Skip to main content
Log in

Comparison of tight junction permeability for albumin in iris pigment epithelium and retinal pigment epithelium in vitro

  • Laboratory Investigation
  • Published:
Graefe's Archive for Clinical and Experimental Ophthalmology Aims and scope Submit manuscript

Abstract

• Background: The degenerative retinal diseases are one of the major causes of visual loss in the western world. Although heterologous RPE transplants rescue the photoreceptors in the dystrophic rat model, rejection remains a major limiting factor. Given the common embryonic origin, iris pigment epithelial (IPE) cells might be able to take over the functions of retinal pigment epithelial (RPE) cells, serving as an autologous graft for transplantation and thereby preventing rejection. One of the main functions of RPE cells is the generation of tight junctions which form the outer blood-retinal barrier. In this study we compared the tight junction permeabilities of IPE and RPE cells isolated from Long Evans rats by measuring their albumin clearances. • Methods: IPE and RPE cells were cultured on semipermeable filter supports with and without the addition of 0.02% ethylene diaminetetraacetic acid (EDTA). At selected intervals, the albumin clearances of the IPE and RPE cells were measured spectrophotometrically and compared. The morphology of the cells was compared using electron microscopy and fluorescent labeling. • Results: IPE and RPE cells both restricted the passage of albumin in vitro. After the modulation of tight junctions with 0.02% EDTA, the clearance increased in both types of cells in a similar fashion. The morphology of tight junctions was visualized with electron microscopy. • Conclusion: These results indicate that the functional barrier for macromolecules is similar in IPE and RPE cells in vitro. This raises the possibility that IPE cells would form tight junctions in the subretinal space, thereby substituting for the blood-retinal barrier normally formed by RPE cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Algvere PV, Berglin L, Gouras P, Sheng Y (1995) Transplantation of human RPE in age-related macular degeneration with subfoveal neovascullarisation. ARVO annual meeting, abstract 966

  2. Berkowitz BA, Sato Y, Wilson CA, Juan E de Jr (1991) Blood retinal barrier function breakdown investigated by real-time MRI following gadolinium-diethylene-triamine pentacetic acid injection. Invest Ophthalmol Vis Sci 32:2854–2860

    Google Scholar 

  3. Bressler NM, Bressler SB, Fine SL (1988) Age-related macula degeneration. Surv Ophthalmol 32:375–411

    Google Scholar 

  4. Burnside B, Laties AM (1979) Pigment movement and cellular contractility in the retinal pigment epithelium. In: Marmor MF, Zinn KM (eds) The retinal pigment epithelium. Harvard University Press, Cambridge, Mass, pp 175–191

    Google Scholar 

  5. Cilluffo MC, Fain MJ, Fain GL (1993) Tissue culture of rabbit ciliary body epithelial cells on permeable supports. Exp Eye Res 57:513–526

    Google Scholar 

  6. Contreras RG, Miller JH, Zamora M, Gonzalez-Mariscal L, Cereijido M (1992) Interaction of calcium with plasma membrane of epithelial (MDCK) cells during junction formation. Am J Physiol 263:313–318

    Google Scholar 

  7. Edwards RB (1977) Culture of rat retinal pigment epithelium. In Vitro 13:301–304

    Google Scholar 

  8. Farrokh-Siar L, Rezai KA, Lappas L, Kohen L, Heimann K (1995) Comparison of phagocytosis activity of iris pigment epithelium and retinal pigment epithelium of newborn rats through a fluorescent vital assay in vitro. ARVO annual meeting, abstract 3785

  9. Garcia JGN, Silfinger-Birnboim A, Bizios R, Del Vecchio PJ, Fenton JW II, Malik AB (1986) Thrombin-induced increase in albumin permeability across the endothelium. J Cell Physiol 128:96–104

    Google Scholar 

  10. Gelanze M, Breipohl W, Wiedemann P, Naib-Majani W, Heimann K (1993) First experimental iris pigment epithelial cell transplantation in subretinal space of RCS rats. ARVO annual meeting, abstract 1942

