PMID- 26530674
OWN - NLM
STAT- MEDLINE
DCOM- 20160906
LR  - 20151104
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 1351
DP  - 2016
TI  - A Single-Cell Resolution Imaging Protocol of Mitochondrial DNA Dynamics in 
      Physiopathology, mTRIP, Which Also Evaluates Sublethal Cytotoxicity.
PG  - 49-65
LID - 10.1007/978-1-4939-3040-1_5 [doi]
AB  - Mitochondria autonomously replicate and transcribe their own genome, which is 
      present in multiple copies in the organelle. Transcription and replication of the 
      mitochondrial DNA (mtDNA), which are defined here as mtDNA processing, are 
      essential for mitochondrial function. The extent, efficiency, and coordination of 
      mtDNA processing are key parameters of the mitochondrial state in living cells. 
      Recently, single-cell analysis of mtDNA processing revealed a large and dynamic 
      heterogeneity of mitochondrial populations in single cells, which is linked to 
      mitochondrial function and is altered during disease. This was achieved using 
      mitochondrial Transcription and Replication Imaging Protocol (mTRIP), a modified 
      fluorescence in situ hybridization (FISH) approach that simultaneously reveals 
      the mitochondrial RNA content and mtDNA engaged in initiation of replication at 
      the single-cell level. mTRIP can also be coupled to immunofluorescence or 
      MitoTracker, resulting in the additional labeling of proteins or active 
      mitochondria, respectively. Therefore, mTRIP detects quantitative and qualitative 
      alterations of the dynamics of mtDNA processing in human cells that respond to 
      physiological changes or result from diseases. In addition, we show here that 
      mTRIP is a rather sensitive tool for detecting mitochondrial alterations that may 
      lead to loss of cell viability, and is thereby a useful tool for monitoring 
      sublethal cytotoxicity for instance during chronic drug treatment.
FAU - Chatre, Laurent
AU  - Chatre L
AD  - Team "Stability of Nuclear and Mitochondrial DNA" CNRS UMR 3525, Paris, France.
AD  - Stem Cells and Development, Department of Developmental & Stem Cell Biology, 
      Institut Pasteur, 25-28 Rue du Dr. Roux, Paris, France.
FAU - Montagne, Benjamin
AU  - Montagne B
AD  - Team "Stability of Nuclear and Mitochondrial DNA" CNRS UMR 3525, Paris, France.
AD  - Stem Cells and Development, Department of Developmental & Stem Cell Biology, 
      Institut Pasteur, 25-28 Rue du Dr. Roux, Paris, France.
FAU - Ricchetti, Miria
AU  - Ricchetti M
AD  - Team "Stability of Nuclear and Mitochondrial DNA" CNRS UMR 3525, Paris, France. 
      mricch@pasteur.fr.
AD  - Stem Cells and Development, Department of Developmental & Stem Cell Biology, 
      Institut Pasteur, 25-28 Rue du Dr. Roux, Paris, France. mricch@pasteur.fr.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Aldehydes)
RN  - 0 (DNA Probes)
RN  - 0 (DNA, Mitochondrial)
RN  - 0 (Membrane Proteins)
RN  - 0 (MitoTracker Red 580)
RN  - 0 (Organic Chemicals)
RN  - 0 (mitotracker green FM)
SB  - IM
MH  - Aldehydes/chemistry
MH  - DNA Probes/genetics
MH  - DNA, Mitochondrial/analysis/*genetics
MH  - Fluorescent Antibody Technique/*methods
MH  - Genome, Mitochondrial/genetics
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Membrane Proteins/*chemistry
MH  - Mitochondria/genetics/metabolism
MH  - Mitochondrial Diseases/genetics
MH  - Mitochondrial Dynamics/*genetics
MH  - Organic Chemicals/chemistry
MH  - Polymerase Chain Reaction/*methods
MH  - Staining and Labeling/methods
OTO - NOTNLM
OT  - Cytotoxicity
OT  - DNA replication
OT  - FISH
OT  - Imaging
OT  - Long-term drug treatment
OT  - Metabolism
OT  - Mitochondrial DNA
OT  - Single-cell
OT  - Transcription
EDAT- 2015/11/05 06:00
MHDA- 2016/09/07 06:00
CRDT- 2015/11/05 06:00
PHST- 2015/11/05 06:00 [entrez]
PHST- 2015/11/05 06:00 [pubmed]
PHST- 2016/09/07 06:00 [medline]
AID - 10.1007/978-1-4939-3040-1_5 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2016;1351:49-65. doi: 10.1007/978-1-4939-3040-1_5.