J Vet Sci.  2012 Dec;13(4):331-338.

Prolonged expression of senescence markers in mice exposed to gamma-irradiation

• Seol MA
• Jung U
• Eom HS
• Kim SH
• Park HR
• Jo SK

Abstract

Although ionizing radiation is known to induce cellular senescence in vitro and in vivo, its long-term in vivo effects are not well defined. In this study, we examined the prolonged expression of senescence markers in mice irradiated with single or fractionated doses. C57BL/6 female mice were exposed to 5 Gy of gamma-rays in single or 5, 10, 25 fractions. At 2, 4, and 6 months after irradiation, senescence markers including mitochondrial DNA (mtDNA) common deletion, p21, and senescence-associated beta-galactosidase (SA beta-gal) were monitored in the lung, liver, and kidney. Increases of mtDNA deletion were detected in the lung, liver, and kidney of irradiated groups. p21 expression and SA beta-gal staining were also increased in the irradiated groups compared to the non-irradiated control group. Increases of senescence markers persisted up to 6 months after irradiation. Additionally, the extent of mtDNA deletion and the numbers of SA beta-gal positive cells were greater as the number of radiation fractions increased. In conclusion, our results showed that ionizing radiation, especially that delivered in fractions, can cause the persistent upregulation of senescence marker expression in vivo. This should be considered when dealing with chronic normal tissue injuries caused by radiation therapy or radiation accidents.

Keyword

mitochondrial DNA deletion; p21; radiation; senescence; senescence-associated beta-galactosidase

MeSH Terms

Aging
Animals
Cell Aging
DNA, Mitochondrial
Female
Humans
Kidney
Liver
Lung
Mice
Radiation, Ionizing
Radioactive Hazard Release
Up-Regulation
beta-Galactosidase

Figure

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  Fig. 1 Schematic illustration of irradiation delivery and experimental schedule. 8-week-old female C57BL/6 mice were irradiated with a single dose (5 Gy × 1) or fractionated doses (0.2 Gy × 25, 0.5 Gy × 10, or 1 Gy × 5) of ionizing radiation (IR). The mice were then sacrificed at 2, 4, and 6 months after the first exposure to radiation.

• 

  Fig. 2 Induction of mitochondrial DNA (mtDNA) common deletion (3867 bp) in the tissues of irradiated C57BL/6 mice. (A) Tissues isolated from young (2-month-old) and old (24-month-old) animals were analyzed for mtDNA common deletion. (B) 8-week-old mice were exposed to IR delivered in a single dose (5 Gy × 1) or fractionated doses (0.2 Gy × 25, 0.5 Gy × 10, or 1 Gy × 5), and sacrificed 2, 4, and 6 months after IR exposure.

• 

  Fig. 3 Increased expression of p21 in tissues of irradiated C57BL/6 mice. (A) Tissues isolated from young (2-month-old) and old (24-month-old) mice were analyzed for p21 protein expression. (B) 8-week-old C57BL/6 mice were exposed to a single dose (5 Gy × 1) or fractionated doses (0.2 Gy × 25, 0.5 Gy × 10, or 1 Gy × 5) of IR, and sacrificed at 2, 4, 6, months after radiation exposure.

• 

  Fig. 4 Increased numbers of senescence-associated β-galactosidase (SA β-gal)-positive cells in the kidney of irradiated C57BL/6 mice. (A and C) Kidneys isolated from young (2-month-old) and old (24-month-old) C57BL/6 mice were stained for SA β-gal. (B and D) Mice were exposed to IR delivered in a single dose (5 Gy × 1) or fractionated doses (0.2 Gy × 25, 0.5 Gy × 10, or 1 Gy × 5), and sacrificed at 2, 4, and 6 months after radiation exposure. Cells stained blue are positive for SA β-gal. Significant differences relative to the control group are indicated (*p < 0.05 and **p < 0.01). Scale bars = 20 µm.

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