A series of experiments was conducted to examine the effect of subcutaneous administration of synthetic testosterone (500 micrograms/100 g body weight) on the acidic and neutral glycolipids. The kidney glycosphingolipids were separated by using Sep-Pak cartridge column. Florisil column, and high performance thin layer chromatography, and analyzed by gas liquid chromatography. Testosterone administration increased the total content of ceramide, acidic and neutral glycolipids. The increase in total neutral glycolipids was mainly due to an increase in glucosyl- and globotetraosylceramide. The increase in total gangliosides was due to the increase in GM3 (hematoside) and "others". The relative percentage of N-glycolylneuraminic acid in GM3 was increased whereas the relative percentage of sphingoside and phytosphingosine were increased and decreased, respectively, in GM3, glucosyl- and globotetraosylceramide. The relative percentage and chain length of hydroxy and nonhydroxy fatty acids of major glycosphingolipids were altered in this organ.