RESUMEN
This study evaluated the effect of prostaglandin F2α (PGF2α) associated with gonadotropin-releasing hormone (GnRH) for ovulation induction in precocious indicus heifers submitted to a fixed-time superovulation (SOV) programme. Precocious Nellore heifers (n = 35), aged 13 months, were subjected to the SOV protocol. On day 0 (D0), all animals received intravaginal insertion of a progesterone (P4) device along with intramuscular administration of 2 mg of oestradiol benzoate, plus 200 IU of follicle-stimulating hormone in decreasing doses, with 12-h intervals between D4 and D7, in addition to 150 µg of D-cloprostenol on D6 and device removal on D7. On D8, the donors received 10.5 µg of buserelin acetate and the treatment group received 300 µg of D-cloprostenol/PGF2α. Artificial insemination was performed 12 h and 24 h after GnRH administration using frozen semen. On D15 of the protocol (i.e., D7 after insemination), the embryos were collected and evaluated. All animals passed through the control and treatment groups. Results were evaluated by analysis of variance using an adjusted mixed-effects model (p < 0.05). There was no difference in the total number of embryos between the control and treatment groups (10.40 ± 1.52 vs. 9.60 ± 1.36; p = 0.63) or viable embryos (6.30 ± 1.22 vs. 4.30 ± 0.71). For precocious indicus heifers, treatment with PGF2α in association with GnRH did not affect embryo production in the fixed-time SOV protocol.
Asunto(s)
Dinoprost , Estradiol , Hormona Liberadora de Gonadotropina , Inseminación Artificial , Inducción de la Ovulación , Progesterona , Superovulación , Animales , Bovinos , Femenino , Dinoprost/farmacología , Dinoprost/administración & dosificación , Hormona Liberadora de Gonadotropina/farmacología , Hormona Liberadora de Gonadotropina/administración & dosificación , Superovulación/efectos de los fármacos , Inseminación Artificial/veterinaria , Inducción de la Ovulación/veterinaria , Inducción de la Ovulación/métodos , Estradiol/farmacología , Estradiol/administración & dosificación , Estradiol/análogos & derivados , Progesterona/farmacología , Progesterona/administración & dosificación , Embarazo , Cloprostenol/farmacología , Cloprostenol/administración & dosificación , Buserelina/farmacología , Buserelina/administración & dosificación , Hormona Folículo Estimulante/farmacología , Hormona Folículo Estimulante/administración & dosificaciónRESUMEN
This study evaluated the use of the GnRH agonist hormone, deslorelin, to control the follicular population before initiating multiple ovulation and embryo transfer (MOET) treatment. Twenty-four cross-bred Santa Inês ewes, aged between 2 and 4 years, were randomly assigned to either a control group (n = 11) or a treated group (n = 13). All ewes received an intravaginal device containing 60 mg of medroxyprogesterone acetate on day 0, and a new device on day 7, which remained in place until day 14. Additionally, the ewes were administered 125 µg of cloprostenol on day 7. The superovulatory treatment involved administering 200 mg of pFSH, divided into eight decreasing doses at 12-h intervals starting on day 12. On day 14, 300 IU of eCG was administered. In the deslorelin group, three doses of 100 µg of deslorelin were administered starting on day 3 after the insertion of the vaginal device, with subsequent doses given at 72-h and 144-h intervals. Natural mating was performed 36 h after the removal of the progesterone implant using males with proven fertility. Embryo collection took place on the 6th day after mating, and the recovered structures were quantified and evaluated for quality and developmental stage. Transrectal ultrasonography was conducted on days 12, 16 and 21 to evaluate the ovaries, specifically to assess the ovarian follicular population and the presence of the corpus luteum. Ewes in the control group had higher embryo recovery rates (p < .01) compared to the treated group (5.2 ± 0.8 vs. 1.1 ± 0.8), with differences observed primarily in the number of morulae. The number of corpus luteum observed during the laparotomy on day 21 was significantly higher (p < .01) in the control group (10.44 vs. 4.5 corpus luteum per ewe). Yet, the treated group had a significantly higher number of follicles (p < .05) on the first day of pFSH application (5.5 vs. 3.0 follicles per ewe). In conclusion, although the inclusion of deslorelin in the superovulation protocol resulted in increased synchronization of oestrus and follicle number, it did not lead to an increase in the number of corpus luteum or harvested embryos.
Asunto(s)
Transferencia de Embrión , Hormona Folículo Estimulante , Superovulación , Pamoato de Triptorelina , Animales , Femenino , Pamoato de Triptorelina/análogos & derivados , Pamoato de Triptorelina/farmacología , Pamoato de Triptorelina/administración & dosificación , Superovulación/efectos de los fármacos , Hormona Folículo Estimulante/farmacología , Hormona Folículo Estimulante/administración & dosificación , Transferencia de Embrión/veterinaria , Cloprostenol/farmacología , Cloprostenol/administración & dosificación , Embarazo , Ovario/efectos de los fármacos , Folículo Ovárico/efectos de los fármacos , Oveja Doméstica , Ovinos/fisiología , Hormona Liberadora de Gonadotropina/farmacología , Hormona Liberadora de Gonadotropina/administración & dosificación , Acetato de Medroxiprogesterona/farmacología , Acetato de Medroxiprogesterona/administración & dosificaciónRESUMEN
Hormonal protocols based on progestogens and equine chorionic gonadotrophin (eCG) are efficient for estrus and ovulation synchronization in ewes. Although eCG is indispensable during seasonal anestrus, it may not be necessary during the breeding season. Thus, we tested the hypothesis that GnRH is effective in replacing eCG during the breeding season allowing satisfactory ovulation rate, luteal function and conception rates after timed artificial insemination (TAI). Ewes (n = 134) with a minimum body condition score of 2.5 (0-5 scale) were treated with intravaginal devices (IVD) containing 60 mg of medroxyprogesterone acetate (MPA) for seven days and received 0.26 mg of sodium cloprostenol at the time of IVD removal. In Exp. 1, at IVD removal, ewes (n = 29) were allocated to three groups: eCG (200 IU at IVD removal; n = 10); eCG+GnRH (200 IU eCG at IVD removal and 4 µg of buserelin 36 h later; n = 10); or GnRH (buserelin 36 h after IVD removal; n = 9). Blood samples were collected 2, 6 and 12 days after TAI moment (54 h after IVD removal), for progesterone (P4) analysis. In Exp 2, the ewes were allocated to eCG (n = 10) or GnRH (n = 10) groups, as above described, and ovulation moment was evaluated 54, 66 and 78 h after IVD removal. In Exp 3, TAI was performed in ewes from eCG (n = 45) and GnRH (n = 40) groups using 100 × 106 motile spermatozoa from a pool of semen collected from four rams. In Exp. 1, based on P4 levels, we confirmed that all the ewes ovulated (29/29) and there was no significant effect of group (P = 0.89) or group x day (P = 0.18) on P4 concentration, being observed a significant effect of day (P = 0.0001). In Exp. 2, the maximum DF diameter (P = 0.26) and ovulation moment (P = 0.69) did not differ between groups. In Exp. 3, pregnancy rate was significantly lower (P = 0.02) in GnRH (22.5 %; 9/40) compared to eCG (46.7 %; 21/45). The results indicate that, although ovulation and luteal function were not altered after eCG, eCG+GnRH or GnRH treatment, GnRH alone before TAI cannot be used to replace eCG treatment during the breeding season.
