Document Detail

Clostridium difficile toxin A causes early damage to mitochondria in cultured cells.
MedLine Citation:
PMID:  10889163     Owner:  NLM     Status:  MEDLINE    
BACKGROUND & AIMS: The mechanism by which Clostridium difficile toxin A causes actin depolymerization and cell rounding involves toxin internalization and subsequent monoglucosylation of the Rho family of proteins. This study explored toxin internalization and effects on mitochondrial function before cell rounding. METHODS: Chinese hamster ovary (CHO) cells were exposed to toxin A, and mitochondrial localization was assayed by confocal microscopy. Mitochondrial function was measured by adenosine triphosphate (ATP) concentration, mitochondrial permeability, and leakage of cytochrome c. RESULTS: Confocal microscopy showed toxin A colocalization with the mitochondrial protein GRP 75 at 5 minutes after toxin exposure. Between 5 and 15 minutes, toxin A caused an 80% diminution in cellular ATP levels; cell rounding and Rho glucosylation commenced between 15 and 30 minutes. Toxin A also resulted in reduction of mitochondrial membrane potential and a 2-3-fold increase in reactive oxygen radicals. Preincubation of CHO cells with the antioxidants butylated hydroxyanisole or butylated hydroxytoluene blocked the toxin A-induced increase in oxygen radicals and diminished cell rounding. Western blot analysis of toxin A-exposed isolated mitochondria showed a direct effect of toxin A on leakage of cytochrome c. CONCLUSIONS: The results show that extensive mitochondrial damage occurs within 15 minutes in CHO cells exposed to toxin A. Diminished ATP concentrations and increased oxygen radicals are likely to contribute to cytotoxicity from this bacterial toxin.
D He; S J Hagen; C Pothoulakis; M Chen; N D Medina; M Warny; J T LaMont
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Gastroenterology     Volume:  119     ISSN:  0016-5085     ISO Abbreviation:  Gastroenterology     Publication Date:  2000 Jul 
Date Detail:
Created Date:  2000-08-03     Completed Date:  2000-08-03     Revised Date:  2005-11-17    
Medline Journal Info:
Nlm Unique ID:  0374630     Medline TA:  Gastroenterology     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  139-50     Citation Subset:  AIM; IM    
Division of Gastroenterology and Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.
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MeSH Terms
Adenosine Triphosphate / antagonists & inhibitors
Bacterial Toxins / metabolism,  pharmacology*
CHO Cells
Cell Membrane / metabolism
Cytochrome c Group / metabolism
Enterotoxins / metabolism,  pharmacology*
Membrane Potentials / drug effects
Microscopy, Electron
Mitochondria / drug effects*,  metabolism,  physiology,  ultrastructure
Reactive Oxygen Species / metabolism
Tissue Distribution
Reg. No./Substance:
0/Bacterial Toxins; 0/Cytochrome c Group; 0/Enterotoxins; 0/Reactive Oxygen Species; 0/tcdA protein, Clostridium difficile; 56-65-5/Adenosine Triphosphate

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