Use of oxytocin to attain cervical dilation for transcervical embryo transfer in sheep.

The aim of this work was to determine whether a cervical dilation protocol (CDP) composed of only oxytocin can be used to perform transcervical (non-surgical) embryo transfer in sheep (NSET) without affecting the viability of the corpus luteum (CL). Likewise, we evaluated whether a cervical transposing test with a Hegar dilator (CT Hegar test), performed at oestrous time, could be used to screen ewes for NSET (greater or lower chances to transpose the cervix). For that, oestrous and ovulation synchronization was performed in 25 Santa Inês ewes to induce the dioestrous condition. Animals went through the following CDP in a crossover design: E + OX, oestradiol benzoate (100 µg intravenously [IV]) and oxytocin (100 IU IV); OX, oxytocin (100 IU IV); and SAL, saline solution (IV). Using a Hegar dilator, cervical transposing attempts were performed at oestrous (D0) and dioestrous time (D8). The viability of the CL (morphology, luteal blood flow and progesterone values) was evaluated by ultrasonography (colour Doppler and B-mode) and by serum progesterone measurement from D7 to D13. The cervical transposing rate was lower for the SAL group (64%; 16/25; p < .05) and did not differ between the E + OX (88%; 22/25, p > .05) and OX (84%; 21/25, p > .05) groups. No treatment affected the CL viability. The CT Hegar test showed a high sensitivity (85.7%-93.3%), satisfactory accuracy (72%-84%), low false-negative rate (6.7%-14.6%), but high false-positive rate (46%-66.7%). In conclusion, a CDP protocol composed exclusively of oxytocin can lead to good cervical transposing rates and does not affect the viability of the CL. In addition, a screening test (CT Hegar) performed at oestrus can identify ewes for which cervical transposing will likely not occur at NSET.

Dose and administration protocol for FSH used for ovarian stimulation affect gene expression in sheep cumulus-oocyte complexes.

The present study evaluated the effect of four ovarian stimulation protocols on the follicular population and molecular status of cumulus-oocyte complexes (COCs). Twelve Santa Inês ewes (in a cross-over design) received 80 or 120mg FSH alone in a multiple-dose (MD80 and MD120) regimen or in combination with 300IU equine chorionic gonadotrophin (eCG) in a one-shot (OS80 and OS120) protocol. The follicular population, COC recovery rate, mean COCs per ewe and the rate of brilliant Cresyl blue-positive (BCB+) COCs were similar among treatments (P>0.05). The expression of markers of oocyte competence (ZAR1, zygote arrest 1; MATER, maternal antigen that embryo requires; GDF9, growth differentiation factor 9; BMP15, bone morphogenetic protein 15; Bcl-2, B-cell lymphoma 2; BAX, Bcl-2 associated X protein) and the steroidogenic pathway (ERα, oestrogen receptor α; LHr, LH receptor; FSHr, FSH receptor; STAR, steroidogenic acute regulatory protein) was affected by stimulation. Specifically, the expression of markers of the steroidogenic pathway was reduced with increasing FSH dose in the OS protocol. FSH at a dose of 80mg reduced the expression of FSHr and ERα in the OS versus MD protocol. Conversely, in MD protocol, only LHr was affected by increasing FSH dose. In conclusion, 80mg FSH in the MD or OS protocol was sufficient to promote the development of multiple follicles and obtain fully grown (BCB+) oocytes. The MD protocol may be more appropriate for the production of better-quality oocytes.