PMID- 3858089
OWN - NLM
STAT- MEDLINE
DCOM- 19850725
LR  - 20131121
IS  - 0196-4763 (Print)
IS  - 0196-4763 (Linking)
VI  - 6
IP  - 3
DP  - 1985 May
TI  - Denaturation and condensation of DNA in situ induced by acridine orange in 
      relation to chromatin changes during growth and differentiation of Friend 
      erythroleukemia cells.
PG  - 195-207
AB  - DNA in situ is progressively denatured when the cells or nuclei are treated with 
      increasing concentration of acridine orange (AO). This transition can be 
      monitored by flow cytometry as a decrease in green fluorescence. The complexes of 
      denatured DNA and AO undergo immediate condensation and aggregation; this step is 
      manifested by appearance of red luminescence and formation of precipitates that 
      can be detected by electron microscopy. The precipitates form preferentially in 
      heterochromatin as well as in ribosomes and polysomes. Their formation and 
      further aggregation affects cellular light scatter properties in both the forward 
      and right-angle direction. The AO-induced DNA denaturation and condensation was 
      studied in nuclei of Friend erythroleukemia cells from exponentially growing, 
      differentiated or quiescent cells. The DNA in nuclei of quiescent cells, from 
      plateau-phase cultures, was the most sensitive to denaturation; it denatured 
      (measured by changes in luminescence) at an AO concentration between 50 and 80 
      microM with the midpoint of the transition (Cd) at 70 microM. DNA in nuclei of 
      differentiated cells (dimethyl-sulfoxide-induced erythroid differentiation) was 
      more resistant (Cd = 77-83 microM), whereas DNA in exponentially growing cells 
      was the most resistant (Cd = 86 microM). Extraction of proteins with 0.1 M HCl at 
      0 degree C abolished the differences between the cells and shifted the transition 
      to a lower AO concentration (Cd = 46 microM). For comparison, the midpoint 
      transitions representing condensation of free, nucleic acids measured as light 
      scatter changes occurred at 13, 22, 31 and 53 microM of AO, for rRNA, tRNA, and 
      denatured and native-calf thymus DNA, respectively. Denaturation and condensation 
      of DNA, which can be induced by AO either in isolated nuclei or viable 
      permeabilized or fixed cells provides a new approach to discriminate cell 
      subpopulations with different chromatin structure by flow cytometry. The 
      molecular mechanisms of this phenomenon are discussed.
FAU - Darzynkiewicz, Z
AU  - Darzynkiewicz Z
FAU - Traganos, F
AU  - Traganos F
FAU - Kapuscinski, J
AU  - Kapuscinski J
FAU - Melamed, M R
AU  - Melamed MR
LA  - eng
GR  - CA 28704/CA/NCI NIH HHS/United States
GR  - CA23296/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Cytometry
JT  - Cytometry
JID - 8102328
RN  - 0 (Chromatin)
RN  - 0 (DNA, Neoplasm)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange/*pharmacology
MH  - Animals
MH  - Cell Differentiation
MH  - Cell Division
MH  - Cell Line
MH  - Chromatin/ultrastructure
MH  - DNA, Neoplasm/*metabolism
MH  - Friend murine leukemia virus
MH  - Leukemia, Erythroblastic, Acute/*metabolism/ultrastructure
MH  - Light
MH  - Mice
MH  - Microscopy, Electron
MH  - Nucleic Acid Conformation/drug effects
MH  - Nucleic Acid Denaturation/*drug effects
MH  - Scattering, Radiation
EDAT- 1985/05/01 00:00
MHDA- 1985/05/01 00:01
CRDT- 1985/05/01 00:00
PHST- 1985/05/01 00:00 [pubmed]
PHST- 1985/05/01 00:01 [medline]
PHST- 1985/05/01 00:00 [entrez]
AID - 10.1002/cyto.990060305 [doi]
PST - ppublish
SO  - Cytometry. 1985 May;6(3):195-207. doi: 10.1002/cyto.990060305.