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).
Villa, A., Fusi, P., Pastori, V., Amicarelli, G., Pozzi, C., Adlerstein, D., et al. (2012). Ethidium bromide as a marker of mtDNA replication in living cells. JOURNAL OF BIOMEDICAL OPTICS, 17(4), 046001 [10.1117/1.JBO.17.4.046001].
Citazione: | Villa, A., Fusi, P., Pastori, V., Amicarelli, G., Pozzi, C., Adlerstein, D., et al. (2012). Ethidium bromide as a marker of mtDNA replication in living cells. JOURNAL OF BIOMEDICAL OPTICS, 17(4), 046001 [10.1117/1.JBO.17.4.046001]. | |
Tipo: | Articolo in rivista - Articolo scientifico | |
Carattere della pubblicazione: | Scientifica | |
Presenza di un coautore afferente ad Istituzioni straniere: | No | |
Titolo: | Ethidium bromide as a marker of mtDNA replication in living cells | |
Autori: | Villa, A; Fusi, P; Pastori, V; Amicarelli, G; Pozzi, C; Adlerstein, D; Doglia, S | |
Autori: | FUSI, PAOLA ALESSANDRA (Secondo) DOGLIA, SILVIA MARIA (Ultimo) | |
Data di pubblicazione: | 2012 | |
Lingua: | English | |
Rivista: | JOURNAL OF BIOMEDICAL OPTICS | |
Digital Object Identifier (DOI): | http://dx.doi.org/10.1117/1.JBO.17.4.046001 | |
Appare nelle tipologie: | 01 - Articolo su rivista |