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Nitric oxide synthesis in retinal photoreceptor cells

Published online by Cambridge University Press:  02 June 2009

Akiko Yoshida,
Nikolay Pozdnyakov,
Loan Dang,
Stephen M. Orselli,
Venkat N. Reddy and
Ari Sitaramayya
Akiko Yoshida
Affiliation:
Eye Research Institute, Oakland University, Rochester
Nikolay Pozdnyakov
Affiliation:
Eye Research Institute, Oakland University, Rochester
Loan Dang
Affiliation:
Eye Research Institute, Oakland University, Rochester
Stephen M. Orselli
Affiliation:
Eye Research Institute, Oakland University, Rochester
Venkat N. Reddy
Affiliation:
Eye Research Institute, Oakland University, Rochester
Ari Sitaramayya
Affiliation:
Eye Research Institute, Oakland University, Rochester
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Abstract

Nitric oxide (NO) is known to be synthesized in several tissues and to increase the formation of cyclic GMP through the activation of soluble guanylate cyclases. Since cyclic GMP plays an important role in visual transduction, we investigated the presence of nitric oxide synthesizing activity in retinal rod outer segments. Bovine rod outer segments were isolated intact and separated into membrane and cytosolic fractions. Nitric oxide synthase activity was assayed by measuring the conversion of L-arginine to L-citrulline. Both membrane and cytosolic fractions were active in the presence of calcium and calmodulin. The activity in both fractions was stimulated by the nitric oxide synthase cofactors FAD, FMN, and tetrahydrobiopterin and inhibited by the L-arginine analog, L-monomethyl arginine. The Km for L-arginine was similar, about 5 μM for the enzyme in both fractions. However, the two fractions differed in their calcium/calmodulin dependence: the membrane fraction exhibited basal activity even in the absence of added calcium and calmodulin while the cytosolic fraction was inactive. But the activity increased in both fractions when supplemented with calcium/calmodulin: in membranes from about 40 to 110 fmol/min/mg of protein and in the cytosol from near zero to about 350 fmol/min/mg of protein in assays carried out at 0.3 μM L-arginine. The two enzymes also responded differently to detergent: the activity of the membrane enzyme was doubled by Triton X-100 while that of the cytosolic enzyme was unaffected. These results show that NO is produced by cytosolic and membrane-associated enzymes with distinguishable properties. Investigations on the purity of isolated ROS showed that about 50% of the NOS activity is endogenous to the outer segments, and that the rest is due to membrane vesicles rich in Na, K-ATPase activity. If and how NO influences the rod outer segment physiology remains to be investigated.

Keywords

Nitric oxide synthaseNADPH-diaphoraseRod outer segmentsPhotoreceptor

Information

Type
Research Articles
Information
Visual Neuroscience , Volume 12 , Issue 3 , May 1995, pp. 493 - 500
Copyright
Copyright © Cambridge University Press 1995

