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Evidence for cytoskeletal changes secondary to plasma membrane functional alterations in the in vitro cell response
Gianfranco Donelli1, Carla Fiorentini, Paola Matarrese
1Laboratory of Ultrastructures, Istituto Superiore di Sanità, Viale Regina Elena, 299-00161 Rome, Italy.
Abstract:
To investigate the mode of action of Clostridium perfringens epsilon-toxin, MDCK cells were treated with purified toxin and incubated at 37 degrees C for up to 24h. Exposure to epsilon-toxin caused a time-dependent decrease in cell-cell and cell-substrate interactions. After 30min of treatment retraction of the cell body and the emission of filopodia were detectable in a number of cells. Longer exposure resulted in cell rounding and cell blebbing which reached a maximum after 5h of toxin treatment. A parallel modification in the cytoskeleton was also detected. Actin marginalization and the entanglement of microtubules and intermediate filaments were observed by fluorescence microscopy after 30min of toxin exposure. Functional alterations of the plasma membrane of MDCK cells were assessed by flow cytometry. After 10 or 30min of intoxication an increase in cell volume was detected, indicating an alteration in plasma membrane permeability. These findings provide evidence for cytoskeletal changes and plasma membrane functional alterations in the in vitro cell response to C. perfringens epsilon-toxin.
Insights
Clostridium perfringens epsilon-toxin disrupts cell interactions and alters the cytoskeleton and plasma membrane in MDCK cells. This study details the toxin's in vitro cellular effects and mode of action.
Area of Science:
- Microbiology
- Cell Biology
- Toxicology
Background:
- Clostridium perfringens epsilon-toxin is a potent cytotoxin.
- Its precise mode of action on host cells requires further elucidation.
Purpose of the Study:
- To investigate the cellular mechanisms and in vitro effects of Clostridium perfringens epsilon-toxin on Madin-Darby canine kidney (MDCK) cells.
Main Methods:
- MDCK cells were treated with purified epsilon-toxin.
- Time-dependent effects on cell morphology, cell-cell/substrate interactions, cytoskeleton, and plasma membrane permeability were analyzed.
- Techniques included fluorescence microscopy and flow cytometry.
Main Results:
- Epsilon-toxin induced a time-dependent decrease in cell adhesion.
- Cytoskeletal alterations, including actin marginalization and filament entanglement, were observed.
- Increased cell volume indicated plasma membrane permeability changes.
Conclusions:
- Clostridium perfringens epsilon-toxin causes significant cytoskeletal rearrangements.
- The toxin functionally alters the plasma membrane, leading to increased permeability.
- These findings clarify the in vitro cellular response to epsilon-toxin.
