DETECTION OF GERM CELL DAMAGE IN RODENTS

SENSITIVE, REPRODUCIBLE AND COST-EFFECTIVE METHODS ARE NEEDED TO EVALUATE THE POTENTIAL OF CHEMICALS TO INDUCE DNA DAMAGE IN MALE AND ESPECIALLY, FEMALE GERM CELLS.THE SINGLE CELL GEL (SCG) ELECTROPHORESIS ASSAY IS A SENSITIVE METHOD FOR DETECTING SINGLE STRAND DNA BREAKS AND ALKALI-LIABILE SITES IN THE DNA OF INDIVIDUAL CELLS. USING SAMPLES CONSISTING OF A FEW THOUSAND CELLS, THIS ASSAY CAN PROVIDE INFORMATION ON THE INTERCELLULAR DISTRIBUTION AND PERSISTENCE OF DNA DAMAGE INDUCED IN MOUSE GERM CELLS (SPERMATOCYTES AND ROUND SPERMATIDS) BY CHEMICAL AGENTS. THE PURPOSE OF THE STUDY IS: (I) TO FURTHER CHARACTERIZE THE SENSITIVITY AND SUITABILITY OF THIS TECHNIQUE FOR DETECTING DIFFERENT CLASSES OF GERM CELL DNA DAMAGE IN MALE MICE EXPOSED TO MUTAGENS AND NONMUTAGENS. BLOOD LEUKOCYTES USED TO PROVIDE INFORMATION ABOUT SOMATIC VERSUS GERM CELL LEVELS OF DAMAGE: (II) TO DETERMINE IF THE TECHNIQUE CAN BEUSED TO DETECT DNA DAMAGE IN OVARIAN CELLS FROM MUTAGEN TREATED FEMALE MICE: (III) TO EXTEND THE DATA BASE TO A SECOND SPECIES (E.G., RAT). THE AVAILABILITY OF A TECHNICALLY SIMPLE MALE/FEMALE GERM CELL ASSAY WOULD GREATLYINCREASE GERM CELL RESEARCH BY GOVERNMENT AGENCIES AND THE PRIVATE SECTOR. AWARDEE'S STATEMENT OF THE POTENTIAL COMMERCIAL APPLICATIONSOF THE RESEARCH: WHEN DEVELOPED, THE SINGLE CELL GEL ASSAY WILL BE USED BY PRIVATE INDUSTRY FOR THE TESTING OF GERM CELL ACTIVITY IN PHARMACEUTICALS AND CHEMICALS. BECAUSE OF ITS SIMPLICITY OF USE, THIS TECHNIQUE WILL BE SUITABLE FOR INTEGRATION INTO EXISTING TOXICOLOGICAL STUDIES, REDUCING ANIMAL USAGE AND PROVIDING FOR A MORE COMPREHENSIVE ANALYSISOF HUMAN RISK.