[Comparative analysis of ionizing radiation and xenobiotics influence on spermatogenic epithelium and dominant lethal mutations output in laboratory animals].

The study covered state of spermatogenic epithelium and dominant lethal mutations output in mice of BALB/c and CBA lines, subjected to total gamma-irradiation and in Wistar rats after intraperitoneal injection of potassium bichromate (K2Cr2,O7) in small and sublethal doses. The BALB/c line mice under low irradiation dose (0.25 Gy) demonstrated stimulation effect on spermatogenic epithelium, but in the CBA line mice no such effect was seen. Both mice lines under irradiation of 0.25 Gy and 1.0 Gy demonstrated increase in pathologic sperm counts and in percentage ofpreimplantation embryonal death. In rats, injection of potassium bichromate in doses of 0.028 mg/kg and 2.8 mg/kg increased number of micronuclear spermatids, larger pathologic sperm counts and percentage of postimplantation deaths. Thus, lower general embryonal deaths under radiation exposure is due to preimplantation embryonal deaths, under exposure to 6-valent chromium--is due to postimplantation losses.

[Evaluation of effects of gamma-irradiation at low doses on repair and meiotic recombination mutants of Drosophila melanogaster].

A comparative study of the effects of gene mutations mus209, mus309, mei-41 and rad54 of Drosophila melanogaster on the sensitivity to low-level exposure of different duration was carried out. Taken into account was the survival rate at different stages of ontogeny, female fecundity, the frequency of dominant lethal mutations (DLM) and the DNA damage. mei-41 and rad-54 mutants were most sensitive to the action of low dose radiation (80 mGy) in terms of survival and DLM. However, at the level of DNA damage, an increased radiosensitivity is observed only at larger doses of low intensity irradiation. Based on these observations, we can conclude about the importance of repair and its genes in the formation of the effect of low level doses of ionizing radiation in Drosophila.

[The influence of gamma-irradiation in low doses on the rate of dominant lethal mutations in drosophila melanogaster].

The dose-rate effect of acute and chronic irradiation in the dose of 0.2 Gy in Drosophila melanogaster repair (mei-41, mus209 [Russian character: see text] mus309) and free radicals detoxication (sod) mutant strains was investigated. Was shown the lack of dose rate effect on the rate of dominant lethal mutations in mei-41, mus209 and sod. However in mus309, that has defect in the main Drosophila pathway of the DNA double strand breack repair, the increase of the mutation rate after chronic irradiation was observed (inverse dose-rate effect). The obtained results suggest the main role of DNA double strand breack repair in dose-rate effect formation in Drosophila.

[Genetic effects in Drosophila melanogaster induced by chronic low-dose irradiation]. / Geneticheskie éffekty u Drosophila melanogaster, indutsirovannye khronicheskim oblucheniem v malykh dozakh.

It was investigated the influence of the chronic gamma-irradiation in the dose rate of 0.17 sGy/h on the rate of genetic variability in the laboratory strains of Drosophila melanogaster with genotypic distinguishes in mobile genetic elements and defects in the DNA repair processes. It was shown that the rates of induction of recessive lethal mutations depended on genotype of investigated strains. In the different strains we have observed an increase as well as a decrease of the mutation rate. Also in was established that irradiation leads to the frequencies of the GD-sterility and mutability of the snw and h(w+) in the P-M and H-E dysgenic crosses. The obtained results suggest that mobile genetic elements play an important role in the forming of genetic effects in response to low dose irradiation.

[Genetic effects in populations of plants growing in the zone of Kyshtym and Chernobyl accidents]. / Geneticheskie éffekty v populiatsiiakh rastenii, proizrastaiushchikh v zonakh Kyshtymskoi i Chernobyl'skoi avarii.

Studies to analyze the genetic processes in natural populations of plants were started on the territory of the East-Ural Radioactive Trace (EURT) in 1962 and in the zone of the Chernobyl accident in May 1986. The main directions of the genetic studies in both radioactive areas were similar: 1) study of the mutation process intensity depending on the dose and dose rate and analysis of dose-effect relationships for different genetic changes (point mutations, chromosome aberrations in mitosis and meiosis) in irradiated plant populations; 2) study of the mutation process dynamics in generations of chronically (prolongly) irradiated populations of plants; 3) analysis of microevolutionary processes in irradiated plant populations. The report presents an analysis of observed dose-effect relationships under the action of radiation on populations of Arabidopsis thaliana, Pinus sylvestris and a number of other plant species. Analysis of the mutation processes dynamics in 8 Arabidopsis populations growing in the zone of the Chernobyl catastrophe has demonstrated that the level of the embryo lethal mutations 10 years after the accident in the irradiated populations significantly exceeds the control level. The following phenomena observed in chronically irradiated populations have also been considered: increased radioresistance of irradiated populations (radioadaptation), the appearance of abnormal karyotypes and selective markers upon chronic irradiation. The authors call attention to the high importance of monitoring of genetic processes in irradiated plant populations for understanding of the action of radiation on human populations.

