Related Experiment Videos
An actin barrier to resealing
K Miyake1, P L McNeil, K Suzuki
1Second Department of Anatomy, School of Medicine, Fukushima Medical University, Fukushima 960-1295, Japan. kmiyake@mail.mcg.edu
Journal of Cell Science
|October 30, 2001
Summary
Cell membrane repair requires calcium and involves actin disassembly. Disassembly of filamentous actin in the cell cortex is crucial for plasma membrane resealing after injury, facilitating necessary fusion events.
Area of Science:
- Cell Biology
- Biophysics
Background:
- Plasma membrane disruption is a common cellular injury.
- Cell survival relies on rapid resealing, dependent on extracellular calcium (Ca2+).
- Vesicle fusion in the cell cortex is key to resealing, but requires overcoming physical barriers.
Purpose of the Study:
- To investigate the role of filamentous actin in the plasma membrane resealing process.
- To determine if actin disassembly is a prerequisite for membrane repair.
Main Methods:
- Used rat gastric epithelial (RGM1) cells.
- Assessed cortical actin using phalloidin staining and flow cytometry.
- Manipulated actin dynamics with DNase1, cytochalasin, phalloidin, and jasplakinolide.
- Evaluated resealing efficiency in response to these treatments.
Main Results:
- Cortical actin staining decreased at disruption sites and in wounded cells.
- Extracellular calcium (Ca2+) dependent reduction in whole cell actin staining was observed.
- Depolymerizing actin with DNase1 enhanced resealing.
- Stabilizing actin inhibited resealing, but this inhibition was reversible.
Conclusions:
- Localized disassembly of filamentous actin removes a barrier to membrane repair.
- Actin disassembly facilitates the membrane-contact events necessary for resealing.
- This process is essential for plasma membrane integrity in mammalian cells.