Asunto(s)
Gonadotropina Coriónica , Sincronización del Estro , Hormona Liberadora de Gonadotropina , Inseminación Artificial , Animales , Femenino , Inseminación Artificial/veterinaria , Hormona Liberadora de Gonadotropina/farmacología , Hormona Liberadora de Gonadotropina/administración & dosificación , Ovinos/fisiología , Gonadotropina Coriónica/farmacología , Gonadotropina Coriónica/administración & dosificación , Embarazo , Sincronización del Estro/métodos , Progesterona/sangre , Progesterona/farmacología , Progesterona/administración & dosificación , Estaciones del Año , Acetato de Medroxiprogesterona/administración & dosificación , Acetato de Medroxiprogesterona/farmacología , Ovulación/efectos de los fármacos , Ovulación/fisiología , Gonadotropinas Equinas/farmacología , Gonadotropinas Equinas/administración & dosificaciónRESUMEN
Two experiments evaluated the effect of different hormonal treatments to synchronize follicle wave emergence on follicle dynamics and pregnancies per AI (P/AI) in estradiol (E2)/progesterone (P4) timed-AI (TAI) protocols in lactating dairy cows. In Experiment 1, lactating, primiparous Holstein cows (n = 36) received a P4 releasing device (Day 0) and were allocated at random to one of the following three treatment groups: Group EB received 2 mg E2 benzoate (EB) intramuscularly (i.m.), Group EB + GnRH received 2 mg EB+20 µg buserelin (GnRH) i.m., or Group EB + P4 received 2 mg EB + 100 mg of injectable P4 (iP4) in oil i.m. All cows received 0.150 mg D-Cloprostenol on Days 7 and 8 followed by P4 device removal, 400 IU eCG and 1 mg ECP on Day 8. Daily ultrasound examinations revealed that although the interval from P4 device removal to ovulation was not affected by treatment, cows that received EB + GnRH had an earlier (P < 0.05) emergence of the new follicular wave (Day 2.6 ± 0.2) than the other two treatment groups (Days 3.5 ± 0.3 and 6.1 ± 0.3, for EB and EB + P4, respectively). In Experiment 2, 808 lactating cows were assigned randomly to the three treatments evaluated in Experiment 1, and all the cows were TAI to determine P/AI. Cows in the EB + GnRH group had greater P/AI (57.4 %, P < 0.01) than those in the EB (44.6 %) or EB + P4 (45.7 %) groups. In conclusion, the administration of GnRH, but not iP4, on the day of insertion of a P4 device improves P/AI in lactating dairy cows synchronized for TAI with an estradiol/P4-based protocol.
Asunto(s)
Estradiol , Sincronización del Estro , Hormona Liberadora de Gonadotropina , Inseminación Artificial , Lactancia , Folículo Ovárico , Progesterona , Animales , Bovinos/fisiología , Femenino , Inseminación Artificial/veterinaria , Inseminación Artificial/métodos , Lactancia/efectos de los fármacos , Folículo Ovárico/efectos de los fármacos , Folículo Ovárico/fisiología , Progesterona/administración & dosificación , Progesterona/farmacología , Estradiol/farmacología , Estradiol/administración & dosificación , Estradiol/análogos & derivados , Sincronización del Estro/métodos , Embarazo , Hormona Liberadora de Gonadotropina/farmacología , Hormona Liberadora de Gonadotropina/administración & dosificación , Buserelina/farmacología , Buserelina/administración & dosificaciónRESUMEN
This study evaluated the effects of dose of equine chorionic gonadotropin (eCG) and its splitting in different days of the synchronization protocol on reproductive performance of primiparous and multiparous Nellore cows. In the present study, 2,536 Nellore cows (1,634 primiparous and 902 multiparous) were assigned to receive in a 2 × 2 factorial design 1) an intravaginal progesterone (P4) device and 2.0 mg of estradiol benzoate (EB) on day -11, 12.5 mg (i.m.) of dinoprost tromethamine (PGF), 300 IU (i.m.) of eCG, 0.6 mg (i.m.) of estradiol cypionate (ECP), and P4 device withdrawal on day -2, followed by TAI on day 0 (n = 632 cows, being 409 primiparous and 223 multiparous; 300-2), 2) 300 IU (i.m) of eCG administered on days -4 and -2 (150 IU of eCG/day; n = 637 cows, being 412 primiparous and 225 multiparous; 300-4-2), 3) 400 IU (i.m.) of eCG administered on day -2 (n = 633 cows, being 406 primiparous and 227 multiparous; 400-2), and 4) 400 IU (i.m) of eCG administered on days -4 and -2 (200 IU of eCG/day; n = 634 cows, being 407 primiparous and 227 multiparous; 400-4-2). Individual cow BCS was assessed on days -11, 0 (timed-AI), and 31 of the study. Body condition score of the animals was classified into LOW or HIGH using the threshold of 2.75 (≤2.75 = LOW; >2.75 = HIGH). For primiparous cows, an eCG splitting effect was observed on follicle size, as cows receiving eCG on days -4 and -2 of the synchronization protocol had a larger follicle than cows administered eCG only on day -2. For day 31 P/AI, primiparous cows receiving 400-4-2, regardless of BCS, had a greater P/AI than cows from other treatments. Administering 400-4-2 to LOW BCS cows also resulted in greater P/AI than all other treatments assigned to LOW BCS cows. For multiparous cows, no treatment effect was observed for follicle size, estrus expression, and day 31 P/AI (P ≥ 0.21). In summary, increasing the dose and splitting the dose of eCG positively impacted the pregnancy rates of primiparous cows under a BCS ≤2.75, but no effects were detected on multiparous cows.