[The mutability of natural populations and laboratory strains of Drosophila under conditions of chronic irradiation at small doses of low intensity]. / Mutabil'nost' prirodnykh populiatsii i laboratornykh linii drozofily v usloviiakh khronicheskogo oblucheniia v malykh dozakh nizkoi intensivnosti.

The frequency of irradiation-induced recessive lethals has been studied in chromosomes 1 and 2 of Drosophila melanogaster. It supposed, that the higher level of mutability may be explained in destabilization of genotype. The essential part in induction of genotype destabilization replies the genome instability mechanisms.

[The genetic monitoring of the house mouse population from areas contaminated by radionuclides as a result of the accident at the Chernobyl Atomic Electric Power Station]. / Geneticheskii monitoring populiatsii domovykh myshei iz raionov, zagriaznennykh radionuklidami v rezul'tate avarii na Chernobyl'skoi AES.

In 1986-1994, genetic effects of radiation on males of wild mice caught in areas of radioactive contamination after the Chernobyl reactor accident were studied. The gamma-radiation dose rates on the soil surface in the areas varied from 0,02 to 200 mR/h. The frequency of reciprocal translocations in mouse spermatocytes was relatively low and increased linearly with increasing of the dose rate. The levels of recessive lethal mutations in mouse populations did not depend on the radiation dose rate in the areas and decreased with a time.

[The demonstration of heritable lethal mutations in the progeny of x-ray-irradiated CHO cells by micronucleus count in clone cells]. / Nachweis vererbbarer Letalmutationen in Nachkommen röntgenbestrahlter CHO-Zellen durch Mikrokernzählung in Klonzellen.

PURPOSE: Low doses of ionizing radiation reduce the growth rates of clones following irradiation of the progenitor cells. Such reductions of clone growth have been proven by means of measurements of clone size distributions. The medians of such distributions can be used to quantify the radiation damage. Prolongations of generation times and cell death as result of heritable lethal mutations have been discussed as causes for the reduction of clone growth. MATERIAL AND METHODS: The cell number of a clone of hypotetraploid CHO-cells was compared to the frequency of micronucleated binucleated cells in the same clone using the cytokinesis-block-micronucleus method. The dose dependent reduction of clone sizes is measured by the difference of the medians (after log transformation) of the clone size distributions. RESULTS: At cytochalasin-B concentrations of 1 microgram/ml and after an incubation time of 16 h a yield of binucleated cells of about 50% was obtained. Median clone size differences as a measure of clonal radiation damage increased linearly with incubation times of 76, 100, 124, and 240 h following irradiation with 3, 5, 7, and 12 Gy. The frequency of binucleated clone cells with micronuclei strongly increased with decreasing clone size by a factor up to 20 following irradiation with 3, 5, and 7 Gy. The frequency of micronucleated binucleated clone cells was found to be independent of incubation time after irradiation. CONCLUSION: Radiation induced clone size reductions result from cell losses caused by intraclonal expression of micronuclei which have its origin in heritable lethal mutations. Measurements of clone size distributions can be done automatically. They can serve as predictive test for determination of median cell loss rates of surviving cell clones.

[Consequences for the intrauterine development of the offspring of the irradiation of male germinal cells at different stages of spermatogenesis]. / Posledstviia dlia vnutriutrobnogo razvitiia potomstva oblucheniia polovykh kletok samtsov na raznykh stadiiakh spermatogeneza.

Single whole-body exposure of adult male rats Wistar in different stages of spermatogenesis to gamma-rays (doses from 0.25 to 5.00 Gy) resulted in violations of antenatal development of the first generation offsprings. The pronouncement of these violations depends on the spermatogenesis stage in the moment of irradiation and exposure dose.

[The effect of low and sublethal doses of ionizing radiation on the function of the spermatogenic epithelium and the yield of dominant lethal mutations in different strains of mice]. / Vliianie malykh i subletal'nykh doz ioniziruiushchego izlucheniia na sostoianie spermatogennogo épiteliia i vykhod dominantnykh letal'nykh mutatsii u myshei raznykh linii.

There was a 30% increase in spermatozoids with microheads and a 3-fold increase in "young" spermatozoids at later times after irradiation of BALB/c mice a dose of 1.0 Gy. With radiation doses of 0.1-1.0 Gy, BALB/c and CBA mice (which differ in viability of spermatogenous cells, the number of Sertoli cells and total embryonal death rate) exhibited a significant increase in the preimplantation death at the stage of spermatogonia and stem cells. The results obtained indicate reduction of the fertilizing capacity of sperm which is probably realized as dominant lethal mutations.

