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The effect of chloroquine on lysosomal function and cell-coat glycoprotein transport in the absorptive cells of cultured human small-intestinal tissue

Summary

The effect of chloroquine, an inhibitor of intralysosomal catabolism, on the synthesis, transport, and degradation of cell-coat glycoproteins in absorptive cells of cultured human small-intestinal tissue was investigated by morphometrical, autoradiographical, and biochemical methods. Neither synthesis nor transport of cell-coat material was affected by the drug, but culturing of the absorptive cells in the presence of chloroquine led to a dose- and time-dependent enlargement of the dense bodies; other cell structures showed no alterations. 3H-fucose-labelled material accumulated in the dense bodies of the absorptive cells of these cultures. Since no increase of β-glucuronidase and acid phosphatase activity (both lysosomal enzymes of glycoprotein nature) was found, this accumulation of radiolabelled material can be explained as a chloroquine-mediated inhibition of the degradation of cell-coat glycoproteins. These macromolecules probably enter the lysosome-like bodies by a crinophagic mechanism, i.e., fusion of these organelles with the apical vesicles and tubules involved in intracellular transport. These findings suggest that the lysosome-like bodies have a function in the regulation of cell-coat glycoprotein transport in human intestinal absorptive cells, i.e., the degradation of excess cell-coat material.

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References

  1. Bennett G (1970) Migration of glycoprotein from Golgi apparatus to cell coat in the columnar cells of the duodenal epithelium. J Cell Biol 45:668–673

  2. Bennett G, Leblond CP (1970) Formation of cell coat material for the whole surface of columnar cells in the rat small intestine, as visualized by radioautography with L-fucose-3H. J Cell Biol 46:409–417

  3. Bennett G, Leblond CP, Haddad A (1974) Migration of glycoprotein from the Golgi apparatus to the surface of various cell types as shown by radioautography after labeled fucose injections into rats. J Cell Biol 60:258–284

  4. Bessey OA, Lowry OH, Brock MJ (1946) A method for the rapid determination of alkaline phosphatase with five cubic millimeters of serum. J Biol Chem 164:321–329

  5. Blok J, Mulder-Stapel AA, Ginsel LA, Daems WTh (1980) Binding of cationized ferritin to the cell-coat glycoproteins of human and rat small-intestinal absorptive cells. Histochemistry 69:131–135

  6. Blok J, Ginsel LA, Mulder-Stapel AA, Onderwater JJM, Daems WTh (1981a) The effect of colchicine on the intracellular transport of 3H-fucose-labelled glycoproteins in the absorptive cells of cultured human small-intestinal tissue. An autoradiographical and biochemical study. Cell Tissue Res 215:1–12

  7. Blok J, Mulder-Stapel AA, Ginsel LA, Daems WTh (1981b) Endocytosis in absorptive cells of cultured human small-intestinal tissue: horseradish peroxidase, lactoperoxidase, and ferritin as markers. Cell Tissue Res 216:1–13

  8. Browning TH, Trier JS (1969) Organ culture of mucosal biopsies of human small intestine. J Clin Invest 48:1423–1432

  9. Buys ChCM, Bouma JMW, Gruber M, Wisse E (1978) Induction of lysosomal storage by suramin. Naunyn-Schmiedeberg's Arch. Pharmacol 304:183–190

  10. Daems WTh, Gemund JJ van, Vio PMA, Willighagen RGJ, Tandt WR den (1973) The use of intestinal suction biopsy material for the study of lysosomal storage diseases. In: Hers HG, Hoof H van (eds) Lysosomal storage diseases. Academic Press, New York, pp 575–598

  11. Dean RT (1975) Multiple forms of lysosomal enzymes. In: Dingle JT, Dean RT (eds) Lysosomes in biology and pathology, vol 4, North Holland Publ Comp, Amsterdam-Oxford, pp 349–382

  12. Drenckhahn D, Kleine L, Lüllmann-Rauch R (1976) Lysosomal alterations in cultured macrophages exposed to anorexigenic and psychotropic drugs. Lab Invest 35:116–123

  13. Duve C de, Barsy T de, Poole B, Trouet A, Tulkens P, Hoof F van (1974) Lysosomotropic agents. Biochem Pharmacol 23:2495–2531

  14. Farquhar MG (1969) Lysosome function in regulation secretion disposal of secretory granules in cells of the anterior pituitary gland. In: Dingle JT, Fell HB (eds) Lysosomes in Biology and Pathology, Vol 2. North-Holland Publ Comp, Amsterdam, pp 462–482

  15. Ginsel LA, Daems WTh, Emeis JJ, Vio PMA, Gemund JJ van (1973) Fine structure and silver-staining patterns of lysosome-like bodies in absorptive cells of the small intestine in normal children and children with a lysosomal storage disease. Virchows Arch Abt B Zellpathol 13:119–144

  16. Ginsel LA, Debets WF, Daems WTh (1975) The effect of colchicine on the distribution of apical vesicles and tubules in absorptive cells of cultured human intestine. In: Borgers M, Brabander M de (eds) Microtubules and Microtubule Inhibitors. North-Holland Publ Comp, Amsterdam, pp 187–198

  17. Ginsel LA, Want JJL van der, Daems WTh (1977) Qualitative and quantitative preservation of the fine structure of absorptive cells in cultured biopsies of human small intestine. Cell Tissue Res 181:143–162

  18. Ginsel LA, Cambier PH, Daems WTh (1978) Fucosidosis and I-cell disease: a fine structural and silver-staining study of abnormal inclusion bodies in small-intestinal cells. Virchows Arch B Cell Pathol 27:99–117