Asunto(s)
Progesterona , Reproducción , Embarazo , Femenino , Bovinos , Animales , Caballos , Progesterona/farmacología , Estradiol/farmacología , Índice de Embarazo , Dinoprost/farmacología , Inseminación Artificial/veterinaria , Inseminación Artificial/métodos , Gonadotropina Coriónica/farmacología , Sincronización del Estro/métodos , Hormona Liberadora de Gonadotropina/farmacologíaRESUMEN
To date, there have been no studies testing the capacity of GnRH analogs and respective doses to induce a LH peak in sheep. In this sense, the present study aimed to evaluate the capacity of different synthetic forms and doses of GnRH in inducing LH release in sheep, and the effect of GnRH administration at timed artificial insemination (TAI) on pregnancy per timed-AI. In experiment 1, ewes (n = 40) received an intravaginal device (IVD) of medroxyprogesterone acetate (MPA; 60 mg) for 7 d and prostaglandin F2α analog on Day 5. On Day 7, the ewes were allocated randomly into one of eight groups (n = 5/group), which received a GnRH analog at a specific dose, as follows: lecirelin (12.5 or 25 µg), gonadorelin (50 or 100 µg), buserelin acetate (4.2 or 8.4 µg), or deslorelin (375 or 750 µg). Blood samples for LH determination were obtained at 0, 2, 4, and 6 h after GnRH and the IVDs were removed after the last blood collection. The maximal LH concentration induced by gonadorelin at doses of 50 µg and 100 µg (12.0 ± 2.4 ng/mL and 28.6 ± 7.1 ng/mL, respectively) was lower (P < 0.05) than serum LH induced by 8.4 µg of buserelin (78.9 ± 12.9 ng/mL), 375 µg and 750 µg of deslorelin (75.6 ± 7.4 ng/mL and 72.1 ± 10.6 ng/mL, respectively) and 12.5 µg and 25 µg of lecirelin (73.3 ± 17.8 ng/mL and 61.6 ± 5.9 ng/mL, respectively). However, the maximal LH concentration induced by 4.2 µg of buserelin (49.4 ± 5.9 ng/mL) was similar (P > 0.05) to the 100 µg of gonadorelin. The total release of LH (area under the curve - AUC) after treatment with 50 µg of gonadorelin (31.7 ± 5.9 ng h/mL) was lower (P < 0.05) than after other agonists. In a second experiment, 330 ewes were treated with IVD containing MPA for 7 d. Simultaneously with IVD removal, 250 µg of cloprostenol and 200 IU of eCG were administered. Then, ewes were assigned randomly to either no further treatment (control); or to receive 4.2 µg of buserelin acetate (GnRH group) at cervical TAI, which was performed with fresh semen 54 h after IVD withdrawal in all the animals. Higher pregnancy per timed-AI was observed for GnRH (50.3 %) compared to control (40.7 %). We conclude that buserelin acetate (8.4 µg), lecirelin (12.5 and 25 µg) and deslorelin (375 and 750 µg) induced a greater stimulatory effect on LH secretion than gonadorelin treatment. Furthermore, buserelin acetate treatment at TAI increased pregnancy per timed-AI in ewes previously treated with MPA and eCG.
Asunto(s)
Buserelina , Sincronización del Estro , Embarazo , Femenino , Ovinos , Animales , Buserelina/farmacología , Hormona Liberadora de Gonadotropina/farmacología , Acetato de Medroxiprogesterona/farmacología , Inseminación Artificial/veterinaria , Prostaglandinas F/farmacología , Progesterona , Dinoprost/farmacologíaRESUMEN
Two experiments were performed to evaluate the effects of GnRH treatment on the fertility of suckled Nelore beef cows treated with an estradiol/progesterone (E2/P4)-based protocol for timed artificial insemination (TAI). Experiment 1 focused on determining the effects of estradiol cypionate (EC) on ovulation in TAI cows treated with GnRH 34 h after removal of the intravaginal P4 device (IPD). Suckled cows (n = 26) were treated with 2 mg estradiol benzoate (EB) and IPD containing 1 g P4. After 8 days, IPDs were removed, and all cows were treated with 150 µg of d-cloprostenol (prostaglandin F2 alpha analog) and 300 IU of equine chorionic gonadotropin (eCG), then separated into two treatment groups consisting of cows who received 1) saline 0.9% i.m. (GnRH34 group) or 2) 0.6 mg i.m. of EC (EC-GnRH34 group). On day 9 (05:00 p.m.), all cows were given GnRH (10.5 µg of buserelin acetate) i.m. No differences were observed between the groups (P > 0.05) in the time of ovulation after IPD removal or in the proportion of cows ovulating. Experiment 2 focused on determining the effects of GnRH34 along with or in the absence of EC on day 8 on pregnancy per AI (P/AI) in postpartum beef cows. Cows (n = 981) were treated similarly to those in Experiment 1, but an additional group, the EC-GnRH48 group, was included, in which cows received EC on day 8 whereas those that did not show estrus received GnRH at TAI. Thus, in this experiment, groups consisted of GnRH34 (n = 322), EC-GnRH34 (n = 335), and EC-GnRH48 (n = 324). A higher rate of estrus expression was observed in cows treated with EC following IPD removal (EC-GnRH34: 69%, EC-GnRH48: 64.8%) than in cows in the GnRH34 group (45.6%). No difference in P/AI was observed between the treatment groups (P = 0.45), but P/AI in cows in the EC-GnRH34 group (64.2%) tended to be greater (P = 0.1) than in cows in the GnRH34 group (58%). In summary, although ovulation synchrony did not differ among the groups, P/AI in cows treated with EC and GnRH 34 h after IPD removal tended to be greater than in cows treated solely with GnRH; this was most likely due to a shorter proestrus/estrus period, considering the lower proportion of cows that displayed estrus in the GnRH34 group. Finally, given that P/AI did not differ between the EC-GnRH34 and EC-GnRH48 groups, our results suggest that, for cows not displaying estrus, administration of EC at the time of IPD removal followed by treatment with GnRH 48 h afterward represents the most cost-efficient TAI strategy for South American Zebu-based beef operations.