[An experimental assessment of the genetic effects of low doses of internal irradiation]. / Eksperimental'naia otsenka geneticheskikh éffektov malykh doz vnutrennego oblucheniia.

Genetic study of rats showed the possibility of registration of the minor doses of the inside and outside irradiation. The postmiosis mutations, number of reciprocal translocations in spermatogonia may be used for indexation of the effects.

Radiobiological studies with the nematode Caenorhabditis elegans. Genetic and developmental effects of high LET radiation.

The biological effects of heavy charged particle (HZE) radiation are of particular interest to travellers and planners for long-duration space flights where exposure levels represent a potential health hazard. The unique feature of HZE radiation is the structured pattern of its energy deposition in targets. There are many consequences of this feature to biological endpoints when compared with effects of ionizing photons. Dose vs response and dose-rate kinetics may be modified, DNA and cellular repair systems may be altered in their abilities to cope with damage, and the qualitative features of damage may be unique for different ions. The nematode Caenorhabditis elegans is being used to address these and related questions associated with exposure to radiation. HZE-induced mutation, chromosome aberration, cell inactivation and altered organogenesis are discussed along with plans for radiobiological experiments in space.

Measurement of neutron-induced genetic damage in mouse immature oocytes.

Recent experimental evidence concerning the nature of radiosensitive targets in mouse immature (resting) oocytes has led to new experimental designs that permit measurement of radiation-induced genetic damage in these important cells. We have previously reported initial results of the detection of genetic damage in mouse immature oocytes using monoenergetic 0.43-MeV neutrons. Here we provide a full report of our data and compare the genetic sensitivity of immature oocytes with those measured by others for maturing oocytes. Until recently, all attempts to detect radiation-induced genetic damage in mouse immature oocytes had failed. This appears to have been because the radiation types and modes of dose delivery used in those studies did not sufficiently spare the hypersensitive lethality target (the plasma membrane) while at the same time deposit enough dose in DNA to produce detectable mutation. Recoil protons from 0.43-MeV neutrons produce short ionization tracks (2.6 micron mean) and can therefore deposit energy in the DNA without simultaneously traversing the plasma membrane. Using these particles, we have obtained dose-response relationships for both chromosome aberrations and dominant lethal mutations in oocytes from females irradiated 8-12 weeks earlier, when oocytes were immature. Results suggest that the intrinsic mutational sensitivity of mouse immature oocytes is not very different from that of maturing oocytes.

Development of a possible nonmammalian test system for radiation-induced germ-cell mutagenesis using a fish, the Japanese medaka (Oryzias latipes).

To develop a specific-locus test (SLT) system for environmental mutagenesis using vertebrate species other than the mouse, we first established a tester stock of the fish medaka (Oryzias latipes) that is homozygous recessive at three loci. The phenotypic expression of these loci can be easily recognized early in embryonic development by observation through the transparent egg membrane. We irradiated wild-type males with 137Cs gamma-rays to determine the dose-response relationships for dominant lethal and specific-locus mutations induced in sperm, spermatids, and spermatogonia. Through observation of 322,666 loci in control offspring and 374,026 loci in offspring obtained from 0.64-, 4.75-, or 9.50-Gy-irradiated gametes, specific-locus mutations were phenotypically detected during early development. These putative mutations, designated "total mutation," can be recognized only in embryos of oviparous animals. The developmental fate of these mutant embryos was precisely followed. During subsequent embryonic development, a large fraction died and thus was unavailable for test-crossing, which was used to identify "viable mutations." Our medaka SLT system demonstrates that the vast majority of total mutations is associated with dominant lethal mutations. Thus far only one spontaneous viable mutation has been observed, so that all doubling calculations involving this endpoint carry a large error. With these reservations, however, we conclude that the quantitative data so far obtained from the medaka SLT are quite comparable to those from the mouse SLT and, hence, indicate the validity of the medaka SLT as a possible nonmammalian test system.

Induction of dominant lethal mutations by combined X-ray-acrylamide treatment in male mice.

The induction of mutations following combined treatment with acrylamide (AA) plus X-rays has been determined using the dominant lethal mutations test in Pzh:SFISS male mice. Combinations of a mutagenic dose of both agents (1.00 Gy, 125 mg/kg b.w.) and a non-mutagenic dose, i.e., a dose that alone does not produce dominant lethals (0.25 Gy, 25 mg/kg b.w.), were used. For the discussion of the effects of combined action of X-rays and acrylamide the term 'enhancement in risk' was used whenever the effects observed after combined exposure significantly exceeded the sum of the effects produced separately by the agents. Such an enhanced risk has been observed in late spermatids after combined action of X-rays and AA at non-mutagenic doses, and in spermatozoa, spermatids and late spermatocytes after exposure to mutagenic doses.

