GMP-gated ion channels of cone photoreceptors. Journal of General Physiology 110, 515–528.Google Scholar
Hackos, D.H. & Korenbrot, J.I. (1999). Divalent cation selectivity is a function of gating in native and recombinant cyclic nucleotide-gated ion channels from retinal photoreceptors. Journal of General Physiology 113, 799–818.Google Scholar
Haynes, L.W. (1995). Permeation and block by internal and external divalent cations of the catfish cone photoreceptor cGMP-gated channel. Journal of General Physiology 106, 507–523.Google Scholar
Haynes, L.W. & Stotz, S.C. (1997). Modulation of rod, but not cone, cGMP-gated photoreceptor channel by calcium-calmodulin. Visual Neuroscience 14, 233–239.Google Scholar
Henry, D., Burke, S., Shishido, E., & Matthews, G. (2003). Retinal bipolar neurons express the cyclic nucleotide-gated channel of cone photoreceptors. Journal of Neurophysiology 89, 754–761.Google Scholar
Hofmann, F., Biel, M., & Kaupp, U.B. (2003). International Union of Pharmacology. XLII. Compendium of voltage-gated ion channels: Cyclic nucleotide-modulated channels. Pharmacological Review 55, 587–589.Google Scholar
Hsu, Y.T. & Molday, R.S. (1994). Interaction of calmodulin with the cyclic GMP-gated channel of rod photoreceptor cells. Modulation of activity, affinity purification, and localization. Journal of Biological Chemistry 269, 29765–29770.Google Scholar
Huttl, S., Michalakis, S., Seeliger, M., Luo, D.G., Acar, N., Geiger, H., Hudl, K., Mader, R., Haverkamp, S., Moser, M., Pfeifer, A., Gerstner, A., Yau, K.W., & Biel, M. (2005). Impaired channel targeting and retinal degeneration in mice lacking the cyclic nucleotide-gated channel subunit CNGB1. Journal of Neuroscience 25, 130–138.Google Scholar
Ivics, Z., Kaufman, C.D., Zayed, H., Miskey, C., Walisko, O., & Izsvak, Z. (2004). The Sleeping Beauty transposable element: Evolution, regulation and genetic applications. Current Issues in Molecular Biology 6, 43–55.Google Scholar
Karpen, J.W., Loney, D.A., & Baylor, D.A. (1992). Cyclic GMP-activated channels of salamander retinal rods: Spatial distribution and variation of responsiveness. Journal Of Physiology 448, 257–274.Google Scholar
Kaupp, U.B., Niidome, T., Tanabe, T., Terada, S., Bonigk, W., Stuhmer, W., Cook, N.J., Kangawa, K., Matsuo, H., Hirose, T., Miyata, T., & Numa, S. (1989). Primary structure and functional expression from complementary DNA of the rod photoreceptor cyclic GMP-gated channel. Nature 342, 762–766.Google Scholar
Kaupp, U.B. & Seifert, R. (2001). Molecular diversity of pacemaker ion channels. Annual Review of Physiology 63, 235–257.Google Scholar
Kaupp, U.B. & Seifert, R. (2002). Cyclic nucleotide-gated ion channels. Physiological Reviews 82, 769–824.Google Scholar
Korenbrot, J.I. & Rebrik, T.I. (2002). Tuning outer segment Ca2+ homeostasis to phototransduction in rods and cones. Advances Experimental Medical Biology 514, 179–203.Google Scholar
Korschen, H.G., Illing, M., Seifert, R., Sesti, F., Williams, A., Gotzes, S., Colvilee, C., Muller, F., Dose, A., Godde, M., Molday, L., Kaupp, U.B., & Molday, R.S. (1995). A 240 kDa protein represents the complete β subunit of the cyclic nucleotide-gated channel from rod photoreceptors. Neuron 15, 627–636.Google Scholar
Krajewski, J.L., Luetje, C.W., & Kramer, R.H. (2003). Tyrosine phosphorylation of rod cyclic nucleotide-gated channels switches off Ca2+/calmodulin inhibition. Journal of Neuroscience 23, 10100–10106.Google Scholar
Leconte, L. & Barnstable, C.J. (2000). Impairment of rod cGMP-gated channel α-subunit expression leads to photoreceptor and bipolar cell degeneration. Investigative Ophthalmology and Visual Science 41, 917–926.Google Scholar
