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Cervical relaxation (CR) was performed in ewes during diestrus, to prospect a feasible protocol for non-surgical embryo transfer (NSET). In Trial 1, naturally mated ewes (n=13) received CR protocols with estradiol benzoate (EB, 1 mg on D6) and oxytocin (OT, 50 IU on D7) only (G-EB+OT) or associated with human chorionic gonadotropin (hCG, 300 IU on D7, G-EB+OT+hCG) and were compared to non-hormonally treated (G-control) ewes. Estradiol concentration increased (P<0.05) from D6 to D7 in G-EB+OT and G-EB+OT+hCG, then decreased (P<0.05) on D8. The G-EB+OT had reduced (P<0.05) progesterone on D8, and 0% (0/5) pregnancy, while G-control had 100% (3/3) conception rate. The treatment with hCG mitigated the progesterone drop on D8 and resulted in 50% (2/4) conception rate. In Trial 2, cervical transposing success after treatment with EB, OT, and hCG on D6 and D7 was assessed in nulliparous (n=25) and multiparous (n=30) ewes. The EB+OT+hCG protocol allowed efficient cervical transposition (~78% of cervical transposition success into ~5 min) in nulliparous and multiparous females and did not alter total and blood perfusion areas of corpora lutea (CL). The use of EB+OT+hCG was efficient to ensure effective cervical transposing in nulliparous and multiparous ewes, with no apparent deleterious effect on CL function, and may contribute to animal welfare, allowing a less invasive NSET procedure.

Background: Commercial embryo flushing of horses has required hormonal management of both the donor and recipient mares throughout the breeding season., Aim: This study aimed to find out the effect of using human chorionic gonadotropin (hCG) and prostaglandin F2α (PG) on the ovarian and uterine dynamics and hemodynamics, estradiol (E2), progesterone, oxidants-antioxidants, and blood biochemicals in embryo donor mares during the hottest months of the year in a subtropical climate., Methods: Three Control estrous cycles of native mares (10-20 years; N = 10) followed by two treated cycles with hCG and PGF2α were examined daily from May to August using Doppler ultrasound with blood sampling. Circulating, progesterone (P4), total cholesterol, total proteins, albumin, haptoglobin, nitric oxide (NO), catalase, alkaline phosphatase, lactate dehydrogenase (LDH), and myeloperoxidase were measured in blood serum., Results: Days during the control estrous cycle impacted the dominant follicle (DF) diameter ( p < 0.0001), antrum diameter ( p < 0.0001), area ( p < 0.0001), antral area ( p < 0.0001), and color area % ( p > 0.05), and corpus luteum (CL) diameter ( p < 0.0001). PG tended to impact DF diameter ( p > 0.05) but influenced its antrum diameter ( p < 0.05), color area ( p < 0.05), CL diameter ( p < 0.01), and area ( p = 0.013). Days after hCG tended to impact DF antrum diameter ( p > 0.05) and the antrum area ( p > 0.05), but influenced CL diameter ( p < 0.0001). PGF2α and hCG increased uterine horn area ( p = 0.016) and color area ( p = 0.023), total cholesterol ( p < 0.0001), and NO ( p < 0.0001) levels but hCG increased the levels of myeloperoxidase ( p < 0.005), total proteins ( p < 0.001), and albumin ( p < 0.0001). Globulins achieved the highest level ( p = 0.054) but the Albumin/globulin ratio reached a minimum value on Day 0 of the control mares ( p < 0.0001). PGF2α increased LDH ( p < 0.0001) and sharply declined ( p = 0.028) progesterone., Conclusion: In conclusion, the treatment protocols of hCG and PGF2α showed minimal effects on the produced ovulating follicles and can be used during the summer season to manage embryo donor mares., Competing Interests: The authors declare that they do not have any conflict of interest.

This study investigated the effect of hCG or GnRH on structural changes of the corpora lutea (CL) and the regulation of the expression of steroidogenic enzymes involved in P 4 secretion in post-ovulatory (po-CL) and accessory CL (acc-CL). Sixty-four ewes were assigned to three groups receiving: 300 IU of hCG (hCG) or 4 µg Buserelin (GnRH) or 1 mL of saline solution (Control) on Day (d) 4 post artificial insemination (FTAI). Laparoscopic ovarian were performed on d 4, 14 and, 21 post-FTAI to determine the numbers of CL. Blood samples were collected for serum LH and P 4 analysis. On d 14 post-FTAI, both CL were removed from the ovary to determine large luteal cell (LLC) number and to evaluate the expression of steroidogenic enzymes (HSD3B1, STAR, CYP11A1). Only hCG and GnRH treated ewes generated acc-CL. The LLC in both po- and acc-CL were significantly greater in the hCG group compared to GnRH and Control groups (P<0.05). Overall, hCG group showed the greatest immunodetection of HSD3B1and STAR in both po- and acc-CL (P<0.05). rnRNA expression of HSD3B1, STAR and CYP11A1 in the acc-CL tended to be greater in hCG group than in GnRH group (P<0.1). The LH concentration was increased in GnRH group (P<0.05) and P 4 concentration was greater in hCG group compared to the other groups (P<0.05). In conclusion, administration of hCG has a notably impact on acc-CL development and the expression of steroidogenic enzymes compared to GnRH treatment in ewes. This leads to elevated P 4 concentration and improved luteal function., Competing Interests: Declaration of Competing Interest This research paper is submitted to be consider for publication as a full text. It has not been published before or simultaneously submitted for publication elsewhere. All authors have approved the final version of the manuscript., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study was conducted in ewes to assess effects of human chorionic gonadotropin (hCG) administration after imposing an estrous induction treatment regimen. Ewes (n = 115) were treated with a 60 mg medroxyprogesterone-intravaginal-sponge for 6 d plus 200 IU of equine chorionic gonadotropin (eCG) im and 37.5 μg d-cloprostenol im 36 h before sponge removal (Day 0). After natural mating, ewes having at least one corpus luteum (CL; n = 108) were administered either 1 mL of saline (G-Control; n = 53) or 300 IU of hCG (G-hCG; n = 55) on Day 7.5 after sponge removal (Day 0). Ovarian ultrasonography and blood collection were performed on Days 7.5, 13.5, 17.5, 21.5, and 30.5. Accessory CL (aCL) were observed in 81.5 % (G-hCG) and 0.0 % (G-Control) of ewes (P = 0.0001). Diameter, area, and volume of luteal tissue were greater (P < 0.05) in G-hCG from Day 13.5 to 30.5. Progesterone (P 4 ) concentrations were greater (P < 0.05) on Days 13.5, 17.5, 21.5 and 30.5 for ewes of the G-hCG group. Pregnancy percentage was similar (P = 0.25) between groups [47.1 % (G-control) compared with 60.0 % (G-hCG)], although total number of lambs produced by estrous synchronized ewes was greater (P = 0.005) in ewes of the G-hCG group (90.9 % compared with 66.0 %). In conclusion, hCG administration 7.5 days after sponge removal from Morada Nova ewes during the non-breeding season is an effective treatment to induce aCL formation, improve luteal tissue biometry and P4 concentrations, and to enhance the total number of lambs born., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Objectives were to evaluate effects of a smaller than typically used dose of equine chorionic gonadotropin (eCG) during a fixed-time artificial insemination (FTAI) treatment regimen. Transrectal ultrasonic (US) examinations were conducted on dairy cows on Day 0 (D0) and the treatment regimen was initiated with administrations of an intravaginal progesterone (P 4 ) implant, estradiol benzoate (im), and prostaglandin F 2α (PGF 2α; im). On D8, the P 4 implant was removed and PGF 2α and estradiol cypionate were administered to all animals. Subsequently, cows were randomly assigned to three groups and eCG was administered to Groups 1, 2, and 3 in doses of 300 (im); 100 (im); and 100 (Baihui acupoint) IUs, respectively. The B-mode and power-flow US cineloops were performed to assess follicular dynamics and evaluate various morphological and vascular characteristics of the corpus luteum. Blood samples were collected to quantify serum P 4 concentrations. There were no differences between the ovulation synchronization treatment regimens for all follicular dynamic variables tested; however, cows in Group 3 differed from Group 2 having a larger follicle diameter (FD) on D10 (P = 0.06) and larger preovulatory FD (P = 0.09), as well as a blood perfusion area of the dominant follicle wall on D8 (P = 0.07). There were no differences in responses to the ovulation synchronization treatment regimens for the luteal variables evaluated subsequent to ovulation. In conclusion, the Baihui acupoint was effective as an alternative route for eCG dose reduction when FTAI treatment regimens were imposed without detrimentally affecting values for reproductive variables., (Copyright © 2020 Elsevier B.V. All rights reserved.)

There was investigation of whether there were ovulations from co-dominant follicles following eCG administration. In all experiments, there was GnRH injection and CIDR insertion on day 0 (D0), CIDR withdrawal on D8, and cloprostenol administration on D8 (Exp. I and II) or D7 and D8 (Exp. III). Females in the control group were not administered any further treatment. Females in other group(s) were treated with eCG (500 IU) on Day 2 in Exp. I, Day 2 (eCG-2) or 8 (eCG-8) in Exp. II and Day 2 (eCG-2) or Days 2 and 6 (eCG-2-6) in Exp. III. Ovaries were examined using ultrasonography. In Experiments I and II, females had follicle emergence on Day 2. At the time of CIDR removal, more eCG-treated heifers (8/9; Exp. I) and cows (5/6; eCG-2; Exp. II) had co-dominant follicles compared to those in the control group (P < 0.05). Occurrence of ovulations from co-dominant for individual cows was minimal. In Experiment III, the time period from CIDR removal to estrus in cows treated with eCG-2 (68 ± 13 h) was longer compared to cows in the control (37±2 h) and eCG-2-6-treated group (38 ± 5 h; P < 0.05). There was a greater proportion of heifers having ovulations and thus greater progesterone concentration in the eCG-2-6 than eCG-2 group (P < 0.05). Administering eCG twice 4 days apart with the initial administration being two days after GnRH administration, at the time of follicle wave emergence, could induce growth of and ovulation from co-dominant follicles and enhance progesterone production in cattle., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The aims of this study were to compare the effects of an intramuscular human chorionic gonadotropin (hCG) administration on corpus luteum (CL) development, plasma progesterone (P4) and estradiol (E2) concentration in ipsilateral (first-wave dominant follicle [W1DF] in the same ovary as the CL) and contralateral (W1DF and CL in opposite ovaries) cattle. Cross-bred beef heifers (Holstein × Japanese black, n=83) with synchronized ovulation were randomly assigned to either treatment with 1,500 IU hCG or no treatment on day 5 post-ovulation and were subdivided into ipsilateral (hCG treatment, n=21; no treatment, n=23) or contralateral (hCG treatment, n=17; no treatment, n=17) groups. Five heifers were excluded from the study, as they presented with double ovulation in response to hCG treatment. The effects of hCG treatment, location (ipsilateral and contralateral), and the number of days post-ovulation (days 5, 7, and 14) were analyzed using three-way ANOVA. hCG treatment significantly increased CL diameter on day 7 and plasma P4 concentration on days 7 and 14 in the contralateral group, but not the ipsilateral group. In contrast, hCG treatment decreased plasma E2 concentration on days 7 and 14 in both groups. In summary, our results indicate that the hCG treatment more significantly promoted CL development and increased plasma P4 concentration in the contralateral than in the ipsilateral group.