Analysis of interactions between mutagens, I. Heat and ultraviolet light in Saccharomyces cerevisiae.

A new mathematical approach to the description of interaction data (Ager and Haynes, 1987) is applied here to the interaction between heat and ultraviolet light (UV) in Saccharomyces cerevisiae. A strong synergism for cell killing is found to be associated with large increases in gene conversion (of up to 8-fold), and mutation (of up to 14-fold). Analysis of the interaction data for both wild-type and repair-deficient strains indicates that the heat-UV synergism arises via the inhibition of two different repair pathways. Unambiguous conclusions regarding the molecular mechanisms by which these repair processes are inhibited cannot be drawn on the basis of dose-response data alone. However, this approach does enable one to make well defined, empirical comparisons of the nature and kinetics of such interactions.

Analysis of interactions between mutagens, II. Ethyl methanesulfonate and ultraviolet light in Saccharomyces cerevisiae.

The results of this study indicate the existence of a strong interaction between ethyl methanesulfonate (EMS) and ultraviolet light (UV) for cell killing in the yeast Saccharomyces cerevisiae. Conversely, mutation and gene conversion frequencies observed for the combined treatment of EMS and UV do not deviate significantly from that expected on the basis of simple additivity. Studies involving repair-deficient mutants (rad mutants) reveal that the synergistic interaction for cell killing depends on RAD52 function (recombinational repair), but not on RAD3 function (excision repair). On the basis of this analysis, the interaction between EMS and UV in S. cerevisiae might arise from the inhibition of double-strand break repair by one, or both agents.

[Genetic damage in domestic mice inhabiting in the areas with elevated background radiation]. / Geneticheskie povrezhdeniia u domovykh myshei, obitaiushchikh v usloviiakh povyshennogo fona radiatsii.

Genetic effects of irradiation in males of wild house mice which were caught in the region of Chernobyl Nuclear Power Station were studied. The dose rate on the ground surface varied from 0.04 to 200 mR/h of gamma-irradiation. The increasing yield of dominant lethal mutations was only observed in males from the most contaminated sector. Reciprocal translocations were observed in spermatocytes of mice at all the levels of contamination. The rate of reciprocal translocations was relatively low and increased linearly with the elevation of the dose rate. The extent of testis damages increased also, as the dose rate grew. The frequency of abnormal sperm heads, the yield of recessive lethal mutations, litter size and radiosensitivity of the first progeny were not changed, depending on the dose rate.

[Segregation of the somatic and germ-cell pathways of development in Drosophila]. / Segregatsiia somaticheskogo i polovogo putei razvitiia u drozofily.

Two groups of lethal mutations that are expressed only in somatic cells were isolated using the gonosomic selective method. The mutations tend to be organized in clusters along the sex chromosome. Analysis of mutation expression allowed to characterize the degree of changes in genetic diversity of somatic and germ lines in development; it is minimal in the III instar larvae and prepupae and maximal in the I and II instar larvae and pupae.

Molecular analysis of two mouse dilute locus deletion mutations: spontaneous dilute lethal20J and radiation-induced dilute prenatal lethal Aa2 alleles.

The dilute (d) coat color locus of mouse chromosome 9 has been identified by more than 200 spontaneous and mutagen-induced recessive mutations. With the advent of molecular probes for this locus, the molecular lesion associated with different dilute alleles can be recognized and precisely defined. In this study, two dilute mutations, dilute-lethal20J (dl20J) and dilute prenatal lethal Aa2, have been examined. Using a dilute locus genomic probe in Southern blot analysis, we detected unique restriction fragments in dl20J and Aa2 DNA. Subsequent analysis of these fragments showed that they represented deletion breakpoint fusion fragments. DNA sequence analysis of each mutation-associated deletion breakpoint fusion fragment suggests that both genomic deletions were generated by nonhomologous recombination events. The spontaneous dl20J mutation is caused by an interstitial deletion that removes a single coding exon of the dilute gene. The correlation between this discrete deletion and the expression of all dilute-associated phenotypes in dl20J homozygotes defines the dl20J mutation as a functional null allele of the dilute gene. The radiation-induced Aa2 allele is a multilocus deletion that, by complementation analysis, affects both the dilute locus and the proximal prenatal lethal-3 (pl-3) functional unit. Molecular analysis of the Aa2 deletion breakpoint fusion fragment has provided access to a previously undefined gene proximal to d. Initial characterization of this new gene suggests that it may represent the genetically defined pl-3 functional unit.