Microtubules are involved in the secretion of proteins at the apical cell surface of the polarized epithelial cell, Madin-Darby canine kidney.
Microtubule-disrupting drugs (nocodazole, colchicine) and cytochalasin D, which inhibits the polymerization of the actin microfilaments, were used to study the role of the cytoskeleton in protein secretion in the polarized Madin-Darby canine kidney (MDCK) epithelial cells. Two proteins were analyzed. The gp 80 glycoprotein complex, which in untreated cells is sorted into the apical pathway and lysozyme, which is released randomly at both cell surfaces in transfected MDCK cells. Our results show that cytochalasin D has no influence on the transport of the gp 80 complex and lysozyme to either cell surface. However, in the presence of nocodazole or colchicine the secretion of both proteins at the apical cell surface is reduced by 50% with a concomitant increase in the basolateral release. These data suggest that microtubules are necessary for an efficient secretion of proteins at the apical cell surface of MDCK cells. In regard to the yet unresolved discrepancy concerning the involvement of microtubules in the transport of membrane proteins to the apical surface of MDCK cells, our results are consistent with the data of Rindler et al. (Rindler, M. J., Ivanov, I. E., and Sabatini, D. D. (1987) J. Cell. Biol. 104, 231-241) who observed a nonpolarized delivery of the influenza virus hemagglutinin in the presence of nocodazole or colchicine.
Institut für Biochemie, Abteilung Molekulare Genetik, Universität Frankfurt/M, Federal Republic of Germany.
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Parczyk
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Haase
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Kondor-Koch
D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology*; D002460:Cell Line; D003078:Colchicine / Q000494:pharmacology*; D015638:Cytochalasin D; D003572:Cytochalasins / Q000494:pharmacology; D004285:Dogs; D004848:Epithelium / Q000502:physiology; D006023:Glycoproteins / Q000378:metabolism; D007668:Kidney; D007700:Kinetics; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D009113:Muramidase / Q000378:metabolism; D015739:Nocodazole; D011506:Proteins / Q000378:metabolism*
D001562:Benzimidazoles; D003572:Cytochalasins; D006023:Glycoproteins; D011506:Proteins; D015638:Cytochalasin D; D009113:Muramidase; D015739:Nocodazole; D003078:Colchicine