Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1993 Apr 2;121(2):335–343. doi: 10.1083/jcb.121.2.335

Apical polarization of N-CAM in retinal pigment epithelium is dependent on contact with the neural retina

PMCID: PMC2200109  PMID: 8468350

Abstract

The retinal pigment epithelium (RPE) is unique among epithelia in that its apical surface does not face a lumen, but, instead, is specialized for interaction with the neural retina. The molecules involved in the interaction of the RPE with the neural retina are not known. We show here that the neural cell adhesion molecule (N-CAM) is found both on the apical surface of RPE in situ and on the outer segments of photoreceptors, fulfilling an important requisite for an adhesion role between both structures. Strikingly, culture of RPE results in rapid redistribution of N-CAM to the basolateral surface. This is not due to an isoform shift, since the N-CAM expressed by cultured cells (140 kD) is the same as that expressed by RPE in vivo. Rather, the reversed polarity of N-CAM appears to result from the disruption of the contact between the RPE and the photoreceptors of the neural retina. We suggest that N-CAM in RPE and photoreceptors participate in these interactions.

Full Text

The Full Text of this article is available as a PDF (2.7 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson D. H., Guérin C. J., Matsumoto B., Pfeffer B. A. Identification and localization of a beta-1 receptor from the integrin family in mammalian retinal pigment epithelial cells. Invest Ophthalmol Vis Sci. 1990 Jan;31(1):81–93. [PubMed] [Google Scholar]
  2. Barbas J. A., Chaix J. C., Steinmetz M., Goridis C. Differential splicing and alternative polyadenylation generates distinct NCAM transcripts and proteins in the mouse. EMBO J. 1988 Mar;7(3):625–632. doi: 10.1002/j.1460-2075.1988.tb02856.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bartles J. R., Feracci H. M., Stieger B., Hubbard A. L. Biogenesis of the rat hepatocyte plasma membrane in vivo: comparison of the pathways taken by apical and basolateral proteins using subcellular fractionation. J Cell Biol. 1987 Sep;105(3):1241–1251. doi: 10.1083/jcb.105.3.1241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bloch R. J. Clusters of neural cell adhesion molecule at sites of cell-cell contact. J Cell Biol. 1992 Jan;116(2):449–463. doi: 10.1083/jcb.116.2.449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Buskirk D. R., Thiery J. P., Rutishauser U., Edelman G. M. Antibodies to a neural cell adhesion molecule disrupt histogenesis in cultured chick retinae. Nature. 1980 Jun 12;285(5765):488–489. doi: 10.1038/285488a0. [DOI] [PubMed] [Google Scholar]
  6. Chambard M., Gabrion J., Mauchamp J. Influence of collagen gel on the orientation of epithelial cell polarity: follicle formation from isolated thyroid cells and from preformed monolayers. J Cell Biol. 1981 Oct;91(1):157–166. doi: 10.1083/jcb.91.1.157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chu P. G., Grunwald G. B. Functional inhibition of retinal pigment epithelial cell-substrate adhesion with a monoclonal antibody against the beta 1 subunit of integrin. Invest Ophthalmol Vis Sci. 1991 May;32(6):1763–1769. [PubMed] [Google Scholar]
  8. Chu P. G., Grunwald G. B. Identification of an adhesion-associated protein of the retinal pigment epithelium. Invest Ophthalmol Vis Sci. 1990 May;31(5):847–855. [PubMed] [Google Scholar]
  9. Chu P., Grunwald G. B. Identification of the 2A10 antigen of retinal pigment epithelium as a beta 1 subunit of integrin. Invest Ophthalmol Vis Sci. 1991 May;32(6):1757–1762. [PubMed] [Google Scholar]
  10. Covault J., Sanes J. R. Distribution of N-CAM in synaptic and extrasynaptic portions of developing and adult skeletal muscle. J Cell Biol. 1986 Mar;102(3):716–730. doi: 10.1083/jcb.102.3.716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cunningham B. A., Hemperly J. J., Murray B. A., Prediger E. A., Brackenbury R., Edelman G. M. Neural cell adhesion molecule: structure, immunoglobulin-like domains, cell surface modulation, and alternative RNA splicing. Science. 1987 May 15;236(4803):799–806. doi: 10.1126/science.3576199. [DOI] [PubMed] [Google Scholar]
