Estradiol-17beta-Oxytocin Induced Cervical Dilation in Sheep: Application to Transcervical Embryo

Experiments were initiated to determine whether exogenous estradiol-17beta (E2) and oxytocin (OT) can be used to dilate the cervix and improve transcervical embryo transfer (ET) procedures for sheep. However, there was concern that the E2-OT treatment may alter luteal function and that embryo quality would decrease as the superovulatory response to FSH increased. In Exp. 1, 32 ewes were assigned to a 2 x 2 factorial array of treatments. On d 7, ewes received an i.v. injection of either 100 micrograms of E2 in 5 mL of 1:1 ethanol:saline or 5 mL of 1:1 ethanol:saline; 12 h later, ewes received i.v. injection of either 400 USP units of OT or saline. Jugular blood was collected on d 7, 8, 9, 10, 12, 14, 16, and 18. Progesterone concentrations were unaffected by the treatments. Experiment 2 was conducted to determine the dose of pFSH needed to induce approximately six corpora lutea (CL). Ten-day Norgestomet implants inserted between d 8-12 of the estrous cycle were used to synchronize estrus in Hampshire and Hampshire x Dorset ewes (n = 23). Ewes received a total of either 0, 18, 27, or 36 mg of pFSH, which was injected i.m. at -24, -12, 0, 12, 24, and 36 h relative to implant removal. The dose at each respective time was 19.4, 19.4, 16.7, 16.7, 13.9, and 13.9% of the total. Ewes received 400 IU of PMSG i.m. at -24 h. The CL were counted laparoscopically on d 6 (d 0 = estrus). Number of CL increased linearly (P < .01) with dose of pFSH; there were 1.8, 3.6, 6.3, and 11.2 CL/ewe, respectively. Experiment 3 was conducted to determine the effect of the E2-OT treatment, mode of transfer or the interaction of E2-OT treatment x mode of transfer on embryo survival and development. Experiment 3 was conducted over two breeding seasons and across two trials. In the first trial ewes were assigned to one of three randomized treatments. Procedural limitations that were later overcome prevented a true 2 x 2 factorial design; therefore, transcervical transfer without hormonal treatment was excluded in the first trial. In the second trial, ewes were assigned to a 2 x 2 factorial array of treatments. On d 6 of pregnancy, embryos rating a fair or better were transferred into recipients either transcervically or laparoscopically. Recipients were administered either an E2 (d 6) - OT (d 7) treatment or an ethanol:saline-saline treatment following the same protocol as in Exp. 1. Embryos were recovered on d 12 in Trial 1 and d 14 in Trial 2. Embryos were evaluated morphologically for development and ranked on a scale of one to four; one represented no development and four represented development to the morphological stages associated with the day of collection. The treatments did not affect the percentage of embryos recovered after transfer or the percentage of embryos that showed some developed. However, there was an effect of mode of transfer on mean rank of embryo development; embryos transferred laporscopically developed further than embryos transferred transcervically (P < .01). This may have been an artifact of a technician effect between trials. There was an effect of E2-OT treatment on transcervical transfer (P < .01), indicating that it may be detrimental to transfer embryos transcervically without dilating the cervix. In conclusion, the E2-OT treatment did not affect luteal function, and the E2-OT treatment can be used to dilate the cervix and enhance success of transcervical transfer of embryos. A 400 IU priming dose of PMSG and a total dose of 27 mg of pFSH can be used to induce the target number of six CL.