The aim of the present study was to determine the effects of one or two doses of prostaglandin F2α (PGF) on the pregnancy rate (PR) in Simmental cows subjected to the Ovsynch or Ovsynch + Controlled internal drug release (CIDR) protocols. On d 0, 100 µg gonadorelin diacetate tetrahydrate (GnRH) was injected into cows in group 1 (n = 50) and group 2 (n = 40) and CIDR was intravaginally inserted. On d 7, 25 mg dinoprost tromethamine (PGF2α, PGF) was administered, and GnRH was injected 56 h later. Fixed-time artificial insemination (TAI) was performed 16 h after GnRH injection. In group 3 (n = 62) and group 4 (n = 63), PGF was administered 7 d following GnRH treatment, and GnRH was injected 56 h later. TAI was performed 16 h after GnRH injection. In groups 2 and 4, a second dose of PGF was injected 24 h after the first dose. Pregnancy was determined by transrectal ultrasonography on d 30 and 60 post-TAI. The pregnancy rate (PR) on d 30 post-TAI was significantly different among the groups: 46%, 55%, 29%, and 36.5%, in groups 1, 2, 3, and 4, respectively (P = 0.048). In addition, the PR was 36.3% and 43.7% in the single PGF and double PGF injection groups, respectively. There was no statistical difference in the PR on d 60 post-TAI (P > 0.05). Pregnancy loss was between 4.34% and 11.11% (P > 0.05). The highest PR in the primiparous cows was observed in group 3 (57.7%). The PR was not significantly different among multiparous cows (P > 0.05). In conclusion, two injections of PGF administered 24 h apart in Simmental cows may increase the PR in the Ovsynch or Ovsynch + CIDR protocols.