PMID- 10366440
OWN - NLM
STAT- MEDLINE
DCOM- 19990702
LR  - 20161122
IS  - 0014-4827 (Print)
IS  - 0014-4827 (Linking)
VI  - 249
IP  - 2
DP  - 1999 Jun 15
TI  - The Schrodinger's cat quandary in cell biology: integration of live cell 
      functional assays with measurements of fixed cells in analysis of apoptosis.
PG  - 404-12
AB  - The existing cytometric methodologies do not allow one to directly correlate, 
      within the same cells, functional cell attributes that are revealed supravitally 
      with features that require cell fixation to be detected or measured. Taking 
      advantage of the "file merge" feature of the laser-scanning cytometer, we have 
      been able to correlate the supravital changes that occur during apoptosis, namely 
      the drop in mitochondrial transmembrane potential (Delta Psim) and generation of 
      the reactive oxygen intermediates (ROIs), with features revealed by analysis of 
      fixed cells: the cell cycle position and DNA fragmentation. The cell cycle 
      position was established based on the cell's stainability with propidium iodide 
      while DNA fragmentation was assessed by in situ DNA strand break labeling using 
      exogenous terminal deoxynucleotidyltransferase. During apoptosis of HL-60 cells 
      induced by the DNA topoisomerase I inhibitor camptothecin (CPT), the dissipation 
      of Delta Psim occurred preferentially in S-phase cells and preceded the 
      appearance of DNA strand breaks. Essentially all cells with DNA strand breaks had 
      dissipated Delta Psim. Compared to the decrease of Delta Psim, the CPT-induced 
      rise in ROIs during apoptosis was less restricted to S-phase cells. Furthermore, 
      no elevation of ROIs was detected in a significant proportion of cells with DNA 
      strand breaks. The data suggest that DNA fragmentation may occur in some cells 
      prior to the increase in ROIs and thus, unlike the dissipation of Delta Psim, the 
      oxidative stress may not be a prerequisite for activation of an endonuclease. 
      Alternatively, the oxidative stress may be a transient event, occupying a narrow 
      "time window" during the apoptotic process. The approach opens a possibility to 
      study direct relationships, within the same cells, between cellular changes 
      (e.g., occurring during apoptosis, mitogenesis, differentiation, etc.) detected 
      by functional assays of live cells and changes that cannot be analyzed 
      supravitally.
CI  - Copyright 1999 Academic Press.
FAU - Li, X
AU  - Li X
AD  - The Brander Cancer Research Institute, New York Medical College, Valhalla, New 
      York, 10595, USA.
FAU - Darzynkiewicz, Z
AU  - Darzynkiewicz Z
LA  - eng
GR  - R01 CA028704/CA/NCI NIH HHS/United States
GR  - R01 CA028704-20/CA/NCI NIH HHS/United States
GR  - CA 28704/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Exp Cell Res
JT  - Experimental cell research
JID - 0373226
RN  - XT3Z54Z28A (Camptothecin)
SB  - IM
MH  - Apoptosis/drug effects/*physiology
MH  - Biological Assay/*methods
MH  - Camptothecin/pharmacology
MH  - HL-60 Cells
MH  - Humans
MH  - Image Cytometry/methods
MH  - Tissue Fixation
EDAT- 1999/06/15 00:00
MHDA- 1999/06/15 00:01
CRDT- 1999/06/15 00:00
PHST- 1999/06/15 00:00 [pubmed]
PHST- 1999/06/15 00:01 [medline]
PHST- 1999/06/15 00:00 [entrez]
AID - S0014-4827(99)94525-1 [pii]
AID - 10.1006/excr.1999.4525 [doi]
PST - ppublish
SO  - Exp Cell Res. 1999 Jun 15;249(2):404-12. doi: 10.1006/excr.1999.4525.