Pasteurella multocida enters polarized epithelial cells by interacting with host F-actin

Mireille J. Rpabier, Nancy K. Tyler, Naomi J. Walker, Lori M. Hansen, Dwight C. Hirsh, Fern Tablin

Research output: Contribution to journalArticle

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Abstract

We investigated the interaction of an avian strain of Pasteurella multocida with the cytoskeleton of MDCK cells, which formed a polarized epithelium when grown on type 1 collagen coated filters. Bacteria were incubated with MDCK cells for 30 min, 2, 4 and 6 hours and their location and association with the cell cytoskeleton determined by double-label immunofluorescence confocal microscopy. Cells were stained with a polyclonal antiserum to the outer-membrane proteins of P. multocida and with rhodamine phalloidin which specifically binds filamentous (F) actin. Confocal microscopy revealed that bacteria entered the cells by 30 min, and that by 6 hours there was a marked alteration in the actin cytoskeleton in which long filaments were reorganized to discrete foci of short actin filaments, within which were one or more bacteria. Electron microscopy demonstrated that by 2 hours, each bacterium was associated with many short 5-6 nm filaments. Treatment of MDCK cells with cytochalasin D for either 30 minutes or 24 hours prior to infection disrupted the actin cytoskeleton and inhibited entry of P. multocida.

Original languageEnglish (US)
Pages (from-to)343-355
Number of pages13
JournalVeterinary Microbiology
Volume54
Issue number3-4
DOIs
StatePublished - 1997

Fingerprint

Pasteurella multocida
Madin Darby Canine Kidney Cells
actin
Actins
epithelial cells
Epithelial Cells
Actin Cytoskeleton
Bacteria
microfilaments
Cytoskeleton
Confocal Microscopy
bacteria
cells
cytoskeleton
Cytochalasin D
Collagen Type I
Fluorescence Microscopy
cytochalasin D
Immune Sera
Electron Microscopy

Keywords

  • actin
  • confocal microscopy
  • cytochalasin D
  • cytoskeleton
  • Pasteurella multocida

ASJC Scopus subject areas

  • Animal Science and Zoology
  • Microbiology
  • veterinary(all)

Cite this

Pasteurella multocida enters polarized epithelial cells by interacting with host F-actin. / Rpabier, Mireille J.; Tyler, Nancy K.; Walker, Naomi J.; Hansen, Lori M.; Hirsh, Dwight C.; Tablin, Fern.

In: Veterinary Microbiology, Vol. 54, No. 3-4, 1997, p. 343-355.

Research output: Contribution to journalArticle

Rpabier, Mireille J. ; Tyler, Nancy K. ; Walker, Naomi J. ; Hansen, Lori M. ; Hirsh, Dwight C. ; Tablin, Fern. / Pasteurella multocida enters polarized epithelial cells by interacting with host F-actin. In: Veterinary Microbiology. 1997 ; Vol. 54, No. 3-4. pp. 343-355.
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