Effect of hydroquinone on meiotic segregation in Drosophila melanogaster females.

The induction of sex chromosomes meiotic nondisjunction (ND) by hydroquinone (HQ) given orally was investigated in Drosophila melanogaster 2-7, 8-22, 24, 48, 72 and 96 h-old females. ND was assessed by a system where exceptional females (XXY) and only 1/4 of the expected regular progeny are viable. Oocytes were treated at different stages of development. 4% HQ tested only in 72 h-old females induced ND in oocytes sampled in brood I (mostly mature oocytes at metaphase I). 6% HQ increased ND in brood I of 8-22 h-old females, while other broods, (including cells treated at early prophase) were also affected in older flies, the highest significance being attained in the 48 h-old series. Newly hatched females (2-7 h-old) were refractory to the treatment, though oocytes sampled in the first three subcultures are comparable to cells showing enhancement of ND in series run with older females. Toxicity of 2, 4 and 6% HQ increased with concentration and females' age: (a) 2% was not toxic; (b) 4% was toxic only to 72 h-old females; (c) 6% was increasingly toxic to females 24, 48 and 72 h-old. The results indicate that age plays a significant role on both chromosomal segregation and toxicity and suggest that in Drosophila HQ is metabolized to its reactive species. The lack of toxic and aneugenic effect in very young females could reflect a more efficient detoxification due to the known high specific activity of glutathione-S-transferase (GST) after eclosion. The decline in GST activity around day 2 of adult life coincides with the high effect of HQ in 48 h-old females.

Genetic damage induced by methylglyoxal and methylglyoxal plus X-rays in Drosophila melanogaster germinal cells.

The effect of methylglyoxal (MG) and MG administered prior to X-irradiation was investigated in Drosophila melanogaster germinal cells using the sex-linked recessive lethal (s.l.r.l.), II-III autosomal translocation (AT) and X-chromosome nondisjunction (ND) tests. For the s.l.r.l. test the males were either injected with MG (0.5 M, 0.75 M or 1.7 M) or fed for 24 h (1 M) and two 24 h broods (A and B) were obtained. For the AT test the males were injected with MG 1.7 M and the same brooding scheme was followed. ND was tested in females fed on MG 1 M. The only effect observed after MG treatment was a significant increase on the yield of s.l.r.l. with MG 1.7 M. In the combined treatments MG was administered prior to irradiation with 20 Gy of X-rays and the induction of s.l.r.l. and AT was assessed. Pre-treatment with MG 0.75 M and 1.7 M enhanced the frequency of s.l.r.l. in cells sampled in brood B, consisting mainly of the rather hypoxic late spermatids. It is suggested that this radiosensitizing effect could be ascribed to a decrease in the level of glutathione due to the metabolization of MG.

Evaluation of the genotoxicity of aniline.HCl in Drosophila melanogaster.

The induction of X-chromosome malsegregation, sex-linked recessive lethals and II-III autosomal translocations by aniline.HCl was investigated in Drosophila melanogaster. Nondisjunction was tested in 2 and 4 d old virgin females fed on aniline.HCl solutions (3, 5, 10 and 15%) using a system where exceptional females (XXY) and only 1/4 of the expected regular progeny are viable. After mating, the females were subcultured daily. Similarly treated 7-day-old wild-type males were used to run classical II-III translocation and recessive lethal tests; for the latter, the solutions were also injected intraabdominally. In all cases, five broods were obtained. A direct correlation was observed between concentration and toxicity. Furthermore, males were more sensitive than females, and the latter's sensitivity was higher at 4-day-old than at 2-day-old. This could be attributed to a decrease with age in the efficiency of a detoxifying mechanism, or to the generation of a toxic metabolite in older flies. Significant increases in nondisjunction were observed with 5, 10 and 15% solutions suggesting the existence of a threshold. No dose effect was detected within the range of the effective concentrations used. The increases were observed in the first subculture (representing mostly stage 14 oocyte, i.e., cells in metaphase I) and in the third subculture, representing cells in which the spindle has not yet formed, thereby pointing to a direct effect of the chemical on the chromosomes and not on the spindle. It is proposed that the second sensitivity peak detected might be the outcome of the transient loss of a protective configuration provided by the karyosome, due to its expansion in stages 9 and 10 of the developing oocytes. No sex-linked lethals or translocations were induced.

Lack of effect of acute acetaldehyde treatment on X chromosome segregation in Drosophila melanogaster females.

