Ii. Ploskonosova et al., PCR assay of DNA damage and repair at the gene level in brain and spleen of gamma-irradiated young and old rats, MUT R-DNA R, 434(2), 1999, pp. 109-117
The PCR amplification of Fragments of transcribed (beta-actin, p53) and non
transcribed (IgE, heavy chain) genes in brain and spleen DNA from gamma-irr
adiated and unirradiated 2- and 28-month-old rats was studied. The amplific
ation levels of fragments of these genes in DNA from old rats were substant
ially lower than those from young rats, which suggested that these gene fra
gments in old-rat DNA contained lesions blocking thermostable polymerase in
PCR. The beta-actin and IgE gene fragments of spleen DNA from old rats exh
ibited a significantly higher level of lesions inhibiting Tth polymerase co
mpared to analogous fragments of brain DNA from the same animals. DNA from
the tissues of gamma-irradiated rats showed the amount of damage inhibiting
amplification to be dependent on animal age and the postirradiation time b
efore DNA isolation. As judged from the changes in the amplification level
of gene fragments, there was no preferential fast repair of lesions in the
actively transcribed gene beta-actin compared to the nontranscribed gene Ig
E (heavy chain) in the brain and spleen of gamma-irradiated young and old r
ats. The amplification results suggest that equal amounts of DNA lesions we
re repaired in the brain of both old and young rats during the first 0.5 h
of the postirradiation time (fast-repair phase). whereas in the subsequent
postirradiation period over 5 h (slow-repair phase), the efficiency of dama
ge elimination in the brain DNA of old rats was markedly lower. As for the
spleen tissue, the elimination of lesions blocking Tth polymerase was much
lower in old gamma-irradiated animals for both of the repair phases. (C) 19
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