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References

Berman, A.L., Azimova, A.M. & Gribakin, F.G. (1977). Localization of Na+, K+ATPase and Ca2+-activated Mg2+-dependent ATPase in retinal rods. Vision Research 17, 527–526.Google Scholar
Bredt, D.S. & Snyder, S.H. (1990). Isolation of nitric oxide synthe-tase, a calmodulin-requiring enzyme. Proceedings of the National Academy of Sciences of the U.S.A. 87, 682–685.Google Scholar
Bredt, D.S. & Snyder, S.H. (1992). Nitric oxide, a novel neuronal messenger. Neuron 8, 3–11.Google Scholar
Cho, H.J., Xie, Q-W., Calaycay, J., Mumford, R.A., Swiderek, K.M., Lee, T.D. & Nathan, C. (1992). Calmodulin is a subunit of nitric oxide synthase from macrophages. Journal of Experimental Medicine 176, 599–604.Google Scholar
Dawson, T.M., Bredt, D.S., Fotuhi, M., Hwang, P.M. & Snyder, S.H. (1991). Nitric oxide synthase and neuronal NADPH diaphorase are identical in brain and peripheral tissues. Proceedings of the National Academy of Sciences of the U.S.A. 88, 7797–7801.Google Scholar
Devries, S.H. & Schwartz, E.A. (1992). Hemi-gap-junction channels in solitary horizontal cells of the catfish retina. Journal of Physiology 445, 201–230.Google Scholar
Ehret-Hilberer, S., Nullans, G., Aunis, D. & Virmaux, N. (1992). Mono ADP-ribosylation of transducin catalyzed by rod outer segment extract. FEBS Letters 309, 394–398.Google Scholar
Feelisch, M. & Noack, E. (1991). The in vitro metabolism of nitrova-sodilators and their conversion into active species. In Heart-Failure Mechanisms and Management, ed. Lewis, B. & Kmchi, A., pp. 241–255. Berlin: Springer-Verlag.Google Scholar
Förstermann, U., Pollock, J.S., Schmidt, H.H.H.W., Heller, M. & Murad, F. (1991). Calmodulin-dependent endothelium-derived relaxing factor/nitric oxide synthase activity is present in the particulate and cytosolic fractions of bovine aortic endothelial cells. Proceedings of the National Academy of Sciences of the U.S.A. 88, 1788–1792.Google Scholar
Furchgott, R.F. (1988). Studies on relaxation of rabbit aorta by sodium nitrite: The basis for the proposal that the acid-activable inhibitory factor from retractor penis is inorganic nitrite and the endothelium derived relaxing factor is nitric oxide. In Vasodilation: Vascular Smooth Muscle, Peptides, Autonomic Nerves and Endothelium, ed. Vanhoutte, P.M., pp. 401–414. New York: Raven Press.Google Scholar
Gever, O., Podos, S.M. & Mittag, T.W. (1993). Nitric oxide synthase: Distribution and biochemical properties of the enzyme in the bovine eye. Investigative Ophthalmology and Visual Science 34, 826.Google Scholar
Goureau, O., Lepoivre, M., Mascarelli, F. & Courtois, Y. (1992). Nitric oxide synthase activity in bovine retina. In Structures and Functions of Retinal Proteins (Collogue INSERM), Vol. 221, ed. Rigaud, J.L., pp. 395–398. London: John Libbey Eurotext Ltd.Google Scholar
Hasan, K.S., Chang, K., Allison, W., Faller, A., Santiago, J.V., Tilton, R.G. & Williamson, J.R. (1993). Glucose-induced increases in ocular blood flow are prevented by aminoguanidine and L-NMMA, inhibitors of nitric oxide synthase. Investigative Ophthalmology and Visual Science 34, 1127.Google Scholar
Hiki, K., Hattorj, R., Kawai, C. & Yin, Y. (1992). Purification of insoluble nitric oxide synthase from rat cerebellum. Journal of Biochemistry 111, 556–558.Google Scholar
Hope, B.T., Michael, G.J., Knigge, K.M. & Vincent, S.R. (1991). Neuronal NADPH diaphorase is a nitric oxide synthase. Proceedings of the National Academy of Sciences of the U.S.A. 88, 2811–2814.Google Scholar
Ignarro, L.J. & Gruetter, C.A. (1980). Requirement of thiols for activation of coronary arterial guanylate cyclase by glyceryl trinitrate and sodium nitrite. Possible involvement of s-nitrosothiols. Biochimica et Biophysica Acta 631, 221–231.Google Scholar
lida, S., Ohshima, H., Oguchi, S., Hata, T., Suzuki, H., Kawasaki, H. & Esumi, H. (1992). Identification of inducible calmodulin-dependent nitric oxide synthase in the liver of rats. Journal of Biological Chemistry 267, 25385–25388.Google Scholar
Kelm, M. & Schrader, J. (1990). Control of coronary vascular tone by nitric oxide. Circulation Research 66, 1561–1575.Google Scholar
Kitamura, Y., Okamura, T., Kani, K. & Toda, T. (1993). Nitric oxide-mediated retinal arteriolar and arterial dilation induced by substance P. Investigative Ophthalmology and Visual Science 34, 2859–2865.Google Scholar
Knowles, R.G., Palacios, M., Palmer, R.M.J. & Moncada, S. (1989). Formation of nitric oxide from L-arginine in the central nervous system: A transduction mechanism for stimulation of the soluble guanylate cyclase. Proceedings of the National Academy of Sciences of the U.S.A. 86, 5159–5162.Google Scholar
Koch, K.-W., Lambrecht, H.-G., Haberecht, M., Redburn, D. & Schmidt, H.H.H.W. (1994). Functional coupling of a Ca2+ calmodulin-dependent nitric oxide synthase and a soluble guanylate cyclase in vertebrate photoreceptor cells. The EMBO Journal 13, 3312–3320.Google Scholar
Koistinaho, J., Swanson, R.A., Vente, J.D. & Sagar, S.M. (1993). NADPH-diaphorase (nitric oxide synthase) reactive amacrine cells of rabbit retina: Stimulation by light and putative target cells. Investigative Ophthalmology and Visual Science 34, 752.Google Scholar
