We conducted the present study to establish a standardized method for cervical stimulation without affecting the endometrium, and to investigate the effect on estrous cycle pattern and concentrations of progesterone, oxytocin and PGF2alpha-metabolite of cervical dilatation in the mare. Six healthy Haflinger mares underwent three different treatments (control, insertion, dilatation) on Days 5 and 7 of the cycles in different orders according to a Latin square design. During dilatation, the balloon of the catheter was inflated stepwise every 30s with warm physiological saline to a maximum of 50 ml. At this stage the size of the balloon was 4.5 cm in diameter and 6 cm length. Estrous cycle length was significantly shortened by dilatation when compared to controls (control: 22.8+/-1.7, insertion: 21.8+/-2.5, dilatation: 20.0+/-1.3 days; P<0.05). Concentrations of progesterone at Days 10, 12 and 14 after ovulation were significantly lower in dilatation cycles. Calculation of the area under the curve (AUC) for progesterone secretion from Day 7 to Day 12 also revealed a significant decrease in progesterone secretion in the dilatation group (dilatation: 34.1+/-7.3, insertion: 35.6+/-7.8, control: 39.1+/-5.9 ng/ml; P<0.05). Cervical insertion and dilatation caused a rapid and pronounced increase in plasma concentrations of oxytocin from basal levels (1.0-6.1 pg/ml) to maximum peaks (insertion: 125.5 pg/ml and dilatation: 305.2 pg/ml). The AUC for oxytocin was significantly higher after insertion (Day 5: 858.4+/-469.9; Day 7: 411.9+/-213 pg/ml/h) and dilatation (Day 5: 1697+/-1725; Day 7: 1078.5+/-764 pg/ml/h) when compared to controls (Day 5: 186+/-98; Day 7: 156+/-23.5 pg/ml/h; P<0.05). Manipulations did not cause considerable changes in plasma PGF2alpha-metabolite concentrations. Because cervical dilatation up to a diameter of 4.5 cm did not cause any immediate PGF2alpha release, the luteolytic pathway is unlikely to be responsible for shortening the length of diestrus and the estrous cycle. The present data suggest an involvement of oxytocin in the shortening of the luteal phase in response to cervical manipulation.