  11. Gonzalez-Mariscal L, Contreras RG, Bolivar JJ, Ponce A, Chavez De Ramirez B, Cereijido M (1990) Role of calcium in tight junction formation between epithelial cells. Am J Physiol 259:978–986

    Google Scholar 

  12. Gumbiner B (1987) Structure, biochemistry, and assembly of epithelial tight junctions. Am Phys Soc 253:749–758

    Google Scholar 

  13. Gumbiner B, Simons K (1987) The role of uvomorulin in the formation of epithelial occluding junctions. In: Stoker M (ed) Junctional complexes of epithelial cells. Wiley, Chichester, UK

    Google Scholar 

  14. Hogan MJ, Alvarado JA, Weddel JA (1971) Histology of the human eye, 1st edn. Saunders, Philadelphia, pp 202–259

    Google Scholar 

  15. Hu DN, Ritch R, McCormicks SA, Pelton-Henrion K (1992) Isolation and cultivation of human iris pigment epithelium. Invest Ophthlmol Vis Sci 33:2443–2453

    Google Scholar 

  16. Jiang LQ, Jarquera M, Streilein JW (1994) Immunologic consequences of intraocular implantation of retinal pigment epithelial allografts. Exp Eye Res 58:719–728

    Google Scholar 

  17. Kartenbeck J, Schmid E, Franke WW, Geiger B (1982) Different modes of internalisation of proteins associated with adhaerens junctions and desmosomes: experimental seperation of lateral contacts induces endocytosis of desmosomal plaque material. EMBO J 1:725–732

    Google Scholar 

  18. Katakam M, Banga AK (1995) Aggregation of proteins and its prevention by carbohydrate excipients: albumins and gamma-globulin. J Pharm Pharmacol 47:103–107

    Google Scholar 

  19. Kohen L, Nishi M, Hall MO, Gabrielian K, Kohen S, Pararajasegaram G, Rao NA (1993) Immunohistochemical study of the rejection of transplanted RPE cells in a rat model. ARVO annual meeting 1993: abstract 1929

  20. LaVail MM, Li L, Turner JE, Yasumura D (1992) Retinal pigment epithelial cell transplantation in RCS rats: normal metabolism in rescued photoreceptors. Exp Eye Res 55:555–562

    Google Scholar 

  21. Lee VHL, Pince KJ, Frambach DA, Martini B (1989) Drug delivery to the posterior segment. In: Ryan SJ, Ogden TE (eds) Retina, vol 1. Mosby, St. Louis, pp 483–498

    Google Scholar 

  22. Levitt DG (1981) Routes of membrane water transport: comparative physiology. Alfred Benzon Symp 15:248–257

    Google Scholar 

  23. Lopez R, Gouras P, Kjeldbye H, Sullivan B, Reppucci V, Brittis M, Wapner F, Goluboff E (1989) Transplanted retinal pigment epithelium modifies the retinal degeneration in the RCS rat. Invest Ophthalmol Vis Sci 30:86–588

    Google Scholar 

  24. Madara JL (1987) Intestinal absorptive cell tight junctions are linked to the cytoskeleton. Am J Physiol 253:171–175

    Google Scholar 

  25. Marmor MF (1990) Control of subretinal fluid: experimental and clinical studies. Eye 4:340–344

    Google Scholar 

  26. Martinez-Palmo A, Meza I, Beaty G, Cereijido M (1980) Experimental modulation of occluding junctions in a cultured transport medium. J Cell Biol 87:736–745

    Google Scholar 

  27. Mayerson PL, Hall MO, Clark V, Abrams T (1985) An improved method for isolation and culture of rat retinal pigment epithelial cells. Invest Ophthalmol Vis Sci 26:1599–1609

    Google Scholar 

  28. Meldolesi J, Castiglioni G, Parma R, Nassivera N, De-Camilli P (1978) Ca2+-dependent disassembly and reassembly of occluding junctions in guinea pig pancreatic acinar cells. J Cell Biol 79:156–172

