Assessment of Nuclear and Mitochondrial DNA, Expression of Mitochondria-Related Genes in Different Brain Regions in Rats after Whole-Body X-ray Irradiation.

Serazhutdin Abdullaev,Tatiana Bulanova,Azhub Gaziev,Nina Gubina

doi:10.3390/ijms21041196

Abstract

Studies of molecular changes occurred in various brain regions after whole-body irradiation showed a significant increase in terms of the importance in gaining insight into how to slow down or prevent the development of long-term side effects such as carcinogenesis, cognitive impairment and other pathologies. We have analyzed nDNA damage and repair, changes in mitochondrial DNA (mtDNA) copy number and in the level of mtDNA heteroplasmy, and also examined changes in the expression of genes involved in the regulation of mitochondrial biogenesis and dynamics in three areas of the rat brain (hippocampus, cortex and cerebellum) after whole-body X-ray irradiation. Long amplicon quantitative polymerase chain reaction (LA-QPCR) was used to detect nDNA and mtDNA damage. The level of mtDNA heteroplasmy was estimated using Surveyor nuclease technology. The mtDNA copy numbers and expression levels of a number of genes were determined by real-time PCR. The results showed that the repair of nDNA damage in the rat brain regions occurs slowly within 24 h; in the hippocampus, this process runs much slower. The number of mtDNA copies in three regions of the rat brain increases with a simultaneous increase in mtDNA heteroplasmy. However, in the hippocampus, the copy number of mutant mtDNAs increases significantly by the time point of 24 h after radiation exposure. Our analysis shows that in the brain regions of irradiated rats, there is a decrease in the expression of genes (ND2, CytB, ATP5O) involved in ATP synthesis, although by the same time point after irradiation, an increase in transcripts of genes regulating mitochondrial biogenesis is observed. On the other hand, analysis of genes that control the dynamics of mitochondria (Mfn1, Fis1) revealed that sharp decrease in gene expression level occurred, only in the hippocampus. Consequently, the structural and functional characteristics of the hippocampus of rats exposed to whole-body radiation can be different, most significantly from those of the other brain regions.

Highlights

Ionizing radiation (IR)-induced brain damage is most often observed after radiotherapy in the management of malignant tumors of the head, neck, nasopharynx, upper jaw, pituitary gland, skull base, or metastatic brain tumors
We studied nuclear DNA damage (nDNA) damage and repair, changes in the copy number of mitochondrial DNA and the level of its heteroplasmy, as well as the expression of genes involved in the regulation of mitochondrial biogenesis and dynamics in three rat brain areas following whole-body X-ray radiation
These data are consistent with the results of a recent study, which showed that a delay in DNA repair in the hippocampus occurs even at significantly low doses of mouse brain irradiation [17]

Summary

Introduction

Ionizing radiation (IR)-induced brain damage is most often observed after radiotherapy in the management of malignant tumors of the head, neck, nasopharynx, upper jaw, pituitary gland, skull base, or metastatic brain tumors. IR induces functional and morphological changes in brain tissues, vascular damage, cerebral radiation necrosis, increased oxidative stress, inhibition of neurogenesis and proliferation, changes in synoptic plasticity, decreased cognitive functions and the development of secondary brain tumors [1,2,3]. Though pronounced damage to brain tissue is usually caused by exposure to relatively high doses of IR used in radiotherapy of tumors, markedly more significant morphological and functional changes in the brain may occur from exposure to moderate- and low-level ionizing radiation [4,5]. Radiation-induced brain injury can be observed in patients who receive radiotherapy used to destroy tumors, and after exposure to IR for different diagnostic and therapeutic applications. It is known that, when total body irradiation is used, radiosensitive hematopoietic systems (primarily bone marrow), the gastrointestinal tract, and the vascular system are damaged [9]

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