Testes determining factor Sry is encoded by the Sry locus on the Y chromosome and may be involved in the regulation of blood pressure. Here we tested the hypothesis that Sry regulates transcription of tyrosine hydroxylase (TH), the rate-limiting enzyme in the biosynthesis of catecholamines. Sry was found to be expressed in catecholaminergic regions, in male but not female rats. Co-transfection of PC12 cells with expression vector for Sry and the reporter construct [p5′TH(−773/+27)/Luc], containing 773 of the proximal nucleotides of the TH promoter directing luciferase reporter activity, led to elevation of reporter activity. The reporter activity of a shorter construct [p5′TH(−272/+27)/Luc] lacking putative Sry sites also responded to Sry. However, mutation of the AP1 site in the TH promoter greatly reduced induction by Sry, indicating that the regulation is primarily at this motif. The remaining, significantly increased expression with the mutated TH promoter construct may reflect Sry function at other sites in addition to the AP1 motif. These results reveal that Sry can regulate TH transcription and suggest that this may be one of the mechanisms of Sry mediated regulation of catecholamine biosynthesis in catecholaminergic neurons in males.