Lee, H.M., Park, Y.S., Kim, W., & Park, C.S. (2001). Electrophysiological characteristics of rat gustatory cyclic nucleotide—gated channel expressed in Xenopus oocytes. Journal of Neurophysiology 85, 2335–2349.Google Scholar
Matulef, K. & Zagotta, W.N. (2003). Cyclic nucleotide-gated ion channels. Annual Review Cell Developmental Biology 19, 23–44.Google Scholar
Miller, J.L. & Korenbrot, J.I. (1993). Phototransduction and adaptation in rods, single cones, and twin cones of the striped bass retina: A comparative study. Visual Neuroscience 10, 653–667.Google Scholar
Misaka, T., Kusakabe, Y., Emori, Y., Gonoi, T., Arai, S., & Abe, K. (1997). Taste buds have a cyclic nucleotide-activated channel, CNGgust. Journal of Biological Chemistry 272, 22623–22629.Google Scholar
Molokanova, E., Krajewski, J.L., Satpaev, D., Luetje, C.W., & Kramer, R.H. (2003). Subunit contributions to phosphorylation-dependent modulation of bovine rod cyclic nucleotide-gated channels. Journal of Physiology 552, 345–356.Google Scholar
Molokanova, E., Savchenko, A., & Kramer, R.H. (1999). Noncatalytic inhibition of cyclic nucleotide-gated channels by tyrosine kinase induced by genistein. Journal of General Physiology 113, 45–56.Google Scholar
Muller, F., Vantler, M., Weitz, D., Eismann, E., Zoche, M., Koch, K.W.& Kaupp, U.B. (2001). Ligand sensitivity of the 2 subunit from the bovine cone cGMP-gated channel is modulated by protein kinase C but not by calmodulin. Journal of Physiology 532, 399–409.Google Scholar
Nakatani, K., Koutalos, Y., & Yau, K.-W. (1995). Ca+ modulation of the cGMP-gated channel of bullfrog retinal rod photoreceptor. Journal of Physiology 484, 69–76.Google Scholar
Ohyama, T., Hackos, D.H., Frings, S., Hagen, V., Kaupp, U.B., & Korenbrot, J.I. (2000). Fraction of the dark current carried by Ca(2+) through cGMP-gated ion channels of intact rod and cone photoreceptors. Journal of General Physiology 116, 735–754.Google Scholar
Ohyama, T., Picones, A., & Korenbrot, J.I. (2002). Voltage-dependence of ion permeation in cyclic GMP-gated ion channels is optimized for cell function in rod and cone photoreceptors. Journal of General Physiology 119, 341–354.Google Scholar
Peng, C., Rich, E.D., Thor, C.A., & Varnum, M.D. (2003). Functionally important calmodulin-binding sites in both NH2- and COOH- terminal regions of the cone photoreceptor cyclic nucleotide-gated channel CNGB3 subunit. Journal of Biological Chemistry 278, 24617–24623.Google Scholar
Peng, C., Rich, E.D., & Varnum, M.D. (2004). Subunit configuration of heteromeric cone cyclic nucleotide-gated channels. Neuron 42, 401–410.Google Scholar
Picones, A. & Korenbrot, J.I. (1992). Permeation and interaction of monovalent cations with the cGMP-gated channel of cone photoreceptors. Journal of General Physiology 100, 647–673.Google Scholar
Picones, A. & Korenbrot, J.I. (1995a). Permeability and interaction of Ca2+ with cGMP-gated ion channels differ in retinal rod and cone photoreceptors. Biophysical Journal 69, 120–127.Google Scholar
Picones, A. & Korenbrot, J.I. (1995b). Spontaneous, ligand-independent activity of the cGMP-gated ion channels in cone photoreceptors of fish. Journal of Physiology 485, 699–714.Google Scholar
Pugh, E.N., Jr. & Lamb, T.D. (2000). Phototransduction in vertebrate rods and cones: molecular mechanisms of amplification, recovery and light adaptation. In Handbook of Biological Physics, ed. Stavenga, D.G., DeGrip, W.J., & Pugh E.N., Jr., pp. 186–255. Amsterdam: Elsevier Science B.V.
