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Volume 159, Issue Pt_2

Disruption of MDCK cell tight junctions by the free-living amoeba Free

Abstract

is the aetiological agent of primary amoebic meningoencephalitis. This parasite invades its host by penetrating the olfactory mucosa. However, the mechanism of epithelium penetration is not well understood. In the present study, we evaluated the effect of trophozoites and the non-pathogenic on Madin–Darby canine kidney (MDCK) tight junction proteins, including claudin-1, occludin and ZO-1, as well as on the actin cytoskeleton. Trophozoites from each of the free-living amoeba species were co-cultured with MDCK cells in a 1 : 1 ratio for 1, 3, 6 or 10 h. Light microscopy revealed that caused morphological changes as early as 3 h post-infection in an epithelial MDCK monolayer. Confocal microscopy analysis revealed that after 10 h of co-culture, trophozoites induced epithelial cell damage, which was characterized by changes in the actin apical ring and disruption of the ZO-1 and claudin-1 proteins but not occludin. Western blot assays revealed gradual degradation of ZO-1 and claudin-1 as early as 3 h post-infection. Likewise, there was a drop in transepithelial electrical resistance that resulted in increased epithelial permeability and facilitated the invasion of trophozoites by a paracellular route. In contrast, did not induce alterations in MDCK cells even at 10 h post-infection. Based on these results, we suggest that trophozoites disrupt epithelial monolayers, which could enable their penetration of the olfactory epithelium and subsequent invasion of the central nervous system.

  • Received:
  • Accepted:
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2013-02-01
2024-04-18
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Disruption of MDCK cell tight junctions by the free-living amoeba Naegleria fowleri
Microbiology 159, 392 (2013); https://doi.org/10.1099/mic.0.063255-0
/content/journal/micro/10.1099/mic.0.063255-0
/content/journal/micro/10.1099/mic.0.063255-0
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