We examined the effect of human chorionic gonadotropin (hCG) treatment 5 days after estrus on ovarian dynamics and plasma progesterone (P 4 ) and estradiol (E 2 ) concentrations when the first-wave dominant follicle (DF) was ipsilateral or contralateral to the corpus luteum (CL) in lactating dairy cows. Seventy cows were divided into two groups: (1) ipsilateral group (IG; n = 37), in which the first-wave DF was ipsilateral to the CL, and (2) contralateral group (CG; n = 33), in which the first-wave DF was contralateral to the CL. IG and CG were further subdivided into two groups: non-treatment group (IG, n = 18; CG, n = 19), and hCG treatment group: administrated 1500 IU of hCG 5 days after estrus (IG, n = 19; CG, n = 14). Blood sampling and ovarian examination were performed at 3, 5, 7, 9, 11, and 13 days after estrus. Mean diameter of the first-wave DF on Day 9 tended (P = 0.067) to be larger in IG than in CG in the non-treatment group. Mean diameter of CL and plasma P 4 and E 2 concentrations did not differ between IG and CG in the non-treatment and hCG treatment groups. Accessory CL development did not differ between IG and CG in the hCG treatment group. Our findings indicate that CL development and plasma P 4 and E 2 concentrations were not affected by the existence of the first-wave DF; however, first-wave DF development was affected by the existence of a CL in the same ovary.

Conventional methods for determining the reproductive performance of sheep bred either after estrus synchronization during the breeding season or after induction of estrus/ovulation during the non-breeding season take a long time and may give misleading results due to the effect of environmental factors. Laparoscopic observations allow real-time monitoring of ovarian activity around estrus or ovulation. This study was aimed at assessing the superovulatory effects of follicle-stimulating hormone (FSH) and equine chorionic gonadotropin (eCG) treatments by laparoscopy during breeding (September-November, n=12) and non-breeding (April-June, n=12) seasons in Akkaraman sheep. In both seasons, after CIDR withdrawal, the ewes were injected either with 600 IU eCG or 300 μl (20 mg/ml) FSH twice at 12 hour intervals. Plasma P4, E2 and LH concentrations were determined at the time of intra-vaginal CIDR insertion (day 0) and then at its withdrawal (day 12), followed by 3 and 6 days of eCG or FSH injections. After 3 (first observation) and 6 (second observation) days of hormone injections, laparoscopy was performed to record ovarian activity in both seasons. The eCG increased (p⟨0.05) the numbers of large follicles (first observation) and CL (first and second observations) in the breeding season compared to FSH treatment. CL, small-moderate and large follicle numbers of eCG treated ewes were higher (p⟨0.05) than those of FSH at both observations in the non-breeding season. In the breeding season, eCG treated ewes had higher (p⟨0.05) plasma P4 (3 and 6 days after hormones injections) and E2 (3 days after hormones injections) concentrations than those of FSH. In conclusion, the results of the present study indicate that treatment with eCG during the non-breeding season can support ovarian activity, and thus increase ovulation rate and plasma hormone concentrations around induced estrus/ovulation in Akkaraman ewes., (Copyright© by the Polish Academy of Sciences.)

Human chorionic gonadotropin (hCG) is a luteotropic hormone that promotes the survival and steroidogenic activity of corpus luteum (CL) by acting through luteinizing hormone receptors (LHRs) expressed on luteinized theca and granulosa cells (GCs). Therefore, it is used to support luteal phase in in vitro fertilization (IVF) cycles to improve clinical pregnancy rates and prevent miscarriage. However, the molecular mechanism underlying this action of hCG is not well characterized. To address this question, we designed an in vitro translational research study on the luteal GCs obtained from 58 IVF patients. hCG treatment at different concentrations and time points activated c-Jun N-terminal kinase (JNK) pathway and significantly increased its endogenous kinase activity along with upregulated expression of steroidogenic enzymes (steroidogenic acute regulatory protein (stAR), 3β-Hydroxysteroid dehydrogenase (3β-HSD)) in a dose-dependent manner in the luteal GCs. As a result, in vitro P production of the cells was significantly enhanced after hCG. When JNK pathway was inhibited pharmacologically or knocked-down with small interfering RNA luteal function was compromised, P4 production was declined along with the expression of stAR and 3β-HSD in the cells. Further, hCG treatment after JNK inhibition failed to correct the luteal defect and promote P4 output. Similar to hCG, luteinizing hormone (LH) treatment improved luteal function as well and this action of LH was associated with JNK activation in the luteal GCs. These findings could be important from the perspective of CL biology and luteal phase in human because we for the first time identify a critical role for JNK signaling pathway downstream LHR activation by hCG/LH in luteal GCs., Summary Sentence: JNK signaling pathway plays a central role in the upregulated expression of the steroidogenic enzymes StAR and 3b-HSD and augmented progesterone production by hCG/LH in human luteal granulosa cells., (© The Author(s) 2020. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The objective was to compare pregnancy per AI (P/AI) between two shortened timed-AI (TAI) protocols in beef cattle. This study also determined whether administration of eCG in heifers and timing of AI in cows would affect P/AI. Cattle were submitted at random to either a modified 5-d Co-synch protocol (Day 0 = progesterone releasing device (CIDR); Day 5 = CIDR removal and 500 μg of cloprostenol (PGF); Day 8 = 100 μg GnRH concurrent with AI) or J-synch protocol (Day 0 = CIDR insertion and 2 mg of estradiol benzoate i.m.; Day 6 = CIDR removal and 500 μg PGF; Day 9 = 100 μg GnRH concurrent with AI). In Experiment 1, 1135 heifers (13-15 mo of age) received an estrus detection patch (Estrotect™) on Day 5 and 579 were selected at random to receive 300 IU of equine chorionic gonadotropin (eCG) at the time of CIDR removal. Patches were scored from 0 to 3 based on color change between initial application and AI; 0 = unchanged, 1 = ≤ 50% change, 2 = > 50% change, 3 = missing. Estrus was defined to have occurred when the patch was scored 2 or 3. In Experiment 2, 399 cyclic, non-lactating beef cows from 1 location were submitted to either the modified 5-d Co-synch or J-synch protocol and within each protocol cows were TAI at either 66 ± 1 (n = 199) or 72 ± 1 h (n = 200) following CIDR removal. Transrectal ultrasonography was used in both experiments to determine presence of a corpus luteum (CL) on Day 0, and to diagnose pregnancy 35 d after TAI. In Experiment 1, eCG increased estrus rate only in heifers without a CL on day 0 that were submitted to the modified 5-d Co-synch protocol (41.9 vs. 69.6%). Heifers submitted to the J-synch protocol had greater (P = 0.03) P/AI compared with those in the modified 5-d Co-synch (48.7 vs. 41.1%) and heifers that expressed estrus before AI had increased (P < 0.0001) P/AI compared to those that did not (53.6 vs. 36.5%). Administration of eCG and presence of a CL tended to affect P/AI (P = 0.13). In Experiment 2, cows submitted to the J-synch protocol tended (P = 0.07) to have greater P/AI compared to those in the modified 5-d Co-synch (74.1 vs. 66.5%). There was no association between P/AI and timing of AI. In summary, the J-synch protocol resulted in greater P/AI than the modified 5-day Co-synch protocol in heifers and cows. Administration of eCG increased estrus rate in heifers without a CL at the start of the protocol and tended to improve P/AI in all heifers. Timing of AI (66 vs. 72 h) had no effect on P/AI in cows subjected to either TAI protocol., (Crown Copyright © 2019. Published by Elsevier Inc. All rights reserved.)

