Luteal Response Research Articles

Pregnancy and Luteal Responses to Embryo Reinsertion following Embryo Flushing in Donor Mares.

The effect of embryo reinsertion immediately after embryo flushing was studied. In Experiment 1, eight mares were used during 32 cycles (8 cycles in each group). For the first two groups, inseminated mares were flushed 8 days after ovulation and prostaglandin F2α was not administered: in group EF-ET (embryo flushing and embryo transfer) the embryo was reinserted in the same donor mare, while in the EF group, no further procedure was performed. In the third group (ET), non-inseminated mares (recipients) received a Day 8 embryo. Progesterone concentration was measured before EF/ET and 72 h after in the three groups. In Experiment 2, twelve mares were used during 17 cycles in two groups, EF-ET (n = 11) and ET (n = 6), as in Experiment 1, except that every mare was flushed 24 h after embryo transfer to retrieve the embryo. Fewer pregnancies resulted after transfer in EF-ET cycles (0/8, 0%) than in the ET group (6/8, 75%). Progesterone concentration decreased significantly (p = 0.05) 72 h after EF-ET but not in EF or ET cycles (p > 0.1). Three mares from the EF-ET showed full luteolysis and signs of endometritis. In Experiment 2, more (5/6; p = 0.08) grade 1 embryos were recovered in the ET compared to the EF-ET group (3/7); 4 embryos were graded 3-4 (were broken or had signs of degeneration) in the EF-ET group but none in the ET group. In both groups, capsule fragments were obtained as indicative of the presence of a recently destroyed embryo in the EF-ET (n = 3) and ET (n = 1) groups. Positive bacterial cultures were obtained in 2/11 and 1/6 embryo flushes from the EF-ET and ET groups, respectively.

Processes involved in prostaglandin F2alpha autoamplification in heifers.

Endometrial and luteal synthesis of prostaglandin F2alpha (PGF2A) occurs before and during luteolysis and is critical for luteal regression. This study demonstrates that PGF2A stimulates further PGF2A synthesis (autoamplification) apparently from the corpus luteum. Understanding the endocrine profile of prostaglandin F2alpha (PGF2A) autoamplification is fundamental to comprehend luteal and endometrial responses to PGF2A. On day 10 of postovulation (preluteolysis), heifers (n = 6/group) were treated intrauterine with saline or PGF2A (0.5 mg; hour 0). A third group received flunixin meglumine + PGF (FM+PGF) to prevent endogenous synthesis of PGF2A. Exogenous PGF2A was metabolized at hour 2 as measured by PGF2A metabolite (PGFM). From hours 5 to 48, concentrations of PGFM were greatest in the PGF group, smallest in the FM+PGF, and intermediate in the control suggesting endogenous synthesis of PGF2A only in PGF group. Progesterone (P4) concentrations decreased transiently between hours 0 and 1 in PGF and FM+PGF groups but rebounded to pretreatment concentrations by hours 6 and 4, respectively. No control or FM+PGF heifers underwent luteolysis during the experimental period. Conversely, in the PGF group, one heifer had complete luteolysis (P4 < 1 ng/mL), two heifers had partial luteolysis followed by P4 and CL resurgence by hour 48, and three heifers did not undergo luteolysis. Endogenous PGF2A appears to be of luteal origin due to the lack of pulsatile pattern of PGFM and lack of endometrial upregulation of oxytocin receptor (typical of endometrial synthesis of PGF2A), whereas luteal downregulation of PGF receptor and HPGD indicates a classic luteal response to PGF2A signaling although other specific mechanisms were not investigated. The hypothesis was supported that a single PGF2A treatment simulating the peak of a natural luteolytic pulse and the uteroovarian transport of PGF2A stimulates measurable endogenous PGF2A production.

