Graf U, Abraham S K, Guzmán-Rincón J, Würgler F E
Institute of Toxicology, Swiss Federal Institute of Technology (ETH) and University of Zurich, Zurich CH-8603, Schwerzenbach, Switzerland.
Mutat Res. 1998 Jun 18;402(1-2):203-9. doi: 10.1016/s0027-5107(97)00298-4.
The fruit fly Drosophila melangaster with its well developed array of genotoxicity test systems has been used in a number of studies on antigenotoxicity of various compounds and mixtures. In recent years, the newly developed Somatic Mutation and Recombination Tests (SMART) have mainly been employed. These one-generation tests make use of the wing or eye imaginal disc cells in larvae and have proven to be very efficient and sensitive. They are based on the principle that the loss of heterozygosity of suitable recessive markers can lead to the formation of mutant clones of cells that are then expressed as spots on the wings or eyes of the adult flies. We have employed the wing spot test with the two markers multiple wing hairs (mwh,3-0.3) and flare (flr,3-38.8). Three-day-old larvae, trans-heterozygous for these markers, are treated chronically or acutely by oral administration with the test compound(s) or complex mixtures. For antigenotoxicity studies, chronic co-treatments can be used, as well as separate pre-treatments with an antigenotoxic agent followed by a chronic treatment with a genotoxin. After eclosion, the wings of the adult flies are scored for the presence of single and twin spots. These spots can be due to different genotoxic events: either mitotic recombination or mutation (deletion, point mutation, specific types of translocation, etc.). The analysis of two different genotypes (one with structurally normal chromosomes, one with a multiply inverted balancer chromosome) allows for a quantitative determination of the recombinagenic activity of genotoxins. Results of two separate studies presented: (1) instant coffee has antirecombinagenic but not antimutagenic activity in the wing spot test; and (2) ascorbic acid and catechin are able to protect against in vivo nitrosation products of methyl urea in combination with sodium nitrite.
黑腹果蝇拥有一套完善的遗传毒性测试系统,已被用于多项关于各种化合物和混合物抗原毒性的研究中。近年来,新开发的体细胞突变和重组试验(SMART)得到了主要应用。这些一代试验利用幼虫翅膀或眼睛的成虫盘细胞,已被证明非常有效且灵敏。它们基于这样的原理:合适的隐性标记杂合性的丧失会导致细胞突变克隆的形成,这些克隆随后会在成年果蝇的翅膀或眼睛上表现为斑点。我们采用了带有多个翅毛(mwh,3 - 0.3)和耀斑(flr,3 - 38.8)这两个标记的翅斑试验。对于这些标记呈反式杂合的三日龄幼虫,通过口服给予受试化合物或复合混合物进行慢性或急性处理。对于抗原毒性研究,可采用慢性联合处理,也可先用抗原毒性剂进行单独预处理,然后再用基因毒素进行慢性处理。羽化后,对成年果蝇的翅膀进行评分,统计单斑和双斑的数量。这些斑点可能是由不同的遗传毒性事件导致的:有丝分裂重组或突变(缺失、点突变、特定类型的易位等)。对两种不同基因型(一种染色体结构正常,一种带有多重倒位的平衡染色体)的分析能够定量测定基因毒素的重组活性。给出了两项独立研究的结果:(1)速溶咖啡在翅斑试验中具有抗重组活性,但不具有抗突变活性;(2)抗坏血酸和儿茶素能够联合抵御甲基脲与亚硝酸钠在体内产生的亚硝化产物。