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Asynchrony and commitment to die during apoptosis.

Authors
  • Messam, C A
  • Pittman, R N
Type
Published Article
Journal
Experimental cell research
Publication Date
Feb 01, 1998
Volume
238
Issue
2
Pages
389–398
Identifiers
PMID: 9473347
Source
Medline
License
Unknown

Abstract

Time lapse video microscopy is used to study the chronology of morphological changes and commitment to die in individual PC12 cells after induction of apoptosis. Cell death is highly asynchronous occurring over a 2- to 3-day period following serum removal; however, all cells go through three characteristic morphological phases irrespective of the time they die following serum removal. During phase 1, which lasts from 2 to 44 h, cells maintain normal morphology. Phase 2 is characterized by plasma membrane bubbling which lasts from 10 min to 40 h. Phase 3 represents the active or execution phase of apoptosis and involves dynamic whole cell body blebbing. Phase 3/execution phase has a restricted duration, lasting 96 +/- 5 min. At the end of the execution phase of apoptosis, cells die. The inherently asynchronous nature of cell death is still present in cells that are synchronized following mitosis. Daughter cells enter phase 2 synchronously but remain in phase 2 for varying periods and die at different times. Addition of serum 24-48 h after initiating apoptosis blocks death of 89% of cells in phase 1, 79% in phase 2, and 0% in phase 3. Serum rescue experiments are consistent with cells committing to die about 2-3 h prior to the onset of phase 3 (execution phase of apoptosis). These studies indicate that although apoptosis is an asynchronous process it can be defined in terms of reproducible morphological changes that can be used to place other events, such as the commitment to die, in a temporal sequence.

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