Efficacy of estradiol or GnRH in combination with progesterone intravaginal devices to control the follicular wave dynamics and resulting fertility in lactating dairy cows.

This study was conducted to compare efficacy of treatments with EB or GnRH and different quantities of exogenous progesterone (P4) for synchronization of time of ovulation on follicular growth and pregnancy in lactating dairy cows. In Experiment 1, 40 cows were treated with EB or GnRH and 1.9 or 3.0 g of P4 via progesterone-containing intravaginal devices (IVPD; D0), two doses of PGF2α on D7, GnRH on D9, and TAI on D10. In Experiment 2, 1,440 cows were treated with EB or GnRH and 1 g IVPD on D0, cloprostenol, eCG and EB on D7. Cows in estrus by 48 h were AIDE, and non-estrous cows were administered GnRH and TAI 60 h after IVPD removal. Non-estrous cows were AIDE 72 h after IVPD removal. In Experiment 1, P4 was greater on D7 for cows treated with GnRH than those treated with EB. The dominant follicle was larger for cows treated with GnRH than those treated with EB. In Experiment 2, for estrous cows, pregnancy per AI was greater in cows AI at 48 h compared to 60 h after IVPD removal for cows treated with GnRH, and greater with AI at 60 h after IVPD removal compared to 48 h in EB-treated cows. In non-estrous cows, there was no effect on pregnancy. In conclusion, treatment with GnRH compared with EB resulted in increased P4 regardless of amount of exogenous P4, and there were differential proportions of estrous cows pregnant depending on time of AI after IVPD removal.

Comparison between the 5-day cosynch and 7-day estradiol-based protocols for synchronization of ovulation and timed artificial insemination in suckled BOS taurus BEEF cows.

The objective was to compare pregnancy per AI and follicular dynamic in suckled Bos taurus beef cows treated with either a 7-day progesterone + estradiol-based protocol or a 5-day progesterone CoSynch protocol for timed artificial insemination (TAI) during four breeding seasons. We hypothesized that estrous cycle status, days postpartum (DPP), fat depth and plasma progesterone concentration differentially modify the effect of treatments. Every year, 9 days before initiation of each breeding season, cows were randomly assigned to one of two groups. Cows in the 7-d P + E group (n = 428) received a progesterone intravaginal device (DIB) and estradiol benzoate on Day -9. On Day -2 the device was removed, and cows received cloprostenol and estradiol cypionate. Forty-eight hours later (Day 0) cows received TAI. Cows in the 5-d P + CoS group (n = 428) received a DIB, and GnRH on Day -8. On Day -3, the device was removed, and cows received cloprostenol. A second dose of cloprostenol was given on Day -2. Cows received GnRH and TAI 72 h after device removal (Day 0). On Day -9, estrous cycle status was determined. In a subset of cows (n = 79) the size of the dominant follicle was determined between Days -2 and 0. In another subset of cows (n = 340), DPP, fat depth (mm) and plasma progesterone concentration (ng/mL) were evaluated on Day -9. Pregnancy per AI was determined 30 d after TAI. Pregnancy per AI was greater for cows in the 5-d P + CoS group than for cows in the 7-d P + E group (50.9% vs. 41.3%, P = 0.01) and was also greater in cyclic than in anestrus cows (54.3% vs. 33.2%, P < 0.0001). There was also a significant effect of breeding season (P = 0.0002) and sire (P = 0.03), and an interaction between treatment group and breeding season (P = 0.03). The dominant follicle was larger (P < 0.0001) in cows in the 5-d P + CoS group than the 7-d P + E group (10.7 ± 0.29 mm vs. 9.0 ± 0.28 mm). Pregnancy per AI was greater in cows with ≥55 DPP (47.0% vs. 29.6%, P = 0.001), fat depth ≥0.50 mm (44.7% vs. 29.7%), and with plasma progesterone concentration ≥1 ng/mL (47.2% vs. 28.7%, P = 0.01). In cows with plasma progesterone ≥1 ng/mL on Day -9, pregnancy per AI was greater in the 5-d P + CoS group (60.5%) than in the 7-d P + E group (34.9%), but there was no difference between treatment groups in cows with plasma progesterone < 1 ng/mL (P = 0.07). In conclusion, the 5-d P + CoS protocol resulted in greater size of the dominant follicle and pregnancy per AI in suckled Bos taurus beef cows subjected to TAI.

Extending the duration of treatment with progesterone and equine chorionic gonadotropin improves fertility in suckled beef cows with low body condition score subjected to timed artificial insemination.

