Direct analysis by small-pool PCR of MS205 minisatellite mutation rates insperm after mutagenic therapies

We have used small-pool PCR to analyse mutation in samples of sperm taken f rom men after mutagenic therapy. Small-pool PCR uses direct analysis of ger mline DNA at a highly unstable tandem-repeated "minisatellite" locus to mea sure rates of length-change mutation in individual sperm samples. The advan tages of this approach are that the normal mutation rate is extremely high (about 0.4% per gamete at the locus analysed here), so that relatively smal l increases in mutation rate can be detectable in individual samples. It is known from work on sperm from untreated individuals that different alleles at this locus have different mutation rates. For this reason, we have anal ysed the germline mutation rates in sperm samples from two men, in each cas e comparing a post-treatment sample with a pre-treatment sample from the sa me individual. We find no evidence for altered mutation in the post-treatme nt sample, suggesting that the repopulation of the germ-cell compartment af ter treatment may be subject to stringent mechanisms for the detection and elimination of germ-cell damage. (C) 1999 Elsevier Science B.V. All rights reserved.