Asunto(s)
Estradiol , Progesterona , Embarazo , Femenino , Bovinos , Animales , Caballos , Progesterona/farmacología , Estradiol/farmacología , Hormona Liberadora de Gonadotropina/farmacología , Buserelina , Inseminación Artificial/veterinaria , Inseminación Artificial/métodos , Sincronización del Estro/métodosRESUMEN
In this study, we aimed to propose changes in the protocol of cultured Astyanax altiparanae hypophysation to increase the maximum ovulation rate of 60% registered previously. To that two consecutive experiments were conducted. In the first experiment, three carp pituitary homogenate (CPH) doses (3, 6, and 9 mg/kg) were administered in a single injection, while in the second experiment, the 6 mg/kg CPH dose was tested either in single or double injections. In the first experiment, a single injection of 3 mg/kg CPH did not induce final oocyte maturation or spawning, while a dose of 6 mg/kg CPH resulted in an increase in the plasma level of prostaglandin (PGF2α) at ovulation. The single higher dose of 9 mg/kg CPH did not improve reproductive performance and even though anticipated the resumption of meiosis it was detrimental to the spawning rate. In the second experiment, the dose of 6 mg/kg CPH fractionated into two injections led to a higher spawning rate, spawning volume per female body mass, frequency of post-ovulatory complexes, and PGF2α concentration at ovulation compared to the single injection. The most effective treatment remained the 6 mg/kg of CPH fractionated into two injections, but still providing very low proportion of ovulated females (â¼40 %). Overall, this study indicates that the spawning protocols for this species need to be improved to induce ovulation in a larger number of females and be more potent in those females that respond positively.
Asunto(s)
Carpas , Dinoprost , Femenino , Animales , Dinoprost/farmacología , Oocitos , Reproducción , Ovulación , Oogénesis , Hormona Liberadora de Gonadotropina/farmacologíaRESUMEN
This study aimed to compare milk production and reproductive performance in high yield Holstein cows that lose BCS early and late in the postpartum period. Lactating dairy cows (n = 76) were received first timed AI at 60 to 75 DIM using the farm-managed estradiol-progesterone-GnRH-based timed AI protocol. The BCS of all cows was daily evaluated by automated BCS cameras. Aiming to evaluate the effect of the days in milk (DIM) in which a cow reached the nadir BCS on the reproductive parameters, cows were separated into two groups: early BCS loss (n = 42), cows that reached the nadir BCS ≤ 34 DIM, and late BCS loss (n = 34), cows that reached the nadir BCS > 34 DIM. The optimal cut-off point for determining the relationship between days to nadir BCS and pregnancy by 150 DIM (P150) was calculated using the receiver operating characteristic (ROC) curve. From the ROC analysis, the cut-off was 34 DIM (Se, 80.9%; Sp, 66.7%; AUC, 0.74; P < 0.01). No differences (P>0.05) were detected between groups on the BCS and milk production. The average of milk production in both groups was 46.65 ± 6.15 Kg/day. Cows that reached the nadir BCS early postpartum presented lower (P < 0.01) calving interval and greater (P < 0.01) pregnancy at first AI and P150. In summary, cows that lost BCS early had better reproductive performance and had similar milk yield compared with cows that lost BCS late in the postpartum period.
Asunto(s)
Lactancia , Reproducción , Embarazo , Femenino , Bovinos , Animales , Leche , Hormona Liberadora de Gonadotropina/farmacología , Periodo Posparto , Inseminación Artificial/veterinariaRESUMEN
This study was designed to evaluate whether the utilization of a second PGF2α treatment at the end of an estradiol/progesterone (E2/P4)-based protocol with or without GnRH at the beginning of the protocol would improve pregnancy rates of lactating Holstein cows assigned to timed embryo transfer. A total of 501 lactating Holstein cows in 5 farms were enrolled in the experiment. Within farm, cows were blocked by parity and, within block, were assigned randomly to (1) insertion of an intravaginal P4 device (controlled internal drug-releasing device; CIDR) and estradiol benzoate on d -11, PGF2α on d -4, CIDR withdrawal and an injection of estradiol cypionate on d -2, and timed embryo transfer on d 7 (1-PGF; n = 164); (2) the same treatments as 1-PGF, but with PGF2α administered on d -4 and -2 (2-PGF; n = 171); and (3) 2-PGF with the addition of a GnRH treatment on d -11 (GnRH+2-PGF; n = 166). Ovaries were scanned by transrectal ultrasonography on d -11, -4, and 7, and blood samples were collected on d -11, -4, 0, and 7 for P4 determination. Treatment comparisons were performed using contrasts. The proportion of cows with a new corpus luteum on d -4 was greater in GnRH+2-PGF cows. Cows in 1-PGF had a greater P4 concentration on d 0 but lesser P4 on d 7 compared with cows in the other groups. Cows assigned to receive 2-PGF (2-PGF and GnRH+2-PGF) had greater estrus expression, and a greater proportion of cows ovulated to estradiol cypionate. No further contrast effects were observed for follicle diameter, double ovulation rate, pregnancy per embryo transfer (P/ET) on d 32 and 60, or pregnancy loss. As P4 concentration on d -4 increased, P/ET on d 60 tended to increase. Cows with P4 ≥3.66 ng/mL on d -4 had greater P/ET on d 32 and 60 than those with P4 below that threshold. Regardless of treatment, cows with P4 concentration ≥3.66 ng/mL also had a greater pregnancy per synchronized protocol (P/SP) on d 60. Also, a P4 concentration on d -4 (low or high) × follicle diameter (continuous) interaction tendency was observed when evaluating P/ET. Although P/ET did not differ among cows with different follicles sizes with reduced P4 concentration on d -4 (<3.66 ng/mL), it increased in cows with larger follicles exposed to increased P4 concentration (≥3.66 ng/mL). When P4 on d 0 was evaluated, P/ET on d 32 and 60 was greater for cows with low (≤0.09 ng/mL) versus high (>0.21 ng/mL) P4; as P4 concentration on d 0 increased, P/ET linearly decreased. In summary, cows with increased P4 concentrations during growth of the ovulatory follicular wave had improved P/ET. Administering a second PGF2α dose reduced P4 concentration on d 0 and increased ovulatory response to the protocol, but no benefits were observed on P/ET or P/SP.
