PMID- 2015848
OWN - NLM
STAT- MEDLINE
DCOM- 19910517
LR  - 20190707
IS  - 0014-4827 (Print)
IS  - 0014-4827 (Linking)
VI  - 194
IP  - 1
DP  - 1991 May
TI  - Impossibility of acridine orange intercalation in nuclear DNA of the living cell.
PG  - 147-53
AB  - The ability of a "vital" dye, acridine orange (AO), to intercalate into the DNA 
      of living cells was investigated by quantitative intensified fluorescence 
      microscopy and digital imaging under various conditions of dye concentration, 
      excitation light intensity, and ionic concentration. Our results demonstrate that 
      the bulk of chromatin DNA is packed in a way that does not allow AO 
      intercalation. At low dye concentrations and very low levels of light intensity, 
      the only fluorescent structures observed inside the nucleus are the nucleoli. 
      This nonpermissive state of the chromatin appears to be a characteristic feature 
      of the nucleus in living cells. AO intercalation into DNA can be mediated by 
      raising the nuclear Na+ concentration. This was achieved here by using a cation 
      carrier, monensin, a procedure which permits the avoidance of cell 
      permeabilization. Furthermore, we show that the discharge of lysosomal enzymes in 
      the living cell, via a targeted photodynamic reaction which occurs at high levels 
      of light intensity, can also release the constraints which impede dye 
      intercalation into nuclear DNA. In conclusion, studies carried out under 
      conditions where intercalative dyes such as AO are able to stain the nuclear DNA 
      have to be interpreted with caution and do not provide direct evidence on the 
      structural state of native chromatin. The molecular origin of the absence of AO 
      intercalation in chromatin of the living cell is discussed with regard to the 
      restrained uncoiling of the double helix which is required for dye intercalation.
FAU - Delic, J
AU  - Delic J
AD  - Institut National de la Sante et de la Recherche Medicale, U219, Biophysique 
      Moleculaire, Institut Curie, Orsay, France.
FAU - Coppey, J
AU  - Coppey J
FAU - Magdelenat, H
AU  - Magdelenat H
FAU - Coppey-Moisan, M
AU  - Coppey-Moisan M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Exp Cell Res
JT  - Experimental cell research
JID - 0373226
RN  - 0 (Chromatin)
RN  - 0 (Intercalating Agents)
RN  - 9007-49-2 (DNA)
RN  - 906O0YJ6ZP (Monensin)
RN  - 9NEZ333N27 (Sodium)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange/*metabolism
MH  - Biological Transport
MH  - Cell Nucleus/chemistry/*metabolism/ultrastructure
MH  - Cells, Cultured
MH  - Chromatin/chemistry/metabolism/ultrastructure
MH  - Cytoplasm/metabolism/physiology/ultrastructure
MH  - DNA/analysis/*metabolism
MH  - Dose-Response Relationship, Drug
MH  - Fibroblasts/*cytology/metabolism/ultrastructure
MH  - Humans
MH  - Intercalating Agents/*metabolism
MH  - Light
MH  - Microscopy, Fluorescence
MH  - Monensin
MH  - Organelles/metabolism/physiology/ultrastructure
MH  - Sodium/pharmacokinetics
EDAT- 1991/05/01 00:00
MHDA- 1991/05/01 00:01
CRDT- 1991/05/01 00:00
PHST- 1991/05/01 00:00 [pubmed]
PHST- 1991/05/01 00:01 [medline]
PHST- 1991/05/01 00:00 [entrez]
AID - 0014-4827(91)90144-J [pii]
AID - 10.1016/0014-4827(91)90144-j [doi]
PST - ppublish
SO  - Exp Cell Res. 1991 May;194(1):147-53. doi: 10.1016/0014-4827(91)90144-j.