PMID- 31763788
OWN - NLM
STAT- MEDLINE
DCOM- 20200729
LR  - 20200729
IS  - 1934-9300 (Electronic)
IS  - 1934-9297 (Linking)
VI  - 91
IP  - 1
DP  - 2019 Dec
TI  - Assessment of DNA Susceptibility to Denaturation as a Marker of Chromatin 
      Structure.
PG  - e65
LID - 10.1002/cpcy.65 [doi]
AB  - The susceptibility of DNA in situ to denaturation is modulated by its 
      interactions with histone and nonhistone proteins, as well as with other 
      chromatin components related to the maintenance of the 3D nuclear structure. 
      Measurement of DNA proclivity to denature by cytometry provides insight into 
      chromatin structure and thus can be used to recognize cells in different phases 
      of the cell cycle, including mitosis, quiescence (G(0) ), and apoptosis, as well 
      as to identify the effects of drugs that modify chromatin structure. Particularly 
      useful is the method's ability to detect chromatin changes in sperm cells related 
      to DNA fragmentation and infertility. This article presents a flow cytometric 
      procedure for assessing DNA denaturation based on application of the 
      metachromatic property of acridine orange (AO) to differentially stain single- 
      versus double-stranded DNA. This approach circumvents limitations of biochemical 
      methods of examining DNA denaturation, in particular the fact that the latter 
      destroy higher orders of chromatin structure and that, being applied to bulk cell 
      populations, they cannot detect heterogeneity of individual cells. Because the 
      metachromatic properties of AO have also found application in other cytometric 
      procedures, such as differential staining of RNA versus DNA and assessment of 
      lysosomal proton pump including autophagy, to avert confusion between these 
      approaches and the use of this dye in the DNA denaturation assay, these AO 
      applications are briefly outlined in this unit as well. (c) 2019 by John Wiley & 
      Sons, Inc. Basic Protocol: Differential staining of single- versus 
      double-stranded DNA with acridine orange.
CI  - (c) 2019 John Wiley & Sons, Inc.
FAU - Darzynkiewicz, Zbigniew
AU  - Darzynkiewicz Z
AD  - Department of Pathology, New York Medical College, Valhalla, New York.
FAU - Halicka, Dorota H
AU  - Halicka DH
AD  - Brander Cancer Research Institute, New York Medical College, Valhalla, New York.
FAU - Zhao, Hong
AU  - Zhao H
AD  - Brander Cancer Research Institute, New York Medical College, Valhalla, New York.
FAU - Li, Jiangwei
AU  - Li J
AD  - Brander Cancer Research Institute, New York Medical College, Valhalla, New York.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Curr Protoc Cytom
JT  - Current protocols in cytometry
JID - 100899351
RN  - 0 (Chromatin)
RN  - 0 (Chromosomal Proteins, Non-Histone)
RN  - 0 (DNA, Single-Stranded)
RN  - 0 (Genetic Markers)
RN  - 9007-49-2 (DNA)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange/chemistry/pharmacology
MH  - Cells, Cultured
MH  - Chromatin/*chemistry/metabolism
MH  - Chromatin Assembly and Disassembly/genetics
MH  - Chromosomal Proteins, Non-Histone/metabolism
MH  - DNA/analysis/chemistry/drug effects
MH  - DNA, Single-Stranded/chemistry/drug effects
MH  - Flow Cytometry/methods
MH  - *Genetic Markers
MH  - *Genetic Techniques
MH  - Genomic Instability/*genetics
MH  - Humans
MH  - Nucleic Acid Conformation
MH  - *Nucleic Acid Denaturation
MH  - Protein Binding
OTO - NOTNLM
OT  - acridine orange
OT  - apoptosis
OT  - autophagy
OT  - cell cycle
OT  - lysosomes
OT  - mitosis
OT  - sperm chromatin fertility assay
EDAT- 2019/11/26 06:00
MHDA- 2020/07/30 06:00
CRDT- 2019/11/26 06:00
PHST- 2019/11/26 06:00 [entrez]
PHST- 2019/11/26 06:00 [pubmed]
PHST- 2020/07/30 06:00 [medline]
AID - 10.1002/cpcy.65 [doi]
PST - ppublish
SO  - Curr Protoc Cytom. 2019 Dec;91(1):e65. doi: 10.1002/cpcy.65.