The objective was to examine the effects of presynchronization and bovine somatotropin (bST) on pregnancy rates to a timed artificial insemination protocol in lactating dairy cows. Lactating Holstein cows (n = 543) were assigned randomly in a 2 x 3 factorial experiment in which cows received a presynchronization treatment or not, and were treated with bST (500 mg) at 63 +/- 3, 73 +/- 3, or 147 +/- 3 d postpartum. The latter group was used as a control. Presynchronization treatment consisted of two injections of PGF2alpha (25 mg) given 14 d apart, with the second injection of PGF2alpha being administered 12 d before initiation of the timed artificial insemination protocol. All cows received GnRH (100 microg) at 63 +/- 3 d postpartum, an injection of PGF2alpha (40 mg) 7 d later, a GnRH injection at 48 h after PGF2alpha and were inseminated 16 to 20 h later. Cows were resynchronized if determined to be nonpregnant at ultrasonography at 32 d after insemination with a GnRH injection (100 microg), an injection of PGF2alpha (40 mg) 7 d later, and a GnRH injection at 48 h after PGF2alpha and were inseminated 16 to 20 h later. Cows were examined for pregnancy at 32 d and reexamined at 74 d after insemination. No differences in pregnancy rates were observed between cows receiving bST treatment at 63 +/- 3 d postpartum or at 73 +/- 3 d postpartum. An interaction between presynchronization and bST treatment indicated that pregnancy rates were increased for cows treated with bST when cows were presynchronized. When anestrous cows were excluded from the analyses, both an effect of bST and of presynchronization were observed, indicating that bST increased pregnancy rates regardless of presynchronization treatment and that presynchronization also increased pregnancy rates independently of bST treatment. Presynchronization and bST treatment may be used to increase first-service pregnancy rates to a timed artificial insemination protocol.