PMID- 15258176
OWN - NLM
STAT- MEDLINE
DCOM- 20040826
LR  - 20131121
IS  - 0022-1554 (Print)
IS  - 0022-1554 (Linking)
VI  - 52
IP  - 8
DP  - 2004 Aug
TI  - Rapid analysis of mitochondrial DNA depletion by fluorescence in situ 
      hybridization and immunocytochemistry: potential strategies for HIV therapeutic 
      monitoring.
PG  - 1011-8
AB  - Nucleoside reverse transcriptase inhibitors (NRTIs) have been a mainstay in the 
      treatment of human immunodeficiency virus since the introduction of 
      azidothymidine (AZT) in 1987. However, none of the current therapies can 
      completely eradicate the virus, necessitating long-term use of anti-retroviral 
      drugs to prevent viral re-growth. One of the side effects associated with 
      long-term use of NRTIs is mitochondrial toxicity stemming from inhibition of the 
      mitochondrial DNA (mtDNA) polymerase gamma, which leads to mtDNA depletion and 
      consequently to mitochondrial dysfunction. Here we report the use of fluorescence 
      in situ hybridization (FISH) and immunocytochemistry (ICC) to monitor mtDNA 
      depletion in cultured fibroblasts treated with the NRTI 2',3'-dideoxycytidine 
      (ddC). These techniques are amenable to both microscopy and flow cytometry, 
      allowing analysis of populations of cells on a single-cell basis. We show that, 
      as mtDNA depletion progresses, a mosaic population develops, with some cells 
      being depleted of and others retaining mtDNA. These techniques could be useful as 
      potential therapeutic monitors to indicate when NRTI therapy should be 
      interrupted to prevent mitochondrial toxicity and could aid in the development of 
      less toxic NRTIs by providing an assay suitable for pharmacodynamic evaluation of 
      candidate molecules.
FAU - Janes, Michael S
AU  - Janes MS
AD  - Molecular Probes, Inc., 29851 Willow Creek Road, Eugene, Oregon 97402, USA. 
      mike.janes@probes.com
FAU - Hanson, Bonnie J
AU  - Hanson BJ
FAU - Hill, Dani M
AU  - Hill DM
FAU - Buller, Gayle M
AU  - Buller GM
FAU - Agnew, Jakyoung Y
AU  - Agnew JY
FAU - Sherwood, Steven W
AU  - Sherwood SW
FAU - Cox, W Gregory
AU  - Cox WG
FAU - Yamagata, Kunihiro
AU  - Yamagata K
FAU - Capaldi, Roderick A
AU  - Capaldi RA
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PL  - United States
TA  - J Histochem Cytochem
JT  - The journal of histochemistry and cytochemistry : official journal of the 
      Histochemistry Society
JID - 9815334
RN  - 0 (DNA, Mitochondrial)
RN  - 0 (Reverse Transcriptase Inhibitors)
RN  - 6L3XT8CB3I (Zalcitabine)
SB  - IM
MH  - Cell Line
MH  - DNA, Mitochondrial/*biosynthesis
MH  - Fibroblasts/drug effects/metabolism
MH  - Humans
MH  - Immunohistochemistry
MH  - In Situ Hybridization, Fluorescence
MH  - Polymerase Chain Reaction
MH  - Reverse Transcriptase Inhibitors/*adverse effects
MH  - Zalcitabine/*adverse effects
EDAT- 2004/07/20 05:00
MHDA- 2004/08/27 05:00
CRDT- 2004/07/20 05:00
PHST- 2004/07/20 05:00 [pubmed]
PHST- 2004/08/27 05:00 [medline]
PHST- 2004/07/20 05:00 [entrez]
AID - 52/8/1011 [pii]
AID - 10.1369/jhc.3A6209.2004 [doi]
PST - ppublish
SO  - J Histochem Cytochem. 2004 Aug;52(8):1011-8. doi: 10.1369/jhc.3A6209.2004.