Response to the sexually active buck effect in Beni Arouss goats primed with progestagens during the anoestrus and breeding seasons

The response to buck effect (BE) was studied in Beni Arouss goats during anoestrus (AS) and breeding season (BS). Prior to AS, bucks were exposed to artificial long days during 75 days followed by a natural photoperiod. Goats of group 1 were treated for 11 days with 20 mg of fluorogestone acetate (FGA) combined to 300 IU of eCG and 50 µg of cloprostenol injected 48 h prior to FGA-removal. Goats of group 2 and 3 were subjected to the same treatments, but eCG injection was replaced by a sexually active buck introduced 0 (group 2) or 48 hours (group 3) before FGA-removal. During the AS, 77% of goats induced by the BE showed an oestrus at [60–74] hr following FGA-removal for group 3 and [71–77] hr for group 2 (p < .05). In group 1, 77% of goats displayed an LH surge [24–40] hr after FGA-removal and 67% developed a luteal phase, but no LH surge or luteal response was detected in groups 2 and 3. During the BS, oestrus response reached 100% in goats synchronised with the BE ([22–68] hr in groups 2 and 3). In all groups, 84% of the goats displayed an LH surge at [30–70] hr post-treatment and 69% displayed luteal phase within 3–8 days post-treatment. After 11–15 days, the occurrence of ovulations followed by normal luteal phase was raised in all groups (91%). It was concluded that photostimulated bucks failed to induce and synchronise ovulation in goats previously treated with 20 mg of FGA and 50 µg of cloprostenol during anoestrus. Highlights The response to the buck effect was studied in Beni Arouss goats during anoestrus and breeding season. The use of sexually active bucks to induce and to synchronise oestrus and ovulation in goats treated with 20 mg of FGA and cloprostenol during the anoestrus season is not efficient in comparison with an hormonal treatment based on administration of FGA, eCG and cloprostenol. The same protocol appears as an adequate alternative for oestrus and ovulation synchronisation during the breeding season.

Endometrial and luteal responses to a prostaglandin F2alpha pulse: a comparison between heifers and mares†.

In heifers and mares, multiple pulses of prostaglandin F2alpha (PGF) are generally associated with complete luteal regression. Although PGF pulses occur before and during luteolysis, little is known about the role of minor PGF pulses during preluteolysis on subsequent luteal and endometrial PGF production that may initiate luteolysis. Heifers (n = 7/group) and mares (n = 6/group) were treated with a single minor dose of PGF (3.0 and 0.5mg, respectively) during mid-luteal phase (12 and 10days postovulation respectively). After treatment, a transient decrease in progesterone (P4) concentrations occurred in heifers between Hours 0 and 2 but at Hour 4 P4 was not different from pretreatment. In mares, P4 was unaltered between Hours 0 and 4. Concentrations of P4 decreased in both species by Hour 24 and complete luteolysis occurred in mares by Hour 48. Luteal and endometrial gene expression were evaluated 4h posttreatment. In heifers, luteal mRNA abundance of PGF receptor and PGF dehydrogenase was decreased, while PTGS2, PGF transporter, and oxytocin receptor were increased. In the heifer endometrium, receptors for oxytocin, P4, and estradiol were upregulated. In mares, luteal expression of PGF receptor was decreased, while PGF transporter and oxytocin receptor were increased. The decrease in P4 between Hours 4 and 24 and changes in gene expression were consistent with upregulation of endogenous synthesis of PGF. The hypotheses were supported that a single minor PGF treatment upregulates endogenous machinery for PGF synthesis in heifers and mares stimulating endogenous PGF synthesis through distinct regulatory mechanisms in heifers and mares.

Research and development of a silicone letrozole-releasing device to control reproduction in cattle

Two experiments were conducted to test the effectiveness of a silicone matrix as an intravaginal drug delivery device for letrozole, an aromatase inhibitor used for synchronization protocols in cattle. A wax dip-coat formulation of the intravaginal device used in previous studies was effective in releasing letrozole but was cumbersome to manufacture and deploy, resulting in unwanted variation in drug delivery and circulating concentrations of letrozole. In Experiment 1, a 3 × 3 design was used to test the release kinetics of letrozole from silicone in vitro. Silicone was mixed with 3 different letrozole drug loads (5%, 10%, 15%) and 3 different mineral oil loads (5%, 10%, 15%), and letrozole release into 62.5% ethanol was compared with the wax dip-coat formulation (positive control) by UV spectrophotometry. Letrozole was released from silicone in a dose-dependent manner, with no effect of mineral oil. Release kinetics were then examined in vivo (Experiment 2) in nulliparous beef heifers assigned randomly to six groups (n = 6/group) and given either a large surface area (LSA) with 5% or 15% drug load, a standard surface area (SSA) intravaginal silicone device impregnated with 10% or 15% drug load, a wax dip-coat device (positive control), or a blank device (negative control). Devices were inserted on Day 3 (Day 0 = ovulation) until Day 11. Blood samples were collected at 0, 30 min, 1, 2, 3, 4, 6, 8, 10, 12, and 24 h, and twice daily until Day 11 to determine letrozole plasma concentrations by LC-MS/MS and estradiol plasma concentrations by radioimmunoassay The ovaries were examined once daily from Day 3–13 by ultrasonography to determine follicular and luteal responses to treatment. Letrozole plasma concentrations were higher in heifers given a device with an LSA vs SSA (P < 0.05). Plasma concentrations of estradiol decreased the most in heifers given the 15% LSA device (P = 0.06). The interval between emergence of successive follicular waves was longest (P < 0.05) in the positive control and the 15% LSA groups. As well, the diameter profiles of the dominant follicle and the corpus luteum were largest (P < 0.01) in the positive control and 15% LSA groups. In conclusion, letrozole was released from a silicone matrix in vitro in a dose-dependent manner, and the 15% LSA devices achieved target effects on ovarian function. Results may be used to manufacture a silicone intravaginal device for delivering aromatase inhibitors in a novel synchronization protocol for cattle.