The objective of this study was to evaluate the effects of plasma progesterone (P 4 ) concentrations during eCG-ovarian follicular superstimulatory treatment performed in early luteal phase and estradiol concentrations during peri-ovulatory period on ovarian response, number and embryo quality. On Day -2, females (n = 75) having a follicle ≥7 mm were treated with GnRH to induce ovulation. On Day 0, females that had ovulations (n = 54) were treated with 1000 IU eCG and were assigned to one of two treatments: (1) intravaginal device (ID) containing 0.5 g P 4 (P 4 group) and (2) no ID (Control group). On Day 5, females were administered PGF 2α and the ID was removed. On Day 7 and 8, females were mated and embryo recovery was performed 7 or 8 days later. Blood samples were collected from Day 0 to 9. Number (± SD) of follicles ≥7 mm on day of mating was greater (P =  0.04) in the control (9.7 ± 4.2) than P 4 -treated (6.7 ± 4.9) group; number of corpora lutea did not differ (5.5 ± 3.1 and 5.2 ± 3.4 respectively). Ovulation rate was greater (P <  0.01) in the P 4 -group (77.4%; 130/168) than control group (53.3%; 135/253). Number of embryos with an excellent grade (grade 1) tended to be greater (P =  0.07) in the P 4 -group (82.4%; 42/51) than control group (65.4%; 36/55). It was concluded that supplementation with exogenous P 4 during eCG treatment in early luteal phase inhibits excessive follicular growth, increases ovulation rate and improves embryo quality., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Objective: To characterize corpora lutea (CL) function after gonadotropin-releasing hormone agonist (GnRHa) trigger with the use of adjuvant human chorionic gonadotropin (hCG)., Design: Secondary analysis of serum from prospective randomized clinical trial., Setting: University-based fertility center., Patient(s): Women under 40 years of age at risk of ovarian hyperstimulation syndrome (OHSS) with serum E 2 level <4,000 pg/mL., Interventions(s): All subjects underwent ovarian stimulation with the use of a GnRH antagonist protocol. Within a larger study, subjects were randomized to receive 1,000 IU hCG at the time of GnRHa trigger and placebo at the time of vaginal oocyte retrieval (VOR) or placebo at the time of GnRHa trigger and 1,500 IU hCG at the time of VOR., Main Outcome Measure(s): Luteal phase and early pregnancy curves of serum prorenin and 17α-hydroxyprogesterone (17OH-P)., Result(s): Thirty subjects enrolled in this secondary analysis. Serum 17OH-P peaked in the early luteal phase, 5 days after GnRHa trigger, with a nadir in the mid-luteal phase 9 days after trigger. Serum prorenin peaked in the luteal phase 2 days after GnRHa trigger, independently from adjuvant hCG timing, and reached a nadir at 9 days after trigger. CL function appears higher when adjuvant hCG is given at VOR compared with adjuvant hCG given at the time of trigger., Conclusion(s): CL function, as interpreted by proxy measures of serum prorenin and 17OH-P with pregnancy, continues despite GnRHa trigger. Both options for adjuvant hCG timing are sufficient for CL rescue and successful pregnancy, so the potential for OHSS risk with increased CL activity after hCG at VOR should be considered., Clinical Trial Registration Number: NCT01815138., (Copyright © 2019 American Society for Reproductive Medicine. All rights reserved.)

We examined the effect of human chorionic gonadotropin (hCG) treatment 5 days after artificial insemination (AI) on conception rate when the first-wave dominant follicle (DF) in the ovaries was either ipsilateral or contralateral to the corpus luteum (CL) in lactating dairy cows. 577 cows from 4 dairy farms were divided into the following two groups 5 days after AI using transrectal ultrasonography: (1) the ipsilateral group (IG; n = 348), in which the DF was ipsilateral to the CL, and (2) the contralateral group (CG; n = 229), in which the DF was contralateral to the CL. IG and CG were further subdivided into two groups: non-treatment groups, which received no treatment (IG, n = 220; CG, n = 128), and hCG treatment group, that was administrated 1500 IU hCG 5 days after AI (IG, n = 143; CG, n = 86). Pregnancy was diagnosed by rectal palpation or transrectal ultrasonography from 53 to 67 days after AI. Conception rate was significantly (P < 0.01) higher in the hCG treatment group of IG (40.6%) than in the non-treatment group of IG (21.4%); however, there was no difference in the non-treatment (51.7%) and hCG treatment (43.0%) groups of CG. Parity, farm, days in milk at AI, interaction between the farm and hCG treatment and interaction between the farm and location of the first-wave DF and CL did not affect conception rate. Our results suggest that conception rate can be improved by administrating hCG only to cows with the first wave DF ipsilateral to the CL.

The objective of this study was to evaluate the effect of equine chorionic gonadotropin (eCG) on the estrous response, ovulatory follicle development, ovulation rate, corpus luteum (CL) diameter, concentrations of progesterone (P4) and pregnancy per AI (P/AI) in CIDR-EB treated lactating anovular Nili-Ravi buffalo during the breeding season. Buffalo (n = 87), 60 to 200 Days postpartum were treated with CIDR at a random stage of the estrous cycle (Day 0) and assigned to two groups; with eCG (+eCG; n = 44), or without eCG (-eCG; n = 43). In both groups, each animal received 2 and 1 mg estradiol benzoate (EB) on Day 0 and then on Day 8, respectively. On Day 6, eCG and PGF 2α were administered. On Day 7, CIDR were removed and FTAI was performed at 48 and 60 h later and pregnancy was diagnosed 35 Days post AI. The results revealed that eCG treatment increased (P <  0.05) the CL diameter on 15, 18, and 21 after the ovulation that resulted from imposing the estrous synchronization protocol than when eCG was not administered. Similarly, treatment with eCG resulted in an increased (P <  0.05) mean concentration of P4 as compared with animals not treated with eCG on Days 18 and 21 after the ovulation that occurred subsequent to imposing the estrous synchronization protocol. The ovulation rate was greater in eCG-treated buffalo as compared with those not treated with eCG (93% compared with 74%, respectively; P < 0.05). The P/AI was greater in buffalo treated with eCG than those not treated with eCG (56% compared with 31%, respectively; P < 0.05). In conclusion, the addition of eCG along with EB and P4 based treatments enhanced the estrous response, follicular and luteal function, ovulation rate, concentrations of P4, and P/AI in lactating anovular Nili-Ravi buffalo., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Progestin concentration in plasma during the early luteal phase is crucial for endometrial function and conceptus development. We hypothesized that periovulatory gonadotrophin treatment via support of luteal function affects endometrial gene expression in horses. Effect of age was analyzed as well. Shetland mares (n = 8, age 4-25 years) were assigned to the following treatments during five consecutive cycles in alternating order following a cross-over design: treatment hCG/-: preovulatory injection of hCG, but no gonadotrophin injection at detection of ovulation, treatment -/hCG: no preovulatory gonadodrophin injection, but injection of hCG at detection of ovulation, treatment eCG/-: preovulatory injection of eCG, but no gonadotrophin injection at detection of ovulation, treatment -/eCG: no preovulatory gonadotrophin injection, but injection of eCG at detection of ovulation, treatment control: no treatment. Concentration of progestin was analyzed by ELISA from the day of ovulation until Day 10. On Day 10, endometrial cells were collected transvaginally by cytobrush technique. Expression of mRNA of cyclooxygenase-2 (COX-2), prostaglandin F2α-synthase, prostaglandin E-synthase, progesterone receptor (PR), estradiol receptor (E2R), acyl-CoA-dehydrogenase (ACAD), uteroglobin (UGB), uteroferrin, and uterocalin was analyzed by RT qPCR. Immunohistological staining of endometrial tissue, obtained via biopsy, was performed for COX-2, PR and UGB. The P4 concentration was influenced by day of cycle (P < 0.01), but not by treatment. No effects of age on gene expression were determined. Neither of the periovulatory gonadotrophin treatments nor age influenced mRNA expression of the genes of interest. Treatment did also not affect immunohistological staining of the endometrium. In contrast, age affected the percentage of PR positive stromal cells (e.g. mare 1 (4 years): 65.5 ± 2.6, mare 2 (24 years): 82.7 ± 2.2%, P < 0.05) and COX-2 positive stained ciliated cells (e.g. mare 1: 15.8 ± 2.9, mare 2: 33.4 ± 6.0%, P < 0.05). In conclusion, no effects of periovulatory gonadotrophin treatment and age on endometrial gene expression in luteal phase pony mares were reported. A lack of treatment effects on luteal function and expression of PRs in the endometrium can at least in part be explained by differences in the reproductive physiology between horses and ponies., (Copyright © 2016 Elsevier Inc. All rights reserved.)

This study was designed to investigate the impact of buserelin acetate (BA) or human chorionic gonadotropin (hCG) administration on the day of first artificial insemination (AI) on subsequent luteal profile (diameter of corpus luteum (CL) and plasma progesterone) and conception rate in Murrah buffalo. The present experiment was carried out at two locations in 117 buffalo that were oestrus-synchronized using cloprostenol (500 μg) administered (i.m.) 11 days apart followed by AI during standing oestrus. Based on treatment (i.m.) at the time of AI, buffalo were randomly categorized (n = 39 in each group) into control (isotonic saline solution, 5 ml), dAI-BA (buserelin acetate, 20 μg) and dAI-hCG (hCG, 3000 IU) group. Out of these, 14 buffalo of each group were subjected to ovarian ultrasonography on the day of oestrus to monitor the preovulatory follicle and on days 5, 12, 16 and 21 post-ovulation to monitor CL diameter. On the day of each sonography, jugular vein blood samples were collected for the estimation of progesterone concentrations. All the buffalo (n = 117) were confirmed for pregnancy on day 40 post-ovulation. The conception rate was better (p < 0.05) in dAI-BA (51.3%) and dAI-hCG (66.7%) groups as compared to their control counterparts (30.8%). Furthermore, the buffalo of dAI-hCG group had improved (p < 0.05) luteal profile, whereas the buffalo of dAI-BA group failed (p > 0.05) to exhibit stimulatory impact of treatment on luteal profile when compared to control group. In brief, buserelin acetate or hCG treatment on the day of first AI leads to an increase in conception rate; however, an appreciable impact on post-ovulation luteal profile was observed only in hCG-treated Murrah buffalo., (© 2016 Blackwell Verlag GmbH.)

CXADR-like membrane protein (CLMP) is a novel cell-cell adhesion molecule. The present study investigated the spatiotemporal expression pattern of CLMP and its regulation in the rat ovary during the periovulatory period. Real-time polymerase chain reaction analysis revealed that Clmp mRNA was rapidly stimulated in intact ovaries by 4h after human chorionic gonadotrophin (hCG) treatment. In situ hybridisation analysis demonstrated that Clmp mRNA expression was stimulated in theca cells at 4h after hCG and remained elevated until 12h. Clmp mRNA was also upregulated in granulosa cells and was present in forming corpora lutea. Our data indicate that the protein kinase A but not the protein kinase C pathway regulates the expression of Clmp mRNA in granulosa cells. Phosphatidylinositol 3 kinase and p38 kinase are also involved in regulating Clmp mRNA expression. The stimulation of Clmp mRNA by hCG requires new protein synthesis. Furthermore, inhibition of epidermal growth factor receptor activation significantly inhibited Clmp mRNA expression, whereas inhibition of prostaglandin synthesis or progesterone action had no effect. The stimulation of CLMP in the rat ovary may be important in cell adhesion events during ovulation and luteal formation such as maintaining the structure and communication of ovarian follicular and luteal cells.