Asunto(s)
Hormona Liberadora de Gonadotropina , Progesterona , Embarazo , Femenino , Bovinos , Animales , Hormona Liberadora de Gonadotropina/farmacología , Lactancia , Dinoprost/farmacología , Sincronización del Estro/métodos , Estradiol , Ovulación , Transferencia de Embrión/veterinaria , Inseminación Artificial/veterinariaRESUMEN
The aim of the present study was to determine the impact of different strategies for increasing the level of serum progesterone (P4) on luteal morphology and function in bovine females. The effects of increasing P4 on pregnancy rate and gestational loss (GL) in Nelore cows subjected to timed artificial insemination (TAI) were also evaluated. A total of 939 cows were divided into three groups: P4LA (n = 305), 150 mg of long-acting injectable P4 7 days after TAI; GnRH (n = 306), 10 µg of buserelin acetate 7 days after TAI; and control (n = 328), no hormone treatment after TAI. Doppler ultrasound assessments and P4 measurements were performed on days 7 and 16 after TAI. The pregnancy rate and GL as a function of treatment were compared using the SAS GLIMMIX procedure. Corpus luteum (CL) vascular perfusion, volume, and plasma P4 concentration were analysed using the SAS PROC MIXED procedure. No significant difference was found among the treatments in terms of volume, number of pixels, and CL intensity or in the serum P4 concentration at 7 days after ovulation. The CL blood flow at 16 days after ovulation was lower in the P4LA and GnRH groups than that in the control group (p < .01). Serum concentrations of P4 at 16 days after ovulation were higher in the P4LA and GnRH groups than those in the control group (p = .04). A difference in the pregnancy rate (p = .003) and a trend in GL (p = .07) as a function of treatment were found. Overall, long-acting injectable P4 supplementation on day 7 after TAI or GnRH administration affected CL vascularization and increased the serum concentrations of P4 16 days after ovulation, promoting better pregnancy rates than the control.
Asunto(s)
Fertilidad , Progesterona , Embarazo , Femenino , Bovinos , Animales , Progesterona/farmacología , Fertilidad/fisiología , Sincronización del Estro/métodos , Cuerpo Lúteo , Ovulación/fisiología , Hormona Liberadora de Gonadotropina/farmacología , Inseminación Artificial/veterinaria , Inseminación Artificial/métodos , Buserelina/farmacología , Lactancia/fisiología , Dinoprost/farmacologíaRESUMEN
This study aimed to evaluate the reproductive performance of ewes submitted to timed AI protocol with 7 (D7) or 9 (D9) days of progesterone. A total of 220 crossbred ewes (Doper × Santa Ines) were distributed in the treatments according to body weight (52.67 ± 11.76 kg), body condition score (2.5 ± 0.8; scale of 1-5), and the female category (multiparous, nulliparous and primiparous). Ewes received an intravaginal device of 0.3 g of progesterone (CIDR® Easy breed) on day 0, keeping in ewes for 7 or 9 days, which were the experimental treatments. At the time of P4 withdrawal, ewes received 300 IU of eCG (Novormon®) and 6.70 mg of dinoprost tromethamine (Lutalyse®). Insemination by laparoscopy was performed between 46 and 59 h after device removal, lasting from 1 to 5 min/ewe, which did not differ between protocols. The estrus detection rate was higher in the 9-day protocol compared to the 7-day protocol (82% and 65%, respectively, P = 0.0096). However, the 7-day protocol tended to increase pregnancy rate on the TAI compared to the 9-day protocol (45% and 33%, respectively, P = 0.09). The plasma concentration of progesterone at device removal was higher in the 7-day protocol than in the 9-day protocol (2.35 and 1.22 ng/mL, respectively, P = 0.04). We conclude that reducing the length of the estrus synchronization protocol to 7 days is recommended, which improved the reproductive response in ewes.
Asunto(s)
Inseminación Artificial , Progesterona , Animales , Dinoprost/farmacología , Detección del Estro , Sincronización del Estro , Femenino , Hormona Liberadora de Gonadotropina/farmacología , Inseminación Artificial/métodos , Inseminación Artificial/veterinaria , Embarazo , Índice de Embarazo , OvinosRESUMEN
This research aimed to determine the effect of temperament on reproductive parameters including cortisol and progesterone (P4) in Nellore cows. Additionally, two methods for increasing plasma progesterone (P4) levels in excitable animals to enhance pregnancy rate (P/AI) and reduce pregnancy loss were investigated. In total, 939 cows were subjected to timed artificial insemination (TAI) and divided into three groups: (P4LA; n = 305) 150 mg of injectable long-acting progestogen 7 days after TAI; (GnRH; n = 306), 10 µg of buserelin acetate on day 7 after TAI; control group (CG; n = 328) without hormonal treatment. In 213 cows, randomly chosen from each group, ultrasound evaluations of the preovulatory follicle (Mode B) were performed on the day of insemination and of the corpus luteum (Color Doppler) 7 and 16 days after TAI. Blood samples were obtained from 20% of the 939 animals, randomly chosen from each group, on the day of insemination and after 7 and 16 days to measure cortisol and progesterone, respectively. At the time of insemination, subjective temperament evaluations were performed with the animals being classified as excitable (EXC) or adequate (ADQ). The SAS GLIMMIX procedure was used to compare the pregnancy rate (P/AI) and gestational loss within each temperament for the three experimental groups. Continuous variables were analyzed utilizing SAS PROC MIXED procedure. Cortisol concentration was higher and POF (preovulatory follicle) and CL (corpus luteum) volumes at the time of insemination and 7 days after AI, respectively, were lower in EXC animals than in ADQ. No significant difference was observed between the number of pixels, CL intensity, and plasma concentration of P4, 7 days after TAI. However, 16 days post-insemination, among the animals classified as EXC, higher concentrations of P4 were observed in the GnRH and P4LA groups than in the control. Regarding P4 concentrations, there was a tendency to be lower in animals classified as EXC than in ADQ within the control group (P = 0.06), while rate of blood flow from the CL was lower in EXC animals than in ADQ animals (P = 0.04). Among the ADQ animals, the GnRH and P4LA groups showed a lower CL flow rate than that observed in the control (P = 0.04), 16 days after the TAI. Among EXC animals, a higher pregnancy rate was observed in the GnRH and P4LA groups than in the control group (P = 0.01). In the control group, the pregnancy rate (P/AI) of the ADQ animals was higher than that of the EXC animals (P = 0.05). No statistically significant differences were observed between gestational losses when the treatments or temperaments were compared. In conclusion, the use of GnRH or P4LA, 7 days after insemination, improves pregnancy rates in excitable animals and is a viable alternative to minimize the negative impact of stress and improve reproductive efficiency in beef cattle.