Evaluation of different hormonal treatments on oestral and ovarian responses in Moroccan Beni Arouss goats during anoestrus and breeding season.

The efficacy of eight combinations of fluorogestone acetate (FGA, 20 or 40mg as intravaginal device during 11days), equine chorionic gonadotropin (eCG, 300 or 500UI injected 48hr before FGA removal) and prostaglandin F2α (cloprostenol, 0 or 50μg injected 48hr before FGA removal) aiming at induction and synchronization of oestrus and ovulation was evaluated during the anoestrus season in spring and during the breeding season in autumn in adult Beni Arouss goats. Oestrous behaviour was recorded between 12 and 60hr after FGA removal. Blood samplings allowing to assess onset of the pre-ovulatory LH surge and increase of progesterone as sign of an active corpus luteum were performed, respectively, between 20 and 60hr and 3, 5, 8 and 15days after FGA removal. No season-related differences (spring vs. autumn) were observed for oestrous response (95% vs. 93%), pre-ovulatory LH surge (94% vs. 84%) and luteal response after 3-8 and 11-15days post-treatment (respectively 92% vs. 66% and 92% vs. 98%). The onset of oestrus (21 [13-53] vs. 32 [12-54]hr) and LH surge (26 [20-60] vs. 38 [22-60]hr) occurred significantly later in autumn. FGA (40 vs. 20mg) in autumn significantly delayed the onset of oestrus (36 [16-54] vs. 23 [12-47]hr) and LH surge (44 [26-58] vs. 33 [22-60]hr). Significant treatment-related differences were recorded for onset of LH surge (earliest for 20mg FGA, 300IU eCG, 50μg PGF2α ) and onset of luteal phase (latest for 40mg FGA, 300IU eCG, 50μg PGF2α ). In conclusion, the hormone combinations tested appeared equally effective in terms of oestrous and ovulation rates. Season has influenced significantly the onset of oestrus and LH surge, and the high dose regimen of FGA delayed the ovarian response in autumn.

Effect of eCG on the follicular dynamics and vascularization of crossbred cows with different circulating progesterone concentrations during synchronization of ovulation in an FTAI protocol

ABSTRACT: This study aimed to evaluate the effect of treatment with equine chorionic gonadotrophin (eCG) on the follicular dynamics and function of crossbred cows with different circulating progesterone (P4) concentrations during synchronization of ovulation in a fixed-time artificial insemination (FTAI) protocol. To this end, 30 crossbred cows were submitted to a pre-synchronization protocol to ensure that all of them presented corpus luteum (CL) at the beginning of the protocol, and were evaluated by transrectal ultrasonography (TRUS) to verify the presence of CL. After that, the animals underwent an ovulation synchronization protocol and evaluation of follicular dynamics and vascularization by B-mode and power-Doppler ultrasound (US). High plasma P4 concentrations at the time of ovulation synchronization negatively influenced follicle diameter on day 10 (D10), preovulatory follicle diameter, and preovulatory follicle wall vascularization area (p&lt;0.05). Cows with high P4 concentration at the time of ovulation synchronization that were treated with eCG showed follicle diameter on D10 and preovulatory follicle diameter and wall vascularization area (p&gt;0.05) similar to those of animals with low P4 concentration at the time of ovulation synchronization. Therefore, high P4 concentrations at the time of ovulation synchronization negatively influence follicular diameter and vascularization, and eCG can be used as a strategy to favor better follicular and luteal response in crossbred cows with high P4 concentrations submitted to an FTAI protocol.