The effect of acute acetaldehyde treatments on X chromosome segregation was tested in germinal cells of Drosophila melanogaster females. The experiments were carried out using a test system where the nondisjunctional females (XXY) and only 1/4 of the expected regular progeny are viable. 24 h old virgin females were exposed for 60 min to 3, 4 and 5% acetaldehyde solutions by means of soaked tissue paper placed at the bottom of regular culture vials. After mating the females were brooded daily. Two additional experiments were performed with 0-2 h old and 4-5 day old virgin females using a 4% acetaldehyde solution. The results obtained show that acetaldehyde did not affect X chromosomal segregation in oocytes. This lack of effect could result from the highly efficient ADH-ALDH dependent detoxifying mechanism operating in Drosophila melanogaster.

Protective effect of ethanol on X-ray-induced mitotic recombination in Drosophila melanogaster.

The effect of ethanol pretreatment on X-ray-induced mitotic recombination in D. melanogaster females was investigated by means of the white/white+ (w/w+) spot test. White females inseminated by yellow males were allowed to oviposit for 8 hr on medium containing 5%, 7.5% and 10% (v/v) ethanol and submitted to 10 Gy of X-rays 52 hr after the beginning of the egg laying period (chronic treatments). For acute treatments 56 +/- 4-hr-old larvae grown in regular medium were held in petri dishes containing filter paper soaked with 50% (v/v) ethanol for 30 min before being irradiated with 10 Gy. The emerging heterozygous w/w+ females were inspected for the presence of white spots (LS) in their eyes. Acute ethanol pretreatments lead to a significant reduction in the frequency of LS. This is suggested to be due to the scavenging by ethanol of free radicals originating during irradiation. If so, the contribution of the indirect action of radiation to mitotic recombination induced by X-rays must be significant. Chronic ethanol pretreatments also resulted in a decrease of LS, though impairment of larval development by ethanol may have partly contributed to the effect observed. At the concentrations tested, ethanol by itself did not modify the frequency of LS observed in the control.

Nondisjunction induced by ethanol in Drosophila melanogaster females.

The effect of ethanol on chromosomal segregation was investigated in Drosophila melanogaster females homozygous for a structurally normal X chromosome marked with the recessive mutation yellow (y/y). For chronic treatments the females were kept from eclosion in food supplemented with 10% or 15% (v/v) ethanol, mated 24 or 48 h later to wild-type males and brooded in freshly prepared ethanol food. For the acute treatments 24- or 48-h-old females were exposed for 60 min to a 75% (v/v) ethanol solution by means of soaked tissue paper placed at the bottom of regular culture vials and brooded daily after mating. The results obtained show that: (1) both treatments significantly increased the frequency of X-chromosome nondisjunction; (2) after acute treatment this effect declined in later broods; (3) the yield of malformed flies in the progeny of acutely treated females was significantly higher than control values and also declined in later broods; (4) ovary analysis showed that chronic ethanol treatments caused a cessation of egg production. The induction pattern of nondisjunction and malformed flies is interpreted as giving support to the assumption that these effects may result from a direct action of ethanol. Ethanol toxicity was assessed by exposing females of different ages to a 50% or a 75% (v/v) solution for 60 min and counting the surviving flies 24 h later. The surviving fraction decreased steeply from 1-day-old (100%) to 5-day-old females (1.8%). It is suggested that toxicity may have been due to the action of a metabolite of ethanol, probably acetaldehyde.

Effect of ethanol pretreatment on genetic damage induced by X-rays in Drosophila melanogaster sperm.

Drosophila melanogaster males were treated with 96% ethanol for 45 min by means of soaked tissue paper placed at the bottom of regular culture vials before being exposed to 2 krad of X-rays. The use of ethanol was dictated by its high efficiency to scavenge hydroxyl radicals that play a substantial role in the indirect effect of ionizing radiation. The data obtained show that the frequency of sex-linked recessive lethals, reciprocal translocations and chromosome losses induced in postmeiotic cells were not reduced by ethanol pretreatment. Rather, in the combined treatments a significant increase in II-III translocations was observed in sperm. This effect declined in late and mid spermatids. Treatment with ethanol alone did not modify the frequencies of the genetic endpoints tested. It is tentatively suggested that: (i) ethanol or ethanol radicals impair the restitution of broken chromosome ends, thereby increasing the chances for rearrangement formation in the egg, or (ii) ethanol given prior to irradiation acts as a weak dose-modifying factor. If so, a slight increase in the effective dose could have resulted in a detectably higher frequency of translocations whose induction, unlike the other genetic damages investigated that increase linearly with dose, follows the slope of a 2-hit kinetic curve.

Effect of glyoxal pretreatment on radiation-induced genetic damage in Drosophila melanogaster.