  19. Ginsel LA, Onderwater JJM, Daems WTh (1979a) Transport of radiolabelled glycoprotein to cell surface and lysosome-like bodies of absorptive cells in cultured small-intestinal tissue from normal subjects and patients with a lysosomal storage disease. Virchows Arch B Cell Pathol 30:245–273

  20. Ginsel LA, Onderwater JJM, Daems WTh (1979b) Resolution of a gold latensification-elon ascorbic acid developer for Ilford L4 emulsion. Histochemistry 61:343–346

  21. Goldstein JL, Brunschede GY, Brown MS (1975) Inhibition of the proteolytic degradation of low density lipoprotein in human fibroblasts by chloroquine, concanavalin A and triton WR 1339. J Biol Chem 250:7854–7862

  22. Goldstone A, Koenig H (1972) Biosynthesis of lysosomal glycoproteins in rat kidney. Life Sci 11:11:511–523

  23. Hauri HP, Kedinger M, Haffen K, Freiburghaus A, Grenier JF, Hadorn B (1977) Biosynthesis of brush border glycoproteins by human small intestinal mucosa in organ culture. Biochim Biophys Acta 467:327–339

  24. Hoof F van, Hers HG (1968) The abnormalities of lysosomal enzymes in mucopolysaccharidoses. Eur J Biochem 7:34–44

  25. Hugon JS, Borgers M (1968) Fine structural localization of three lysosomal enzymes and nonspecific alkaline phosphatase in the villus of the human duodenum. Gastroenterol 55:608–618

  26. Jersild RA, Crawford RW (1978) The distribution and mobility of anionic sites on the brush border of intestinal absorptive cells. Amer J Anat 152:287–306

  27. Karsten U, Wollenberger A (1977) Improvements in the ethidium bromide method for direct fluorometric estimation of DNA and RNA in cell and tissue homogenates. Anal Biochem 77:464–470

  28. Kedinger M, Hauri HP, Haffen K, Green JR, Grenier JF, Hadorn B (1979) Turnover studies of human intestinal brush border membrane glycoproteins in organ culture. Enzyme 24:96–106

  29. Lie SO, Schofield B (1973) Inactivation of lysosomal function in normal cultured human fibroblasts by chloroquine. Biochem Pharmacol 22:3109–3114

  30. Lloyd JB (1973) Experimental support for the concept of lysosomal storage disease. In: Hers HG, Hoof H van (eds) Lysosomes and storage diseases. Academic Press, New York, pp 173–195

  31. Lüllmann-Rauch R (1979) Drug-induced Lysosomal Storage Disorders. In: Dingle JT, Jacques PJ, Shaw IH (eds) Lysosomes in Applied Biology and Therapeutics, Vol 6. North-Holland Publ Comp, Amsterdam New York Oxford, pp 49–130

  32. Mego JL, Chung CH (1979) Effects of some antimalarials and related substances on intralysosomal proteolysis. Biochem Pharmacol 28:465–470

  33. Michaels JE, Leblond CP (1976) Transport of glycoprotein from Golgi apparatus to cell surface by means of “carrier vesicles”, as shown by radioautography of mouse colonic epithelium after injection of 3H-fucose. J Microscopie Biol Cell 25:243–248

  34. Ohkuma S, Poole B (1978) Fluorescence probe measurement of the intralysosomal pH in living cells and the pertubation of pH by various agents. Procs Natl Acad Sci USA 75:3327–3331

  35. Plaice CHJ (1961) A note on the determination of serum beta-glucuronidase. J Clin Pathol 14:661–665

  36. Reijngoud DJ, Tager JM (1976) Chloroquine accumulation in isolated rat liver lysosomes. FEBS Letters 64:231–235

  37. Ridout RM, Decker RS, Wildenthal K (1977) Chloroquine-induced lysosomal abnormalities in cultured foetal mouse hearts. J Mol Cell Cardiol 10:175–183

  38. Schmitz J, Preiser H, Maestracci D, Ghosh BK, Cerda JJ, Crane RK (1973) Purification of the human intestinal brush border membrane. Biochim Biophys Acta 323:98–112

  39. Seglen PO, Grinde B, Solheim AE (1979) Inhibition of the lysosomal pathway of protein degradation in isolated rat hepatocytes by ammonia, methylamide, chloroquine and leupeptin. Eur J Biochem 95:215–225

  40. Wibo M, Poole M (1974) Protein degradation in cultured cells. II. The uptake of chloroquine by rat fibroblasts and the inhibition of cellular protein degradation and cathepsin B1. J Cell Biol 63:430–440

  41. Wildenthal K, Wakeland JR, Morton PC, Griffin EE (1978) Inhibition of protein degradation in mouse hearts by agents that cause lysosomal dysfunction. Circulation Res 42:787–792

  42. Williams MA (1973) Electron microscopic autoradiography: Its application to protein biosynthesis, Campbell PN, Sargent JR (eds) Academic Press, London, pp 125–191

  43. Wisse E, Tates AD (1968) A gold latensification-elon ascorbic acid developer for Ilford L4 emulsion. In: Bocciarelli DS (ed) Electron microscopy, 4th European Region. Conf on Electron Microscopy, Vol II, Rome Tipografia Poliglotta, Vaticana, pp 465–466

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Correspondence to J. Blok.

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Blok, J., Mulder-Stapel, A.A., Ginsel, L.A. et al. The effect of chloroquine on lysosomal function and cell-coat glycoprotein transport in the absorptive cells of cultured human small-intestinal tissue. Cell Tissue Res. 218, 227–251 (1981). https://doi.org/10.1007/BF00210340

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Key words

  • Chloroquine
  • Human enterocyte
  • Glycoprotein transport
  • Lysosomal degradation