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Effect of hydroxylamine on the subcellular distribution of arrestin (S-antigen) in rod photoreceptors

Published online by Cambridge University Press:  02 June 2009

Nancy J. Mangini ,
Grady L. Garner ,
Tinging L. Okajima ,
Larry A. Donoso  and
David R. Pepperberg
Nancy J. Mangini
Affiliation:
Department of Ophthalmology and Visual Sciences, Lions of Illinois Eye Research Institute, University of Illinois at Chicago College of Medicine, Chicago
Grady L. Garner
Affiliation:
Department of Ophthalmology and Visual Sciences, Lions of Illinois Eye Research Institute, University of Illinois at Chicago College of Medicine, Chicago
Tinging L. Okajima
Affiliation:
Department of Ophthalmology and Visual Sciences, Lions of Illinois Eye Research Institute, University of Illinois at Chicago College of Medicine, Chicago
Larry A. Donoso
Affiliation:
Wills Eye Hospital, Philadelphia
David R. Pepperberg
Affiliation:
Department of Ophthalmology and Visual Sciences, Lions of Illinois Eye Research Institute, University of Illinois at Chicago College of Medicine, Chicago
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Abstract

The immunocytochemical labeling of arrestin (S-antigen) in photoreceptors of the ovine retina was examined following incubation of the retina with hydroxylamine (NH2OH), an agent known to inhibit the phosphorylation of photoactivated rhodopsin. Intact, isolated retinas bathed in medium containing 20 mM NH2OH, or in control medium lacking NH2OH, were maintained in darkness or exposed to bright light for 3 min (dark-adapted and light-adapted conditions, respectively); further incubated in darkness for 10 min; and then fixed and prepared for cryosectioning. Cryosections were incubated with anti-S-antigen monoclonal antibody MAb A2G5; with secondary antibodies that were conjugated with horseradish peroxidase; and with either 3–amino-9–ethyl carbazole or diaminobenzidine as chromogen. Anti-arrestin labeling in cryosections was then analyzed densitometrically using a light-microscopic image processing system. In dark-adapted control retinas, labeling density of the photoreceptor outer segment (OS) layer (0.061 ± 0.004; average ± S.e.m.) was less than that of the inner segment (IS) layer (0.138 ± 0.011). In light-adapted control retinas, OS labeling density (0.139 ± 0.007) exceeded IS labeling density (0.095 ± 0.005). Incubation with NH2OH eliminated this light-dependent increase in labeling of the OS relative to that of the IS, i.e. eliminated the increase in relative OS/IS labeling. Densities of labeling were 0.110 ± 0.006 (OS) and 0.183 ± 0.006 (IS) in NH2OH-treated dark-adapted retinas vs. 0.078 ± 0.004 (OS) and 0.182 ± 0.008 (IS) in NH2OH-treated light-adapted retinas. Anti-arrestin labeling was also examined in retinas that were exposed to 3 min or 13 min of bright light and then immediately fixed. Among retinas incubated in the absence of NH2OH, an increase in OS/IS labeling density was evident after 3 min of illumination, and retinas illuminated for 13 min exhibited an even larger increase in OS/IS labeling. An increase in OS/IS labeling was also exhibited by NH2OH-treated retinas that had been illuminated for 3 min; by comparison with dark-adapted NH2OH-treated controls (average value of OS/IS labeling: 0.60), OS/IS labeling in these illuminated retinas was 0.97. However, OS/IS labeling in NH2OH-treated retinas that had been illuminated for 13 min (average value: 0.35) was lower than that of the dark-adapted controls. The results indicate that, within intact rods, NH2OH inhibits the light-dependent increase in OS/IS anti-arrestin labeling that is ordinarily expressed at long times (~10 min) after major bleaching of the visual pigment. Among the possible bases for the effect of NH2OH are a reduction in the driving force for the movement of arrestin from the inner to the outer segment and/or a facilitation of the degradation of arrestin in the outer segment.

Keywords

ArrestinHydroxylamineLight adaptationPhotoreceptorS-antigen
Type
Research Articles
Information
Visual Neuroscience , Volume 11 , Issue 3 , May 1994 , pp. 561 - 568
Copyright
Copyright © Cambridge University Press 1994