The aim was to examine the effect of a single intramuscular (i.m.) injection of equine chorionic gonadotrophin (eCG) on Day 3 after oestrus on corpus luteum (CL) development, circulating progesterone and conceptus development in cross-bred beef heifers. In Experiment 1, heifers received: (1) saline, or a single i.m. injection of eCG on Day 3 at (2) 250IU (3) 500IU (4) 750IU or (5) 1000IU. Administration of eCG resulted in increased luteal tissue area and progesterone and oestradiol concentrations compared with controls. In Experiment 2, heifers received (1) a progesterone-releasing intravaginal device (PRID Delta) from Day 3 to 5 or (2) a PRID Delta from Day 3 to 5 plus a single injection of 750IU eCG on Day 3. In vitro-produced blastocysts (n=10 per recipient) were transferred on Day 7 and heifers were slaughtered on Day 14 to assess conceptus development. Administration of eCG reduced the number of short cycles (6.3% vs 31.3%) and increased mean luteal tissue weight (P=0.02). Insertion of a PRID Delta on Day 3 resulted in an elevation (P<0.05) in serum progesterone until removal on Day 5. Administration of eCG at the time of PRID Delta insertion resulted in higher progesterone levels (P<0.05) from Day 10 onwards. Conceptus dimensions were not affected. In conclusion, a single injection of eCG on Day 3 increased CL size and progesterone concentrations and, when given in conjunction with a progesterone-releasing device, appeared to reduce the number of short cycles, presumably due to its luteotrophic nature. The implications of the elevated oestradiol concentrations for embryo quality require further study.

The present study aimed to establish the impact of buserelin acetate or hCG administration on day 5 post-ovulation on subsequent luteal profile and conception rate in buffalo. The buffalo (n=45) were subjected to an estrous synchronization protocol (synthetic analog of PGF2α administered, through intramuscular route, 11 days apart), followed by artificial insemination (AI) during mid to late estrus. On day 5 post-ovulation, buffalo were administered (i.m.) normal saline (Control, n=14), buserelin acetate (20μg, d5-BA, n=14) or human chorionic gonadotropin (3000IU, d5-hCG, n=17). Ovarian ultrasonography was conducted on the day of induced estrus and on days 0, 5, 12, 16 and 21 post-ovulation to assess preovulatory follicle or corpus luteum (CL) diameter. Also, on these days, jugular vein blood sampling was conducted for the estimation of plasma progesterone. First service conception rate was greater (χ(2)=5.18, P>0.05) in d5-BA and d5-hCG groups (71.4% and 47.1%, respectively) as compared to control (28.6%). Both treatment groups had a greater (P<0.05) CL diameter and plasma progesterone during the post-treatment period in comparison to that control treatment group. Treatment-induced accessory CL formation was observed in 92.9% and 76.5% buffalo of d5-BA and d5-hCG groups, respectively. In conclusion, buserelin acetate and hCG administration on day 5 post-ovulation leads to accessory CL formation that may have a role in enhancing conception rate., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This study aimed to evaluate the morphological changes in microvascular density and corpus luteum (CL) vascularization in cows treated with eCG during stimulatory and superovulatory protocols. Sixteen cows were synchronized and divided into three groups: control (n = 6), stimulated (n = 4) and superovulated (n =6), one was submitted to estrous synchronization (ES) and received no eCG (control), and those that were submitted to ES and received eCG before or after follicular deviation (superovulation and stimulation of the dominant follicle, respectively). Ovulation was synchronized using a progesterone device-based protocol. After six days of ovulation, the cows were slaughtered and the ovaries and CL were collected. The CLs were processed and photomicrographs were taken under light microscopy to assess the vascular volume density (Vv) by stereology, and scanning electron microscopy (SEM) was used to perform ultrastructural analysis of the microvasculature. The Vv in stimulated and superovulated cows significantly increased (P ≤ 0.0001) when compared to control, indicating that the eCG is able to induce angiogenic activity in bovine CL. However, no significant differences were observed between stimulated and superovulated cows. The SEM demonstrated ratings indicative of angiogenesis, marked by several button-shaped projections in the capillaries, and the presence of more dilated capillaries in CL treated with eCG. These morphological findings are evidence of an angiogenic effect of the eCG treatment in CL of cows., (© 2015 Wiley Periodicals, Inc.)

Early gestation is a critical period when implantation and placental vascularization are established, processes influenced by progesterone (P4). Although human chorionic gonadotropin (hCG) is not endogenously synthesized by livestock, it binds the LH receptor, stimulating P4 synthesis. We hypothesized treating pregnant ewes with hCG would increase serum P4, number of corpora lutea (CLs) and concepti, augment steroidogenic enzymes, and increase membrane P4 receptors (PAQRs) and angiogenic factors in reproductive tissues. The objective was to determine molecular alterations induced by hCG in pregnant sheep that may promote pregnancy. Ewes received either 600 IU of hCG or saline i.m. on day 4 post mating. Blood samples were collected daily from day 0 until tissue collection for serum P4 analysis. Reproductive tissues were collected on either day 13 or 25 of gestation and analyzed for PAQRs, CXCR4, proangiogenic factors and steroidogenic enzymes. Ewes receiving hCG had more CL and greater serum P4, which remained elevated. On day 25, StAR protein production decreased in CL from hCG-treated ewes while HSD3B1 was unchanged; further, expression of CXCR4 significantly increased and KDR tended to increase. PAQR7 and CXCR4 protein was increased in caruncle tissue from hCG-treated ewes. Maternal hCG exposure influenced fetal extraembryonic tissues, as VEGFA, VEGFB, FLT1, and ANGPT1 expression increased. Our results indicate hCG increases serum P4 due to augmented CL number per ewe. hCG treatment resulted in greater PAQR7 and CXCR4 in maternal endometrium and promoted expression of proangiogenic factors in fetal extraembryonic membranes. Supplementing livestock with hCG may boost P4 levels and improve reproductive efficiency., (© 2015 Society for Reproduction and Fertility.)

The luteal structure that develops postovulation is critical to the facilitation and maintenance of pregnancy in dairy cattle. The objectives of this experiment were to determine if the induction of an accessory corpus luteum (CL), via human chorionic gonadotropin, altered blood perfusion of CL, peripheral concentrations of progesterone, or hepatic steroid-inactivating enzymes. Twenty-eight late-lactation Holstein cows were synchronized using the Ovsynch protocol and randomly assigned to 1 of 2 treatment groups. Cows received either an injection of human chorionic gonadotropin (1,000IU, i.m.) to induce an accessory CL (cows had exactly 2CL in 1 ovary) or no treatment (cows had exactly 1CL). Corpora lutea were examined daily from d 10 to 18 (d 0 was induced ovulation) via Doppler ultrasonography and a blood sample was collected. Volume of the CL was recorded, as well as images and videos of each CL, which were analyzed for blood perfusion. On d 13, a liver biopsy was performed to analyze hepatic steroid-inactivating enzymes. Cows with 1 or 2CL had similar peripheral concentrations of progesterone. Cows with 2CL had similar luteal volumes to cows with 1CL but cows with 2CL had greater total luteal blood perfusion. Hepatic enzyme [cytochrome P450 (CYP) 1A, 3A, and 2C, aldo-keto reductase 1C, and uridine diphosphate glucuronosyltransferase] activities did not differ between cows with 1 and 2CL. Overall, the observed increase in total luteal blood perfusion in cows with 2CL did not correspond to differences in peripheral concentrations of progesterone or clearance of progesterone measured by the hepatic enzyme activity. This could indicate that induction of an accessory CL would not affect concentrations of progesterone necessary to maintain pregnancy., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Objective: To ultrasonographically investigate the effect of administration of exogenous progesterone using a progesterone-releasing intravaginal device (PRID), with or without the addition of equine chorionic gonadotropin (eCG), to cows with cystic ovarian disease (COD) in the first month postpartum., Material and Method: Thirty-three dairy cows (n = 11/group) diagnosed with COD between day 20 and 30 postpartum were randomly assigned to treatment groups (TG) PRID/PGF and PRID/PGF+eCG or a control group (CG). The CG remained untreated during the 20-day study period, which started on the day the diagnosis was made (day 0). Cows of group PRID/PGF received a PRID, which was removed on day 10, at which time PGF2α was given intravenously. Cows of the PRID/PGF+eCG group additionally received eCG on day 7 intravenously. Ovarian ultrasonography and blood sampling for measurement of plasma progesterone (P4) and estradiol-17β (E2) were done on days 0, 7, 10, 13, and 20 in the TG and on days 0, 10, and 20 in the CG., Results: On day 0, all cows had follicular cysts, six of which developed into luteal cysts in the CG. The mean cyst diameter remained unchanged in the CG and decreased in the TG by day 20 (p < 0.01). All treated cows ovulated and formed at least one corpus luteum (CL) after PRID removal, but only two control cows ovulated (p < 0.05). Of 12 ovulatory follicles in the TG dominant on day 7, eight formed a CL with a cavity, whereas of 10 ovulatory follicles in the same groups, dominant on day 10, only one formed a CL with a cavity (p < 0.05). The diameters of follicles, that formed a CL with a cavity, were larger on day 10 (p < 0.01). Mean P4 concentrations on day 0 were ≤ 1 ng/ml in all groups. P4 concentration increased during treatment and decreased below 1 ng/ml after PRID removal (p < 0.05)., Conclusion and Clinical Relevance: Treatment with PRID alone led to resolution of COD in all cows and therefore the additional benefit of eCG could not be critically assessed. CL with a cavity more often originate from larger and older dominant ovulatory follicles.

The objective of this study was to evaluate the effect of equine chorionic gonadotropin (eCG) administration associated to fixed-time AI (FTAI) on follicular dynamics, ovulation, corpus luteum (CL) development and serum progesterone concentrations. Multiparous suckled Hereford cows (n=46) in anestrus with 60-75 days postpartum were used. Females received an intravaginal device containing 0.5g of progesterone during 8 days and 2mg of estradiol benzoate i.m. at device insertion. At device removal 500μg of cloprostenol and 0.5mg of estradiol cypionate were administered i.m., and FTAI was performed 52-56h later. Cows were divided into two experimental groups to receive 400IU of eCG i.m. at device removal (n=23), while control group did not receive eCG (n=23). Daily ovarian ultrasonography (7.5MHz transducer) and progesterone concentrations determined by RIA were assayed from device removal until 30 or 14 days after FTAI, respectively. Treatment with eCG increased ovulation rate [65.2% (15/23) vs. 30.4% (7/23); P=0.018], ovulatory follicle diameter (14.5±0.4 vs. 13.1±0.7mm, mean±SEM; P=0.081), CL area from 6 to 14 days after FTAI (344.3±25.1 vs. 274.2±23.9mm(2); P=0.045) and mean serum progesterone concentrations from FTAI to 14 days later (3.0±0.2 vs. 1.8±0.2ng/ml; P=0.001), in comparison with control cows. In conclusion, the addition of eCG to a progesterone and estradiol' based treatment for FTAI improves ovulation rate and luteal function in anestrous cows. These findings have implications in order to increase pregnancy rates in FTAI treatments in Bos taurus beef cattle., (Copyright © 2014 Elsevier B.V. All rights reserved.)