Asunto(s)
Enfermedades de los Bovinos , Hormona Liberadora de Gonadotropina , Aborto Veterinario , Animales , Buserelina/farmacología , Bovinos , Dinoprost/farmacología , Sincronización del Estro/métodos , Femenino , Fertilidad/fisiología , Hormona Liberadora de Gonadotropina/farmacología , Hidrocortisona , Inseminación Artificial/métodos , Inseminación Artificial/veterinaria , Lactancia/fisiología , Embarazo , Índice de Embarazo , Progesterona/farmacología , Progestinas/farmacologíaRESUMEN
Two experiments determined whether the addition of GnRH at the beginning of an estradiol (E2)/progesterone (P4)- based synchronization protocol and/or a second dose of prostaglandin F2α (PGF2α) the day before P4 device removal improves pregnancy rate in lactating dairy cows. On Day 0, all cows received a CIDR-B device and 2 mg i.m. estradiol benzoate, and half received 200 µg i.m. gonadorelin acetate (GnRH). On Day 7, cows were further subdivided to receive PGF2α (500 µg i.m. cloprostenol) or no PGF2α treatment. On Day 8, CIDR-B were removed, and all cows received PGF2α, 1 mg estradiol cypionate and 400 IU eCG i.m., and an estrus detection aid. Experiment 1 was designed to evaluate the effect of treatments on follicular development from P4 device removal to ovulation, expression of estrus, time of ovulation and serum P4 concentrations. Cows (n = 76) were examined by ultrasonography and bled for serum P4 determinations every 12 h from the time of P4 device removal but were not inseminated. In Experiment 2, all cows (n = 1036) were inseminated based on estrus detection using tail-paint. Cows with >50% of the paint rubbed-off by 48 h after P4 device removal were inseminated at that time, whereas those not in estrus received 100 µg i.m. of GnRH and were inseminated 12 h later. In Experiment 1, the interval from P4 device removal to ovulation was 71.7 ± 1.5 h and did not differ among groups. However, cows that received 2 injections of PGF2α had a greater (P < 0.01) estrus rate and lower (P < 0.01) P4 concentrations at 48 h after P4 device removal than those that received 1 PGF2α (estrus rate: 86.8% vs 68.4% and P4 concentration: 0.12 ± 0.01 vs 0.36 ± 0.07, for 2 and 1 PGF2α, respectively). In Experiment 2, estrus rate was also influenced by the number of PGF2α treatments, regardless of whether cows received or did not receive GnRH on Day 0 (2 PGF2α: 84.7%, 438/517 vs 1 PGF2α: 65.7%, 341/519; P < 0.01). Furthermore, there was a GnRH treatment by number of PGF2α treatments interaction (P < 0.05) on P/AI that was attributed to greater (P < 0.05) P/AI in cows that received GnRH on Day 0 and 2 PGF2α than in the other three treatment groups (EB+1 PGF2α: 45.2%, 119/263; EB+2 PGF2α: 45.8%, 119/260; EB + GnRH + 1 PGF2α: 45.7%, 117/256 and EB + GnRH + 2 PGF2α: 57.2%, 147/257). It was concluded that the addition of GnRH on Day 0 and a second dose of PGF2α the day before P4 device removal improves P/AI in lactating dairy cows synchronized with an estradiol/P4-based protocol.
Asunto(s)
Dinoprost , Progesterona , Animales , Bovinos , Dinoprost/farmacología , Estradiol/farmacología , Sincronización del Estro/métodos , Femenino , Hormona Liberadora de Gonadotropina/farmacología , Inseminación Artificial/métodos , Inseminación Artificial/veterinaria , Lactancia , EmbarazoRESUMEN
Three experiments evaluated the effects of insemination based on estrus expression or treatment with GnRH, on the time of ovulation and pregnancy rates to AI (P/AI) in dairy cows. All cows were treated with a P4 device for 8 d, estradiol benzoate at device insertion and prostaglandin F2α, estradiol cypionate and eCG at device removal. In Experiment 1, P/AI was less (P < 0.01) when all cows were inseminated at 48 h after device removal (31%) than when cows showing estrus at 48 h were inseminated at that time and those not in estrus received GnRH and were inseminated 12 h later (53%). In Experiment 2, cows in estrus by 48 h ovulated earlier than cows that were not in estrus but received GnRH and those that did not receive GnRH. Cows that were in estrus and inseminated at 48 h (47.8%) and those not in estrus at 48 h but inseminated at 60 h (53.4%) had greater P/AI (P < 0.05) than cows that were in estrus at 48 h but were inseminated at 60 h (29.7%) or inseminated at 48 h in the absence of estrus (19.4%). In Experiment 3, GnRH treatment of cows not showing estrus at 48 h and inseminated at 60 h had greater P/AI (58.5%; P < 0.05) than those not treated with GnRH but inseminated at 60 h (39.5%). Artificial insemination based on estrus expression and the inclusion of GnRH in those not in estrus increased P/AI in lactating dairy cows.
