Comparison of Insulin-Like Growth Factor Receptors in Human Retinal Cells

  • Joyce F. Haskell
  • Linda E. Haws
  • Alberta Davis
  • Richard Hunt


Specific receptors for insulin and insulin-like growth factors have been identified in a variety of tissues, including brain and retinal cells. Covalent crosslinking analysis using disuccinimidyl suberate has enabled us to determine altered molecular weight forms of both the insulin and Type I IGF receptors in human retinal tissue. Autopsy eye donations were frozen and collected until adequate tissue was available for tissue to be solubilized using Triton-X-100. Retinal pigment epithelial cells were also identified and similarly treated. In addition, retinal pigment epithelial cells were cultured following transformation with plasmid pSV2Neo. Using these cells or tissues, we have analyzed the binding of [125I] insulin, IGF I and IGF II, and have determined the molecular weight for the insulin and IGF receptors. We have also examined the effect of mannose-6-phosphate (M6P) on the binding of IGF I to adult rat tissue. M6P had a dramatic effect on the crosslinking of IGF I binding to the Type II IGF/M6P receptor in detergent solubilized tissue preparations; however, no effect of M6P has been seen on autoradiographs of 125I-IGF I crosslinked to solubilized human retinal pigment epithelial cells or human neural retinal membranes. This agrees with our finding that human retinal cells have few Type II IGF/M6P receptors.


