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Published on: November 20, 2011
Temporal dynamics of receptor-induced apoptosis in an affinity microdevice
Randall D Reif1, Charmaine Aguas, Michelle M Martinez
1Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX 79409, USA.
Analytical and Bioanalytical Chemistry
|March 3, 2010
Summary
This study reveals the precise timing of Fas-induced apoptosis in Jurkat cells using a microdevice. Real-time monitoring of caspase activity provides insights into the cell death pathway dynamics.
Area of Science:
- Cell Biology
- Molecular Biology
- Biophysics
Background:
- Apoptosis, or programmed cell death, is crucial for development and homeostasis.
- The extrinsic pathway of apoptosis is initiated by death receptors like Fas (CD95).
- Understanding the temporal dynamics of apoptosis is key to deciphering cellular signaling.
Purpose of the Study:
- To elucidate the temporal dynamics of Fas-induced apoptosis.
- To establish a microfluidic method for real-time monitoring of apoptosis.
- To investigate caspase activity timing during apoptosis.
Main Methods:
- Jurkat cells were immobilized on anti-CD95 coated microdevices.
- Continuous staining with L-bisaspartic acid rhodamine 110 (D(2)R) caspase probe.
- Real-time fluorescence microscopy monitored caspase activity for 6 hours.
Main Results:
- Normalized temporal dynamics of apoptosis induction per cell.
- Quantified individual cell caspase activity and cleavage rates.
- Demonstrated that caspase inhibitor z-VAD-FMK significantly slows apoptosis.
Conclusions:
- The microdevice approach enables precise, real-time measurement of apoptosis dynamics.
- Caspase activity timing and rates are consistent within cell populations.
- Further studies can utilize advanced probes and techniques to explore apoptosis heterogeneity.
