RESUMO
The third instar larvae of Drosophila were irradiated with X rays, and the somatic mutation frequency in their wings was measured after their eclosion. In the flies with normal DNA repair and apoptosis functions, 0.2 Gy irradiation at 0.05 Gy/min reduced the frequency of the so-called small spot (mutant cell clone with reduced reproductive activity) compared with that in the sham-irradiated flies. When apoptosis was suppressed using the baculovirus p35 gene, the small spot frequency increased four times in the sham-irradiated control group, but the reduction by the 0.2-Gy irradiation was still evident. In a non-homologous end joining-deficient mutant, the small spot frequency was also reduced by 0.2 Gy radiation. In a mutant deficient in single-strand break repair, no reduction in the small spot frequency by 0.2 Gy radiation was observed, and the small spot frequency increased with the radiation dose. Large spot (mutant cell clone with normal reproductive activity) frequency was not affected by suppression of apoptosis and increased monotonically with radiation dose in wild-type larvae and in mutants for single- or double-strand break repair. It is hypothesized that some of the small spots resulted from single-strand damage and, in wild-type larvae, 0.2 Gy radiation activated the normal single-strand break repair gene, which reduced the background somatic mutation frequency.
Assuntos
Quebras de DNA de Cadeia Simples/efeitos da radiação , Reparo do DNA/efeitos da radiação , Drosophila melanogaster/genética , Drosophila melanogaster/efeitos da radiação , Mutagênese/efeitos da radiação , Animais , Apoptose/efeitos da radiação , Quebras de DNA de Cadeia Dupla/efeitos da radiação , Relação Dose-Resposta à Radiação , Drosophila melanogaster/anatomia & histologia , Drosophila melanogaster/citologia , Feminino , Larva/anatomia & histologia , Larva/citologia , Larva/genética , Larva/efeitos da radiação , Modelos Lineares , Masculino , Mutação/efeitos da radiação , Asas de Animais/anatomia & histologia , Asas de Animais/citologia , Asas de Animais/metabolismo , Asas de Animais/efeitos da radiação , Raios XRESUMO
We reported previously that low-dose X irradiation of DNA repair-proficient immature sperm of wild-type Drosophila melanogaster at a low dose rate (50 mGy/min) resulted in a mutation frequency that was lower than that in the sham-irradiated group. Therefore, a U-shaped dose-response relationship was suggested. Here we show that the dose-response curve is actually U-shaped by carrying out a large-scale sex-linked recessive lethal assay using Drosophila. No reduction of the mutation frequency was observed in a strain mutant for the nucleotide excision repair gene mei-9a (Drosophila homologue of human XPF). Introduction of a chromosome fragment containing mei-9+ into the mei-9a mutant strain restored the reduction of the mutation frequency in the low-dose-irradiated group. These results showed that DNA repair was responsible for the U-shaped dose-response relationship in Drosophila.
Assuntos
Drosophila melanogaster/genética , Genes Letais , Genes Recessivos , Mutação , Espermatozoides/efeitos da radiação , Animais , Relação Dose-Resposta a Droga , Drosophila melanogaster/efeitos da radiação , Feminino , Marcação In Situ das Extremidades Cortadas , MasculinoRESUMO
To determine whether the linear no-threshold (LNT) model for stochastic effects of ionizing radiation is applicable to very low-dose radiation at a low dose rate, we irradiated immature male germ cells of the fruit fly, Drosophila melanogaster, with several doses of (60)Co gamma rays at a dose rate of 22.4 mGy/h. Thereafter, we performed the sex-linked recessive lethal mutation assay by mating the irradiated males with nonirradiated females. The mutation frequency in the group irradiated with 500 microGy was found to be significantly lower than that in the control group (P < 0.01), whereas in the group subjected to 10 Gy irradiation, the mutation frequency was significantly higher than that in the control group (P < 0.03). A J-shaped dose-response relationship was evident. Molecular experiments using DNA microarray and quantitative reverse transcription PCR indicated that several genes known to be expressed in response to heat or chemical stress and grim, a positive regulator of apoptosis, were up-regulated immediately after irradiation with 500 microGy. The involvement of an apoptosis function in the non-linear dose-response relationship was suggested.
