Related Experiment Video
Updated: Jul 29, 2026
06:26
Monitoring Dynamic Changes In Mitochondrial Calcium Levels During Apoptosis Using A Genetically Encoded Calcium Sensor
Published on: April 1, 2011
24.0K
Monitoring Calcium Fluxes and Lysosome Exocytosis During Pyroptosis
Wendy P Loomis1, Tessa Bergsbaken2
1Department of Laboratory Medicine and Pathology, University of Washington, Seattle, WA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|April 19, 2023
Summary
Inflammasome activation triggers pyroptosis, a proinflammatory cell death. Methods are presented to measure gasdermin D pore formation, calcium influx, and lysosome exocytosis following inflammatory caspase activation.
Area of Science:
- Cellular biology
- Immunology
- Molecular mechanisms of cell death
Background:
- Inflammasomes activate inflammatory caspases, initiating pyroptosis, a programmed inflammatory cell death.
- Gasdermin D cleavage by caspases forms pores, releasing IL-1β and IL-18 and causing membrane disruption.
- These pores also facilitate calcium influx, leading to lysosome exocytosis.
Purpose of the Study:
- To outline methods for assessing cellular events downstream of inflammatory caspase activation.
- To provide techniques for measuring calcium flux, lysosome exocytosis, and membrane integrity.
Main Methods:
- Measurement of calcium influx using fluorescent indicators.
- Assessment of lysosome exocytosis via marker release or imaging.
- Quantification of membrane disruption and pore formation.
Main Results:
- Established protocols for quantifying key pyroptosis events.
- Demonstrated links between caspase activation, gasdermin D pore formation, and downstream cellular responses.
- Provided a framework for studying inflammasome-mediated cell death.
Conclusions:
- The presented methods enable detailed analysis of pyroptosis.
- Understanding these mechanisms is crucial for studying inflammatory diseases.
- This work facilitates research into inflammasome signaling pathways.