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References

Bennett, N. & Sitaramayya, A. (1988). Inactivation of photoexcited rhodopsin in retinal rods: The roles of rhodopsin kinase and 48–kDa protein (arrestin). Biochemistry 27, 1710–1715.CrossRefGoogle ScholarPubMed
Bowes, C., van Veen, T. & Farber, D.B. (1988). Opsin, G-protein and 48–kDa protein in normal and rd mouse retinas: Developmental expression of mRNAs and proteins and light/dark cycling of mRNAs. Experimental Eye Research 47, 369–390.Google Scholar
Bridges, C.D.B. (1962). Studies on the flash-photolysis of visual pigments – IV. Dark reactions following the flash-irradiation of frog rhodopsin in suspensions of isolated photoreceptors. Vision Research 2, 215–232.CrossRefGoogle Scholar
Brin, K.P. & Ripps, H. (1977). Rhodopsin photoproducts and rod sensitivity in the skate retina. Journal of General Physiology 69, 97–120.CrossRefGoogle ScholarPubMed
Broekhuyse, R.M., Tolhuizen, E.F.J., Janssen, A.P.M. & Winkens, H.J. (1985). Light induced shift and binding of S-antigen in retinal rods. Current Eye Research 4, 613–618.Google Scholar
Broekhuyse, R.M., Janssen, A.P.M. & Tolhuizen, E.F.J. (1987). Effect of light-adaptation on the binding of 48–kDa protein (S-antigen) to photoreceptor cell membranes. Current Eye Research 6, 607–610.Google Scholar
Craft, C.M., Whitmore, D.H. & Donoso, L.A. (1990). Differential expression of mRNA and protein encoding retinal and pineal S-antigen during the light/dark cycle. Journal of Neurochemistry 55, 1461–1473.Google Scholar
Dartnall, H. J.A. (1968). The photosensitivities of visual pigments in the presence of hydroxylamine. Vision Research 8, 339–358.Google Scholar
Dolph, P.J., Ranganathan, R., Colley, N.J., Hardy, R.W., Socolich, M. & Zuker, C.S. (1993). Arrestin function in inactivation of G protein-coupled receptor rhodopsin in vivo. Science 260, 1910–1916.Google Scholar
Dua, H.S., Hossain, P., Brown, P.A.J., McKinnon, A., Forrester, J.V., Gregerson, D.S. & Donoso, L.A. (1991). Structure-function studies of S-antigen: Use of proteases to reveal a dominant uveito-genic site. Autoimmunity 10, 153–163.CrossRefGoogle ScholarPubMed
Farber, D.B., Danciger, J.S. & Organisciak, D.T. (1991). Levels of mRNA encoding proteins of the cGMP cascade as a function of light environment. Experimental Eye Research 53, 781–786.CrossRefGoogle ScholarPubMed
Gurevich, V.V. & Benovic, J.L. (1992). Cell-free expression of visual arrestin. Truncation mutagenesis identifies multiple domains involved in rhodopsin interaction. Journal of Biological Chemistry 267, 21919–21923.Google Scholar
Gurevich, V.V. & Benovic, J.L. (1993). Visual arrestin interaction with rhodopsin. Sequential multisite binding ensures strict selectivity toward light-activated phosphorylated rhodopsin. Journal of Biological Chemistry 268, 11628–11638.Google Scholar
Hofmann, K.P., Pulvermüller, A., Buczyłko, J., Van Hooser, P. & Palczewski, K. (1992). The role of arrestin and retinoids in the regeneration pathway of rhodopsin. Journal of Biological Chemistry 267, 15701–15706.Google Scholar
Knospe, V., Donoso, L.A., Banga, J.P., Yue, S., Kasp, E. & Gregerson, D.S. (1988). Epitope mapping of bovine retinal S-antigen with monoclonal antibodies. Current Eye Research 7, 1137–1147.Google Scholar
Kühn, H., Hall, S.W. & Wilden, U. (1984). Light-induced binding of 48–kDa protein to photoreceptor membranes is highly enhanced by phosphorylation of rhodopsin. FEBS Letters 176, 473–478.CrossRefGoogle ScholarPubMed
Liebman, P.A. & Pugh, E.N., Jr. (1980). ATP mediates rapid reversal of cyclic GMP phosphodiesterase activation in visual receptor membranes. Nature 287, 734–736.Google Scholar
Mangini, N.J. (1991). Selective degradation of arrestin in rod outer segments (ROS): A possible role for cytosolic proteases in down-regulating phototransduction processes. Journal of Cell Biology 115, 210a.Google Scholar
Mangini, N.J. & Garner, G.L. (1991). Arrestin is a PEST protein. Investigative Ophthalmology and Visual Sciences (ARVO Suppl.) 32, 1150.Google Scholar
Mangini, N.J., Garner, G.L., Okajima, T.-I.L., Donoso, L.A. & Pep-perberg, D.R. (1992). Hydroxylamine inhibits light-dependent changes in arrestin immunolabeling in isolated retina. Investigative Ophthalmology and Visual Sciences (ARVO Suppl.) 33, 1005.Google Scholar
Mangini, N.J. & Pepperberg, D.R. (1987). Localization of retinal “48K” (S-antigen) by electron microscopy. Japanese Journal of Ophthalmology 31, 207–217.Google Scholar