Assuntos
Drosophila melanogaster/citologia , Drosophila melanogaster/genética , Raios gama/efeitos adversos , Células Germinativas/efeitos da radiação , Mutagênese/efeitos da radiação , Mutação/efeitos da radiação , Doses de Radiação , Animais , Apoptose/efeitos da radiação , Relação Dose-Resposta à Radiação , Drosophila melanogaster/efeitos da radiação , Feminino , Perfilação da Expressão Gênica , Genes Recessivos/genética , Genes Recessivos/efeitos da radiação , Masculino , Análise de Sequência com Séries de OligonucleotídeosRESUMO
We investigated the mechanism underlying the radioadaptive response that rescues mice from hematopoietic failure. C57BL/6 mice were irradiated with low-dose acute X rays (0.5 Gy) for priming 2 weeks prior to a high-dose (6 Gy) challenge irradiation. Bone marrow cells, erythrocytes and platelets in low-dose-preirradiated mice showed earlier recovery after the challenge irradiation than those in mice subjected only to the challenge irradiation. This suggests that hematopoiesis is enhanced after a challenge irradiation in preirradiated mice. The rapid recovery of bone marrow cells after the challenge irradiation was consistent with the proliferation of hematopoietic progenitors expressing the cell surface markers Lin-, Sca-1- and c-Kit+ in low-dose-preirradiated mice. A subpopulation of myeloid (Mac-1+/Gr-1+) cells, which were descendants of Lin-, Sca-1- and c-Kit+ cells, rapidly recovered in the bone marrow of low-dose-preirradiated mice, whereas the number of B-lymphoid (CD19+/B220+) cells did not show a statistically significant increase. Plasma cytokine profiles were analyzed using antibody arrays, and results indicated that the concentrations of several growth factors for myelopoiesis after the challenge irradiation were considerably increased by low-dose preirradiation. The rapid recovery of erythrocytes and platelets but not leukocytes was observed in the peripheral blood of preirradiated mice, suggesting that low-dose preirradiation triggered the differentiation to myelopoiesis. Thus the adaptive response induced by low-dose preirradiation in terms of the recovery kinetics of the number of hematopoietic cells may be due to the rapid recovery of the number of myeloid cells after high-dose irradiation.
Assuntos
Adaptação Fisiológica/fisiologia , Citocinas/metabolismo , Células Mieloides/fisiologia , Células Mieloides/efeitos da radiação , Recuperação de Função Fisiológica/efeitos da radiação , Irradiação Corporal Total/métodos , Adaptação Fisiológica/efeitos da radiação , Animais , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Células Mieloides/citologia , Tolerância a RadiaçãoRESUMO
A sex-linked recessive lethal mutation assay was performed in Drosophila melanogaster using immature spermatocytes and spermatogonia irradiated with X rays at a high or low dose rate. The mutation frequency in the sperm irradiated with a low dose at a low dose rate was significantly lower than that in the sham-irradiated group, whereas irradiation with a high dose resulted in a significant increase in the mutation frequency. It was obvious that the dose-response relationship was not linear, but rather was U-shaped. When mutant germ cells defective in DNA excision repair were used instead of wild-type cells, low-dose irradiation at a low dose rate did not reduce the mutation frequency. These observations suggest that error-free DNA repair functions were activated by low dose of low-dose-rate radiation and that this repaired spontaneous DNA damage rather than the X-ray-induced damage, thus producing a practical threshold.
Assuntos
Drosophila/genética , Drosophila/efeitos da radiação , Mutação , Espermatócitos/efeitos da radiação , Espermatozoides/efeitos da radiação , Raios X , Animais , Mapeamento Cromossômico , Cruzamentos Genéticos , Dano ao DNA , Reparo do DNA , Relação Dose-Resposta à Radiação , Feminino , Genes Recessivos , MasculinoRESUMO
An adaptive response induced by long-term low-dose-rate irradiation in mice was evaluated in terms of the amount of DNA damage in the spleen analyzed by a comet assay. C57BL/ 6N female mice were irradiated with 0.5 Gy of (137)Cs gamma rays at 1.2 mGy/h; thereafter, a challenge dose (0.4, 0.8 or 1.6 Gy) at a high dose rate was given. Less DNA damage was observed in the spleen cells of preirradiated mice than in those of mice that received the challenge dose only; an adaptive response in terms of DNA damage was induced by long-term low-dose-rate irradiation in mice. The gene expression of catalase and Mn-SOD was significantly increased in the spleen after 23 days of the low-dose-rate radiation (0.5 Gy). In addition, the enzymatic activity of catalase corresponded to the gene expression level; the increase in the activity was observed at day 23 (0.5 Gy). These results suggested that an enhancement of the antioxidative capacities played an important role in the reduction of initial DNA damage by low-dose-rate radiation.
