暴露于低水平γ辐射的雄性小鼠后代DNA变化的AP-PCR测定

Galina V Vasil’eva, Vladimir G Bezlepkin, Milena G Lomaeva, Nikolai P Sirota, Azhub I Gaziev
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引用次数: 11

摘要

通过对任意引物聚合酶链式反应(AP-PCR)产物指纹图谱的比较分析,研究了长期暴露于低剂量γ辐射的雄性小鼠后代(F1代)体细胞的DNA变化。暴露于10-50 cGy的雄性BALB/c小鼠在辐照后15天与未辐照的雌性交配。DNA是从2个月大的后代的尾部尖端进行的活组织检查中分离出来的。用代表微卫星和/或小卫星核心序列或其侧翼寡核苷酸的17个引物进行初步AP PCRs。用这些引物中的一个获得了基因组DNA的最佳定量复制AP-PCR指纹,该引物是位于小鼠11号染色体上的微卫星位点Atplb2侧翼的20聚体寡核苷酸。对在辐照和完整雄性后代的DNA模板上获得的AP-PCR产物的个体指纹的比较分析显示,交配前15天长期暴露于γ辐射的雄性后代的微卫星相关序列的变异性增加,DNA指纹中“非亲代带”的频率增加(在精子发生的减数分裂后阶段)。结果表明,增加的微卫星不稳定性可以由雄性亲本的辐射引发,随后产生或传递到F1代的胞体。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
AP-PCR assay of DNA alterations in the progeny of male mice exposed to low-level γ-radiation

By comparative analysis of fingerprints of arbitrarily primed polymerase chain reaction (AP-PCR) products, DNA alterations in somatic cells of the progeny (F1 generation) of male mice chronically exposed to low-doses of γ-radiation was investigated. Male BALB/c mice exposed to 10–50 cGy were mated with unirradiated females 15 days after irradiation. DNA was isolated from biopsies taken from tail tips of 2-month-old progeny. Preliminary AP-PCRs were carried out with 17 primers representing core sequences of micro- and/or minisatellites or their flanking oligonucleotides. Best quantitatively reproduced AP-PCR fingerprints of genomic DNA were obtained with one of these primers, a 20-mer oligonucleotide flanking the micro-satellite locus Atplb2 on mouse chromosome 11. Comparative analysis of individual fingerprints of AP-PCR products obtained on DNA templates from the progeny of irradiated and intact males revealed an increased variability of micro-satellite-associated sequences and an increased frequency of “non-parental bands” in DNA-fingerprints from the progeny of males chronically exposed to γ-radiation 15 days before mating (at the postmeiotic stage of spermatogenesis). The results show that increased micro-satellite instability can be initiated by irradiation of the male parent to subsequently arise or be transmitted to the soma of the F1 generations.

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