The objective of this study was to evaluate the effect of an extended progesterone treatment on follicular development and fertility in postpartum, suckled beef cows subjected to timed artificial insemination (TAI). In experiment 1, cows (n = 24) with body condition score (BCS) ≥4.5 received either a 2-g progesterone intravaginal device on Day -23 or a 0.558-g progesterone intravaginal device on Day -9. Then, all cows received 2 mg of estradiol benzoate on Day -9; removal of the device, 1-mg estradiol cypionate, and PGF2α on Day -2; and TAI on Day 0. Metabolic status was assessed between Days -9 and -2. Ovarian structures and plasma progesterone were determined weekly from Day -23 to -9, daily from Day -9 to 0, and weekly until Day 28. In experiment 2, cows (n = 302) with BCS ≥4.5 received identical treatment to cows in experiment 1, but on Day -2, cows received 400 IU of two different commercial preparations of equine chorionic gonadotropin (eCG). Ovarian structures were determined on Days -23 and -9 on a subset of cows (n = 40). Pregnancy was determined 39 days after TAI. In experiment 3, multiparous cows (n = 244) with BCS <5.0 received identical treatment as cows in experiment 1 initiated on Day -18, and on Day -2, cows received 400 IU of eCG or no treatment. Ovarian structures were determined in a subset of cows (n = 31) on Days -3, -2, -1, 0, 1, and on Day 10. Pregnancy was determined 39 days after TAI. The results indicated that in experiment 1, plasma progesterone was higher in treated than nontreated (control cows) during the first 14 days (P = 0.0001). The extended progesterone treatment increased the size of the largest follicle between Days -23 and Day -5 (Group by Day, P = 0.04) and tended to increase the size of the dominant follicle from Day -5 to Day -1 (Group by Day, P = 0.06). There was no effect of metabolic status or interaction between metabolic status and day on follicular growth. In experiment 2, extended progesterone treatment tended to increase the size of the largest follicle between Day -23 and -9 (P = 0.06). There was no effect of Group, eCG, BCS, and parity on pregnancy per AI. In experiment 3, extended progesterone treatment combined with eCG increased the size of the dominant follicle (P = 0.01). Both extended progesterone treatment (P = 0.02) and eCG (P = 0.03) increased pregnancy per AI. In conclusion, an extended progesterone treatment stimulated follicular growth postpartum and improved fertility only in cows with low BCS.

Differential effects of leptin on ovarian metalloproteinases and their tissue inhibitors between in vivo and in vitro studies.

In this study, we investigated the effect of leptin on the ovarian metalloproteinase system in the rat during the ovulatory process. Ovulation was induced in immature rats primed with gonadotropins. In both in vitro and in vivo experiments, we measured i) the protein expression of the ovarian metalloproteinases (matrix metalloproteinases, MMPs) and their tissue inhibitors (TIMPs) by western blot; ii) the gelatinase activity of the ovarian MMPs by zymography; and iii) the inhibitory action of TIMPs by reverse zymography. Using cultures of ovarian explants, leptin increased the activity but not the protein expression of MMP-2 and MMP-9 in both culture medium and ovarian tissue, and the protein expression of TIMPs, without a higher inhibitory action of the gelatinase activity. These results suggest either that the increase in TIMP proteins was not sufficient or that the inhibitory actions of TIMPs were impaired to suppress the MMP activity when the ovaries were directly exposed to leptin. To study the in vivo effect, rats received an acute treatment with high doses of leptin to inhibit ovulation. This treatment increased the expression of both the latent and the active forms of MMP-2 but did not result in a greater activity of MMP-2. In addition, the inhibitory action of TIMP-2 was also increased by this treatment. These results suggest that the administration of high doses of leptin could be regulating the follicle wall degradation, at least in part, by increasing the action of the ovarian TIMP-2 as a result of an extraovarian mechanism or signaling pathway.

Leptin modulates the expression of its receptors in the hypothalamic-pituitary-ovarian axis in a differential way.

To investigate the expression of leptin receptors (Ob-R) in the rat hypothalamus-pituitary-ovarian axis, immature rats were treated with eCG/hCG and Ob-R expression was evaluated by western blot analysis. The Ob-R expression increased 24 h after eCG administration in all the tissues assayed. In the hypothalamus, these levels immediately decreased to those obtained without treatment. In the pituitary, the Ob-R expression continued to be elevated 48 h after eCG administration, whereas the hCG injection did not modify these levels. Similar results were obtained with the ovarian long isoform. To assess the effect of leptin on its receptors, Ob-R was assessed in hypothalamus, pituitary and ovarian explants cultured in the presence or absence of leptin (0.3-500 ng/ml). In the hypothalamus, we found a biphasic effect: the Ob-R expression was either reduced or increased at low or high concentrations of leptin respectively. LH-releasing hormone secretion increased at 1 ng/ml. In the pituitary, Ob-R increased at 10 or 30 ng/ml of leptin for the long and short isoforms respectively. Leptin also induced an increase in LH release at 30 ng/ml. In the ovarian culture, the presence of leptin produced an increase in Ob-R expression at different ranges of concentrations and a dose-dependent biphasic effect on the progesterone production. In conclusion, all these results clearly suggest that leptin is able to modulate the expression of its own receptors in the reproductive axis in a differential way. Moreover, the positive or negative effect that leptin exerts on the ovulatory process may be dependent on this regulation.

Expression of different abscisic acid-responsive genes during somatic embryogenesis in sugarcane (Saccharum officinarum).

We have examined the expression of four genes during somatic embryogenesis in two cultivars of sugarcane, Saccharum officinarum, one drought resistant (JA-605) and the other one sensitive (C-8751), as well as in embryogenic and nonembryogenic tissues treated with abscisic acid (ABA) or after drought stress. Three of the cDNAs probed, from plasmids pMA1049, pMA2005 and CM1, were lea genes and the other one, from a barley hemoglobin gene, was induced in other species under stress conditions. Only transcripts homologous to the pMA2005 and CM1 clones were differentially accumulated during somatic embryogenesis; moreover, the CM1 clone only appeared in somatic embryos of the non-resistant variety. All the lea cDNAs were dramatically increased in the embryogenic tissues treated with ABA, but we observed differences among the accumulation of these mRNAs in the nonembryogenic tissues treated with ABA, and only the pMA2005 transcript appeared. The mRNA homologous to the barley hemoglobin gene appeared in the nonembryogenic tissue and the expression increased after ABA treatment. The desiccation treatments had different effects on the varieties, and the pMA2005 transcripts only appeared in the sensitive variety.