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Loss of microtubules and alteration of glycoprotein migration in organ cultures of mouse intestine exposed to nocodazole or colchicine

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Summary

Explants from mouse jejunum were cultured for 3–7 h in the absence (control) or presence of colchicine (100 gm/ml) or nocodazole (10 μg/ml). In recovery experiments, expiants were cultured in fresh medium for an additional period. To label glycoproteins, 3H-fucose was added during the last 3 or 6 h of the initial culture or recovery period. Subcellular fractionation studies revealed that colchicine and nocodazole inhibited migration of labelled glycoproteins to the brush border (P2) by 40–45%. Radioautographic studies of absorptive cells showed that colchicine and nocodazole inhibited labelling of the microvillous border by 67% and 87%, while labelling of the basolateral plasma membrane increased by 114% and 275%. Immunocytochemical studies revealed that both colchicine and nocodazole caused the virtual disappearance of the microtubular network in the absorptive cells. It is possible that some glycoproteins normally destined for the microvillous border are rerouted to the basolateral membrane. The observed loss of microtubules after drug treatment suggests that microtubules may play a role in the intracellular migration of membrane glycoproteins. Additional support for this concept is provided by the fact that in recovery experiments the distribution of label returned to control values after the microtubular network became re-established.

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References

  • Aitchison WA Brown DL (1984) Duplication of the flagellar apparatus and the development of cell morphology during cell division in alga Polytomella. J Cell Biol 99:188a

    Google Scholar 

  • Azhar S, Hwang SF, Reaven EP (1983) Effect of antimicrotubule agents on terminal glycosyltransferases and other enzymes associated with rat liver subcellular fractions. Biochem J 212:721–731

    Google Scholar 

  • Bennett G (1983) Autoradiographic analysis of glycoprotein synthesis and secretion of hepatocytes. In: Structural Carbohydrates of the Liver. Falk Symposium No. 34. MTP Press, Lancaster England, pp 147–162

    Google Scholar 

  • Bennett G, Hugon JS (1982) Effect of colchicine on migration of 3H-fucoselabelled glycoproteins in mouse jejunal absorptive cells in organ culture: an electron microscope radioautographic study. J Cell Biol 95:401a

    Google Scholar 

  • 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 labelled fucose injection into rats. J Cell Biol 60:258–284

    Google Scholar 

  • Bennett G, Parsons S, Carlet E (1984a) Influence of colchicine and vinblastine on the intracellular migration of secretory and membrane glycoproteins. I. Inhibition of glycoprotein migration in various rat cell types as shown by light microscope radioautography after injection of 3H-fucose. Am J Anal 170:521–530

    Google Scholar 

  • Bennett G, Carlet E, Wild G, Parsons S (1984b) Influence of colchicine and vinblastine on the intracellular migration of secretory and membrane glycoproteins. III. Inhibition of intracellular migration of membrane glycoproteins in rat intestinal columnar cells and hepatocytes as visualized by light and electron-microscope radioautography after 3H-fucose injection. Am J Anat 170:545–566

    Google Scholar 

  • Bernier-Valentin F, Aunis D, Rousset B (1983) Evidence for tubulin-binding sites on cellular membranes: plasma membranes, mitochondrial membranes, and secretory granule membranes. J Cell Biol 97:209–216

    Google Scholar 

  • Berteloot A, Hugon JS (1982) Studies on the incorporation of radioactive amino acids into tissue and brush border membrane proteins of mouse jejunum in organ culture. Comp Biochem Physiol 73B:491–499

    Google Scholar 

  • Bhattacharyya B, Wolff J (1976) Polymerization of membrane tubulin. Nature (London) 264:576–577

    Google Scholar 

  • Birkett CR, Coulson C, Pogson CI, Gull K (1981) Inhibition of secretion of proteins and triacylglycerol from isolated rat hepatocytes mediated by benzimidazole carbamate antimicrotubule agents. Biochem Pharmacol 30:1629–1633

    Google Scholar 

  • Bloch RJ (1983) Acetylcholine receptor clustering in rat myotubes: requirement for Ca2+ and effects of drugs which depolymerize microtubules. J Neurosci 3:2670–2680

