Micronuclei and chromosome aberrations in Xenopus laevis spermatocytes and spermatids exposed to adriamycin and colcemid

doi:10.1016/0027-5107(91)90030-R

Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis

Volume 247, Issue 1, March 1991, Pages 29-38

https://doi.org/10.1016/0027-5107(91)90030-R Get rights and content

Abstract

Cultured testes and spermatocytes from the frog Xenopus laevis have been incubated (40–42 h) with adriamycin or colcemid followed by quantitation of chromosome aberrations in secondary spermatocytes and quantitation of micronuclei in secondary spermatocytes, early round spermatids, and round spermatids with acrosomal vacuoles (AV) at 18–162 h of culture. Micronucleus frequencies were consistently higher in secondary spermatocytes relative to round spermatids after exposuse to either adriamycin or colcemid due to a higher rate of micronucleus formation during meiosis I compared to meiosis II. Also, some of the micronuclei formed during meiosis I did not survive meiosis II to form micronucleated spermatids. Micronucleus formation occurred in 3–7% of secondary spermatocytes with detectable chromosome aberrations, depending upon drug treatment. Thus, the ratio of micronuclei to total chromosome aberrations in secondary spermatocytes was always higher in colcemid-treated cells compared to adriamycin-treated cells following 18- and 42-h treatment periods. Adriamycin induced significant increases in micronuclei in both secondary spermatocytes and spermatids after 162 h of culture, the time for initial pachytene stages to develop into secondary spermatocytes and spermatids. The data show that cultured testes and spermatocytes from Xenopus may be used to quantify specific meiotic chromosome aberrations induced by both clastogens and spindle poisons using either a rapid secondary spermatocyte micronucleus assay or meiotic chromosome analysis.

