Many marine fish in polar waters produce antifreeze proteins (AFPs) to depress their serum freezing point to below that of seawater. Winter flounder from the east coast of North America contain multiple AFP gene copies organized both as tandem repeats and as linked but irregularly spaced genes, with the tandemly repeated genes encoding the bulk of the circulating AFPs. We report here on AFP gene organization in winter flounder from nine locations ranging from Long Island, NY to Conception Bay, Nfld. There are clear differences in AFP gene copy number and arrangement between some of the populations. The greatest variation is seen in the size of the tandem component in fish from the warmer, deeper locations. This contrasts to the conservation of organization in the dispersed, β-tubulin multigene family used for comparative purposes. We suggest that variation in AFP gene family size and organization reflects a relaxation of selection in some geographical areas in the postglacial period.Key words: gene dosage, multigene family, tandem repeats, intrapopulation variation, glaciation.