Effects of Injecting GnRH 48 Hours after PGF2α on the Dynamic Follicular and Luteal Endocrine Cells in Post - Pubertal Holstein Heifers

Eighteen cycling Holstein heifers were allotted at random by weight and body condition score to one of two treatments to evaluate the effects of GnRH on luteal response when injected 48 hours (h) after the first injection in a 10 day interval between two i njections of PGF2α . Heifers in the control group (n=9) received an injection of saline 48 h after the first injection of PGF2α; however, heifers in the μg) 48treatmenthafter group n=9) received an injection of GnRH (100 the first injection of PGF2α. Heife rs were checked for estrus 3 times daily for 60 minutes each time. Blood samples were collected for analysis of progesterone on d ays 0 (first injection of PGF2α), 2 (48 h after the fi rst injection), 10 ( at the second injection of PGF2) and at day 17 (7 days a fte r the second injection of PGF2α ). Plasma samples were analyzed fo r concentration of progesterone via radioimmunoassay to evaluate luteal cell function. Concentrations of progesterone did not differ between the control heifers and treatment animals at any day of the study. However, heifers treated with GnRH showed a significant decline in concentration of progesterone from day 0 to day 2 in a luteal response to the injection; nevertheless, concentrations of progesterone increased significantly from day 2 t o day 10. Thus, this data demonstrated that injecting GnRH 48 h after PGF2α either speeds up formation of new corpus luteum or prevents full regression of the corpus luteum present prior to the first injection of PGF2α.

198 Superovulatory response in cows undergoing aromatase inhibitor treatment

The aromatase inhibitor letrozole has been used in the treatment of infertility in women by inducing mild ovarian superstimulation or augmenting the ovarian response to FSH treatment. The effect has been attributed to an apparent up-regulation of FSH receptors on granulosa cells as a result of increased androgens (Weil et al. 1999 J. Clin. Endocrinol. Metab. 84, 2951-2956). The objective of this study was to determine whether letrozole will augment the superstimulatory response in FSH-treated cattle. Mature, non-lactating Holstein cows (n=30) were given 2 luteolytic doses of PGF2α 12h apart and scanned daily by transrectal ultrasonography to detect ovulation. Transvaginal ultrasound-guided ablation of follicles ≥5mm was performed in all cows as a group, at 5 to 8 days after ovulation. On the day of ablation, cows were assigned randomly to 2 groups (n=15 per group) and given either an intravaginal letrozole-releasing device or a blank device for 5 days. Coinciding with expected wave emergence (Day 0), cows in both groups were given 8 doses of 50mg of pFSH (Folltropin; Vetoquinol, Lure, France) IM at 12-h intervals, and PGF2α on Days 3 and 3.5. At the time of the second PGF2α, vaginal devices were removed and paint was applied to the tailhead to facilitate detection of oestrus. On Day 5, cows were given gonadotropin-releasing hormone (100mg gonadorelin) IM and artificially inseminated 12 and 24h later. Ova/embryos were collected by transcervical uterine flush on Day 12. The ovaries were examined by ultrasonography on Day 0, 3.5, 5, 6.5, and 12 to record the follicular and luteal response. Nominal data were compared between groups by t-test and by ANOVA for repeated-measures and are expressed as mean±s.e.m. Binomial data were compared by chi-squared test. The number of follicles at wave emergence did not differ between letrozole and control groups (24.6±3.1 and 26.5±3.3 respectively; P=0.68). There was no treatment effect or treatment×day interaction in the number of follicles &amp;gt;8mm on Day 3.5, 5, and 6.5. Letrozole-treated cows exhibited oestrus later than controls (50.3±1.1h v. 40.7±2.0h after first PGF2α; P&amp;lt;0.001) and had less variance in interval to oestrus (residuals, 3.1±0.48h v. 6.7±0.87 h; P&amp;lt;0.01). The number of CL on Day 6.5 was lower in the letrozole group than in the control group (9.1±1.1v. 12.3±1.1; P=0.05). The proportion of ovulations (number of CL on Day 12 over number of follicles ≥3mm on Day 0) was lower in the letrozole group than in the control group (0.65±0.05v. 0.82±0.08; P=0.02). The total number of ova/embryos collected per cow did not differ between letrozole and control groups (5.0±1.9v. 5.4±1.8; P=0.75), nor did the number of transferable embryos differ (3.0±1.2v. 4.3±1.5; P=0.56). In conclusion, treatment with a letrozole-releasing device during ovarian superstimulation did not improve the superovulatory response or embryo collection rate in mature Holstein cows, but letrozole treatment resulted in more synchronous oestrus, which may be used in the design of a fixed-time AI protocol following superstimulatory treatment in cattle. This research was supported by the Alberta Livestock and Meat Agency.

