The present study aimed at instigating the buffalo ovarian and uterine response to ovsynch protocol when initiated 21 days postpartum (pp). Animals were arranged into a treated (n = 3) and a control (n = 3) group. The treated group was given the first dose of GnRH on day 0 (day 21 pp) followed by PGF2 alpha on day 7 and the second dose of GnRH on day 9. All animals were allowed a daily ultrasound examination from day 1 to day 9 and then every 12 h until detection of ovulation. The ovarian response of the treated group clearly showed the organized events of follicular growth ended by ovulation in one animal (33.3%) of the treated group; the changes in the small, medium, large and total follicle population and area did not differ significantly between the two groups; luteinization of the dominant follicle following the first GnRH injection in two animals (66.6%) indicated the reliability of the ovsynch strategy to improve the ovarian function; the uterine response was clearly evidenced by increasing the uterine wall homogeneity, decreasing the uterine lumen (less than 0.3 cm) and clearing contents by day 7 - 8 in the treated group. In conclusion, the ovsynch protocol has a beneficial effect on the ovarian function and uterine involution when applied 21 days pp in buffaloes.