Mangini, N.J. & Pepperberg, D.R. (1988). Immunolocalization of 48K in rod photoreceptors: Light and ATP increase OS labeling. Investigative Ophthalmology and Visual Sciences 29, 1221–1234.Google ScholarPubMed
McGinnis, J.F., Whelan, J.P. & Donoso, L.A. (1992). Transient, cyclic changes in mouse visual cell gene products during the light-dark cycle. Journal of Neuroscience Research 31, 584–590.Google Scholar
Miller, J.L., Fox, D.A. & Litman, B.J. (1986). Amplification of phosphodiesterase activation is greatly reduced by rhodopsin phosphorylation. Biochemistry 25, 4983–4988.Google Scholar
Nir, I. & Ransom, N. (1993). Ultrastructural analysis of arrestin distribution in mouse photoreceptors during dark/light cycle. Experimental Eye Research 57, 307–318.Google Scholar
Organisciak, D.T., Xie, A., Wang, H.-M., Jiang, Y.-L., Darrow, R.M. & Donoso, L.A. (1991). Adaptive changes in visual cell transduction protein levels: Effect of light. Experimental Eye Research 53, 773–779.Google Scholar
Palczewski, K., Hargrave, P.A., McDowell, J.H. & Ingebritsen, T.S. (1989 a). The catalytic subunit of phosphatase 2A dephosphor-ylates phosphoopsin. Biochemistry 28, 415–419.Google Scholar
Palczewski, K., McDowell, J.H., Jakes, S., Ingebritsen, T.S. … Hargrave, P.A. (1989 b). Regulation of rhodopsin dephosphorylation by arrestin. Journal of Biological Chemistry 264, 15770–15773.Google Scholar
Palczewski, K., Buczylko, J., Imami, N.R., McDowell, J.H. … Hargrave, P.A. (1991 a). Role of the carboxyl-terminal region of arrestin in binding to phosphorylated rhodopsin. Journal of Biological Chemistry 266, 15334–15339.CrossRefGoogle ScholarPubMed
Palczewski, Pulvermüller A., Buczyłko, J. & Hofmann, K.P. (1991 b). Phosphorylated rhodopsin and heparin induce similar conformational changes in arrestin. Journal of Biological Chemistry 266, 18649–18654.Google Scholar
Palczewski, K. & Hargrave, P.A. (1991). Studies of ligand binding to arrestin. Journal of Biological Chemistry 266, 4201–4206.CrossRefGoogle ScholarPubMed
Palczewski, K., Riazance-Lawrence, J.H. & Johnson, W.C. Jr (1992). Structural properties of arrestin studied by chemical modification and circular dichroism. Biochemistry 31, 3902–3906.CrossRefGoogle ScholarPubMed
Pepperberg, D.R. & Okajima, T.-I.L. (1992). Hydroxylamine-dependent inhibition of rhodopsin phosphorylation in the isolated retina. Experimental Eye Research 54, 369–376.Google Scholar
Philp, N.J., Chang, W. & Long, K. (1987). Light-stimulated protein movement in rod photoreceptor cells of the rat retina. FEBS Letters 225, 127–132.Google Scholar
Schleicher, A., Kühn, H. & Hofmann, K.P. (1989). Kinetics, binding constant, and activation energy of the 48–kDa protein-rhodopsin complex by extra-metarhodopsin II. Biochemistry 28, 1770–1775.Google Scholar
Seckler, B. & Rando, R.R. (1989). Schiff-base deprotonation is mandatory for light-dependent rhodopsin phosphorylation. Biochemical Journal 264, 489–493.CrossRefGoogle ScholarPubMed
Tacha, D., Brown, M. & Galey, W.T. (1990). A two-step immuno-peroxidase detection system with repeated use of diluted antibodies for bulk staining of slides. Journal of Histotechnology 13, 15–20.Google Scholar
Vanden, Hoek T.L., Goossens, W. & Knepper, P.A. (1987). Fluorescence-labeled lectins, glycoconjugates, and the development of the mouse AOP. Investigative Ophthalmology and Visual Sciences 28, 451–458.Google Scholar
Wald, G. & Brown, P.K. (1953). The molar extinction of rhodopsin. Journal of General Physiology 37, 189–200.Google Scholar
Whelan, J.P. & McGinnis, J.F. (1988). Light-dependent subcellular movement of photoreceptor proteins. Journal of Neuroscience Research 20, 263–270.CrossRefGoogle ScholarPubMed
Wilden, U., Hall, S.W. & Kühn, H. (1986). Phosphodiesterase activation by photoexcited rhodopsin is quenched when rhodopsin is phosphorylated and binds the intrinsic 48–kDa protein of rod outer segments. Proceedings of the National Academy of Sciences of the U.S.A. 83, 1174–1178.CrossRefGoogle ScholarPubMed
Wilden, U. & Kühn, H. (1982). Light-dependent phosphorylation of rhodopsin: Number of phosphorylation sites. Biochemistry 21, 3014–3022.Google Scholar
Williams, M.A. & Mangini, N.J. (1991). Immunolocalization of arrestin (S-antigen) in rods of pearl mutant and wild-type mice. Current Eye Research 10, 457–462.Google Scholar
Zuckerman, R. & Cheasty, J.E. (1986). A 48 kDa protein arrests cGMP phosphodiesterase activation in retinal rod disk membranes. FEBS Letters 207, 35–41.Google Scholar