Kukreja, R.C., Wei, E.P., Kontos, H.A. & Bates, J.N. (1993). Nitric oxide and S-nitroso-L-cysteine as endothelium derived relaxing factors from acetycholine in cerebral vessels in cats. Stroke 24, 2010–2014.Google Scholar
Margulis, A., Sharma, R.K. & Sitaramayya, A. (1992). Nitroprusside-sensitive and insensitive guanylate cyclases in retinal rod outer segments. Biochemical and Biophysical Research Communications 185, 909–914.Google Scholar
Mathews, W.R. & Kerr, S.W. (1993). Biological activity of S-nitrosothiols: The role of nitric oxide. Journal of Pharmacology and Experimental Therapeutics 267, 1529–1537.Google Scholar
Osborne, N.N., Barnett, N.L. & Herrera, A.J. (1993). NADPH diaphorase localization and nitric oxide synthetase activity in the retina and anterior uvea of the rabbit eye. Brain Research 610, 194–198.Google Scholar
Palmer, R.M.J., Ashton, D.S. & Moncada, S. (1988). Vascular endothelial cells synthesize nitric oxide from L-arginine. Nature 333, 664–666.Google Scholar
Palmer, R.M.J., Ferrige, A.G. & Moncada, S. (1987). Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature 327, 524–526.Google Scholar
Papermaster, D.S. (1982). Preparation of retinal rod outer segments. Methods in Enzymology 81, 48–52.Google Scholar
Pozdnyakov, N., Lloyd, A., Reddy, V.N. & SiTaramayya, A. (1993). Nitric oxide-regulated endogenous ADP-ribosylation of rod outer segment proteins. Biochemical and Biophysical Research Communications 192, 610–615.Google Scholar
Provis, J.M. & Mitrofanis, J. (1990). NADPH-diaphorase neurones of human retinae have a uniform topographical distribution. Visual Neuroscience 4, 619–623.Google Scholar
Pugh, E.N. & Lamb, T.D. (1993). Amplification and kinetics of the activation steps in phototransduction. Biochimica et Biophysica Acta 1141, 111–149.Google Scholar
Radomski, M.W., Palmer, R.M.J. & Moncada, S. (1990). An L-arginine/nitric oxide pathway present in human platelets aggregation. Proceedings of the National Academy of Sciences of the U.S.A. 87, 5193–5197.Google Scholar
Rees, D.D., Palmer, R.M.J., Hodson, H.F. & Moncada, S. (1989). A specific inhibitor of nitric oxide formation from L-arginine attenuates endothelium-dependent relaxation. British Journal of Pharmacology 96, 418–424.Google Scholar
Schmidt, H.H.H.W., Warner, T.D., Nakane, M., Förstermann, U. & Murad, F. (1992 a). Regulation and subcellular location of nitrogen oxide synthases in RAW264.7 macrophages. Molecular Pharmacology 41, 615–624.Google Scholar
Schmidt, K-F., Nöll, G.N. & Yamamoto, Y. (1992 b). Sodium nitro-prusside alters dark voltage and light responses in isolated retinal rods during whole-cell recording. Visual Neuroscience 9, 205–209.Google Scholar
Schnetkamp, P.P.M., Klompmakers, A.A. & Daemen, F.J.M. (1979). The isolation of stable cattle rod outer segments with an intact plasma membrane. Biochimica et Biophysica Acta 552, 379–389.Google Scholar
Sedmak, J.J. & Grossberg, S.E. (1977). A rapid, sensitive, and versatile assay for protein using Coomassie Brilliant Blue G250. Analytical Biochemistry 79, 544–552.Google Scholar
Shiells, R. & Falk, G. (1992). Retinal on-bipolar cells contain a nitric oxide-sensitive guanylate cyclase. Neuro Report 3, 845–848.Google Scholar
Stahl, W.L. & Baskin, D.G. (1984). Immunocytochemical localization of Na+, K+ adenosine triphosphatase in the rat retina. Journal of Histochemistry and Cytochemistry 32, 248–250.Google Scholar
Stamler, J.S., Simon, D.I., Osborne, J.A., Mullins, M.E., Jaraki, O., Michel, T., Singel, D.J. & Loscalzo, J. (1992). S-nitrosylation of proteins with nitric oxide: Synthesis and characterization of biologically active compounds. Proceedings of the National Academy of Sciences of the U.S.A. 89, 444–448.Google Scholar
Tsuyama, Y., Noll, G.N. & Schmidt, K.-F. (1993). L-arginine and nicotinamide adenine nucleotide phosphate alter dark voltage and accelerate light response recovery in isolated retinal rods of the frog (Rana temporaria). Neuroscience Letters 149, 95–98.Google Scholar
Venturini, C.M., Knowles, R.G., Palmer, R.M.J. & Moncada, S. (1991). Synthesis of nitric oxide in the bovine retina. Biochemical and Biophysical Research Communications 180, 920–925.Google Scholar
Winkler, B.S. & Riley, M.V. (1977). Na+ -K+ and HCO3-ATPase activity in retina: Dependence on calcium and sodium. Investigative Ophthalmology and Visual Science 16, 1151–1154.Google Scholar
Winkler, B.S. (1994). A quantitative assessment of glucose metabolism in the isolated rat retina. In Vision and Adaptation, Les Seminaires Ophthalmologiques d'Ipsen, tome 6, ed. Christen, Y., Doly, M., & Droy-Lefaix, M.T.Paris: Elsevier (in press).Google Scholar
Wolff, D.J. & Datto, G.A. (1992). Identification and characterization of a calmodulin-dependent nitric oxide synthase from GH3 pituitary cells. Biochemical Journal 285, 201–206.Google Scholar
Yamamoto, R., Bredt, D.S., Snyder, S.H. & Stone, R.A. (1993). The localization of nitric oxide synthase in the rat eye and related cranial ganglia. Neuroscience 54, 189–200.Google Scholar
Yau, K.-W. & Nakatani, K. (1985). Light-induced reduction of cytoplasmic free calcium in retinal rod outer segment. Nature 311, 661–663.Google Scholar
Zimmerman, W.F. & Godchaux, W. III. (1982). Preparation and characterization of sealed bovine rod cell outer segments. Methods in Enzymology 81, 52–57.Google Scholar