CONFERENCE PROCEEDINGS
Advanced Search
Home > Journals > J. Biomed. Opt. > Volume 17 > Issue 4 > Article
Open Access
6 April 2012 Ethidium bromide as a marker of mtDNA replication in living cells
Anna Maria Villa, Paola Fusi, Valentina Pastori, Giulia Amicarelli, Chiara Pozzi, Silvia Maria Doglia, Daniel Adlerstein
Author Affiliations +
Journal of Biomedical Optics, Vol. 17, Issue 4, 046001 (April 2012). https://doi.org/10.1117/1.JBO.17.4.046001
Abstract
Mitochondrial DNA (mtDNA) in tumor cells was found to play an important role in maintaining the malignant phenotype. Using laser scanning confocal fluorescence microscopy (LSCFM) in a recent work, we reported a variable fluorescence intensity of ethidium bromide (EB) in mitochondria nucleoids of living carcinoma cells. Since when EB is bound to nucleic acids its fluorescence is intensified; a higher EB fluorescence intensity could reflect a higher DNA accessibility to EB, suggesting a higher mtDNA replication activity. To prove this hypothesis, in the present work we studied, by LSCFM, the EB fluorescence in mitochondria nucleoids of living neuroblastoma cells, a model system in which differentiation affects the level of mtDNA replication. A drastic decrease of fluorescence was observed after differentiation. To correlate EB fluorescence intensity to the mtDNA replication state, we evaluated the mtDNA nascent strands content by ligation-mediated real-time PCR, and we found a halved amount of replicating mtDNA molecules in differentiating cells. A similar result was obtained by BrdU incorporation. These results indicate that the low EB fluorescence of nucleoids in differentiated cells is correlated to a low content of replicating mtDNA, suggesting that EB may be used as a marker of mtDNA replication in living cells.
© 2012 Society of Photo-Optical Instrumentation Engineers (SPIE) 0091-3286/2012/$25.00 © 2012 SPIE
Citation Download Citation
Anna Maria Villa, Paola Fusi, Valentina Pastori, Giulia Amicarelli, Chiara Pozzi, Silvia Maria Doglia, and Daniel Adlerstein "Ethidium bromide as a marker of mtDNA replication in living cells," Journal of Biomedical Optics 17(4), 046001 (6 April 2012). https://doi.org/10.1117/1.JBO.17.4.046001
Published: 6 April 2012
JOURNAL ARTICLE
 10 PAGES
DOWNLOAD PAPER SAVE TO MY LIBRARY
GET CITATION
Lens.org Logo
CITATIONS
Cited by 7 scholarly publications and 3 patents.
Advertisement
Advertisement
RIGHTS & PERMISSIONS
Get copyright permission  Get copyright permission on Copyright Marketplace
KEYWORDS
Luminescence
Confocal microscopy
Tumors
Microscopy
Image analysis
Laser scanners
Molybdenum
RELATED CONTENT
Subscribe to Digital Library
Receive Erratum Email Alert
Back to Top