Assuntos
Antioxidantes/metabolismo , Dano ao DNA/fisiologia , DNA/efeitos da radiação , Enzimas/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Irradiação Corporal Total , Adaptação Fisiológica/fisiologia , Adaptação Fisiológica/efeitos da radiação , Animais , Células Cultivadas , Dano ao DNA/efeitos da radiação , Ativação Enzimática/efeitos da radiação , Feminino , Raios gama , Regulação Enzimológica da Expressão Gênica/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Doses de Radiação , Tolerância a Radiação/efeitos da radiação , Baço/enzimologia , Baço/efeitos da radiaçãoRESUMO
Bacteria that selectively kill males ("male-killers") were first characterized more than 50 years ago in Drosophila and have proved to be common in insects. However, the mechanism by which sex specificity of virulence is achieved has remained unknown. We tested the ability of Spiroplasma poulsonii to kill Drosophila melanogaster males carrying mutations in genes that encode the dosage compensation complex. The bacterium failed to kill males lacking any of the five protein components of the complex.
Assuntos
Mecanismo Genético de Compensação de Dose , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Drosophila melanogaster/microbiologia , Spiroplasma/patogenicidade , Acetiltransferases/genética , Acetiltransferases/fisiologia , Animais , Proteínas Cromossômicas não Histona/genética , Proteínas Cromossômicas não Histona/fisiologia , DNA Helicases/genética , DNA Helicases/fisiologia , Proteínas de Ligação a DNA , Proteínas de Drosophila/fisiologia , Drosophila melanogaster/embriologia , Drosophila melanogaster/fisiologia , Feminino , Genes de Insetos , Heterozigoto , Histona Acetiltransferases , Homozigoto , Masculino , Mutação , Proteínas Nucleares/genética , Proteínas Nucleares/fisiologia , Caracteres Sexuais , Fatores de Transcrição/genética , Fatores de Transcrição/fisiologia , Transcrição Gênica , Cromossomo X/metabolismoRESUMO
To assess the possibility that strong static magnetic fields cause DNA damage and mutation, we examined the genotoxic effects of magnetic field exposure by using the somatic mutation and recombination test system in DNA repair-proficient and -deficient strains of Drosophila melanogaster. A postreplication repair-defective mutation mei-41D5 and/or a nucleotide excision repair-defective mutation mei-9(a) was introduced into the conventional loss of the heterozygosity assay system by the use of mwh +/ + flr transheterozygotes, and were exposed to static magnetic fields of up to 14 Tesla (T). We found that exposure to 2, 5, or 14 T fields for 24 h caused a statistically significant enhancement in somatic recombination frequency in the postreplication repair-deficient flies, whereas the frequency remained unchanged in the nucleotide excision repair-deficient flies and in the DNA repair-proficient flies after exposure. An increase linearly dependent on the flux density was observed between 0.5 T and 2 T, but it was saturated at exposure levels over 2 T. These findings suggest that exposure to high-density magnetic fields induce somatic recombination in Drosophila and that the dose-response relationship is not linear.
Assuntos
Dano ao DNA , Reparo do DNA/genética , Drosophila melanogaster/genética , Magnetismo , Mutação , AnimaisRESUMO
The dose-response relationship of ionizing radiation and its stochastic effects has been thought to be linear without any thresholds. The basic data for this model were obtained from mutational assays in the male germ cells of the fruit fly Drosophila melanogaster. However, it is more appropriate to examine carcinogenic activity in somatic cells than in germ cells. Here the dose-response relationship of X irradiation and somatic mutation was examined in Drosophila. A threshold at approximately 1 Gy was observed in DNA repair-proficient flies. In the repair-deficient siblings, the threshold was smaller and the inclination of the dose-response curve was much steeper. These results suggest that the dose-response relationship between X irradiation and somatic mutation has a threshold and that the DNA repair function contributes to its formation.