Microtubules and Actin Filaments Are Not Critically Involved in the Biogenesis of Epithelial Cell Surface Polarity

• Published in: The Journal of cell biology Vol. 102; no. 5; pp. 1853 - 1867
• Main Authors: Pedro J. I. Salas, Misek, David E., Vega-Salas, Dora E., Gundersen, Doris, Cereijido, Marcelino, Rodriguez-Boulan, Enrique
• Format: Journal Article
• Language: English
• Published: New York, NY Rockefeller University Press 01-05-1986; The Rockefeller University Press
• Subjects: Actin Cytoskeleton - physiology; Actins - physiology; Alkaloids - pharmacology; Animals; Benzimidazoles - pharmacology; Biological and medical sciences; Budding; Cell coat. Cell surface; Cell Compartmentation - drug effects; Cell Line; Cell lines; Cell Membrane - metabolism; Cell membranes; Cell structures and functions; Cells; Colchicine - pharmacology; Cytochalasin D; Cytochalasins; Cytochalasins - pharmacology; Cytoskeleton - physiology; Dogs; Epithelial cells; Epithelium - ultrastructure; Exocytosis; Fundamental and applied biological sciences. Psychology; Hemagglutinins, Viral - metabolism; Kidney - cytology; Kidney cells; Membrane Glycoproteins; Membrane Proteins - metabolism; Microscopy, Electron; Microtubules; Microtubules - physiology; Molecular and cellular biology; Nocodazole; Orthomyxoviridae; Paclitaxel; Tubulin - physiology; Vesicular stomatitis Indiana virus; Viral Envelope Proteins; Viral Proteins - metabolism; Virus Replication; Viruses; Vertebrata; Cell line; Mammalia; Transmission electron microscopy; Investigation method; Epithelial cell; Cytoskeleton; Polarity; Microtubule; Cell surface
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.102.5.1853

We have studied the role of microtubules and actin filaments in the biogenesis of epithelial cell surface polarity, using influenza hemagglutinin and vesicular stomatitis G protein as model apical and basolateral proteins in infected Madin-Darby canine kidney cells. Addition of colchicine or nocodazole to confluent monolayers at concentrations sufficient to completely disassemble microtubules did not affect the asymmetric budding of influenza or vesicular stomatitis virus and only slightly reduced the typical asymmetric surface distribution of their envelope proteins, despite extensive cytoplasmic redistribution of the Golgi apparatus. Alteration of microtubular function by taxol or dissociation of actin filaments by cytochalasin D also failed to have a significant effect. Furthermore, neither colchicine nor cytochalasin D pretreatment blocked the ability of subconfluent Madin-Darby canine kidney cells to sustain polarized budding of influenza virus a few hours after attachment to the substrate. Our results indicate that domain-specific microtubule or actin filament "tracks" are not responsible for the vectorial delivery of apically or basolaterally directed transport vesicles. In conjunction with currently available evidence, they are compatible with a model in which receptors in the cytoplasmic aspect of apical or basolateral regions provide vectoriality to the transport of vesicles carrying plasma membrane proteins to their final surface localization.