In the present study, we report the chromosomal localization of the human 5-HT(1F) receptor gene (HTR1F) by the analysis of somatic cell hybrids. Based upon the HTR1F cDNA sequence, a primer set that reacted with human genomic DNA but not mouse or hamster genomic DNA was derived from the relatively non-conserved 5'-untranslated and coding region. Using monochromosomal hybrid cell lines of the NIGMS Mapping Panel 2 we localized the HTR1F to human chromosome 3. To confirm the localization on chromosome 3 and to further sublocalize the HTR1F gene, a set of human cell hybrids regionally separating chromosome 3 into 7 regions was similarly analysed. Analysis of this regional panel showed that the HTR1F gene was located proximal to the 3p14.1 breakpoint in hybrid APH14 and distal to the breakpoint in 3p13 in hybrid APH13. This localizes the HTR1F gene to human chromosome 3p13-p14.1.