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Published on: June 3, 2010
Atomic force microscopy study of early morphological changes during apoptosis
Jessica A Hessler1, Andrew Budor, Krishna Putchakayala
1Department of Chemistry, and the Michigan Nanotechnology Institute for Medicine and Biological Sciences, University of Michigan, Ann Arbor, MI 48109-1055, USA.
Abstract:
Apoptosis is defined by a distinct set of morphological changes observed during cell death including loss of focal adhesions, the formation of cell membrane buds or blebs, and a decrease in total cell volume. Recent studies suggest that these dramatic morphological changes, particularly apoptotic volume decrease (AVD), are an early prerequisite to apoptosis and precede key biochemical time-points. Here we use atomic force microscopy to observe early stage AVD of KB cells undergoing staurosporine-induced apoptosis. After a 3-h exposure to 1 microM staurosporine, a 32% decrease in total cell height and a 50% loss of total cell volume is observed accompanied by only a 15% change in cell diameter. The observed AVD precedes key biochemical hallmarks of apoptosis such as loss of mitochondrial membrane potential, phosphatidyl serine translocation, nuclear fragmentation, and measurable caspase-3 activity. This suggests that morphological volume changes occur very early in the induction of apoptosis.
Insights
Apoptotic volume decrease (AVD), a key cell death morphology, occurs before biochemical markers in staurosporine-induced apoptosis. Atomic force microscopy revealed significant cell shrinkage preceding caspase activity and nuclear fragmentation.
Area of Science:
- Cell Biology
- Biophysics
Background:
- Apoptosis involves distinct morphological changes, including apoptotic volume decrease (AVD).
- Recent evidence suggests AVD precedes biochemical hallmarks of programmed cell death.
Purpose of the Study:
- To investigate early-stage AVD in KB cells during staurosporine-induced apoptosis using atomic force microscopy.
- To determine if morphological changes precede biochemical markers of apoptosis.
Main Methods:
- Utilized atomic force microscopy (AFM) to observe KB cells.
- Induced apoptosis using 1 microM staurosporine for 3 hours.
- Quantified changes in cell height, volume, and diameter.
Main Results:
- Observed a 32% decrease in cell height and a 50% loss in cell volume.
- Noted minimal change (15%) in cell diameter during AVD.
- AVD occurred before loss of mitochondrial membrane potential, phosphatidylserine translocation, nuclear fragmentation, and caspase-3 activation.
Conclusions:
- Morphological changes, specifically AVD, are early events in apoptosis.
- Atomic force microscopy is effective in observing early apoptotic morphological changes.
- Cell shrinkage is a critical early indicator of apoptosis induction.
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