    Google Scholar 

  • Blök J, Ginsel LA, MulderStapel AA, Ounderwater JJM, Daems WTh (1981) The effect of colchicine on the intracellular transport of 3H-fucose-labellcd glycoproteins in the absorptive cells of cultured human small-intestinal tissue. An autoradiographical and biochemical study. Cell Tissue Res 215:1–12

    Google Scholar 

  • De Brabander M, Geuens G, De Mey J, Joniau M (1981) Nucleated assembly of mitotic microtubules in living PTK2 cells after release from nocodazole treatment. Cell Motil 1:469–483

    Google Scholar 

  • Dustin P (1978) “Microtubules”. Springer Verlag, Berlin, Heidelberg New York

    Google Scholar 

  • Ellinger A, Pavelka M, Gangl A (1983) Effect of colchicine on rat small intestinal absorptive cells. II. Distribution of label after incorporation of [3H]fucose into plasma membrane glycoproteins. J Ultrastruct Res 85:260–271

    Google Scholar 

  • Ferland S, Hugon JS (1979) Organ culture of adult mouse intestine. I. Morphological and cytochemical results after 24 and 48 hr of culture. In vitro 15:278–287

    Google Scholar 

  • Ginsei LA, Debets WF, Daems WTh (1975) The effect of colchicine on the distribution of apical vesicles and tubules in absorptive cells of cultured human small inestine. In: Borgers M, Brabander M de (eds) Microtubules and microtubule inhibitors. North Holland Publishing Company, Amsterdam, pp 187–198

    Google Scholar 

  • Hauri HP, Roth J, Sterchi E, Fransen JAM (1984) Biosynthesis and transport to the cell surface of intestinal microvillar hydrolascs. J Cell Biol 99:6a

    Google Scholar 

  • Karim A, Warshawsky H (1979) The effect of colcemid on the structure and secretory activity of ameloblasts in the rat incisor as shown by radioautography after injection of 3H-proline. Anat Rec 195:587–610

    Google Scholar 

  • Kelly WG, Passaniti A, Woods JW, Daiss JL, Roth TF (1983) Tubulin as a molecular component of coated vesicles. J Cell Biol 97:1191–1199

    Google Scholar 

  • Matlin KS, Simons K (1984) Sorting of an apical plasma membrane glycoprotein occurs before it reaches the cell surface in cultured epithelial cells. J Cell Biol 99:2131–2139

    Google Scholar 

  • Michacls JE (1983) The effects of colchicine on the distribution of glycoprotein-containing vesicles in epithelial cells of the murine colon. Cell Tissue Res 228:323–336

    Google Scholar 

  • 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 Microsc Biol Cell 25:243–248

    Google Scholar 

  • Mitranic MM, Boggs JM, Moscarello MA (1981) An effect of colchicine on galactosyl and sialytransferase of rat liver Golgi membranes. Biochim Biophys Acta 672:57–64

    Google Scholar 

  • Pavelka M, Ellinger A (1981) Effect of colchicine on the Golgi apparatus and on GERL of rat jejunal absorptive cells. Ultrastructural localization of thiamine pyrophosphatase and acid phosphatase activity. Eur J Cell Biol 24:53–61

    Google Scholar 

  • Pavelka M, Gangl (1983) Effects of colchicine on the intestinal transport of endogenous lipid. Ultrastructural, biochemical, and radiochemical studies in fasting rats. Gastroenterology 84:544–555

    Google Scholar 

  • Pavelka M, Ellinger A, Gangl A (1983) Effect of colchicine on rat intestinal absorptive cells. I. Formation of basolateral microvillus borders. J Ultrastruct Res 85:249–259

    Google Scholar 

  • Quaroni A, Kirsch K, Weiser MM (1979) Synthesis of membrane glycoproteins in rat smallintestinal villus cells. Effect of colchicine on the redistribution of L-[1,5,6-3]fucoselabelled membrane glycoproteins among Golgi, lateral basal and microvillus membranes. Biochem J 182:213–221

    Google Scholar 

  • Ranscht B, Moss D, Thomas C (1984) A neuronal surface glycoprotein associated with cytoskeleton. J Cell Biol 99:1803–1813

    Google Scholar 

  • Rassatt J, Robenek J, Themann H (1982) Alterations of tight and gap junctions in mouse hepatocytes following administration of colchicine. Cell Tissue Res 223:187–200

