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Lectin-ferritin binding on dystrophic and normal retinal pigment epithelial membranes

  • Published:
Journal of Neurocytology

Summary

Phagocytosis in the rat retina is a process which involves uptake of shed photoreceptor outer segments by the overlying retinal pigment epithelium (RPE). In rats with inherited retinal degeneration, there is a defect in phagocytosis. One aspect of this phagocytic defect may be an alteration in glycoconjugate-containing membrane components on RPE membranes which mediate this phagocytic interaction. Lectin binding sites have been studied in order to localize the distribution of glycoconjugates on pigment epithelial microvilli in normal and dystrophic retinas and to determine if there are differences in the dystrophic retinas which would provide a clue about the defect.

The following ferritin-conjugated lectins were used in this study: Concanavalin A (Con A-fe) or lens culinaris haemagglutinin (LcH-fe) for mannosyl-containing glycoconjugates; and wheat germ agglutinin (WGA-fe) forn-acetylglucosamine and sialic acid-containing glycoconjugates. Control tissue was incubated in the lectin in the presence of its competitor sugar. The number of ferritin particles or lectin-ferritin binding sites per micrometre of microvillous membrane was quantified from electron micrographs using a computer and a digitizing tablet. The number of WGA-fe binding sites on normal RPE microvillous membranes (56.0/μm) was statistically equivalent to the dystrophic membranes (48.8/μm). The number of Con A-fe binding sites on normal (27.3/m) and dystrophic RPE (26.7/μm) was also the same. A dramatic difference in LcH-fe binding sites was demonstrated on normal (1.5/μm) as compared to dystrophic RPE (19.1/μm). Our results indicate that more mannosyl residues are accessible on dystrophic microvillous membranes and, based on what is currently known about LcH binding, that these residues belong to glycoconjugates having fucosyl-containing carbohydrate cores. The data also suggest that in normal animals without a phagocytic defect such fucosyl-containing glycoconjugates are not as accessible and may be masked by other sugar residues in the oligosaccharide chain.

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References

  • Bhavanandan, V. &Katlic, A. (1979) The interaction of wheat germ agglutinin with sialoglycoproteins.Journal of Biological Chemistry 254, 4000–7.

    PubMed  Google Scholar 

  • Bok, D. &Hall, M. O. (1971) The role of the pigment epithelium in the etiology of inherited retinal dystrophy in the rat.Journal of Cell Biology 49, 664–82.

    PubMed  Google Scholar 

  • Bridges, C. D. B. (1981a) Lectin receptors on rods and cones. Visualization of fluorescent label,Investigative Ophthamology and Visual Science 20, 8–16.

    Google Scholar 

  • Bridges, C. D. B. (1981b) Agglutination of isolated rod outer segments by lectins.Investigative Ophthalmology and Visual Science 20, 17–23.

    PubMed  Google Scholar 

  • Bridges, C. D. B. &Fong, S-L. (1979) Different distribution of receptors for peanut and ricin agglutinins between inner and outer segments of rod cells.Nature 282, 513–15.

    PubMed  Google Scholar 

  • Bridges, C. D. B. &Fong, S-L. (1980) Lectins as probes of glycoprotein and glycolipid oligosaccharides in rods and cones.Neurochemistry 1, 255–67.

    Google Scholar 

  • Chaitin, M. H. &Hall, M. O. (1983) Defective ingestion of rod outer segments by cultured dystrophic rat pigment epithelial cells.Investigative Ophthalmology and Visual Science 24, 812–20.

    PubMed  Google Scholar 

  • Custer, N. V. &Bok, D. (1975) Pigment epithelium-photoreceptor interactions in the normal and dystrophic rat retina.Experimental Eye Research 21, 153–66.

    PubMed  Google Scholar 

  • De Camilli, P., Harris, S. M., Huttner, W. B. &Greengard, P. (1983) Synapsin I (Protein I), a nerve terminal-specific phosphoprotein. II. Its specific association with synaptic vesicles demonstrated by immunocytochemistry in agarose-embedded synaptosomes.Journal of Cell Biology 96, 1355–73.

    PubMed  Google Scholar 

  • Dowling, J. E. &Sidman, R. L. (1962) Inherited retinal dystrophy in the rat.Journal of Cell Biology 14, 73–109.

    PubMed  Google Scholar 

  • Edwards, R. B. (1981) The isolation and culturing of retinal pigment epithelium of the rat.Vision Research 21, 147–50.

    PubMed  Google Scholar 

  • Edwards, R. &Szamier, B. (1977) Defective phagocytosis of isolated rod outer segments by RCS rat retinal pigment epithelium in culture.Science 197, 1001–3.

    PubMed  Google Scholar 

  • Feeney, L. &Mixon, R. N. (1976) Anin vitro model of phagocytosis in bovine and human retinal pigment epithelium.Experimental Eye Research 22, 533–48.

    PubMed  Google Scholar 

  • Goldman, A. I. &O'brien, P. (1978) Phagocytosis in the retinal pigment epithelium of the RCS rat.Science 201, 1023–5.

    PubMed  Google Scholar 

  • Griffin, F. M., Griffin, J. A., Leider, J. E. &Silverstein, S. C. (1975) Studies on the mechanisms of phagocytosis. I. Requirements for circumferential attachment of particle bound ligands to specific receptors on the macrophage plasma membrane.Journal of Experimental Medicine 142, 1263–82.

    PubMed  Google Scholar 

  • Griffin, F. M., Griffin,J. A. &Silverstein, S. C. (1976) Studies on the mechanism of phagocytosis. II. The interaction of macrophages with anti-immunoglobulin IgG coated bone marrow derived lymphocytes.Journal of Experimental Medicine 144, 788–809.