Asunto(s)
Hormona Liberadora de Gonadotropina , Progesterona , Animales , Bovinos , Dinoprost/farmacología , Estradiol/farmacología , Estro , Sincronización del Estro/métodos , Femenino , Hormona Liberadora de Gonadotropina/farmacología , Inseminación Artificial/métodos , Inseminación Artificial/veterinaria , Lactancia , Embarazo , Índice de Embarazo , Progesterona/farmacologíaRESUMEN
This experiment was designed to evaluate the effects of increasing doses of estradiol cypionate (ECP) on reproductive function of lactating dairy cows during the summer. In Exp. 1, 643 lactating Holstein cows were blocked by parity and assigned to receive 1) an intravaginal P4 device (1.9 g of P4) and 2.0 mg of estradiol benzoate on day -11, 25 mg (i.m.) of dinoprost tromethamine on day -4, 1.0 mg (i.m.) of estradiol cypionate and CIDR withdrawal on day -2, followed by TAI on day 0 (n = 326; ECP-1) or 2) the same synchronization protocol with 2.0 mg of ECP on day -2 (n = 317; ECP-2). In both treatments, cows were TAI on day 0 of the protocol, and cow rectal temperature was measured on days -2, 0, and 7. In Exp. 2, 608 lactating crossbred Holstein × Gir dairy cows were blocked by parity and enrolled to the same treatments as in Exp. 1, but on day 7, cows received one viable embryo into the uterine horn. In Exp. 1, a greater percentage of ECP-2 cows were detected on estrus (81.3 vs. 91.1%, respectively). A treatment × body condition score (BCS) interaction was observed on day 60 pregnancy per AI (P/AI), as ECP-2 cows with a BCS <2.75 had a greater P/AI vs. ECP-1, but an opposite result was observed in cows with a BCS ≥2.75. Regardless of treatment, there were effects of mean rectal temperature and heat stress events on P/AI. Treatment affected the diameter of the ovulatory follicle at TAI (ECP-1 = 15.3 mm vs. ECP-2 = 14.8 mm) and a greater percentage of ECP-1 cows had larger follicles (≥16.5 mm), but ECP-2 resulted in a greater incidence of early ovulatory cows (ovulating before day 0). Therefore, follicle diameter at TAI affected P/AI on day 60 in cows receiving ECP-2 and tended to affect ECP-1 cows. A treatment effect was observed on time to estrus following ECP treatments and a reduced proportion of ECP-2 cows showed estrus at TAI. Regardless of treatment, cows detected on estrus 48 h after ECP administration had a greater P/AI on day 60 vs. cows detected on estrus 24 h. In Exp. 2, a greater percentage of ECP-2 cows were detected on estrus (68.3 vs. 81.4%, respectively). In summary, a greater dose of ECP increased the percentage of animals expressing estrus, but it did not benefit the reproductive function of lactating dairy cows during the summer, regardless if animals were assigned to a TAI or timed-embryo transfer (TET) protocol.
Asunto(s)
Inseminación Artificial , Lactancia , Animales , Bovinos , Dinoprost/farmacología , Estradiol/análogos & derivados , Estradiol/farmacología , Sincronización del Estro/métodos , Femenino , Hormona Liberadora de Gonadotropina/farmacología , Inseminación Artificial/métodos , Inseminación Artificial/veterinaria , Embarazo , Progesterona/farmacología , Estaciones del AñoRESUMEN
This study evaluated (a) the efficacy of an association between injectable antibiotic therapy and sealant (ATBS) on milk yield (MY), somatic cell count (SCC), and prevalence of intramammary infections (IMI); and (b) the efficacy of gonadotropin-releasing hormone (GnRH) on follicular cyst (FCs) resolution (cyclicity at the 45th day in milk; DIM) and cumulative pregnancy rate (CPR) in heifers submitted to a lactation induction protocol (LIP). A total of 114 crossbred (Holstein × Jersey) heifers, with 34.7 ± 4.8 months and 439 ± 56.35 kg were submitted to LIP. On the 5th day of the LIP, the heifers were assigned to (i) ATBS (n = 57) with 7 mg/kg of norfloxacin associated with sealant and (ii) Control 1 (n = 57; CONT1) with no treatments. Lactation began on the 21st day of LIP and the 15th DIM, FCs were diagnosed and 106 heifers were randomized into two treatment groups with 53 heifers each: (i) GnRH (5 mL injectable GnRH) and (ii) Control 2 (CONT2; no treatment). Of the 114 heifers initially induced, 83.33% (n = 95) responded to LIP with an average MY of 15.19 kg/milk/day during 22 weeks of lactation. In the first 14 DIM, the IMI prevalence was 18% and 28% for heifers ATBS and CONT1 treated, respectively. Additionally, coagulase-negative Staphylococcus was the most frequently isolated group of pathogens. Mammary quarters that received ATBS treatment had a lower risk of IMI and SCC than CONT1. The cyclicity at 45 DIM was 68% (ATBS) and 35% (CONT1), and 57% and 46% for animals in the GnRH and CONT2. CPR was 60% in the ATBS group and 89% in CONT1, but GnRH treatment did not affect the CPR. In conclusion, LIP was effective in stimulating MY in heifers, and the IMI prevalence decreased with ATBS treatment. Also, the use of GnRH did not affect the FC regression, cyclicity at 45 DIM, and CPR.(AU)
Este estudo avaliou a (i) eficácia da associação entre antibioticoterapia injetável e selante interno de tetos (ATBS) na produção de leite (PL), contagem de células somáticas (CCS), e prevalência de infecções intramamárias (IIM); e (ii) eficácia do hormônio liberador de gonadotrofina (GnRH) na resolução de cistos foliculares (CFs), ciclicidade ao 45º dia em lactação (DEL) e taxa de prenhez cumulativa (TPC) em novilhas submetidas a um protocolo de indução de lactação (PIL). Um total de 114 novilhas mestiças (Holandês × Jersey), com 34,7 ± 4,8 meses e 439 ± 56,35 kg foram submetidas ao PIL. No 5º dia do PIL, as novilhas receberam: (i) ATBS (n = 57) com 7 mg/kg de norfloxacina associada ao selante interno de tetos e (ii) Controle 1 (n = 57; CONT1) sem tratamento. A lactação teve início no 21º dia do PIL e no 15º DEL, foram diagnosticados CFs e 106 novilhas foram agrupadas em dois grupos de tratamento com 53 novilhas em cada: (i) GnRH (5 mL de GnRH injetável) e (ii) Controle 2 (CONT2; sem tratamento). Das 114 novilhas inicialmente induzidas, 83,33% (n = 95) responderam ao PIL com PL média de 15,19 kg/leite/d durante 22 semanas de lactação. Nos primeiros 14 DEL a prevalência de IIM foi de 18% e 28% para as novilhas tratadas com ATBS e CONT1, respectivamente. Além disso, estafilococos coagulase negativa foram o grupo de patógenos mais frequentemente isolados. Quartos mamários tratados com ATBS tiveram menor risco (0,56) de IIM e menor CCS do que CONT1. A ciclicidade a 45 DEL foi de 68% (ATBS) e 35% (CONT1), e 57% e 46% para os animais no GnRH e CONT2. A TPC foi de 60% no grupo ATBS e 89% no CONT1, porém o tratamento com GnRH não afetou a TPC. Em conclusão, o PIL foi eficaz em estimular a PL em novilhas tardias e a prevalência de IIM diminiuiu com o tratamento ATBS. Além disso, o uso de GnRH não afetou a regressão de CF, ciclicidade em 45 DEL e a TPC.(AU)
Asunto(s)
Animales , Masculino , Embarazo , Lactancia/fisiología , Hormona Liberadora de Gonadotropina/farmacología , Mastitis Bovina/epidemiología , Antiinfecciosos/uso terapéutico , Quiste Folicular/diagnóstico , Bovinos/fisiología , Norfloxacino/administración & dosificación , Recuento de Células/veterinaria , Leche/fisiologíaRESUMEN
BACKGROUND: Nitric oxide and GnRH are biological factors that participate in the regulation of reproductive functions. To our knowledge, there are no studies that link NO and GnRH in the sympathetic ganglia. Thus, the aim of the present work was to investigate the influence of NO on GnRH release from the coeliac ganglion and its effect on luteal regression at the end of pregnancy in the rat. METHODS: The ex vivo system composed by the coeliac ganglion, the superior ovarian nerve, and the ovary of rats on day 21 of pregnancy was incubated for 180 min with the addition, into the ganglionic compartment, of L-NG-nitro arginine methyl ester (L-NAME), a non-selective NO synthase inhibitor. The control group consisted in untreated organ systems. RESULTS: The addition of L-NAME in the coeliac ganglion compartment decreased NO as well as GnRH release from the coeliac ganglion. In the ovarian compartment, and with respect to the control group, we observed a reduced release of GnRH, NO, and noradrenaline, but an increased production of progesterone, estradiol, and expression of their limiting biosynthetic enzymes, 3ß-HSD and P450 aromatase, respectively. The inhibition of NO production by L-NAME in the coeliac ganglion compartment also reduced luteal apoptosis, lipid peroxidation, and nitrotyrosine, whereas it increased the total antioxidant capacity within the corpora lutea. CONCLUSION: Collectively, the results indicate that NO production by the coeliac ganglion modulates the physiology of the ovary and luteal regression during late pregnancy in rats.