    Google Scholar 

  29. Meza I, Ibarra G, Sabanero M, Martinez-Palomo A, Cereijido M (1980) Occluding junctions and cytoskeletal components in a cultured transporting epithelium. J Cell Biol 87:746–754

    Google Scholar 

  30. Milton SG, Knutson VP (1990) Comparison of the function of the tight junctions of endothelial cells and epithelial cells in regulating the movement of electrolytes and macromolecules across the cell monolayer. J Cell Physiol 144:498–504

    Google Scholar 

  31. Murray MA, Heistad DD, Mayhan WG (1991) Role of protein kinase C in bradykinin-induced increase in microvascular permeability. Circ Res 68:1341–1348

    Google Scholar 

  32. Orgtil S, Reuter U, Kain HL (1993) Osmotic stress in an in vitro model of the outer blood-retinal barrier. German J Ophthalmol 2:436–443

    Google Scholar 

  33. Pitelka DR, Taggart BN, Hamamoto ST (1983) Effect of extracellular calcium depletion on membrane topography and occluding junctions of mammary epithelial cells in culture. J Cell Biol 96:606–612

    Google Scholar 

  34. Plehwe WE, Mcrobbie DW, Lerski RA, Kohner EM (1988) Quantitative magnetic resonance imaging in assessment of the blood-retinal barrier. Invest Ophthalmol Vis Sci 29:663–670

    Google Scholar 

  35. Raviola G (1977) The structural basis of the blood-ocular barrier. Exp Eye Res 25:27–63

    Google Scholar 

  36. Rizzolo LJ, Li ZQ (1993) Diffusible retinal factors stimulate the barrier properties of junctional complexes in the retinal pigment epithelium. J Cell Sci 109:859–867

    Google Scholar 

  37. Sato Y, Berkowitz BA, Wilson CA, Juan E de Jr (1992) Blood-retinal barrier breakdown caused by diode vs argon laser endophotocoagulation. Arch Ophthalmol 110:277–281

    Google Scholar 

  38. Sedar AW, Forte JG (1964) Effects of calcium depletion on the junctional complex between oxyntic cells of gastric glands. J Cell Biol 22:173–188

    Google Scholar 

  39. Sheedlo HJ, Li L, Turner JE (1990) Photoreceptor cell rescue at early and late RPE-cell transplantation periods during retinal disease in RCS dystrophic rat. J Neural Transplant 2:55–63

    Google Scholar 

  40. Spitznas M (1986) Pathogenesis of central serous retinopathy: a new working hypothesis. Graefe's Arch Clin Exp Ophthalmol 224:321–324

    Google Scholar 

  41. Stevenson BR, Anderson JM, Bullivant S (1988) The epilthelial tight junction: structure, function and preliminary biochemical characterisation. Mol Cell Biochem 83:129–145

    Google Scholar 

  42. Tidball CS (1964) Magnesium and calcium as regulators of intestine permability. Am J Physiol 209:243–246

    Google Scholar 

  43. Volberg T, Geiger B, Kartenbeck J, Franke WW (1986) Changes in membrane-microfilament interaction in intercellular adherens junctions upon removal of extracellular Ca2+ ions. J Cell Biol 102:1832–1842

    Google Scholar 

  44. Volk T, Geiger B (1986) A-CAM: a 135-kD receptor of intercellular adherens junctions. II. Antibody-mediated modulation of junction formation. J Cell Biol 103:1451–1464

    Google Scholar 

  45. Zinn KM, Benjamin-Henkind JV (1979) Anatomy of the human retinal pigment epithelium. In: Marmor ME, Zinn KM (eds) The retinal pigment epithelium. Harvard University Press, Cambridge, Mass, pp 3–25

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rezai, K.A., Lappas, A., Kohen, L. et al. Comparison of tight junction permeability for albumin in iris pigment epithelium and retinal pigment epithelium in vitro. Graefe's Arch Clin Exp Ophthalmol 235, 48–55 (1997). https://doi.org/10.1007/BF01007837

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01007837

Keywords

Navigation