This study examined the effect of a single administration of human chorionic gonadotrophin (hCG) on Day 1 to 4 after oestrus on corpus luteum (CL) development and circulating progesterone (P4). Oestrus-synchronized heifers (n=43) were administered a single intramuscular injection of saline on Day 1 (control) or 3000IU hCG on Day 1, 2, 3 or 4 after oestrus. Administration of hCG on Day 1 had no effect on CL area, on Day 2 increased CL area from Day 6 to 12 (P<0.05), on Day 3 increased CL area from Day 9 to 11, while on Day 4 increased CL size on Days 9 and 10 (P<0.05). Administration of hCG on Day 4 induced the formation of an accessory CL in 89% of heifers, resulting in a significant increase in total luteal tissue area on the ovaries compared with all other groups. Consistent with the effects on the CL, hCG on Day 1 did not affect P4 concentrations, on Day 2 significantly increased P4 compared with the control from Day 6 to 11 (P<0.05), on Day 3 resulted in a non-significant increase in P4 while hCG on Day 4 increased P4 from Day 8 to 13 compared with the control (P<0.05). In conclusion, administration of hCG as early as Day 2 after oestrus results in increased P4 in circulation from Day 6, which should have beneficial downstream effects in terms of uterine receptivity and conceptus elongation.

Study Question: Does a GnRH agonist (GnRHa) trigger followed by a bolus of 1.500 IU hCG in a group of patients at risk of ovarian hyperstimulation syndrome (OHSS) reduce the OHSS incidence compared with hCG trigger?, Summary Answer: A GnRHa trigger followed by early luteal hCG support with one bolus of 1.500 IU hCG appears to reduce OHSS in patients at risk of OHSS; however, in a low-risk group a second bolus of 1.500 IU hCG induced two cases of late onset OHSS., What Is Known Already: A GnRHa trigger is an alternative to hCG in GnRH antagonist co-treated cycles., Study Design, Size, Duration: Two RCTs were performed in four Danish IVF units. A total of 446 patients were assessed for eligibility and 390 patients were enrolled in the study from January 2009 until December 2011. The primary outcome of the study was OHSS incidence in the group at risk of OHSS., Participants/materials, Setting, Methods: Patients received a fixed dose of recombinant human FSH for the first 4 days. On the day of triggering, patients were assessed for their risk of OHSS based on the total number of follicles ≥11 mm diameter, and were classified as being at risk of OHSS when the total number of follicles ≥11 mm was between 15 and 25 and at low risk of OHSS when the total number of follicles ≥11 mm was ≤14. Two separate randomization lists were used for each of the OHSS risk groups. Women at risk of OHSS were allocated (RCT 1) to either: Group A (n = 60), ovulation triggering with a bolus of 0.5 mg buserelin (GnRHa) s.c. followed by a single bolus of 1.500 IU hCG s.c. after the oocyte retrieval-or: Group B (n = 58): 5.000 IU hCG. Similarly, women at low risk of OHSS were allocated (RCT 2) to receive either: Group C (n = 125), a bolus of 0.5 mg buserelin s.c., followed by a bolus of 1.500 IU hCG s.c. after oocyte retrieval and a second bolus of 1.500 IU hCG on the day of oocyte retrieval +5-or: Group D (n = 141), 5.000 IU hCG. Groups C and D were included in order to obtain preliminary data., Main Results and the Role of Chance: In women at risk of OHSS (RCT 1) (15-25 follicles) no OHSS case was seen in Group A (GnRHa trigger and one bolus of 1.500 IU hCG), whereas two cases of moderate late-onset OHSS occurred in group B (3.4%), (P = 0.24). In contrast, in women at a low risk of OHSS (RCT 2) (≤14 follicles) two cases of late-onset OHSS occurred in Group C (GnRHa trigger and two boluses of 1.500 IU hCG), whereas no OHSS case was encountered in Group D (P = 0.22)., Limitations, Reasons for Caution: Although the first RCT was powered to include 168 patients at risk of OHSS (15-25 follicles ≥11 mm) randomized to either GnRHa trigger or hCG trigger, the trial was prematurely discontinued when a total of 118 patients at risk of OHSS were randomized. In addition the second RCT in the OHSS low-risk group was designed as a feasibility study to assess the incidence of OHSS after GnRHa trigger and dual hCG administration versus 5.000 IU hCG. No power calculation was performed for this trial. In addition, there was a lack of blinding in the RCTs., Wider Implications of the Findings: Although a non-significant result, one bolus of 1.500 IU hCG after GnRHa trigger tended to reduce the OHSS rate in patients with 15-25 follicles ≥11 mm as well as secure the ongoing pregnancy rate. In contrast, in patients at low risk of OHSS the administration of two boluses of 1.500 IU hCG after GnRHa trigger should be avoided as it may induce OHSS.

Study Question: Is severe early ovarian hyperstimulation syndrome (OHSS) completely prevented with the GnRH agonist trigger and 1500 IU hCG luteal rescue protocol?, Summary Answer: Severe early OHSS can occur even after the GnRH agonist trigger and 1500 IU hCG luteal rescue protocol., What Is Known Already: Prior studies including over 200 women who received the GnRH agonist trigger and 1500 hCG luteal rescue protocol have reported complete prevention of severe early OHSS. Only a few late OHSS cases have been reported and it has been suggested that this protocol can be safely applied to any women under risk., Study Design, Size, Duration: This retrospective cohort study included all women who were at high risk of OHSS and were given the GnRH agonist trigger plus hCG luteal rescue protocol between December 2008 and August 2012 in the two participating centers., Participants/materials, Setting, Methods: There were 23 women with a mean estradiol level of 4891 ± 2214 pg/ml and a mean number of >12 mm follicles of 20 ± 6 on the day of ovulation triggering. OHSS was categorized according to the Golan criteria., Main Results and the Role of Chance: Overall 6 of the 23 (26%) women developed severe OHSS. Five women had severe early OHSS requiring ascites drainage and hospitalization and three of these women did not undergo embryo transfer. The number of follicles measuring 10-14 mm on the day of triggering was significantly different between women who developed severe early OHSS and those who did not., Limitations, Reasons for Caution: The small number of women with severe early OHSS may have prevented identification of other significant risk factors., Wider Implications of the Findings: Although the GnRH agonist plus 1500 IU hCG luteal rescue protocol significantly decreases the risk of severe OHSS, this life threatening complication can still occur in high-risk patients. It would be prudent to avoid hCG luteal rescue and freeze all embryos for future transfer in such women particularly when there are ≥18 follicles with 10-14 mm diameters even with few larger follicles.

Using a novel in vivo model considering a low developmental competence embryo (demi-embryo) and a subnormal fertility recipient (lactating high-yielding dairy cow), this experiment evaluated the effect of human chorionic gonadotrophin (hCG) treatment at embryo transfer (ET) on embryonic size at implantation, embryonic survival and recipient plasma progesterone (P4 ) and bovine pregnancy-specific protein B (PSPB) concentrations until day 63 of pregnancy. Embryos were bisected and each pair of demi-embryos was bilaterally transferred to recipients (n = 61) on day 7 of the oestrous cycle. At ET recipients were randomly assigned to treatment with 1500 IU hCG or to untreated controls. Higher (p < 0.01) pregnancy rates on days 25, 42 and 63, and embryo survival rate on day 63 were observed in hCG-treated cows with secondary CL than in hCG-treated cows without secondary CL and in untreated cows. Pregnancy rates and embryo survival rate were similar in hCG-treated cows without secondary CL and untreated cows. Embryonic size on day 42 was not affected by treatment with hCG, presence of secondary CL and type of pregnancy (single vs twin). Presence of secondary CL increased (p < 0.05) plasma P4 concentrations of pregnant cows on days 14, 19 and 25 but not thereafter and of non-pregnant cows on days 14-21. Treatment with hCG and presence of secondary CL had no effect on plasma PSPB concentrations, which were higher (p < 0.05) in twin than in single pregnancies. In conclusion, secondary CL induced by hCG treatment at ET significantly increased plasma P4 concentrations, the survival rate of demi-embryos and the pregnancy rate of high-yielding lactating dairy cows. Embryos were rescued beyond maternal recognition of pregnancy, but later embryonic survival, growth until implantation and placental PSPB secretion until day 63 of pregnancy were not affected by treatment or presence of secondary CL., (© 2013 Blackwell Verlag GmbH.)