Raymond, P.A., Barthel, L.K., Rounsifer, M.E., Sullivan, S.A., & Knight, J.K. (1993). Expression of rod and cone visual pigments in goldfish and zebrafish: A rhodopsin-like gene is expressed in cones. Neuron 10, 1161–1174.Google Scholar
Rebrik, T.I. & Korenbrot, J.I. (1998). In intact cone photoreceptors, a Ca2+-dependent, diffusible factor modulates the cGMP-gated ion channels differently than in rods. Journal of General Physiology 112, 537–548.Google Scholar
Rebrik, T.I. & Korenbrot, J.I. (2004). In intact mammalian photoreceptors, Ca2+-dependent modulation of cGMP-gated ion channels is detectable in cones but not in rods. Journal of General Physiology 123, 63–76.Google Scholar
Rebrik, T.I., Kotelnikova, E.A., & Korenbrot, J.I. (2000). Time course and Ca(2+) dependence of sensitivity modulation in cyclic GMP-gated currents of intact cone photoreceptors. Journal of General Physiology 116, 521–534.Google Scholar
Root, M.J. & Mackinnon, R. (1993). Identification of an external divalent cation-binding site in the pore of a cGMP-activated channel. Neuron 11, 459–466.Google Scholar
Sagoo, M.S. & Lagnado, L. (1996). The action of cytoplasmic calcium on the cGMP-activated channel in salamander rod photoreceptors. Journal of Physiology 497, 309–319.Google Scholar
Seifert, R., Eismann, E., Ludwig, J., Baumann, A., & Kaupp, U.B. (1999). Molecular determinants of a Ca2+-binding site in the pore of cyclic nucleotide-gated channels: S5/S6 segments control affinity of intrapore glutamates. EMBO Journal 18, 119–130.Google Scholar
Subramaniam, S. (1998). The biology workbench—a seamless database and analysis environment for the biologist. Protein 32, 1–2.Google Scholar
Sugita, M., Ohishi, H., Iwasa, Y., Hirono, C., & Shiba, Y. (2004). Extracellular proton sensing of the rat gustatory cyclic nucleotide-gated channel. Biochemical Biophysical Research Communication 319, 369–374.Google Scholar
Tanaka, J.C. & Furman, R.E. (1993). Divalent effects on cGMP-activated currents in excised patches from amphibian photoreceptors. Journal of Membrane Biology 131, 245–256.Google Scholar
Trudeau, M.C. & Zagotta, W.N. (2003). Calcium/calmodulin modulation of olfactory and rod cyclic nucleotide-gated ion channels. Journal of Biological Chemistry 278, 18705–18708.Google Scholar
Trudeau, M.C. & Zagotta, W.N. (2004). Dynamics of Ca2+-calmodulin-dependent inhibition of rod cyclic nucleotide-gated channels measured by patch-clamp fluorometry. Journal of General Physiology 124, 211–223.Google Scholar
Weitz, D., Ficek, N., Kremmer, E., Bauer, P.J., & Kaupp, U.B. (2002). Subunit stoichiometry of the CNG channel of rod photoreceptors. Neuron 36, 881–889.Google Scholar
Wells, G.B. & Tanaka, J.C. (1997). Ion selectivity predictions from a two-site permeation model for the cyclic nucleotide-gated channel of retinal rod cells. Biophysical Journal 72, 127–140.Google Scholar
Wohlfart, P., Haase, W., Molday, R.S., & Cook, N.J. (1992). Antibodies against synthetic peptides used to determine the topology and site of glycosylation of the cGMP-gated channel from bovine rod photoreceptors. Journal of Biological Chemistry 267, 644–648.Google Scholar
Wohlfart, P., Muller, H., & Cook, N.J. (1989). Lectin binding and enzymatic deglycosylation of the cGMP-gated channel from bovine rod photoreceptors. Journal of Biological Chemistry 264, 20934–20939.Google Scholar
Yokoyama, S. (2002). Molecular evolution of color vision in vertebrates. Gene 300, 69–78.Google Scholar
Zagotta, W.N. & Siegelbaum, S.A. (1996). Structure and function of cyclic nucleotide-gated channels. Annual Review of Neuroscience 19, 235–263.Google Scholar
Zheng, J., Trudeau, M.C., & Zagotta, W.N. (2002). Rod cyclic nucleotide-gated channels have a stoichiometry of three CNGA1 subunits and one CNGB1 subunit. Neuron 36, 891–896.Google Scholar
Zhong, H., Molday, L.L., Molday, R.S., & Yau, K.W. (2002). The heteromeric cyclic nucleotide-gated channel adopts a 3A:1B stoichiometry. Nature 420, 193–198.Google Scholar
Zimmerman, A.L. & Baylor, D.A. (1992). Cation interactions within the cyclic GMP-activated channel of retinal rods from the tiger salamander. Journal of Physiology 449, 759–783.Google Scholar
Zufall, F. & Munger, S.D. (2001). From odor and pheromone transduction to the organization of the sense of smell. Trends Neuroscience 24, 191–193.Google Scholar