In ruminants, superovulatory treatments started at the time of follicular wave emergence result in greater and less variable ovulatory responses and embryo yields compared with the treatments begun in the presence of a large growing antral follicle(s) from the previous waves. The progesterone-oestradiol treatment is routinely used for follicular wave synchronization in cattle. The main objective of this study was to characterize the ovarian responses, hormonal profiles and in vivo embryo production in anoestrous Rideau Arcott ewes (May-June), which were superovulated after pretreatment with medroxyprogesterone acetate (MAP)-releasing intravaginal sponges and a single dose of oestradiol-17beta (E(2)-17beta). Six days after insertion of MAP sponges, eight ewes were given an i.m. injection of 350 microg of E(2)-17beta (E(2)-17beta-treated ewes); 10 ewes were given an i.m. injection of vehicle (control ewes). Multiple-dose Folltropin-V treatment, followed by the bolus injection of GnRH (50 microg i.m.), began 6 days after E(2)-17beta/vehicle injection. Transrectal ovarian ultrasonography revealed that: (i) the interval between E(2)-17beta/vehicle injection and regression of all follicles > or =5 to 3 mm in diameter was shorter (p < 0.01; 2.6 +/- 0.4 vs 4.8 +/- 0.6 days respectively); and (ii) the interval between injection and emergence of the next follicular wave was longer (p < 0.05; 5.4 +/- 0.3 vs 1.2 +/- 0.4 days, respectively) in E(2)-17beta-treated than in control ewes. During the 6 days after injection, the mean FSH peak concentration and basal FSH concentration were lower (p < 0.01) in E(2)-17beta-treated ewes. The mean ovulation rate and the number of recovered embryos did not differ (p > 0.05) between the two groups of ewes. However, the number of luteinized unovulated follicles per ewe, and the variability in the number of luteal structures and overall embryo yield were less (p < 0.05) in E(2)-17beta-treated compared with control ewes. In conclusion, the MAP-E(2)-17beta pretreatment significantly reduced the variability in ovarian responses and embryo yields, without affecting the embryo production in superovulated anoestrous ewes.
The presence of endothelin (ET) immunoreactivity and binding sites in hypothalamus and pituitary gland suggests potential neuroendocrine actions of this family of vasoactive peptides. ET-3, the predominant member of the ET family in brain, exerted significant dose-related (1, 10, and 100 nM) inhibitory effects on PRL release from dispersed anterior pituitary cells in static incubations. The effect was not dependent on voltage-sensitive calcium channels, since the dihydropyridine calcium channel antagonist nifedipine failed to block this action. Nifedipine did, however, significantly reduce the transient acute stimulatory effect of ET-3 on PRL release in cultured cells incubated in dynamic perifusion. The longer lasting inhibitory effect on PRL release that followed the brief stimulatory action was not affected by nifedipine. ET-3 also stimulated a transient but significant release of LH from cells harvested from random cycle female rats, an effect that was not antagonized by a LHRH antagonist, but was blocked by nifedipine, suggesting the mobilization of extracellular calcium as a mechanism of action of ET-3. Nifedipine also reversed the acute stimulatory effect of ET-3 on GH secretion from these cells. Cerebroventricular injections of ET-3 (6 or 60 ng) failed to significantly alter PRL or LH secretion in conscious rats, suggesting that brain-derived ET does not act within the hypothalamus to alter the release of these two hormones. Similarly, iv infusion of even pressor doses of ET-3 (10, 30, or 300 ng) failed to significantly alter PRL, LH, or GH release; thus, it is unlikely that ET of peripheral origin acts within the gland. Our results suggest that locally produced ET may act as a neuroendocrine or paracrine factor controlling pituitary function in the rat.
The synchronization of follicular waves with medroxyprogesterone acetate (MAP) and oestradiol-17beta (E(2)-17beta) prior to ovarian superstimulation in anoestrous ewes reduces the variability in superovulatory responses by an unknown mechanism. Follicle stimulating hormone (FSH) is a primary promoter of antral follicular development, but the relevance of circulating FSH concentrations to the superovulation performance in ewes has not been examined. Eighteen anoestrous Rideau Arcott ewes (May-June) were superovulated with Folltropin-V (porcine FSH), with (n = 8; treated ewes) or without (n = 10; control ewes) a single i.m. dose of 350 microg of E(2)-17beta, given on the sixth day of a 14-day treatment with MAP-releasing intravaginal sponges (60 mg). The superovulatory treatment, begun 6 days after E(2)-17beta injection, consisted of six i.m. applications of Folltropin-V given twice daily (at 08:00 and 16:00 h), followed by an i.m. injection of GnRH (50 microg). Blood samples collected every 8 h throughout the 3-day treatment, were analysed by radioimmunoassays for concentrations of ovine and porcine FSH, using species-specific standards and primary antibodies. Serum concentrations of oFSH were greater (p < 0.05) in the controls compared to treated ewes at 40, 64 and 72 h and the variability in mean oFSH concentrations was greater (p < 0.05) in control ewes at 40, 48, 64 and 72 h after the 1(st) Folltropin-V injection. There were no differences (p > 0.05) between the two groups in serum concentrations of pFSH. Significant correlations were recorded between the number of corpora lutea (CL) and oFSH concentrations at 8 h (r = 0.72, p < 0.05), 16 h (r=0.63, p < 0.05) and 64 h (r = 0.84, p < 0.01) after the 1(st) Folltropin-V injection. The total number of recovered embryos was positively correlated to oFSH concentrations at 56 h (r = 0.69, p < 0.05). We concluded that changes in endogenous FSH concentrations during ovarian superstimulation with pFSH might contribute to the variability in superovulatory responses in ewes.
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