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Cadmium but not lead exposure affects Xenopus laevis fertilization and embryo cleavage
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Citation Excerpt :
We can, however, mention the development and validation of an inhibition of sperm motility test of Xenopus laevis using divalent zinc ion as reference substance (Christensen et al., 2004). Risley and Pohorenec developed a culture of explants of Xenopus testicles to study the formation of micronuclei (genotoxicity marker) and to screen potentially genotoxic agents on germ cells (Risley and Pohorenec, 1991). Concerning the oocytes, in their review, Venturino et al. reported that exposure of Bufo arenarum oocytes to dieldrin or azinphosmethyl could reduce the fertilization rate, increase the phosphoinositide turnover or block the response of phospholipase C (Venturino et al., 2003).
Among the toxicological and ecotoxicological studies, few have investigated the effects on germ cells, gametes or embryos, while an impact at these stages will result in serious damage at a population level. Thus, it appeared essential to characterize consequences of environmental contaminant exposures at these stages. Therefore, we proposed to assess the effects of exposure to cadmium and lead ions, alone or in a binary mixture, on early stages of Xenopus laevis life cycle. Fertilization and cell division during segmentation were the studied endpoints.
Cadmium ion exposures decreased in the fertilization rates in a concentration-dependent manner, targeting mainly the oocytes. Exposure to this metal ions induced also delays or blockages in the embryonic development. For lead ion exposure, no such effect was observed. For the exposure to the mixture of the two metal ions, concerning the fertilization success, we observed results similar to those obtained with the highest cadmium ion concentration.
Flow cytometric assay for in vivo genotoxic effects of pesticides in Green frogs (Rana clamitans)
1997, Aquatic Toxicology
Frogs from farming regions in Quebec suffer a suite of physical and physiological problems associated with the use of agricultural pesticides. Flow cytometry was used to compare incidence of abnormal DNA profile, half-peak coefficient of variation (CV), and variation in genome size (pg DNA per haploid nucleus) between Green frogs (Rana clamitans) from such farming areas (corn and potato fields) and control sites dissociated from agricultural practices, to infer possible genomic manifestations of pesticide use. There was a significant increase in abnormal DNA profile in individuals from corn fields relative to the control sites (P<0.05). In all comparisons, adult frogs showed greater CVs than did juveniles (P<0.0001). Among adults, CVs were higher for samples taken from both potato and corn fields relative to the control samples (P<0.005), while in juveniles, only individuals from corn plots showed elevated CVs relative to controls (P<0.05). Juveniles showing physical deformity had significantly higher CVs than normal individuals (P<0.05), although there were no similar correlations with physiological disruption. Mean C-value (variation in DNA content) was different between adults and juveniles in all treatments (P<0.0001), but there were no significant differences in mean C-value and variance of such among similar age classes between treatments. The different classes of DNA damage found in this study are reflective of either acute or cumulative pesticide toxicity, and are exhibited by both sick and apparently normal individuals. We therefore believe flow cytometry to be a powerful technique for the measurement of pesticide-induced genomic disruption in amphibian populations.
Comparison of spontaneous structural chromosome aberration frequency in 48 h-cultured human lymphocytes mitotically arrested by different colcemid treatments
1996, Mutation Research - Environmental Mutagenesis and Related Subjects
Alterations in mitotic index and cell cycle kinetics are reported to be dependent on both the culture conditions and the ability of the lymphocytes of each individual to respond to phytohaemagglutinin stimulus. Thus, the frequency of structural chromosome aberrations (CA) could prove to be affected to some degree by these parameters. CA frequency and cell proliferation index (PI) were assessed in a group of healthy subjects after adding colcemid to cultured lymphocytes 3 h (standard) and 22 h (modified) before cell fixation at 48 h. All control cultures treated with colcemid for 22 h consisted exclusively of first metaphases, whereas the proportion of second-division lymphocytes in standard cultures (3 h colcemid) ranged from 4% to 49%. In addition, CA frequencies with and without gaps were always elevated in modified cultures as compared to the standard ones, and the difference between CA percentages obtained with the two methods was found to be significantly related with increasing PI values.
Evaluation of the genotoxic potential of selected anti-AIDS treatment drugs at clinical doses in vivo in mice
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Six anti-AIDS drugs were assessed for in vivo genotoxicity and cytotoxicity at human clinical doses with the mouse bone marrow micronucleus assay. These included four dideoxynucleosides (azidothymidine, dideoxycytidine, dideoxyadenosine, and dideoxyinosine), an anthracycline antibiotic (doxorubicin), and a chelating agent (d-penicillamine). Cytological analysis of the mouse bone marrow cells revealed: (i) The dideoxynucleosides and d-penicillamine failed to induce significant number of micronuclei, and except for one of the five doses of dideoxyinosine, none of the dideoxynucleosides were cytotoxic at the doses tested. (ii) Doxorubicin induced micronuclei in a dose-dependent manner which was statistically significant at 4-times the clinical dose but was not cytotoxic at any of the doses tested.
Effect of captopril on the cytological and biochemical changes induced by adriamycin
1993, Food and Chemical Toxicology
Captopril, an angiotensin-converting enzyme inhibitor, was evaluated for its antimutagenic potential. Male Swiss albino mice (6–8 wk old) were treated orally with different doses of captopril dissolved in water for 7 days. Some of the mice in each group were injected ip with adriamycin (ADM; 15 mg/kg body weight) and killed after 30 hr. Femoral cells of mice were collected and studied for reduction of micronuclei. Proteins, RNA and DNA were determined in hepatic cells. Captopril pretreatment was found to reduce ADM-induced micronuclei in polychromatic cells and increase the quantity of protein, RNA and DNA in hepatic cells. The inhibition of clastogenicity observed may be due to free-radical scavenging action of captopril.
Inhibition of adriamycin-induced micronuclei by desferrioxamine in Swiss albino mice
1993, Mutation Research Letters
Swiss albino male mice, 6–8 weeks old, were treated i.p. with different doses of desferrioxamine dissolved in water for 7 days. Some of the mice in each group were injected i.p. with adriamycin (15 mg/kg) and killed after 30 or 24, 48 and 72 h. The femoral cells of mice in different groups were collected and studied. Desferrioxamine treatment failed to affect the incidence of micronuclei at doses of 125–250 mg/kg/day. Pretreatment with desferrioxamine was found to provide significant protection against adriamycin-induced micronuclei without interfering with its cytotoxic potential.

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Micronuclei and chromosome aberrations in Xenopus laevis spermatocytes and spermatids exposed to adriamycin and colcemid

Abstract

Mutation Res.

Mutation Res.

J. Theor. Biol.

Mutation Res.

Mutation Res.

Mutation Res.

Mutation Res.

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Mutation Res.

Mutation Res.

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Mutation Res.