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Release of lysosomal protease from retinal pigment epithelium and fibroblasts during mechanical stresses

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Abstract

• Background: Mechanical expansion in tissues or in cells may occur under physiological and pathological conditions and is accompanied by increased activity of proteolytic enzymes. In traction detachment, retinal cells are subjected to mechanical strain. The purpose of this study was to ascertain whether retinal pigment epithelium (RPE) cells and fibroblasts in tissue culture release proteases due to mechanical stress and to investigate the importance of the cytoskeleton for mechanotransduction to the lysosomes during cellular stress reactions. • Methods: Cell layers were grown on siliconerubber membranes and subjected to mechanical stresses by expansion of the membrane. Concentrations of N-acetyl-β-glucosaminidase (NAG), acid phosphatase (AP), and lactate dehydrogenase (LDH) were determined in extracellular fluid. Colchicine 0.5 × 10−5 M was used to disrupt the cytoskeleton prior to expansion. • Results: RPE cells and fibroblasts separated during mechanical expansion, which was accompanied by extracellular release of proteolytic activity of NAG (RPE 37.50%; fibroblasts 23.22% above control value), but not of AP. LDH activity did not increase, indicating preserved integrity of the cell membranes during stretching. Colchicine caused immediate detachment of fibroblasts, and RPE did not release significant activity of NAG under subsequent extension. • Conclusion: In traction detachment, RPE may release in vivo proteases to cut intercellular adhesions in order to escape mechanical strain. Our results indicate that release of proteases from RPE may be involved in the pathophysiology of traction detachment, facilitating by their degradative action the disconnection between RPE and outer segments. Similarly, fibroblasts may respond to changes in tension of scleral or corneal tissue. Release of proteases following mechanical stress seems to require an intact cytoskeleton.

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References

  1. Abercrombie MA, Heaysman SEM, Pegrum MS (1971) The locomotion in fibroblasts in culture. IV. Electron microscopy of the leading edge. Exp Cell Res 67:359–367

    Google Scholar 

  2. Adler AJ (1989) Selective presence of acid hydrolases in the interphotoreceptor matrix. Exp Eye Res 49:1067–1077

    Google Scholar 

  3. Adler AJ, Martin KJ (1983) Lysosomal enzymes in the interphotoreceptor matrix: acid protease. Curr Eye Res 2:359–366

    Google Scholar 

  4. Beloussov LV, Dorfman JG, Cherdantzev VG (1975) Mechanical stresses and morphological patterns in amphibian embryos. J Embryol Exp Morphol 34:559–574

    Google Scholar 

  5. Bock G, Clark S (eds) (1987) Junctional complexes of epithelial cells. (Ciba Foundation Symposium) Wiley, Chichester

    Google Scholar 

  6. Chang B, Tuchler RE, Siebert JW, Longaker MT, Burd DA (1992) The effect of tissue expansion on dermal fibroblast contraction. Ann Plast Surg 28:315–319

    Google Scholar 

  7. Charkraborty A, Eghbali M (1991) An immunoassay for qualitative estimation of collagenase activity in mammalian tissues. Clin Biochem 24:455–461

    Google Scholar 

  8. Chen WT, Singer SJ (1982) Immunoelectron microscopic studies of the sites of cell-substratum adhesion and cell-cell contacts in cultured fibroblasts. J Cell Biol 1982;95:205–222

    Google Scholar 

  9. Collot M, Louvard D, Singer SJ (1984) Lysosomes are associated with microtubules and not with intermediate filaments in cultured fibroblasts. Proc Natl Acad Sci USA 81:788–792

    Google Scholar 

  10. Curtis ASG, Sehar GM (1978) The control of cell division by tension or diffusion. Nature 274:53–55

    Google Scholar 

  11. deWitt MT, Handley CJ, Oakes BW, Lowther DA (1984) In vitro response of chondrocytes to mechanical loading. The effect of short term mechanical stress. Connect Tissue Res 12:97–104

    Google Scholar 

  12. Edelman GM (1983) Cell adhesion molecules. Science 219:450–457

    CAS  PubMed  Google Scholar 

  13. Emonard H, Grimaud J-A (1990) Matrix metalloproteinases. A review. Cell Mol Biol 36:131–153

    Google Scholar 

  14. Geiger B, Tokuyasu KT, Dutton AH, Singer SJ (1980) Viculin, an intracellular protein localized at specialized sites where microfilament bundles terminate at cell membranes. Proc Natl Acad Sci USA 77:4127–4131

    Google Scholar 

  15. Gospodarowicz P, Delgado D, Vlodavsky I (1980) Permissive effect of the extracellular matrix on cell proliferation in vitro. Proc Natl Acad Sci USA 77:4094–4098

    Google Scholar 

  16. Hayasaka S, Hara S, Mizuno K (1976) Lysosomal enzymes in subretinal fluid. Graefe's Arch Clin Exp Ophthalmol 200:13–20

    Google Scholar 

  17. Heath JP, Dunn GA (1978) Cell to substratum contacts of chick fibroblasts and their relation to the microfilament system. A correlated interference-reflection and high-voltage electron microscopic study. J Cell Sci 29:197–212

    Google Scholar 

  18. Hollyfield JG, Varner HH, Rayborn ME, Osterfeld AM (1989) Retinal attachment to the pigment epithelium. Linkage through an extracellular sheath surrounding cone photoreceptors. Retina 9:59–68