The present study investigated the impact of gonadotropic hormone administration on day 12 post-ovulation on subsequent luteal profile and conception rate in buffaloes. All the buffaloes (n=48) were estrus synchronized by a synthetic analogue of prostaglandin F(2α) (PGF(2α)), administered 11 days apart, followed by insemination during mid to late estrus. To examine the effect of mid-luteal phase hormonal treatment, buffaloes were randomly divided into control (normal saline, n=14), d12-BA (buserelin acetate, 20μg, n=17) and d12-hCG (hCG, 3000IU, n=17) groups. Ovaries were scanned on the day of induced estrus to measure the preovulatory follicle (POF) diameter and on days 5, 12, 16 and 21 post-ovulation to examine the alterations in corpus luteum (CL) diameter. On the day of each sonography, blood samples were collected for the estimation of plasma progesterone. In treatment groups, luteal profile (CL diameter and plasma progesterone) on day 16-21 post-ovulation was better (P<0.05) as well as first service conception rate was higher (52.9% in each treatment group vs. 28.6%, P>0.05) compared to controls. All the pregnant buffaloes exhibited higher (P<0.05) plasma progesterone on various post-ovulation days than their respective non-pregnant counterparts. Treatment-induced accessory corpus luteum (ACL) formation was observed in 58.8 per cent and 70.6 per cent buffaloes of d12-BA and d12-hCG group, respectively, that also had higher (P<0.05) plasma progesterone compared to controls. Compared to the spontaneous CL, the diameter of ACL was less (P<0.05) in the treatment groups. In conclusion, buserelin acetate and hCG administration on day 12 post-ovulation leads to accessory CL formation, improves luteal profile and consequently increases conception rate in buffaloes., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Two independent experiments were performed on cyclic (Experiment I) and pregnant (Experiment II) gilts to examine the effect of human Chorionic Gonadotropin (hCG) administration on day 12 of the estrous cycle/pregnancy on ovarian and endometrial secretory function. Animals were divided into hCG Group (injection of 750 IU hCG) and Control Group (injection of saline). In Experiment I, the prolonged lifespan of the corpus luteum (CL), extended progesterone (P4) production (P < 0.05) and delayed luteolysis were found. In hCG Group increased ratio of PGE2:PGFM during 12 hrs period on day 15 (P < 0.05) of the estrous cycle was observed. In both experiments, higher concentrations of E2 in hCG treated gilts (P < 0.05) on days 14-15 of the estrous cycle/pregnancy were found. In Experiment II, hCG injection did not affect P4, PGE2 and PGFM concentrations in blood plasma, but reduced the number of resorbed embryos on day 30 of pregnancy. In the pregnant hCG treated gilts the immunostaining against von Willebrand Factor (vWF) demonstrated an enhanced (P < 0.05) angiogenesis in CLs and endometrium. Furthermore, the flow cytometry revealed an increased (P < 0.05) viability of cells in CLs of hCG Group. An augmented expression of Steroidogenic Acute Regulatory Protein (STAR; P < 0.05) and LH/hCG receptor mRNA (P < 0.05) in CLs of hCG Group were observed, but an elevated concentration of protein was confirmed only for STAR (P < 0.05). Our studies revealed, for the first time, that administration of hCG affects PGE2:PGFM ratio during the estrous cycle as well as the development of conceptuses through enhanced angiogenesis and decreased luteal apoptosis in early pregnant pigs.

As the only domesticated species known to exhibit both induced and spontaneous ovulation, the cat is a model for understanding the nuances of ovarian control. To explore ovarian sensitivity to exogenous gonadotropins and the influence of progestin priming, we conducted a study of queens that were down-regulated with oral progestin or allowed to cycle normally, followed by low or high doses of equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG). Our metrics included 1) fecal steroid metabolite profiles before and after ovulation induction, 2) laparoscopic examination of ovarian follicles and corpora lutea (CL) on Days 2 and 17 (Day 0 = hCG administration), and 3) ovariohysterectomy (Day 17) to assess CL progesterone concentrations, morphometrics, and histology. Reproductive tracts from time-matched, naturally mated queens (n = 6) served as controls. Every progestin-primed cat (n = 12) produced the desired response of morphologically similar, fresh CL (regardless of eCG/hCG dose) by Day 2, whereas 41.7% of unprimed counterparts (n = 12) failed to ovulate or had variable-aged CL suggestive of prior spontaneous ovulation (P < 0.05). The ovarian response to low, but not high, eCG/hCG was improved (P < 0.05) in primed compared to unprimed cats, indicating increased sensitivity to gonadotropin in the progestin-primed ovary. Progestin priming prevented hyperelevated fecal steroid metabolites and normalized CL progesterone capacity, but only when combined with low eCG/hCG. However, priming failed to prevent ancillary CL formation, smaller CL mass, or abnormal luteal cell density, which were common to all eCG/hCG-treated cats. Thus, the domestic cat exposed to eCG/hCG produces CL with structural and functional aberrations. These anomalies can be partially mitigated by progestin priming, possibly due to a protective effect of progestin associated with enhanced ovarian sensitivity to gonadotropins.

The objective of this study was to evaluate if treatment of equine chorionic gonadotrophin (eCG)-superovulated Sanjabi ewes with repeated administration of human chorionic gonadotropin (hCG) would increase the number of normal corpus luteum (CL) and serum progesterone concentrations and decrease the number of persistent follicles. The superovulated ewes were divided into four groups on day 0 (day of sponge removal); the ewes were treated by an intramuscular administration of 500 IU hCG on day 0 (Group I: n = 10), on days 0 and 1 (Group II: n = 10), or on days 0, 1, and 2 (Group III: n = 10) and no treatment for control group (n = 10). Blood samples were collected on days 0, 1, 2, 5, and 8 (day of slaughter), and serum progesterone concentrations were determined. According to progesterone concentrations, 50 (4/8) and 0 % of the ewes underwent premature luteal regression in the control group and the hCG groups, respectively. There were more CLs in Group III than in Group II and the control group. Ewes treated with hCG had a greater number of normal-looking CL. CL diameter was significantly greater in Group II and Group III than other groups. Total CL weight was significantly (P < 0.05) higher in Group III than in Group I and the control group. Number of persistent follicle and persistent follicle diameter were lower in control group compared to the other groups. Eight days after sponge removal, serum progesterone concentration was significantly higher in Group III than in Group I and the control group. The present results indicate that repeated administration of hCG supported CL formation, increased serum progesterone concentration, and prevented premature luteal regression in eCG-superovulated Sanjabi ewes.

Our objectives were to determine characteristics (size, number, and stayability) of luteal structures formed in response to human chorionic gonadotropin (hCG) administered on d 7 after timed artificial insemination (AI) and the influence of hCG on returns to estrus and pregnancy outcome. Holstein cows (n=328), milked 3 times daily, previously inseminated at first service were assigned randomly to a completely randomized design consisting of 2 treatments when at least 1 corpus luteum (CL) was detected on d 7 after AI. Treatment consisted of 1,000 IU hCG or 1 mL of saline (control) administered i.m. Blood was collected and luteal structures were mapped and sized by transrectal ultrasonography on d 7, 14, 21, 28, and 32 after AI. Blood also was collected on d 60 in all pregnant cows. Treatment with hCG induced new luteal structures in 70% of cows, regardless of pregnancy status or number of pretreatment CL. Cows producing greater than the median 46 kg of energy-corrected milk per day were less likely to respond to hCG. The number of total luteal structures per cow, original CL volume, and total luteal volume (original CL + new luteal structures) were increased by hCG. Progesterone concentration was greater in pregnant than nonpregnant cows on d 14 unless cows responded to hCG by forming new luteal structures. Concentrations of progesterone were greatest in pregnant, hCG-treated cows. Pregnancy per AI at d 32 or 60 after first AI was less in hCG- than saline-treated cows because pregnancy outcome for hCG cows that had only 1 pretreatment CL and failed to respond to hCG was only 55 to 61% of that observed in controls. Proportions of cows returning to estrus from 18 to 25 d after AI were less in hCG than control cows but greater for cows returning >25 d. Regardless of treatment, 25% of cows in both treatments retained at least 1 original CL to d 28 after AI and were not pregnant on d 32. Progesterone concentrations in these nonpregnant cows with retained CL between d 14 and 28 after AI were intermediate between nonpregnant cows that returned to estrus by d 25 and all pregnant cows. Concentrations of pregnancy-specific protein B were elevated in some of these nonpregnant, CL-retained cows, indicating early pregnancy loss. Retention of original luteal tissue in nonpregnant cows to d 28 after AI indicated that pregnancy had been initiated but failed, as verified by concentrations of progesterone and pregnancy-specific protein B., (Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The present study was carried out to elucidate the effect of progesterone (P4) from the induced corpus luteum (CL) on the characteristics of the dominant follicle (DF) in dromedary camels (Camelus dromedarius). Ovarian follicular and induced CL dynamics were monitored by transrectal ultrasonography in eight camels during the peak breeding season. The characteristics of the DF were monitored daily from the day of emergence into a wave, until it appeared to lose its dominance and the DF of a subsequent wave grew to a diameter of 13-17 mm. At this stage ovulation was induced by hCG and the DF was monitored every 8 h for 48 h. After ovulation, CL dynamics and follicular development (emergence of a new wave, growth and mature phase of the selected DF) were monitored daily. Blood samples were collected during each ultrasound examination to study the P4 profile in these animals. The CL developed to a maximum size (22.55 ± 3.24 mm) with a peak concentration of P4 (4.60 ± 2.57 ng/ml) 7 days after ovulation. The size of the CL was positively correlated with the P4 concentration (r = 0.612) during the different stages of the CL dynamics. The presence of CL did not affect the linear growth rate, duration of growth and mature phases of the DF. The development of the DF to its maximum size during its mature phase and inter-wave interval were not affected by the P4 secreted by the induced CL. In conclusion, there is no evidence from this study to suggest that P4 from induced CL altered the characteristics of a DF in dromedary camels., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The aim of the present study was to test the hypothesis that elevated concentrations of progesterone (P4) resulting from the induction of an accessory corpus luteum (CL) by human chorionic gonadotrophin (hCG) administration on day 5 after oestrus would lead to advanced conceptus elongation on day 14 following embryo transfer on day 7. The oestrous cycles of cross-bred beef heifers were synchronised and animals were randomly assigned to receive either of two treatments: (1) intramuscular injection of 3000 IU hCG on day 5 after oestrus (n=14); or (2) intramuscular injection of saline on day 5 after oestrus (n=13). Ovaries were scanned daily by transrectal ultrasonography to assess CL development. Serum concentrations of P4 were determined from daily blood samples collected from the jugular vein. In vitro-produced bovine blastocysts were transferred to synchronised recipients on day 7 after oestrus (n=15 blastocysts per recipient). Heifers were killed on day 14 after oestrus and the uterus was flushed to recover the embryos. Injection of hCG on day 5 induced ovulation of the dominant follicle in all treated heifers and increased the total area of luteal tissue on the ovary, which was associated with a significant increase (P<0.001) in serum concentrations of P4 from day 7 to day 14. Positive associations were detected between circulating P4 with CL area (within-day correlations ranging from r=0.45 to r=0.67) and total area of luteal tissue (within-day correlations ranging from r=0.65 to r=0.86) Administration of hCG did not affect the proportion of day 14 conceptuses recovered. However, compared with the control group, hCG-treated heifers had increased conceptus length (3.91±1.23 vs. 5.57±1.02 mm, respectively; P=0.06), width (1.00±0.06 vs. 1.45±0.05 mm, respectively; P=0.002) and area (5.71±0.97 vs. 8.31±0.83, respectively; P=0.02). Although numerically greater, mean interferon-τ (IFNT) production in vitro did not differ significantly (P=0.54) between embryos recovered from hCG-treated and control heifers. In contrast, there was a strong positive correlation between individual embryo length (r=0.76; P<0.001) and individual embryo area (r=0.72; P<0.001) and IFNT production. In conclusion, administration of hCG on day 5 after oestrus resulted in the formation of an accessory CL and hypertrophy of the original CL, the result of which was an increase in P4 concentrations from day 7 onwards. These elevated P4 concentrations were associated with an increased conceptus area. Furthermore, conceptus size was highly correlated with IFNT secretion in vitro.