Luteal response and follicular dynamics induced with equine chorionic gonadotropin (eCG) administration after insemination in sheep

The objective of this study was to evaluate the ovarian response to equine chorionic gonadotropin (eCG) administrated during the luteal phase after insemination in sheep. Thirty cyclic multiparous ewes received a fixed-time intrauterine insemination (Day 0) and were assigned to three experimental groups that received 400 IU of eCG on Day 5 (n=10) or Day 10 (n=10), while another group remained as control group without eCG (n=10). Corpus luteum area and follicular diameter were daily measured by transrectal ultrasonography and serum progesterone concentrations were daily determined by radioimmunoassay during 30days after insemination in pregnant ewes or until the following ovulation in non pregnant ewes. The administration of eCG on Day 10, but not on Day 5, increased corpus luteum size and serum progesterone concentrations (P<0.05). Follicular dynamics were influenced by both eCG treatments on Day 5 and Day 10, promoting the growth of the largest follicle present during treatment or increasing the length of the wave that emerged after treatment (P<0.05). In conclusion, the administration of 400 IU of eCG 10 days after insemination in sheep increased corpus luteum size and progesterone concentrations, as well as promoted follicular development. This information could be useful to develop new pharmacological strategies to improve the establishment of pregnancy in sheep.

The effect of repeated administrations of llama ovulation-inducing factor (OIF/NGF) during the peri-ovulatory period on corpus luteum development and function in llamas

The objective of the study was to test the hypothesis that repeated administrations of OIF/NGF during the peri-ovulatory period (pre-ovulatory, ovulatory, early post-ovulatory), will enhance the luteotrophic effect in llamas. Female llamas were examined daily by transrectal ultrasonography in B- and Doppler-mode using a scanner equipped with a 7.5-MHz linear-array transducer to monitor ovarian follicle and luteal dynamics. When a growing follicle ≥7mm was detected, llamas were assigned randomly to one of the three groups and given 1mg of purified OIF/NGF im (intramuscular) (a) pre-ovulation (single dose; n=12), (b) pre-ovulation and at the time of ovulation (2 doses, n=10), or (c) pre-ovulation, at the time of ovulation, and 24h after ovulation (3 doses, n=10). The pre-ovulatory follicle diameter at the time of treatment, ovulation rate and the first day of CL detection did not differ (P=0.3) among groups. However, maximum CL diameter was greatest (P=0.003) in llamas in the 2-dose group, and smallest in the 3-dose group. Accordingly, the 2 dose-group had the largest day-to-day profile for CL diameter (P<0.01), area of CL vascularization (<0.01), and plasma progesterone concentration (P=0.01) compared to the other groups. Interestingly, the luteal response to 3-doses of OIF/NGF during the peri-ovulatory period was not different from a single dose. In conclusion, OIF/NGF isolated from llama seminal plasma is luteotrophic and the effect on CL size and function is affected by the number and timing of treatments during the peri-ovulatory period.

Follicular characteristics and luteal development after follicle-stimulating hormone induced multiple ovulations in heifers1

