Four activities of key microsomal steroidogenic enzymes (3 beta-hydroxysteroid dehydrogenase-isomerase, microsomal cytochrome P45017 beta, and 17-ketosteroid reductase) were compared in Leydig cells of six inbred strains of mice: A/He, CBA/Lac, C57BL/6J, DD, YT, and PT. The activities of the enzymes were found to vary considerably from one strain to another, depending on the genotype. Analysis of testosterone biosynthesis revealed a decreased activity of 17-ketosteroid reductase as compared to the activities of other microsomal steroidogenic enzymes. The metabolism of pregnenolone and progesterone demonstrated features dependent on the genotype and caused by genotypic differences in the activities of microsomal steroidogenic enzymes. These activities demonstrated a correlative interstrain variation indicative of their coordinated genetic control. The correlative variation may be related to a major gene effect on the activities of microsomal steroidogenic enzymes.