The effects of glyoxal and of glyoxal pretreatments on radiation-induced genetic damage were investigated in Drosophila melanogaster mature sperm, by means of sex-linked recessive and dominant lethality, reciprocal translocation and chromosome loss tests. In addition, the possible mutagenic effect of glyoxal was assessed in postmeiotic cells up to 7 days after treatment. The results obtained show: (1) the frequencies of recessive lethals after glyoxal treatment were within control values, (2) no clastogenic effect of glyoxal was observed, (3) glyoxal pretreatment did not modify the frequency of recessive lethals induced by X-rays, (4) after pretreatment with glyoxal a consistent, though not significant, increase was seen in the frequency of reciprocal translocations in 3 replicate experiments, (5) the yield of dominant lethals and of complete and partial chromosome loss induced by radiation was significantly increased by pretreatments with glyoxal. It is suggested that the increase of the frequency of genetic endpoints resulting from chromosome breakage, when glyoxal was administered prior to irradiation, could be ascribed to: (a) a sensitizing action of glyoxal to the clastogenic effect of ionizing radiation; (b) the formation of reactive species by the interaction of glyoxal with radiation; and/or (c) interference of glyoxal with the normal handling of radiation-induced lesions in mature postmeiotic male cells.

Spontaneous second-chromosome recessive lethals in a Drosophila melanogaster repair-deficient (mei-9a) mutant.

The frequency of spontaneous second-chromosome recessive lethal mutations was determined in a Drosophila melanogaster strain kept homozygous for the X-linked mei-9 mutant for 50 generations. The data show that the rate of lethals of independent origin is 9 times higher and that of semilethals 4 times higher than the control values. The mei-9 mutant, originally isolated on the ground of its abnormal behaviour during meiosis, was later found to be defective in excision repair in somatic cells and to yield, after exposure to different physical and chemical agents, higher frequencies of genetic damage than mei-9+ flies. Our results are taken as evidence that the normal functioning of the mei-9 locus plays an important role in the repair of spontaneous lesions in the second chromosome and give support to positive findings in the X chromosome. Monitoring the accumulation of mutations in an experimental population could be a useful methodological approach to detect slight, though relevant, genetic damage induced by long-term exposure to environmental mutagens, whose detection is beyond the resolving power of conventional tests in Drosophila.

Comparative study of the clastogenic efficiency of ethyl methanesulfonate and diethyl sulfate in Drosophila melanogaster mature sperm.

Chromosome loss and translocation tests were carried out in Drosophila melanogaster sperm, stored in untreated females for up to 24 days, to compare the clastogenicity of ethyl methanesulfonate (EMS) and diethyl sulfate (DES). The sex-linked recessive lethal test was used as a "biological dosimeter" and the following results were obtained: The yield of 2-3 translocations induced by both mutagens increased steadily with storage, being significantly higher after EMS than after DES treatment. The frequencies of partial losses induced by EMS and DES were similar and increased with storage. With up to 11 days' storage, the frequency of complete loss induced by DES was higher than that induced by EMS and remained unchanged when storage was extended to 24 days. Complete loss induced by EMS increased significantly with further storage (12-24 days). With DES, complete (but not partial) loss was detected with a dose at which EMS failed to modify the control values. These data suggest that the lower recovery of II-III translocations after treatment with DES does not result from a low breaking capacity but from a diminished or delayed rejoining of the induced breaks. This could be due to a physiological impairment of the treated cells by the high toxicity of DES or to an actual lower rejoinability of the broken ends. The differential recovery of complete and partial losses after DES treatment further suggests that the mechanisms leading to the fixation of both types of damage are somehow different, and that processes intervening in the recovery of partial losses are less affected, or not at all, by the proposed reduced rejoining of chromosome breaks.

Modification of radiation-induced genetic damage in Drosophila melanogaster male germ cells by butylated hydroxytoluene.

The effects of butylated hydroxytoluene (BHT) on genetic damage induced by ionizing radiation were studied in post-meiotic male cells of Drosophila melanogaster. Prior to submitting Samarkand males to 2 krad of X-rays, BHT was administered (a) with the food (0.2 per cent final concentration) from emergence of the imago for 7 days or (b) by intra-abdominal injection (0.05 per cent) to 7-day-old adults. Dominant lethality (embryonic and total), II-III translocations and sex-linked recessive lethals were recorded. The only effect of BHT observed was a decrease in the frequency of recessive lethals induced, detected in early spermatids. Since oxygen plays an important role in the high radiosensitivity of these cells, it is suggested that the sparing action of BHT was due to its antioxidative and radical scavenging properties.