The aim of this study was to investigate the protective effect of melatonin (2.5mg/kg/day, given to rats five times by intra-peritoneal injection) against damage in bone-marrow chromosomes induced by a single dose of gamma radiation (4.0 Gy whole-body irradiation; WBI). Ninety-six male albino rats were divided into four equal groups of 24 rats each. They were designated as I-non-irradiated, non-treated control rats, II-non-irradiated rats treated with melatonin for five successive days, III-whole-body gamma-irradiated rats and IV-rats injected with melatonin daily for five successive days, then subjected to whole-body gamma irradiation 2h after the final melatonin injection. Six rats from each group were sacrificed at days 1, 3, 7 and 10 following treatment and/or irradiation and their bone marrows were flushed out for micronuclei scoring and chromosomal analysis. WBI resulted in significant elevations in bone-marrow polychromatic erythrocytes containing micronuclei in their cytoplasm, and caused a significant decrease in the mitotic index of bone-marrow cells; in addition, there was a significant increase in the frequency of aberrant bone-marrow cells and in the different types of structural chromosomal aberration. Melatonin injection prior to WBI significantly reduced the mean frequencies of micro-nucleated polychromatic erythrocytes and of aberrant cells, as well as the incidence of structural chromosomal aberrations in bone-marrow cells; it also caused a highly significant elevation in the value of the mitotic index of bone-marrow cells. This investigation clarifies the protective and/or ameliorative role played by melatonin against deleterious effects of gamma radiation.