    PubMed  Google Scholar 

  • Hall, M. O. (1978) Phagocytosis of light- and dark-adapted rod outer segments by cultured pigment epithelium.Science 202, 526–8.

    PubMed  Google Scholar 

  • Hall, M. O. &Nir, I. (1976) The binding of Concanavalin A to the rod outer segments and pigment epithelium of normal and RCS rats.Experimental Eye Research 22, 469–76.

    PubMed  Google Scholar 

  • Hall, M. O. &Quon, D. S. (1980) Plant lectins inhibit phagocytosis of ROS by cultured PE cells.Investigative Ophthalmology and Visual Science (Suppl.),19, 162.

    Google Scholar 

  • Herron, W. L., Riegel, B. W., Myers, O. E. &Rubin, M. L. (1969) Retinal dystrophy in the rat- a pigment epithelial disease.Investigative Ophthalmology and Visual Science 8, 595–604.

    Google Scholar 

  • Irimura, T. &Nicolson, G. L. (1984) Carbohydrate chain analysis of lectin binding to mixtures of glycoproteins, separated by polyacrylamide slab-gel electrophoresis, within situ chemical modifications.Carbohydrate Research 115, 209–20.

    Google Scholar 

  • Kornfeld, K., Reitman, M. L. &Kornfeld, R. (1981) The carbohydrate-binding specificity of PEA and lentil lectins.Journal of biological Chemistry 256, 6633–40.

    PubMed  Google Scholar 

  • LaVail, M. M., Sidman, R. L. &O'neil, D. (1972) Photoreceptors-pigment epithelial cell relationships in rats with inherited retinal degeneration.Journal of Cell Biology 53, 185–209.

    PubMed  Google Scholar 

  • Maylie-Pfenninger, M. F. &Jamieson, J. D. (1979) Distribution of cell surface saccharides on pancreatic cells.Journal of Cell Biology 80, 69–76.

    PubMed  Google Scholar 

  • McLaughlin, B. J. &Wood, J. G. (1980) The localization of lectin binding sites on photoreceptor outer segments and pigment epithelium of dystrophic retinas.Investigative Ophthalmology and Visual Science 19, 728–42.

    PubMed  Google Scholar 

  • Mullen, R. J. &LaVail, M. M. (1976) Inherited retinal dystrophy: primary defect in pigment epithelium determined with experimental rat chimeras.Science 192, 799–801.

    PubMed  Google Scholar 

  • Nir, I. &Hall, M. O. (1979) Ultrastructural localization of lectin binding sites on the surface of retinal photoreceptors and pigment epithelium.Experimental Eye Research 29, 181–94.

    PubMed  Google Scholar 

  • O'brien, P. J. (1976) Rhodopsin as a glycoprotein: a possible role for the oligosaccharide in phagocytosis.Experimental Eye Research 23, 127–37.

    PubMed  Google Scholar 

  • Philp, N. J. &Bernstein, M. H. (1980) Receptor specificities in RPE phagocytosis.Journal of Cell Biology 87, 91 (abstract).

    Google Scholar 

  • Sarthy, P. C., Bridges, C. D. B., Kretzer, F. L. &Lam, D. M. K. (1981) Lectin receptors on cells isolated from the turtle retina.Journal of Comparative Neurology 202, 561–9.

    PubMed  Google Scholar 

  • Seyfried, R. S. &McLaughlin, B. J. (1982) Phagocytosis of sugar coated and lectin coated beads by dystrophic and normal RPE.Investigative Ophthalmology and Visual Science (Suppl.)22, 142.

    Google Scholar 

  • Seyfried-Williams, R. &McLaughlin, B. J. (1983) The use of sugar-coated beads to study phagocytosis in normal and dystrophic retina.Vision Research 23, 485–94.

    PubMed  Google Scholar 

  • Silverstein, S. C. &Loike, D. J. (1980)Mononuclear Phagocytes-Functional Aspects, Part II (edited byVan Furth, R.), pp. 895–917. The Hague, Netherlands: Martinus Nijhoff Publishers.

    Google Scholar 

  • Stein, M. D., Howard, I. K. &Sage, H. J. (1971) Studies on a phytohemagglutinin from the lentil. IV. Direct binding studies of lens culinaris hemagglutinin with simple saccharides.Archives of Biochemistry and Biophysics 146, 353–5.

    PubMed  Google Scholar 

  • Stossel, T. P. (1974a) Phagocytosis.New England Journal of Medicine 290, 717–23.

    PubMed  Google Scholar 

  • Stossel, T. P. (1974b) Phagocytosis.New England Journal of Medicine 290, 774–80.

    Google Scholar 

  • Young, N. M. &Leon, M. A. (1974) The affinity of Concanavalin A and lens culinaris hemagglutinin for glycopeptides.Biochemica et Biophysica Acta 365, 418–24.

    Google Scholar 

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Authors and Affiliations

  1. Department of Anatomy, University of Tennessee Center for the Health Sciences, 875 Monroe Avenue, 38163, Memphis, Tennessee, USA

    Barbara J. McLaughlin, Lou G. Boykins & R. B. Caldwell

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  1. Barbara J. McLaughlin
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  2. Lou G. Boykins
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  3. R. B. Caldwell
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McLaughlin, B.J., Boykins, L.G. & Caldwell, R.B. Lectin-ferritin binding on dystrophic and normal retinal pigment epithelial membranes. J Neurocytol 13, 467–480 (1984). https://doi.org/10.1007/BF01148335

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  • DOI: https://doi.org/10.1007/BF01148335

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