Pore-forming epsilon toxin causes membrane permeabilization and rapid ATP depletion-mediated cell death in renal collecting duct cells

C. Chassin, M. Bens, J. De Barry, Raphael Jean Courjaret, J. L. Bossu, F. Cluzeaud, S. Ben Mkaddem, M. Gibert, B. Poulain, M. R. Popoff, A. Vandewalle

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Clostridium perfringens epsilon toxin (ET) is a potent pore-forming cytotoxin causing fatal enterotoxemia in livestock. ET accumulates in brain and kidney, particularly in the renal distal-collecting ducts. ET binds and oligomerizes in detergent-resistant membranes (DRMs) microdomains and causes cell death. However, the causal linkage between membrane permeabilization and cell death is not clear. Here, we show that ET binds and forms 220-kDa insoluble complexes in plasma membrane DRMs of renal mpkCCDcl4 collecting duct cells. Phosphatidylinositol-specific phospholipase C did not impair binding or the formation of ET complexes, suggesting that the receptor for ET is not GPI anchored. ET induced a dose-dependent fall in the transepithelial resistance and potential in confluent cells grown on filters, transiently stimulated Na + absorption, and induced an inward ionic current and a sustained rise in [Ca2+]i. ET also induced rapid depletion of cellular ATP, and stimulated the AMP-activated protein kinase, a metabolic-sensing Ser/Thr kinase. ET also induced mitochondrial membrane permeabilization and mitochondrial-nuclear translocation of apoptosis-inducing factor, a potent caspase-independent cell death effector. Finally, ET induced cell necrosis characterized by a marked reduction in nucleus size without DNA fragmentation. DRM disruption by methyl-β-cyclodextrin impaired ET oligomerization, and significantly reduced the influx of Na+ and [Ca2+]i, but did not impair ATP depletion and cell death caused by the toxin. These findings indicate that ET causes rapid necrosis of renal collecting duct cells and establish that ATP depletion-mediated cell death is not strictly correlated with the plasma membrane permeabilization and ion diffusion caused by the toxin.

Original languageEnglish
JournalAmerican Journal of Physiology - Renal Physiology
Volume293
Issue number3
DOIs
Publication statusPublished - 1 Sep 2007
Externally publishedYes

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Cell Death
Adenosine Triphosphate
Kidney
Membranes
Detergents
Necrosis
Enterotoxemia
Cell Membrane
Apoptosis Inducing Factor
Membrane Microdomains
Phosphoinositide Phospholipase C
AMP-Activated Protein Kinases
Cytotoxins
Cyclodextrins
Mitochondrial Membranes
Livestock
DNA Fragmentation
Caspases
Cause of Death
Phosphotransferases

Keywords

  • Ion diffusion
  • Lipid raft
  • Oncosis

ASJC Scopus subject areas

  • Physiology
  • Urology

Cite this

Pore-forming epsilon toxin causes membrane permeabilization and rapid ATP depletion-mediated cell death in renal collecting duct cells. / Chassin, C.; Bens, M.; De Barry, J.; Courjaret, Raphael Jean; Bossu, J. L.; Cluzeaud, F.; Ben Mkaddem, S.; Gibert, M.; Poulain, B.; Popoff, M. R.; Vandewalle, A.

In: American Journal of Physiology - Renal Physiology, Vol. 293, No. 3, 01.09.2007.

Research output: Contribution to journalArticle

Chassin, C. ; Bens, M. ; De Barry, J. ; Courjaret, Raphael Jean ; Bossu, J. L. ; Cluzeaud, F. ; Ben Mkaddem, S. ; Gibert, M. ; Poulain, B. ; Popoff, M. R. ; Vandewalle, A. / Pore-forming epsilon toxin causes membrane permeabilization and rapid ATP depletion-mediated cell death in renal collecting duct cells. In: American Journal of Physiology - Renal Physiology. 2007 ; Vol. 293, No. 3.
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AU - Bens, M.

AU - De Barry, J.

AU - Courjaret, Raphael Jean

AU - Bossu, J. L.

AU - Cluzeaud, F.

AU - Ben Mkaddem, S.

AU - Gibert, M.

AU - Poulain, B.

AU - Popoff, M. R.

AU - Vandewalle, A.

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KW - Oncosis

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