Measurement of caspase activity in individual cells reveals differences in the kinetics of caspase activation between cells.
Most forms of apoptosis involve activation of caspases and it is likely that differences between cells in their ability to activate caspases contributes to the responsiveness of any given cell within a population to apoptotic stimuli. To study the molecular mechanisms that underlie such differences, it is necessary to measure caspase activity in individual cells. Here, we describe a method that allows the continuous monitoring of caspase activity in individual, living mammalian cells. This approach allows studies of the kinetics of caspase activation to be performed in individual cells within a population. We demonstrate that in a group of cells where some cells die and some cells survive in response to the same stimulus, the cells that die can be differentiated from those that survive based on the amount of caspase activity in each cell several hours before death occurs.
Morgan
M J
MJ
Huntsman Cancer Institute, Department of Oncological Sciences, University of Utah, 2000 Circle of Hope, Salt Lake City, Utah 84112, USA.
Thorburn
A
A
D017209:Apoptosis / Q000187:drug effects; D001426:Bacterial Proteins / Q000235:genetics; D001681:Biological Assay / Q000379:methods*; D020169:Caspases / Q000032:analysis / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D004735:Energy Transfer; D004789:Enzyme Activation / Q000187:drug effects; D005434:Flow Cytometry; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008164:Luminescent Proteins / Q000235:genetics; D008353:Mannitol / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D009447:Neuroblastoma / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; C409504:blue fluorescent protein, Aequorea victoria; C054562:yellow fluorescent protein, Bacteria; D008353:Mannitol; D020169:Caspases