    Google Scholar 

  • Reaven EP, Reaven GM (1977) Distribution and content of microtubules in relation to the transport of lipid. An ultrastructural quantitative study of the absorptive cell of the small intestine. J Cell Biol 75:559–572

    Google Scholar 

  • Rindler MJ, Ivanov IE, Plesken H, Sabatini DD (1985) Polarized delivery of viral glycoproteins to the apical and basolateral plasma membranes of Madin-Darby canine kidney cells infected with temperature-sensitive viruses. J Cell Biol 100:136–151

    Google Scholar 

  • Robenek H, Gebhardt R (1983) Primary cultures of rat hepatocytes as a model system of canalicular development, biliary secretion, and intrahepatic cholestasis. IV. Desintegration of bile canaliculi and disturbance of tight junction formation caused by vinblastine. Europ J Cell Biol 31:283–289

    Google Scholar 

  • Rodriguez-Boulan E, Paskiet KT, Salas PJI, Bard E (1984) Intracellular transport of influenza virus hemaglutinin to the apical surface of Madin-Darby canine kidney cells. J Cell Biol 98:308–319

    Google Scholar 

  • Rogalski AA, Singer SJ (1984) Association of elements of the Golgi apparatus with microtubules. J Cell Biol 99:1092–1100

    Article  CAS  PubMed  Google Scholar 

  • Sandoval IV, Bonifacino JS, Klausner RD, Henkart M, Wehland J (1984) Role of microtubules in the organization and localization of the Golgi apparatus. J Cell Biol 99:113s-118s

    Article  CAS  PubMed  Google Scholar 

  • 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

    Google Scholar 

  • Stadier J, Franke WW (1974) Characterization of the colchicine binding of membrane fractions from rat and mouse liver. J Cell Biol 60:297–303

    Google Scholar 

  • Steiner M (1983) Membrane-bound tubulin in human platelets. Biochim Biophys Acta 729:17–22

    Google Scholar 

  • Stephens RE (1977) Major membrane protein differences in cilia and flagella: Evidence for a membrane associated tubulin. Biochemistry 16:2047–2058

    Google Scholar 

  • Stephens RE (1983) Reconstitution of ciliary membranes containing tubulin. J Cell Biol 96:68–75

    Google Scholar 

  • Tarone G, Ferracini R, Galetto G, Comoglio P (1984) A cell surface integral membrane glycoprotein of 85,000 mol wt (gp85) associated with Triton-Xinsoluble cell skeleton. J Cell Biol 99:512–519

    Google Scholar 

  • Thyberg J, Piasek A, Moskalewski S (1980) Effects of colchicine on the Golgi complex and GERL of cultured rat peritoneal macrophages and epiphyseal chondrocytes. J Cell Sci 45:42–58

    Google Scholar 

  • Wehland J, Willingham MC (1983) A rat monoclonal antibody reacting specifically with the tyrosylated form of atubulin. II. Effects on cell movement, organization of microtubules, and intermediate filaments, and arrangement of Golgi elements. J Cell Biol 97:1476–1490

    Google Scholar 

  • Wild G, Bennett G (1984) Influence of colchicine and vinblastine on the intracellular migration of secretory and membrane glycoproteins. II. Inhibition of secretion of thyroglobulin in rat thyroid follicular cells as visualized by light and electron microscope radioautography after 3-Hfucose injection. Am J Anat 170:531–543

    Google Scholar 

  • Wolff J, Bhattacharyya B (1975) Microtubules and thyroid hormone mobilization. Ann NY Acad Sci USA 253:763–770

    Google Scholar 

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Author notes

  1. Z. Ngoma

    Present address: Faculty of Medicine, University of Zaire, Kinshasa

Authors and Affiliations

  1. Department of Pathology, Medical Center, University of Sherbrooke, Sherbrooke, Quebec, Canada

    J. S. Hugon, P. Pothier & Z. Ngoma

  2. Department of Anatomy, Faculty of Medicine, McGill University, Montreal, Quebec, Canada

    G. Bennett

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  1. J. S. Hugon
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  2. G. Bennett
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  3. P. Pothier
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  4. Z. Ngoma
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Hugon, J.S., Bennett, G., Pothier, P. et al. Loss of microtubules and alteration of glycoprotein migration in organ cultures of mouse intestine exposed to nocodazole or colchicine. Cell Tissue Res. 248, 653–662 (1987). https://doi.org/10.1007/BF00216496

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