  12. Daniloff J. K., Chuong C. M., Levi G., Edelman G. M. Differential distribution of cell adhesion molecules during histogenesis of the chick nervous system. J Neurosci. 1986 Mar;6(3):739–758. doi: 10.1523/JNEUROSCI.06-03-00739.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. DiFiglia M., Marshall P., Covault J., Yamamoto M. Ultrastructural localization of molecular subtypes of immunoreactive neural cell adhesion molecule (NCAM) in the adult rodent striatum. J Neurosci. 1989 Dec;9(12):4158–4168. doi: 10.1523/JNEUROSCI.09-12-04158.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dol'nikova A. E., Sologub A. A., Stroeva O. G., Iamskova V. P. Rol' kal'tsii-zavisimogo i kal'tsii-nezavisimogo mekhanizmov v adgezii kletok setchatki i pigmentnogo épiteliia kurinykh zarodyshei. Ontogenez. 1985 Mar-Apr;16(2):149–155. [PubMed] [Google Scholar]
  15. Edelman G. M. Cell adhesion molecules in the regulation of animal form and tissue pattern. Annu Rev Cell Biol. 1986;2:81–116. doi: 10.1146/annurev.cb.02.110186.000501. [DOI] [PubMed] [Google Scholar]
  16. Fliesler S. J., Cole G. J., Adler A. J. Neural cell adhesion molecule (NCAM) in adult vertebrate retinas: tissue localization and evidence against its role in retina-pigment epithelium adhesion. Exp Eye Res. 1990 May;50(5):475–482. doi: 10.1016/0014-4835(90)90035-s. [DOI] [PubMed] [Google Scholar]
  17. Griffiths G., Simons K. The trans Golgi network: sorting at the exit site of the Golgi complex. Science. 1986 Oct 24;234(4775):438–443. doi: 10.1126/science.2945253. [DOI] [PubMed] [Google Scholar]
  18. Gundersen D., Orlowski J., Rodriguez-Boulan E. Apical polarity of Na,K-ATPase in retinal pigment epithelium is linked to a reversal of the ankyrin-fodrin submembrane cytoskeleton. J Cell Biol. 1991 Mar;112(5):863–872. doi: 10.1083/jcb.112.5.863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hammerton R. W., Krzeminski K. A., Mays R. W., Ryan T. A., Wollner D. A., Nelson W. J. Mechanism for regulating cell surface distribution of Na+,K(+)-ATPase in polarized epithelial cells. Science. 1991 Nov 8;254(5033):847–850. doi: 10.1126/science.1658934. [DOI] [PubMed] [Google Scholar]
  20. Hoffman S., Friedlander D. R., Chuong C. M., Grumet M., Edelman G. M. Differential contributions of Ng-CAM and N-CAM to cell adhesion in different neural regions. J Cell Biol. 1986 Jul;103(1):145–158. doi: 10.1083/jcb.103.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Lagunowich L. A., Grunwald G. B. Expression of calcium-dependent cell adhesion during ocular development: a biochemical, histochemical and functional analysis. Dev Biol. 1989 Sep;135(1):158–171. doi: 10.1016/0012-1606(89)90166-8. [DOI] [PubMed] [Google Scholar]
  23. Le Bivic A., Real F. X., Rodriguez-Boulan E. Vectorial targeting of apical and basolateral plasma membrane proteins in a human adenocarcinoma epithelial cell line. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9313–9317. doi: 10.1073/pnas.86.23.9313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Le Bivic A., Sambuy Y., Mostov K., Rodriguez-Boulan E. Vectorial targeting of an endogenous apical membrane sialoglycoprotein and uvomorulin in MDCK cells. J Cell Biol. 1990 May;110(5):1533–1539. doi: 10.1083/jcb.110.5.1533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lindner J., Rathjen F. G., Schachner M. L1 mono- and polyclonal antibodies modify cell migration in early postnatal mouse cerebellum. 1983 Sep 29-Oct 5Nature. 305(5933):427–430. doi: 10.1038/305427a0. [DOI] [PubMed] [Google Scholar]
  26. Matter K., Brauchbar M., Bucher K., Hauri H. P. Sorting of endogenous plasma membrane proteins occurs from two sites in cultured human intestinal epithelial cells (Caco-2). Cell. 1990 Feb 9;60(3):429–437. doi: 10.1016/0092-8674(90)90594-5. [DOI] [PubMed] [Google Scholar]
  27. Mayerson P. L., Hall M. O., Clark V., Abrams T. An improved method for isolation and culture of rat retinal pigment epithelial cells. Invest Ophthalmol Vis Sci. 1985 Nov;26(11):1599–1609. [PubMed] [Google Scholar]
  28. McNeill H., Ozawa M., Kemler R., Nelson W. J. Novel function of the cell adhesion molecule uvomorulin as an inducer of cell surface polarity. Cell. 1990 Jul 27;62(2):309–316. doi: 10.1016/0092-8674(90)90368-o. [DOI] [PubMed] [Google Scholar]
  29. Mostov K., Apodaca G., Aroeti B., Okamoto C. Plasma membrane protein sorting in polarized epithelial cells. J Cell Biol. 1992 Feb;116(3):577–583. doi: 10.1083/jcb.116.3.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nabi I. R., Mathews A. P., Cohen-Gould L., Gundersen D., Rodriguez-Boulan E. Immortalization of polarized rat retinal pigment epithelium. J Cell Sci. 1993 Jan;104(Pt 1):37–49. doi: 10.1242/jcs.104.1.37. [DOI] [PubMed] [Google Scholar]
