Alteration of mtDNA copy number, mitochondrial gene expression and extracellular DNA content in mice after irradiation at lethal dose

Abstract

High steady-state transcriptional activity is essential for normal mitochondrial function. The requisite transcription rate is satisfied in part by high copy number of mitochondrial DNA (mtDNA). In the present study, we analyze mtDNA copy number by real-time PCR in nucleated blood cells from control mice and mice exposed to 1- or 10-Gy X-radiation. Transcription of the oxidative phosphorylation-associated genes cytb, atp6, nd4, nd2 and d-loop region was monitored in these nucleated blood cells similarly by real-time PCR. We observed a 50% decrease in the ratio of mitochondrial to nuclear DNA (mtDNA/nDNA) in blood cells, while the mtDNA/nDNA ratio in serum increased. After a lethal 10-Gy dose of X-irradiation, we observed an 80% decrease in the number of circulating lymphocytes. In response to a 10-Gy radiation dose, we observed transiently increased mtDNA/nDNA ratio and transcription within the initial 5 h post-treatment. At 24-72 h, the mtDNA/nDNA ratio in surviving cells was reduced to the level observed in blood cells irradiated with 1 Gy. We observed a decrease in the serum mtDNA/nDNA ratio due to an increase in nDNA content rather than a decrease in mtDNA. Taken together, results presented herein suggest that the mtDNA/nDNA ratio may be of clinical value potentially as a diagnostic tool, particularly in oncology patients undergoing radiation therapy.