The effects of administering hCG on subsequent ovarian structure dynamics and concentrations of progesterone in prepubertal heifers were evaluated. Forty-seven purebred Angus heifers were assigned randomly to 1 of 3 treatments: 1) 100 μg of injection of GnRH (GnRH; n = 16); 2) 1,000 IU injection of hCG (H1000; n = 16); and 3) 500-IU injection of hCG (H500; n = 15). From d -1 to 9 relative to treatment (d 0), daily blood samples were collected to determine concentrations of progesterone and ovaries of each heifer were examined daily by transrectal ultrasonography. Diameter of all follicles ≥ 4 mm and all luteal structures were mapped. Disappearance of the largest follicle occurred within 2 d in a greater percentage (P < 0.05) of all heifers in the H1000 treatment (87.5%) compared with GnRH heifers (43.8%), whereas H500 heifers (73.7%) were intermediate. A new luteal structure formed after follicle disappearance in a greater (P < 0.05) percentage of all heifers treated with H1000 (87.5%) and H500 (73.7%) heifers compared with that in GnRH-treated heifers (18.8%). The largest follicle present in ovaries of H1000 and H500 heifers was smaller (P < 0.05) than that of GnRH heifers from d 2 to 5. Heifers treated with H1000 (1.72 ± 0.19 ng/mL) had peak concentrations of progesterone on d 6 that were greater (P < 0.05) than H500 heifers (1.34 ± 0.20 ng/mL), which were greater than heifers treated with GnRH (0.31 ± 0.19 ng/mL). The mean volume of luteal tissue was greater (P < 0.05) in H1000 heifers (1.54 ± 0.15 cm(3)) than in H500 heifers (1.15 ± 0.15 cm(3)), which was greater (P < 0.05) than in heifers treated with GnRH (0.23 ± 0.15 cm(3)). We conclude that hCG was more effective than GnRH in its ability to cause disappearance of the largest follicle, increase volume of luteal tissue in the subsequent developing luteal structures, and increase concentrations of progesterone in prepubertal heifers. In addition, hCG seems to be more effective when administered at 1,000 IU than at 500 IU.

Flunixin meglumine (FM), a prostaglandin synthetase inhibitor, causes ovulatory failure in the mare. However, the effect of the FM treatment relative to the time of hCG administration on the ovulation failure has not been determined nor has its effect on the luteal function of treated mares. Estrous mares with a follicle ≥32 mm (range of 32-38 mm) were treated with 1.7 mg/kg b.w. of FM iv at zero, 12, 24 and 36 h (n=6), at 24 and 36 h (n=6), at 28 and 36 h (n=6), at 24h (n=6) or at 30 h (n=6) after treatment with 1500 IU hCG. One group received no FM (control, n=6). Progesterone concentrations were determined using RIA. Mares treated with FM 0-36 h and 24-36 h had higher (P<0.05) incidence of ovulatory failure (83 and 80%, respectively) than mares treated twice at 28 and 36 h, or once at 24 or at 30 h after hCG (16.7, 0 and 0%, respectively). The anovulatory follicles of FM treated mares luteinized and produced progesterone (>2 ng/ml). The progesterone concentration was lower in mares treated with FM at zero to 36 h and at 24-36 h after hCG than in the other groups. In conclusion, the FM administration was effective in blocking ovulation only when the treatment began ≤24 h after hCG and was continued every 12 h until ≥36 h. In addition, the FM-induced anovulatory follicles underwent luteinization of follicular cells with active production of progesterone., (Copyright © 2011 Elsevier B.V. All rights reserved.)

We recently reported that HIF-1α plays a critical role in the regulation of vascular endothelial growth factor (VEGF) expression in the developing letual cells (LCs) and VEGF-dependent angiogenesis is essential for normal luteal development. Although it is believed that hypoxia is the primary inducer of VEGF, recent reports have also shown that human chorionic gonadotrophin (hCG) up-regulates VEGF expression in developing corpus luteum (CL). Therefore the present study was designed to test the induced effects of hCG on the expression of VEGF and HIF-1α in LCs under normoxic and hypoxic conditions. In addition, we also investigated whether the signaling pathways such as phosphatidylinositol 3-kinase (PI3K)/mammalian target of rapamycin (mTOR) and mitogen-activated protein kinase (MAPK) are involved in hCG-induced VEGF in LCs. A significant increase of VEGF mRNA was found in LCs treated with hCG, which was consistent with the changes of HIF-1α protein, even under hypoxic conditions. However, there was no obvious changes of HIF-1α mRNA in hCG-treated LCs between normoxic and hypoxic conditions, indicating hCG induces VEGF expression by increasing transcription of HIF-1α, while hypoxia mainly increases HIF-1α protein stability. When LCs were pretreated with inhibitors, we found that the PI3K/mTOR signaling pathway is required for HIF-1α and VEGF expression induced by hCG, while the MAPK pathway is not required. Together, these results suggest that activation of IP3K/mTOR signaling pathway contributes to the induction of VEGF and HIF-1α in hCG-treated LCs. To our knowledge this will provide a new insight into the important mechanism of hCG/LH-induced VEGF-dependent angiogenesis in the bovine ovary., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Hypoxia inducible factors (HIFs) are transcription factors that mediate physiological responses to hypoxia. Hypoxia is established as the major inducer of HIFs, but stimuli such as transition metals and hormones also induce HIF target genes. Whilst the ovarian granulosa cell layer is known to be avascular and the follicle is vascularised via the thecal cell layer, little is known about the role of hypoxia or HIFs in regulating ovarian function. In this study, we hypothesized that hypoxia as well as non-hypoxic stimuli cooperate in promoting follicle differentiation and luteinization via HIF activity and resultant gene regulation. We quantitatively measured the HIF1alpha protein response to hCG in ovarian granulosa cell cultures and in vivo and developed a transgenic (HRE(4)-SV40-EGFP) HIF reporter mouse line. We observed a time-dependent increase of HIF1alpha protein levels in granulosa cells post-hCG in vivo, maximal around time of ovulation. hCG alone was unable to promote HIF1alpha protein accumulation in cultured granulosa cells, but increased protein abundance was observed when combined with a hypoxic stimulus. HRE-EGFP ovaries showed no follicular EGFP in stages prior to antrum formation. However, HIF regulated EGFP was maximally induced in granulosa cells around the time of ovulation and readily observed in corpora lutea. There was also an increase in HIF regulated EGFP activity in the corpora lutea from functional to regressing stages. Taken together, these observations establish the notion that HIFs play a role during follicular differentiation and luteinization., (Copyright 2010 Elsevier Ireland Ltd. All rights reserved.)

Context: The corpus luteum is a dynamic endocrine structure with periodic development and regression during menstrual cycles. Its lifespan can be prolonged by human chorionic gonadotropin (hCG). However, the signal mechanisms of this phenomenon remain unclear., Objective: Our objective was to investigate the molecular mechanisms of hCG in the maintenance of the viability of granulosa-lutein cells., Design: Granulosa-lutein cells were obtained from women undergoing in vitro fertilization. We examined the effects of hCG on the survival of cultured granulosa-lutein cells. The signal pathway inducing antiapoptotic protein was investigated., Results: hCG enhanced viability of granulosa-lutein cells through antiapoptosis but not proliferation, because the apoptotic marker of annexin V was decreased, but the proliferative markers of Ki67 and proliferating cell nuclear antigen were not increased. Myeloid cell leukemia-1 (Mcl-1) protein, but not B-cell lymphoma protein-2 or B-cell lymphoma protein-xL, was significantly induced by hCG and LH. The granulosa-lutein cells secreted vascular endothelial growth factor that induced endothelial permeability. Mcl-1 small interfering RNA increased DNA fragmentation and diminished the antiapoptotic effect of hCG. hCG induced Mcl-1 expression through the LH/hCG receptor, adenylate cyclase, protein kinase A, and cAMP response element-binding protein signal pathway. Flavopiridol inhibited Mcl-1 production, released cytochrome c, and induced apoptosis of granulosa-lutein cells., Conclusions: We first demonstrate that hCG prevents apoptosis of granulosa-lutein cells through the induction of Mcl-1 protein via the LH/hCG receptor and a cAMP response element-binding protein-dependent pathway. We may have found the molecular mechanism for luteal rescue during early pregnancy. Mcl-1 prevents apoptosis and increases cell viability but not proliferation as mechanisms for luteal rescue. Mcl-1 is a key molecule of hCG signaling.