  31. Neill J. M., Barnstable C. J. Expression of the cell surface antigens RET-PE2 and N-CAM by rat retinal pigment epithelial cells during development and in tissue culture. Exp Eye Res. 1990 Nov;51(5):573–583. doi: 10.1016/0014-4835(90)90088-c. [DOI] [PubMed] [Google Scholar]
  32. Nelson W. J. Cytoskeleton functions in membrane traffic in polarized epithelial cells. Semin Cell Biol. 1991 Dec;2(6):375–385. [PubMed] [Google Scholar]
  33. Nitsch L., Wollman S. H. Ultrastructure of intermediate stages in polarity reversal of thyroid epithelium in follicles in suspension culture. J Cell Biol. 1980 Sep;86(3):875–880. doi: 10.1083/jcb.86.3.875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Papermaster D. S., Dreyer W. J. Rhodopsin content in the outer segment membranes of bovine and frog retinal rods. Biochemistry. 1974 May 21;13(11):2438–2444. doi: 10.1021/bi00708a031. [DOI] [PubMed] [Google Scholar]
  35. Persohn E., Schachner M. Immunoelectron microscopic localization of the neural cell adhesion molecules L1 and N-CAM during postnatal development of the mouse cerebellum. J Cell Biol. 1987 Jul;105(1):569–576. doi: 10.1083/jcb.105.1.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Podbilewicz B., Mellman I. ATP and cytosol requirements for transferrin recycling in intact and disrupted MDCK cells. EMBO J. 1990 Nov;9(11):3477–3487. doi: 10.1002/j.1460-2075.1990.tb07556.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Powell S. K., Cunningham B. A., Edelman G. M., Rodriguez-Boulan E. Targeting of transmembrane and GPI-anchored forms of N-CAM to opposite domains of a polarized epithelial cell. Nature. 1991 Sep 5;353(6339):76–77. doi: 10.1038/353076a0. [DOI] [PubMed] [Google Scholar]
  38. Rodriguez-Boulan E., Nelson W. J. Morphogenesis of the polarized epithelial cell phenotype. Science. 1989 Aug 18;245(4919):718–725. doi: 10.1126/science.2672330. [DOI] [PubMed] [Google Scholar]
  39. Rodriguez-Boulan E. Polarized assembly of enveloped viruses from cultured epithelial cells. Methods Enzymol. 1983;98:486–501. doi: 10.1016/0076-6879(83)98176-4. [DOI] [PubMed] [Google Scholar]
  40. Rodriguez-Boulan E., Powell S. K. Polarity of epithelial and neuronal cells. Annu Rev Cell Biol. 1992;8:395–427. doi: 10.1146/annurev.cb.08.110192.002143. [DOI] [PubMed] [Google Scholar]
  41. Rutishauser U., Acheson A., Hall A. K., Mann D. M., Sunshine J. The neural cell adhesion molecule (NCAM) as a regulator of cell-cell interactions. Science. 1988 Apr 1;240(4848):53–57. doi: 10.1126/science.3281256. [DOI] [PubMed] [Google Scholar]
  42. Rutishauser U., Jessell T. M. Cell adhesion molecules in vertebrate neural development. Physiol Rev. 1988 Jul;68(3):819–857. doi: 10.1152/physrev.1988.68.3.819. [DOI] [PubMed] [Google Scholar]
  43. Sargiacomo M., Lisanti M., Graeve L., Le Bivic A., Rodriguez-Boulan E. Integral and peripheral protein composition of the apical and basolateral membrane domains in MDCK cells. J Membr Biol. 1989 Mar;107(3):277–286. doi: 10.1007/BF01871942. [DOI] [PubMed] [Google Scholar]
  44. Simons K., Wandinger-Ness A. Polarized sorting in epithelia. Cell. 1990 Jul 27;62(2):207–210. doi: 10.1016/0092-8674(90)90357-k. [DOI] [PubMed] [Google Scholar]
  45. Takeichi M. The cadherins: cell-cell adhesion molecules controlling animal morphogenesis. Development. 1988 Apr;102(4):639–655. doi: 10.1242/dev.102.4.639. [DOI] [PubMed] [Google Scholar]
  46. Thiery J. P., Duband J. L., Rutishauser U., Edelman G. M. Cell adhesion molecules in early chicken embryogenesis. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6737–6741. doi: 10.1073/pnas.79.21.6737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Tokuyasu K. T. A technique for ultracryotomy of cell suspensions and tissues. J Cell Biol. 1973 May;57(2):551–565. doi: 10.1083/jcb.57.2.551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wang A. Z., Ojakian G. K., Nelson W. J. Steps in the morphogenesis of a polarized epithelium. I. Uncoupling the roles of cell-cell and cell-substratum contact in establishing plasma membrane polarity in multicellular epithelial (MDCK) cysts. J Cell Sci. 1990 Jan;95(Pt 1):137–151. doi: 10.1242/jcs.95.1.137. [DOI] [PubMed] [Google Scholar]
  50. Zurzolo C., Le Bivic A., Quaroni A., Nitsch L., Rodriguez-Boulan E. Modulation of transcytotic and direct targeting pathways in a polarized thyroid cell line. EMBO J. 1992 Jun;11(6):2337–2344. doi: 10.1002/j.1460-2075.1992.tb05293.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES