Serotonin (5-HT) plays an important role in the pathophysiology of migraine. Tryptophan hydroxylase 2 (TPH2) is the rate-limiting enzyme in the biosynthesis of 5-HT in the brain and is therefore a major factor in the availability of 5-HT and its rate of production. We hypothesized that functional TPH2 gene variants are associated with migraine. In a case-control study approach, two proven functional and two putatively functional variants of the TPH2 gene were investigated in 266 migraine patients and 153 controls. Genotype, allele, and haplotype frequencies did not differ between healthy subjects and migraineurs. A subgroup analysis for the occurrence of aura or clinical characteristics, including the number of attacks, did not reveal a positive association for the investigated polymorphisms. Our data argue against a major influence of the TPH2 gene promoter region in migraine.