Retinal Pigment Epithelial Insulin Receptor Retinal Pigment Epithelial Cell Human Retinal Pigment Epithelial Cell Retinal Homogenate 
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  1. 1.
    G.L. King, Cell Biology as an approach to the study of the vascular complications of diabetes, Metabolism 34:17 (1985).PubMedCrossRefGoogle Scholar
  2. 2.
    S. Salardi, E. Cacciari, D. Ballardini, F. Righetti, M. Capello, A. Cicognani, S. Zucchini, G. Natali, and D. Tassinari, Relationships between growth factors (somatomedin-C and growth hormones) and body development, metabolic control, and retinal changes in children and adolescents with IDDM. Diabetes 35:832 (1986).PubMedCrossRefGoogle Scholar
  3. 3.
    S.L. Hyer, P.S. Sharp, R.A. Brooks, J.M. Burrin, and E.M. Kohner, Serum IGF-1 concentration in diabetic retinopathy, Diabetic Med. 5:356 (1986).CrossRefGoogle Scholar
  4. 4.
    M. Grant, B. Russell, C. Fitzgerald, and T.J. Merimee, Insulin-like growth factors in vitreous. Studies in control and diabetic subjects with neovascularization, Diabetes 35:416 (1986).PubMedCrossRefGoogle Scholar
  5. 5.
    J.F. Haskell, E. Meezan, and D.J. Pillion, Identification of the insulin receptor of cerebral microvessels, Amer. J. Physiol. 248:E115 (1985).PubMedGoogle Scholar
  6. 6.
    A. McElduff, P. Poronnik, R.C. Baxter, and P. Williams, A comparison of the insulin-like growth factor 1 receptors from rat brain and liver, Endocrinology 122:1933 (1988).PubMedCrossRefGoogle Scholar
  7. 7.
    R.A. Roth, D.O. Morgan, J. Beaudoin, and V. Sara, Purification and characterization of the human brain insulin receptor, J. Biol. Chem. 261: 3753 (1986).PubMedGoogle Scholar
  8. 8.
    L. Bassas, F. DePablo, M.A. Lesniak, and J. Roth, Ontogeny of receptors for insulin-like peptides in chick embryo tissues: early dominence of insulin-like growth factor over insulin receptors in brain, Endocrinology 117:2321 (1985).PubMedCrossRefGoogle Scholar
  9. 9.
    C.C. Yip, M.L. Moule, and W.T. Yeung, Characterization of insulin receptor subunits in brain and other tissues by photo affinity labeling, Biochem. Biophys. Res. Commun. 96:1671 (1980).PubMedCrossRefGoogle Scholar
  10. 10.
    K.A. Heidenrich, P.R. Gilmore, D. Brandenburg, and E. Hatda, Peptide mapping on Northern blot analyses of insulin receptors in brain and adipocytes, Mol. Cell. Endocrinology 56:255 (1988).CrossRefGoogle Scholar
  11. 11.
    S. Gammeltoft, R. Ballotti, A. Kowalski, B. Westermark, and E. Van Obberghen, Expression of two types of receptors for insulin-like growth factors in human malignant glioma, Cancer Research 48:1233 (1988).PubMedGoogle Scholar
  12. 12.
    M.K. Raizada, J. Shemer, J.J. Judkins, D.W. Clarke, B.A. Masters, and D. LeRoith, Insulin receptors in the brain: structural and physiological characterization, Neurochemical Research 13:297 (1988).PubMedCrossRefGoogle Scholar
  13. 13.
    D. LeRoith, W.L. Lowe, J. Shemer, M.K. Raizada, and A. Ota, Development of brain insulin receptors, Int. J. Biochem. 20:225 (1988).PubMedCrossRefGoogle Scholar
  14. 14.
    R.G. Rosenfeld, H. Phain, B.T. Keller, R.T. Borchardt and W.M. Pardridge, Demonstration and structural comparison of receptors for insulin-like growth factor-I and -II (IGF-I and -II) in brain and blood-brain barrier, Biochem Biophys Res Commun 149:159 (1987).PubMedCrossRefGoogle Scholar
  15. 15.
    R.J. Waldbillig and G.J. Chader, G. Anomalous insulin binding activity in the bovine neural retina: a possible mechanism for regulation of receptor binding specificity, Biochem Biophys Res Commun 151:1105 (1988).PubMedCrossRefGoogle Scholar
  16. 16.
    Y. Zick, A.M. Spiegel, and R. Eisenberg, Insulin-like growth factor I receptors in retinal rod outer segments, J Biol Chem 262:10259 (1987).PubMedGoogle Scholar
  17. 17.
    R.S. Bar, L.C. Harrison, R.C. Baxter, M Boes, B.L. Dake, B. Booth, and A. Cox, Production of IGF-binding proteins by vascular endothelial cells, Biochem Biophys Res Commun 148:734 (1987).PubMedCrossRefGoogle Scholar
  18. 18.
    R.G. Elgin, W. H. Busby, and D.R. Clemmons, An insulin-like growth factor (IGF) binding protein enhances the biologic response to IGF-I, Proc Natl Acad Sci USA 84:3254 (1987).PubMedCrossRefGoogle Scholar
  19. 19.
    B. Pfeifle, H. Hamann, R. Fussganger, and H. Ditschuneit H. Ditschuneit, Insulin as a growth regulator of arterial smooth muscle cells: effect of insulin of IGF I, Diabete Metab 13:326 (1987).PubMedGoogle Scholar
  20. 20.
    R.S. Bar, B.L. Dake, and S. Stueck, Stimulation of proteoglycans by IGF-I and II in microvessel and large vessel endothelial cells, Am J Physiol 253:E21 (1987).PubMedGoogle Scholar
