AP-PCR assay of DNA alterations in the progeny of male mice exposed to low-level gamma-radiation

By comparative analysis of fingerprints of arbitrarily primed polymerase ch ain reaction (AP-PCR) products, DNA alterations in somatic cells of the pro geny (F-1 generation) of male mice chronically exposed to low-doses of gamm a -radiation was investigated. Male BALB/c mice exposed to 10-50 cGy were m ated with unirradiated females 15 days after irradiation. DNA was isolated from biopsies taken from tail tips of 2-month-old progeny. Preliminary AP-P CRs were carried out with 17 primers representing core sequences of micro- and/or minisatellites or their flanking oligonucleotides. Best quantitative ly 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 II. Comparative analysis of individual fingerpri nts of AP-PCR products obtained on DNA templates from the progeny of irradi ated 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 gamma -ra diation 15 days before mating (at the postmeiotic stage of spermatogenesis) . The results show that increased micro-satellite instability can be initia ted by irradiation of the male parent to subsequently arise or be transmitt ed to the soma of the F-1 generations. (C) 2001 Elsevier Science B.V. All r ights reserved.