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Cuerpo Lúteo/inervación , Cuerpo Lúteo/metabolismo , Hormona Liberadora de Gonadotropina/metabolismo , Óxido Nítrico/metabolismo , Animales , Interacciones Farmacológicas , Femenino , Ganglios Simpáticos/efectos de los fármacos , Ganglios Simpáticos/metabolismo , Edad Gestacional , Hormona Liberadora de Gonadotropina/farmacología , Sistema Nervioso/efectos de los fármacos , Sistema Nervioso/metabolismo , Vías Nerviosas/efectos de los fármacos , Vías Nerviosas/metabolismo , Óxido Nítrico/farmacología , Ovario/inervación , Ovario/metabolismo , Embarazo , RatasRESUMEN
Two experiments evaluated the effects of gonadotropin releasing hormone (GnRH) treatment on fertility of suckled Nelore beef cows treated with an estradiol/progesterone (P4)-based protocol for timed artificial insemination (TAI). Experiment 1 was designed to determine the effect of GnRH administered 34 h after P4 insert removal (GnRH34) on time of ovulation. Suckled cows (n = 34) were treated with 2 mg estradiol benzoate (EB) and an intravaginal insert containing 1.9 g of P4. Eight days later, P4 inserts were removed, and all cows received 150 µg of d-cloprostenol (prostaglandin F2 alpha analogue), 300 IU of eCG, and 1 mg of estradiol cypionate (ECP). On Day 9 (05:00 p.m.), cows were randomly distributed, according to the diameter of the pre-ovulatory follicle, in two treatments: 1) GnRH (n = 17) cows that received 10.5 µg of buserelin acetate, or 2) no further treatment (control, n = 17). Cows treated with GnRH 34 h after P4 insert removal ovulated earlier (P = 0.02) than control cows (66 ± 0.0 and 77.2 ± 4.3 h). Experiment 2 was designed to determine the effect of GnRH34 on the fertility of suckled beef cows. Nelore cows (n = 506) were treated as Experiment 1. On Day 8, cows were painted in the sacrocaudal region to identify cows that displayed estrus. On Day 9 (05:00 p.m.), cows were randomized to receive same treatment as Experiment 1, control (n = 252), or GnRH (n = 254). All cows were TAI 48 h after P4 insert removal. At TAI, estrus was evaluated, and deemed to have occurred in cows without a tail-head chalk mark (>75% paint loss). Cows treated with GnRH 34 h after P4 insert removal had greater (P = 0.01) pregnancy per AI (P/AI) than cows that only received ECP (63.0% and 50.4%). No difference (P = 0.5) was observed in the proportion of cows that displayed estrus between treatments. Furthermore, cows that displayed estrus had greater (P < 0.01) P/AI than cows that did not. Treatment with GnRH, given at 34 h after P4 insert removal, increased (P < 0.05) P/AI in cows that did not show estrus at TAI. In summary, treatment with GnRH 34 h after P4 insert removal was associated with earlier ovulation and resulted in greater P/AI in suckled Nelore cows treated with an estradiol/P4-based protocol for TAI.
Asunto(s)
Inseminación Artificial , Progesterona , Animales , Buserelina , Bovinos , Dinoprost , Estradiol , Estro , Sincronización del Estro , Femenino , Hormona Liberadora de Gonadotropina/farmacología , Inseminación Artificial/veterinaria , Lactancia , EmbarazoRESUMEN
Intracerebroventricular (icv) administration of oxytocin (OT) induces robust lordosis behavior (lordosis quotient and lordosis intensity) in estrogen-primed rats. The present study explored the hypothesis that the OT-Prostaglandin E2-GnRH pathway (a pathway produced in astrocytes) is involved in the facilitation of lordosis behavior by icv infusion of OT (2 µg). In Experiment 1, we tested the involvement of the OT receptor (OTR) by infusion of the OTR antagonist, atosiban (ATO). OT-induced lordosis was significantly reduced at both 30 and 120 min by prior infusion of ATO. In Experiment 2, we studied the effects of aspirin (COX2 inhibitor) and ONO-AE3-208 (ONO; EP4 prostaglandin receptor antagonist) on OT-induced lordosis. Infusions of both compounds diminished OT-induced lordosis at both 120 and 240 min. In Experiment 3, the involvement of the GnRH-1 receptor inhibitor antide on OT-induced lordosis was evaluated. Antide significantly inhibited OT-induced lordosis at all times tested. These data indicate that the OT/PGE2/GnRH pathway is involved in the expression of OT-induced lordosis behavior, an effect that may be occurring directly in hypothalamic astrocytes.