Nakano Eliana, Watanabe Liz Cristina, Ohlweiler Fernanda Pires, Pereira Carlos Alberto de Bragança, Kawano Toshie
Laboratório de Parasitologia/Malacologia, Instituto Butantan, Avenida Vital Brasil, 1500, CEP 05503-900, São Paulo, Brazil.
Mutat Res. 2003 Apr 20;536(1-2):145-54. doi: 10.1016/s1383-5718(03)00042-1.
Mutagens in the environment may represent a long-term risk for ecosystems. The reproductive potential of populations can be affected by alterations in the fecundity and offspring viability caused by germ cell mutations. Despite the ecological relevance of these effects, there are few studies on germ cell mutagenicity in natural populations. Biomphalaria glabrata was chosen for this study because of the scarcity of data on freshwater invertebrates and the ecological importance of this group. The aim of this study was to establish a germ cell mutagenicity test in B. glabrata by using a similar approach to that used in the dominant lethal test in rodents. Mitomycin C was used as a direct mutagen and cyclophosphamide as a mutagen that requires metabolic activation. Wild-type snails were exposed for 10 days to three concentrations of each agent and crossed with non-exposed albino snails at the end of the treatment. The total frequencies of malformations were analyzed in the offspring of wild-type snails; among the offspring of albino snails, only the heterozygous wild-type embryos were analyzed for malformations. Both agents induced germ cell mutations. The analysis of the offspring of the wild-type snails showed an effect of the exposure up to approximately 5 days after the end of the treatment with cyclophosphamide; the effect of mitomycin C was observed until 45 days after the end of the exposure. There was an increase in the frequencies of malformations in the wild-type offspring of the non-exposed albino snails crossed with the wild-type snails exposed to both agents. The dominant lethal test in B. glabrata proposed in this work is easy to perform, efficient, specific and sensitive in the evaluation of germ cell mutations induced by reference mutagens. The possibility of expanding its use to environmental biomonitoring studies seems very promising and worth trying.
环境中的诱变剂可能对生态系统构成长期风险。种群的生殖潜力可能会受到生殖细胞突变引起的繁殖力和后代活力改变的影响。尽管这些影响具有生态相关性,但关于自然种群中生殖细胞致突变性的研究却很少。本研究选择光滑双脐螺是因为淡水无脊椎动物的数据稀缺以及该类群的生态重要性。本研究的目的是通过采用与啮齿动物显性致死试验类似的方法,在光滑双脐螺中建立生殖细胞致突变性试验。丝裂霉素C用作直接诱变剂,环磷酰胺用作需要代谢激活的诱变剂。将野生型蜗牛暴露于每种药剂的三种浓度下10天,并在处理结束时与未暴露的白化蜗牛交配。分析野生型蜗牛后代中畸形的总频率;在白化蜗牛的后代中,仅分析杂合野生型胚胎的畸形情况。两种药剂均诱导生殖细胞突变。对野生型蜗牛后代的分析表明,在用环磷酰胺处理结束后约5天内暴露有影响;在暴露结束后45天仍观察到丝裂霉素C的影响。与暴露于两种药剂的野生型蜗牛交配的未暴露白化蜗牛的野生型后代中畸形频率增加。本研究中提出的光滑双脐螺显性致死试验在评估参考诱变剂诱导的生殖细胞突变方面易于实施、高效、特异且灵敏。将其扩展应用于环境生物监测研究的可能性似乎非常有前景且值得一试。