A protocol based on small doses of FSH was examined for the induction of double or triple (multiple) ovulations in cattle. Ovulation rate, follicular characteristics, and luteal responses were determined. In Exp. 1, three groups of estrous-synchronized, cyclic Holstein heifers were treated once daily, on d 3 to 6 of the cycle, with a FSH product (Folltropin-V): large FSH dose (total of 150 mg; n=18), medium FSH dose (total of 130 mg, n=12), and small FSH dose (total of 80 mg; n=7). Controls received saline (n=6). Prostaglandin F(2α) was injected on d 6, ultrasound-guided aspiration of surplus follicles (if needed) was performed on d 7, and GnRH was injected on d 8 to induce ovulation. The large FSH dose induced growth of more (2.6±0.3, P<0.05) large follicles than controls on d 8; medium and small FSH doses insufficiently stimulated growth of <2 large follicles. Ovulation rates were determined in subgroups of heifers (n=10, 13, 4, and 6, respectively). The large FSH dose induced greater rates (P<0.01) of mostly double and triple ovulations (90% multiple ovulations, 70% double ovulations), most of which (89%) were bilateral, with only 2 out of 10 heifers requiring aspiration of surplus follicles. Medium and small FSH doses induced fewer multiple ovulations (38% and 25%, respectively). Estradiol concentrations on d 8 did not differ among treatments, but the concentration per large follicle in controls was greater (P<0.05) than in FSH treatments. Mean corpus luteum (CL) volume in single-ovulation controls was greater (P<0.05) than that of multiple ovulations in the large FSH group and total CL volume and progesterone concentrations were numerically greater in multiple ovulations. In Exp. 2, the characteristics of follicles aspirated on d 7 from large FSH (n=11) and control heifers (n=10) were compared. Based on estradiol-to-progesterone ratio, 57% of the large FSH-treated follicles were classified as codominant/healthy follicles and 43% as subordinate/early atretic. Although concentrations of estradiol and androstenedione in FSH-treated codominant follicles were less (P<0.05) than in controls, estradiol-to-progesterone ratio indicated that those follicles were steroidogenically active. Finely tuned small doses of FSH administered during the first follicular wave can induce a large incidence of double/triple, mainly bilateral, ovulations in cattle, which may serve as a basis for treatment aimed at promoting twinning in beef cattle.

Life or Death Decisions in the Corpus Luteum

The corpus luteum (CL) is an ephemeral endocrine organ. During its lifespan, it undergoes a period of extremely rapid growth that involves hypertrophy, proliferation and differentiation of the steroidogenic cells, as well as extensive angiogenesis. The growth phase is followed by a period in which remodelling of the tissue ceases, but it engages in unparalleled production of steroids, resulting in extraordinarily high metabolic activity within the tissue. It is during this stage that a critical juncture occurs. In the non-fertile cycle, uterine release of prostaglandin (PG)F(2α) initiates a cascade of events that result in rapid loss of steroidogenesis and destruction of the luteal tissue. Alternatively, if a viable embryo is present, signals are produced that result in rescue of the CL. This review article summarizes the major concepts related to the fate of the CL, with particular focus on recent insights into the mechanisms associated with the ability of PGF(2α) to bring about complete luteolysis. It has become clear that the achievement of luteolysis depends on repeated exposure to PGF(2α) and involves coordinated actions of heterogeneous cell types within the CL. Together, these components of the process bring about not only the loss in progesterone production, but also the rapid demise of the structure itself.

Luteotrophic effects of relaxin, chorionic gonadotrophin and FSH in common marmoset monkeys (Callithrix jacchus)

In early pregnant primates, relaxin (RLX) is highly upregulated within the corpus luteum (CL), suggesting that RLX may have an important role in the implantation of the blastocyst. Therefore, the aim of the present study was to investigate the local effects of RLX and gonadotrophins on the maintenance of the CL using an in vitro microdialysis system. CLs of common marmoset monkeys were collected by luteectomy during different stages of the luteal phase and early pregnancy. Each CL was perfused with either Ringer's solution alone or Ringer's solution supplemented with either porcine RLX (250, 500 and 1000 ng/ml) or gonadotrophins (50 IU/ml). Application of RLX provoked a significant luteal response of progesterone (P(4)) and oestradiol (E(2)) secretions during the mid-luteal phase (500 ng/ml: P(4) 54+/-42%, E(2) 24+/-11%; 1000 ng/ml: E(2) 16+/-13%), and especially during the late luteal phase (250 ng/ml: P(4) 53+/-10%; 500 ng/ml: P(4) 44+/-15%; 1000 ng/ml: P(4) 62+/-15%, E(2) 18+/-7%). The effects of RLX on steroid secretion were irrespective of the RLX dosages. While treatment with human chorionic gonadotrophin did not affect luteal steroid or RLX secretion, the application of FSH resulted in a significant increase in the secretion of both P(4) (20+/-8%) and E(2) (37+/-28%), and a prominent rise in RLX during early pregnancy. In conclusion, our results demonstrate that RLX and FSH have a luteotrophic function in the marmoset monkeys; moreover, FSH has a function beyond its traditional role just as a follicle-stimulating hormone.