    Google Scholar 

  19. Hunt RC, Fox A, Al Pakalnis A, Sigel MM, Kosnosky W, Choudhury P, Black EP (1993) Cytokines cause cultured retinal pigment epithelial cells to secrete metalloproteinases and to contract collagen gels. Invest Ophthalmol Vis Sci 34:3179–3186

    Google Scholar 

  20. Jones JCR, Asmuth J, Baker SE, Langhofer M, Roth SI, Hopkinson SB (1994) Hemidesmosomes. extracellular matrix/intermediate filament connectors. (Special article) Exp Cell Res 213:1–11

    Google Scholar 

  21. Kain HL (1982) Experimental retinal detachment on the isolated retinasclera preparation and measurement of the subretinal pressure. Klin Monatsbl Augenheilkd 180:155–157

    Google Scholar 

  22. Kain HL (1984) A new model for examining chorioretinal adhesion experimentally. Arch Ophthalmol 102:608–611

    Google Scholar 

  23. Kain HL (1984) Retinal adhesion. Trans Ophthalmol Soc UK 103:486–493

    Google Scholar 

  24. Kain HL (1990) Adhesion between sensory retina and retinal pigment epithelium. Invest Ophthalmol Vis Sci 31 [Suppl] p:26

    Google Scholar 

  25. Kaufman PL, Podos SM (1973) The subretinal fluid in primary rhegmatogenous retinal detachment. Surv Ophthalmol 18:100–116

    Google Scholar 

  26. Kolega J (1986) Effects of mechanical tension on protrusive activity and microfilament and intermediate filament organisation in an epidermal epithelium moving in culture. J Cell Biol 102:1400–1411

    Google Scholar 

  27. Machemer R (1984) The importance of fluid absorption, traction, intraocular currents, and chorioretinal scars in the therapy of rhegmatogenous retinal detachment. (XLI. Edward Jackson Memorial Lecture) Am J Ophthalmol 98:681–693

    Google Scholar 

  28. McGuire PG, Seeds NW (1990) Degradation of underlying extracellular matrix by sensory neurons during neurite outgrowth. Neuron 4:633–642

    Google Scholar 

  29. McGuire PG, Orkin RW (1992) Urokinase activity in the developing avian heart: a spatial and temporal analysis. Dev Dyn 193:24–33

    Google Scholar 

  30. Mignatti P, Mazzieri R, Rifkin DB (1991) Expression of urokinase receptor in vascular endothelial cells is stimulated by basic fibroblast growth factor. J Cell Biol 113:1193–1201

    Google Scholar 

  31. Moscatelli D, Rifkin DB (1988) Membrane and matrix localization of proteinases: a common theme in tumor cell invasion and angiogenesis. Biochem Biophys Acta 984:67–85

    Google Scholar 

  32. Odell G, Oster G, Burnside B, Albrech P (1981) Mechanical basis of morphogenesis. I. Epithelial folding and invagination. Dev Biol 85:446–462

    Google Scholar 

  33. Plantner JJ (1992) The presence of neutral metalloproteolytic activity and metalloproteinase inhibitors in the interphotoreceptor matrix. Curr Eye Res 11:91–101

    Google Scholar 

  34. Reddi AH (1984) Extracellular matrix and development. In: Piez KA, Reddi AH (eds) Extracellular matrix biochemistry. Elsevier, New York, pp 399–401

    Google Scholar 

  35. Robert L (ed) (1986) Structural glycoproteins in cell-matrix interactions. (Frontiers in Matrix Biology, vol 11) Karger, Basel

  36. Sawaguchi S, Yue BYJT, Augar J, Gilboy JE (1989) Lysosomal enzyme activities in keratoconus. Arch Ophthalmol 107:1507–1510

    Google Scholar 

  37. Stopak D, Harris AK (1982) Connective tissue morphogenesis by fibroblast traction. I. Tissue culture observations. Dev Biol 90:383–389

    Google Scholar 

  38. Timmenga EJ, Andreassen TT, Houthoff HJ, Klepper PJ (1991) The effect of mechanical stress on healing skin wounds: an experimental study in rabbits using tissue expansion. Br J Plast Surg 44:514–519

    Google Scholar 

  39. Vandenburgh H, Kaufmann S (1990) In vitro model for stretch-induced hypertrophy of skeletal muscle. Science 203:265–268

    Google Scholar 

  40. Wang N, Butler JP, Ingber DE (1993) Mechanotransduction across the cell surface and through the cytoskeleton. Science 260:1124–1127

    Google Scholar 

  41. Wilcox DK (1987) Extracellular release of acid hydrolases from cultured retinal pigment epithelium. Invest Ophthalmol Vis Sci 28:76–82

    Google Scholar 

  42. Wolpert L (1977) Introductory remarks. In: Brinkely BR, Porter KR (eds) International cell biology. Rockefeller University Press, New York, pp 31–35

    Google Scholar 

  43. Zauberman H, de Guillebon H (1972) Retinal traction in vivo and post mortem. Arch Ophthalmol 86:549–554

    Google Scholar 

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  1. Eye Clinic, University of Basel, Mittlere Strasse 90, CH-4095, Basel, Switzerland

    Hermann L. Kain & Ute Reuter

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  1. Hermann L. Kain
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  2. Ute Reuter
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Kain, H.L., Reuter, U. Release of lysosomal protease from retinal pigment epithelium and fibroblasts during mechanical stresses. Graefe's Arch Clin Exp Ophthalmol 233, 236–243 (1995). https://doi.org/10.1007/BF00183598

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  • DOI: https://doi.org/10.1007/BF00183598

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