We determined the effects of administering hCG 7 d after a fixed-time AI (TAI) on ovarian response, concentrations of progesterone, and pregnancy rates in postpartum suckled beef cows. Cows at 6 locations received 100 microg of GnRH (Fertagyl, Intervet Animal Health, Millsboro, DE) and a controlled internal drug release (CIDR) device (CIDR EAZI-Breed, Pfizer Animal Health, New York, NY), followed in 7 d by 25 mg of PGF(2alpha) (Lutalyse, Pfizer Animal Health) and CIDR removal. At 64 h after CIDR removal, cows received an injection of GnRH and AI (d 0), and then were stratified by days postpartum and parity and assigned randomly to 2 treatments administered 7 d after TAI: 1) 1 mL of saline (saline; n = 252); or 2) 1,000 IU of hCG (Chorulon, Intervet Animal Health; n = 254). Blood samples were collected on d -21, -10, and 33 relative to TAI (d 0) at all locations, on d 7 and 68 at 5 locations, and on d 14 at 1 location to determine concentrations of progesterone. Transrectal ultrasonography was used to determine pregnancy status on d 33 and 68 at all locations, to monitor response of follicles and corpora lutea (CL) in response to treatment at 1 location (n = 106) on d 7 and 14, and to determine the number of CL present in pregnant cows on d 33 in 3 locations (n = 130). Pregnant cows had greater (P < 0.05) concentrations of progesterone at the time of treatment (d 7) compared with nonpregnant cows (3.7 +/- 0.1 vs. 2.6 +/- 0.2 ng/mL, respectively). On d 14, hCG-treated cows had a greater (P < 0.05) volume of luteal tissue (12.1 +/- 0.5 vs. 7.3 +/- 0.5 cm(3), respectively) and greater concentrations of progesterone (6.8 +/- 0.4 vs. 5.4 +/- 0.5 ng/mL, respectively) compared with saline-treated cows. A greater (P < 0.01) percentage of hCG-treated cows (90.6%) had multiple CL on d 14 compared with saline-treated cows (0%), and a greater percentage of pregnant cows treated with hCG (74.6%) had multiple CL on d 33 compared with saline-treated cows (3.0%). Pregnancy rates of hCG-treated cows (56.3%) tended (P = 0.07) to differ from those of saline-treated cows (50.0%). Concentrations of progesterone in pregnant hCG-treated cows were greater (P < 0.05; 7.7 +/- 0.3 vs. 5.8 +/- 0.3 ng/mL, respectively) on d 33 than for pregnant saline-treated cows, but were similar between treatments on d 68 (7.2 +/- 0.3 vs. 6.7 +/- 0.4 ng/mL, respectively). We conclude that treatment with hCG increased the volume of luteal tissue on d 14 and concentrations of progesterone on d 14 and 33 after TAI. Treatment with hCG tended to increase pregnancy rates at 5 of 6 locations from 1.1 to 27 percentage points (average = 10.2) compared with saline, but cumulative pregnancy rates determined on d 68 after TAI were similar between treatments.

Unlabelled: Our aim was to compare Corpus luteum (CL) development and blood plasma concentration of progesterone ([P4]) in thoroughbred mares after spontaneous (, Control: C) or human chorionic gonadotrophin (hCG)-induced ovulation. Lactating mares (C=12; hCG=21) were daily teased and mated during second oestrus post-partum. Treated mares received 2500 IU hCG i.v. at first day of behavioural oestrus when dominant follicular size was >35, or=45 mm. Dominant follicle before ovulation, CL and gestational sac were measured by ultrasound and [P4] by radioimmunoassay (RIA). Blood sampling and ultrasound CL exams were done at days 1, 2, 3, 4, 8, 12, 16, 20, 25, 30, 35, 40, 45, 60 and 90 after ovulation and gestational sac from day 12 after ovulation in pregnant (P) mares; non-pregnant (NP) were followed until oestrus returned. Data analyses considered four subgroups: hCG-P, hCG-NP, C-P and C-NP. Preovulatory follicular size was smaller in hCG mares than in C: 39.2+/-2.7 mm vs 51.0+/-1.8 mm (p<0.0001). All hCG mares ovulated 24-48 h after treatment and presented similar oestrus duration as controls. C. luteum size in P mares showed the same pattern of development through days 4-35, presenting erratic differences during initial establishment. Thus, on days 1 and 3, CL was smaller in hCG-P (p<0.05); while in hCG-NP, CL size was greater than in C-NP on day three (p=0.03). Corpus luteum size remained stable until day 90 in hCG-P mares, while in C-P a transient and apparently not functional increase was detected on days 40 and 45 (p<0.05) and the decrease from day 60 onwards, made this difference to disappear. No differences were observed in [P4] pattern between P, or between NP subgroups, respectively. So, hCG-induced ovulation does not affect CL development, neither [P4] during early pregnancy. One cycle pregnancy rate tended to be lower in hCG mares while season pregnancy rates were similar to controls.

The objectives of this study were to determine the effect of a single intramuscular injection of 200IU hCG (Chorulon) on Day 12 post-mating on ovarian function and subsequent lambing performance in ewes and ewe lambs bred at synchronised oestrus during the breeding season and on the lambing performance of ewes induced to breed during late anoestrus. All animals were mated to rams at synchronised oestrus and on Day 12 post-mating given normal saline or 200IU hCG. In Experiment 1, laparoscopic results showed that hCG treatment induced accessory corpora lutea in ewes (control=0/7; hCG=5/7) but not in ewe lambs (control=0/7; hCG=0/7). In Experiment 2, hCG treatment did not improve the lambing rate (control=50; hCG=57) or the litter size (control=1.80; hCG=1.96) in ewes (control=100; hCG=91). However, hCG treatment significantly (P>0.05) improved the lambing rate (control=29; hCG=58; P<0.05) in ewes conceiving at the first oestrus after treatment. hCG treatment (control=42; hCG=42) also failed to improve the lambing rate in ewe lambs (control=48; hCG=41). In Experiment 3, hCG treatment had no significant (P>0.05) effect on the lambing rate (control=72; hCG=62) or the litter size (control=1.59; hCG=1.58) in ewes (control=111; hCG=115) induced to breed during anoestrus or on ewes returning to oestrus and conceiving after treatment (lambing rate: control=86; hCG=72; litter size: control=1.44; hCG=1.35). In conclusion, the data obtained in this study suggest that during the breeding season hCG may, by stimulating ovarian function, improve embryo survival in ewes conceiving at the first post-treatment oestrus. This effect, however was not observed in ewe lambs.

To check human chorionic gonadotropin (hCG) effects on luteal blood flow (LBF) and progesterone (P(4)) synthesis, six cows received either 3000 IU hCG or saline (NaCl) on Day 7 (Day 1=ovulation) during two estrous cycles. Plasma P(4) and LBF were measured before (0h) and up to 48h after treatment. Luteal blood flow increased by 51% (P<0.05) at 1h after hCG administration and returned to baseline levels thereafter. Plasma P(4) levels were increased from pretreatment levels by 30% at 1h (P=0.05) and 81% at 48h (P=0.02) after hCG treatment. In contrast, NaCl did not cause changes in LBF and P(4) (P>0.05). Additionally, central and peripheral parts of 14 abattoir-derived corpora lutea of the mid-luteal phase (Day 8 to 12) were perfused with Ringer solution in an in vitro microdialysis system, supplemented with 50 or 150 IU/mL hCG for 1h. Application of 50 IU/mL hCG showed no influence on P(4) response (P>0.05) in both central and peripheral parts, whereas 150 IU/mL hCG resulted in an increase of P(4) synthesis (P=0.002) in the central parts only. In vivo, hCG provoked an immediate and long-term rise in P(4) but only a temporary elevation of LBF. Luteal blood flow itself does not seem to be the exclusive cause for an increase in P(4), because the in vitro data clearly showed direct effects of hCG on P(4) secretion. Interestingly, different P(4) secretion patterns could be found between central and peripheral parts of the corpus luteum in both control and hCG perfused corpora lutea.

The present study was conducted to evaluate the endocrinological effects of the pituitary on luteal maintenance and regression in the cyclic golden hamster (Mesocritus auratus). After hypophysectomy (Hypox) at 0900 h on day 1 of the estrous cycle (the day of ovulation), the animals received injection of prolactin (PRL) or PRL plus equine chorionic gonadotropin (eCG). They were decapitated at 1500 h on day 3 of the cycle, and trunk blood was collected for measurement of progesterone (P4). Corpora lutea (CLs) were dissected from one ovary for DNA ladder detection by electrophoresis, determination of DNA fragmentation ratio by fluorometric measurement method and measurement of P4. The other ovary was used for histological observation. After the Hypox, the daily injection of 1 mg ovine PRL restrained the DNA fragmentation ratio and number of apoptotic cell in the CLs. The PRL treatment maintained the luteal morphology and increased the luteal P4 concentration, but not in the plasma P4 concentration. In addition to PRL, injection of 2 IU eCG after the Hypox also restrained the DNA fragmentation ratio and number of apoptotic cells in the CLs to the level of a pregnant animal. The PRL plus eCG treatment maintained the luteal morphology in the same manner as the PRL only treatment and increased not only the luteal but also the plasma P4 concentration. These results suggest that PRL restrains luteal apoptosis and maintains luteal morphology and that the combination of PRL and eCG restrains not only structural but also functional luteal regression in the cyclic hamster.

Vascular endothelial growth factor (VEGF)-dependent angiogenesis is essential for normal luteal development. Although it is believed that hypoxia is the primary inducer of VEGF, in the corpus luteum it is up-regulated by human chorionic gonadotrophin (hCG). As hypoxia-inducible factor (HIF)1A has been shown to regulate VEGFA under ligand-stimulated conditions, we hypothesized that the effect of hCG on luteal VEGFA was mediated through HIF1A. We studied the effect of hCG on VEGFA and HIF1A expression in human luteinized granulosa cells in vitro and in human corpora lutea in vivo. HCG up-regulated VEGFA (P < 0.05) and HIF1A (P < 0.001) in vitro and VEGFA (P < 0.05) and HIF1A (P < 0.05) in vivo. There was a correlation between HIF1A and VEGFA in vivo (P < 0.005) and in vitro (P < 0.05). Nuclear HIF1A in granulosa-lutein cells was highest during luteal formation and absent from the fully functional corpus luteum (P < 0.05). Both VEGFA (P < 0.001) and HIF1A (P < 0.01) were up-regulated by dibutyryl-cAMP, through a PKA pathway. Hypoxia increased VEGFA (P < 0.001) and HIF1A (P < 0.05) expression and hCG further augmented VEGFA (P < 0.001) and HIF1A (P < 0.01) under hypoxic conditions. However, progesterone increased hCG-stimulated VEGFA but had no effect on HIF1A expression. The expression of HIF1A is therefore hormonally regulated in luteal cells in vitro and in vivo and may regulate VEGFA expression under normoxic and hypoxic conditions. However, the differential effects of progesterone suggest that not all regulation of VEGFA is associated with an up-regulation of HIF1A.