The Harderian gland of golden hamster (Mesocricetus auratus) secretes copious lipids, most of which is 1-alkyl-2,3-diacylglycerol (ADG). We previously reported that the composition of ADG shows marked sexual dimorphism [Seyama et al. (1995) J. Biochem. 117, 661-670]. Male ADG contains only straight chain alkyl and acyl groups, but female ADG contains a lot of branched chain ones too. In this study, we investigated the metabolism of branched chain amino acids (BCAAs) and analyzed the incorporation of the metabolites into lipids in the Harderian gland. Golden hamsters were injected intraperitoneally with [U-14C]BCAAs, and Harderian glands were obtained at 3, 6, 9, and 24 h after injection. Lipids were then extracted from the glands and analyzed. Thin layer chromatography revealed that the ADG was labeled in both sexes, but the profile depended on the sex. The cholesterol fraction was labeled only in the male gland. The alkyl and acyl groups of ADG were subjected to radio-gas liquid chromatography. As for the alkyl groups, radioactivity was detected in straight-C16 and -C18 chains in males, while branched-C17 and -C19 chains were labeled in females. As for the acyl groups, straight-C14, -C15, and -C16 chains were labeled in males, while in females, branched-C17 and -C19 chains were labeled as well as a straight-C16 chain. These results suggest that the BCAA metabolism should be regulated as to the sex at the step of branched chain acyl-CoA degradation in the Harderian gland of golden hamster, which causes the sexual dimorphism in the lipid composition in this gland.