  21. 21.
    M. Grant, J. Jerdan, T.J. Merimee, Insuiin-like growth factor-I modulates endothelial cell Chemotaxis, J Clin Endocrinol Metab 65:370 (1987).PubMedCrossRefGoogle Scholar
  22. 22.
    H.L. Hachiya, J.L. Carpentier, and G.L. King, Comparative studies on insuiin-like growth factor II and insulin processing by vascular endothelial cells, Diabetes 35:1065 (1986).PubMedCrossRefGoogle Scholar
  23. 23.
    H.J. Frank, W.M. Pardridge, W.L. Morris, R.G. Rosenfeld, T.B. Choi, Binding and internalization of insulin and insulin-like growth factors by isolated brain microvessels, Diabetes 35:654 (1986).PubMedCrossRefGoogle Scholar
  24. 24.
    R.S. Bar, M. Boes, M. Yorek, M. Yorek, Processing of insulin-like growth factors I and II by capillary and large vessel endothelial cells, Endocrinology 118:1072 (1986).PubMedCrossRefGoogle Scholar
  25. 25.
    K.F. Hanssen, K. Dahl-Jorgensen, T. Lauritzen, B. Feldt-Rasmussen, O. Brinchmann-Hansen, and T. Deckert, REVIEW: Diabetic control and microvascular complications: the near-normoglycaemic experience, Diabetologia 29:677 (1986).PubMedCrossRefGoogle Scholar
  26. 26.
    W. Kiess, J.F. Haskell, L. Lee, L.A. Greenstein, B.E. Miller, A.L. Aarons, M.M. Reckler, and S.P. Nissley, An antibody that blocks insulin-like growth factor (IGF) binding to the Type II IGF receptor is neither an agonist nor an inhibitor of several IGF stimulated biologic responses in L6 myoblasts, J Biol Chem 262:12745 (1987).PubMedGoogle Scholar
  27. 27.
    D.O. Morgan, J.C. Edman, D.N. Standring, V.A. Fried, M.C. Smith, R.A. Roth, W.J. Rutter, Insulin-like growth factor II receptor as a multifunctional binding protein, Nature 329:301 (1987).PubMedCrossRefGoogle Scholar
  28. 28.
    J. Haskell, D. J. Pillion, and E. Meezan, Specific high-affinity receptors for insulin-like growth factor II (IGF-II) in rat kidney glomerulus, Endocrinology 123:774 (1987).CrossRefGoogle Scholar
  29. 29.
    D.J. Pillion, J.F. Haskell, and E. Meezan, Distinct receptors for insulin-like growth factor I in rat renal glomeruli and tubules, Amer J Physiol 255:E504 (1988)PubMedGoogle Scholar
  30. 30.
    R.C. Hunt, A.A. Davis, A. Dewey, J.F. Haskell, Transferrin and insulin-like growth factor-I receptors on retinal pigment epithelia cells, J. Cell Biology 105:236A (1987).Google Scholar
  31. 31.
    C. Polychronakos and R. Piscina, Endocytosis of receptor-bound insulin-like growth factor II is enhanced by mannose-6-phosphate in IM9 cells, Endocrinology 126:2943 (1988).CrossRefGoogle Scholar
  32. 32.
    J. Haskell, L. Haws, and T. Lin, Insulin-like growth factor receptors in rat testicular vascular tissue, Endocrinology 70th Annual Meeting Abst. 519 (1988).Google Scholar
  33. 33.
    T. Braulke, C. Causin, A. Waheed, U. Junghans, A. Hasilik, P. Maly, R.E. Humbel, and K. von Figura, Mannose-6-phosphate/insulin-like growth factor II receptor: Distinct binding sites for mannose 6-phosphate and insulin-like growth factor II, Biochem Biophys Res Commun 150:1287 (1988).PubMedCrossRefGoogle Scholar
  34. 34.
    R.G. Rosenfeld and R. Pham, Production of monoclonal antibodies to the rat insulin-like growth factor II (IGF-II receptor), Biochem Biophys Res Commun 146:717 (1987).PubMedCrossRefGoogle Scholar
  35. 35.
    R.A. Roth, C. Stover, J. Hari, D.O. Morgan, M.C. Smith, V. Sara, V.A. Fried, Interaction of the receptor for insulin-like growth factor II with mannose-6-phosphate and antibodies to the mannose-6-phosphate receptor, Biochem Biophys Res Commun 149:600 (1987).PubMedCrossRefGoogle Scholar
  36. 36.
    P.Y. Tong, S.E. Tollefsen, and S. Korafeld, The cation-independent mannose-6-phosphate receptor binds insulin-growth factor II, J Biol Chem 263:2585 (1988).PubMedGoogle Scholar
  37. 37.
    W. Kiess, G. Blickenstaff, M. Sklar, C. Thomas, S.P. Nissley, and G. Sahagian, Biochemical evidence that the Type II IGF receptor is identical to the cation-independent mannose-6-phosphate receptor, J Biol Chem 263:9339 (1988).PubMedGoogle Scholar
  38. 38.
    G. Todderud, and G. Carpenter, Presence of mannose phosphate on the epidermal growth factor receptor in A-431 cells. J Biol Chem 263:17893 (1988).PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1989

Authors and Affiliations

  • Joyce F. Haskell
    • 1
  • Linda E. Haws
    • 1
  • Alberta Davis
    • 2
  • Richard Hunt
    • 2
  1. 1.Research Service, Veterans’ Administration Medical Center and Department of Obstetrics and GynecologyUniversity of Alabama at BirminghamBirminghamUSA
  2. 2.Department of Microbiology and ImmunologyUniversity of South CarolinaUSA

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