Combination of genomic DNA fingerprinting into the medaka specific-locus test system for studying environmental germ-line mutagenesis.

A specific-locus test (SLT) system has been established using the medaka fish (Oryzias latipes), where recessive visible mutations (at the b, lf or gu loci) detected during early embryonic stages (TM) or after hatching (VM), and dominant lethals (DL) can be examined in the same individual F1 progeny of treated parents. It was found that approximately 90% of the F1 embryos with gamma-ray-induced specific-locus mutations were concomitantly accompanied by dominant lethals irrespective of doses and germ-cell stages at the time of exposure, suggesting that DNA alterations in such mutants might include both the marker loci and region(s) responsible for dominant lethals. In contrast, embryonic lethality of the ENU (ethylnitorosourea)-induced specific-locus mutants considerably varied among ENU concentrations as well as germ-cell stages treated. Further, synergistic effect of combined treatments with gamma-rays and ENU on induction of mutations were suggested in postmeiotic male germ cells, while in spermatogonia no synergistic effect was found. DNA alterations at the 87 arbitrarily primed polymerase chain reaction (AP-PCR) markers spread over the genome were examined for individual dominant lethal embryos from 4.75 Gy-irradiated sperm or spermatids. It was found that, 14 out of 20 dominant lethal embryos lost more than one AP-PCR markers, including multiple markers located on the identical linkage group (average genetic distances, approximately 11 cM). Also found was that frequency of loss of the AP-PCR markers in the severely malformed dominant lethal embryos was higher (approximately 4.5%) than that in the slightly malformed lethal embryos (approximately 1.6%). Here, results of these studies, including previously unpublished work, are presented to illustrate the potential usefulness of the medaka SLT system for monitoring environmental mutagens.