Relationships between luteal activity, fertility, blood metabolites and body condition score in multiparous Estonian Holstein dairy cows under different management

The objective was to compare the relationships between luteal activity and fertility, and relate these parameters to metabolic indices and body condition changes in multiparous Estonian Holstein cows on two commercial dairy farms under different management and levels of production and nutrition (higher, H, n=54 (71 lactations) and lower, L, n=39 (39 lactations)). For statistical analysis cows were categorized according to their milk progesterone (P4) profiles as follows: normal ovarian function; delayed start of cyclicity (DC) (interval from calving to first luteal response (P45 ng/ml up to and more than 50 d respectively, followed by regular cyclicity); cessation of luteal activity (prolonged interluteal interval, P4<5 ng/ml, with a duration of 14 d between two adjacent luteal phases); prolonged luteal activity (P4 levels 5 ng/ml for 20 d without preceding insemination). The Mixed procedure of the SAS system was used to compare milk production traits, blood metabolites (ketone bodies, non-esterified fatty acids, total cholesterol) and aspartate aminotransferase, body condition scores (BCS) and fertility parameters between the two farms, and also fertility parameters between the farms within P4 categories. Differences in milk fat/protein ratio, ketone body levels and BCS indicated a deeper negative energy balance (NEB) during the first month after calving on farm L. On both farms nearly 50% of the recently calved dairy cows suffered from ovarian dysfunction during the post-partum period. Delayed start of cyclicity was the most prevalent abnormal P4 profile, 25% and 28% on farms H and L, respectively. Prolonged luteal activity accounted for one-third of atypical ovarian patterns on farm H, and cessation of luteal activity on farm L. On farm L, DC cows had lower BCS values from day 10 to day 90 after calving compared with normal cows (P<0.01) and cows lost more BCS (1.2 units) during the 40 d after calving than normal resumption cows (0.75 units; P<0.05). On farm H with moderate NEB the delayed start of ovulation post partum did not impair subsequent reproductive performance.

Developmental Programming: Deficits in Reproductive Hormone Dynamics and Ovulatory Outcomes in Prenatal, Testosterone-Treated Sheep1

Prenatal testosterone excess leads to neuroendocrine, ovarian, and metabolic disruptions, culminating in reproductive phenotypes mimicking that of women with polycystic ovary syndrome (PCOS). The objective of this study was to determine the consequences of prenatal testosterone treatment on periovulatory hormonal dynamics and ovulatory outcomes. To generate prenatal testosterone-treated females, pregnant sheep were injected intramuscularly (days 30-90 of gestation, term=147 days) with 100 mg of testosterone-propionate in cottonseed oil semi-weekly. Female offspring born to untreated control females and prenatal testosterone-treated females were then studied during their first two breeding seasons. Sheep were given two injections of prostaglandin F2alpha 11 days apart, and blood samples were collected at 2-h intervals for 120 h, 10-min intervals for 8 h during the luteal phase (first breeding season only), and daily for an additional 15 days to characterize changes in reproductive hormonal dynamics. During the first breeding season, prenatal testosterone-treated females manifested disruptions in the timing and magnitude of primary gonadotropin surges, luteal defects, and reduced responsiveness to progesterone negative feedback. Disruptions in the periovulatory sequence of events during the second breeding season included: 1) delayed but increased preovulatory estradiol rise, 2) delayed and severely reduced primary gonadotropin surge in prenatal testosterone-treated females having an LH surge, 3) tendency for an amplified secondary FSH surge and a shift in the relative balance of FSH regulatory proteins, and 4) luteal responses that ranged from normal to anovulatory. These outcomes are likely to be of relevance to developmental origin of infertility disorders and suggest that differences in fetal exposure or fetal susceptibility to testosterone may account for the variability in reproductive phenotypes.

Field Trial on the Relationship of Blood Metabolites and Body Condition Score with the Recurrence of Luteal Activity in Estonian Holstein Cows

Associations of body condition scores and blood metabolites, measured before calving and at different periods during early lactation, with recurrence of luteal activity were investigated in a 250-head commercial dairy farm during a 4-year period (1999-2002). The study was conducted on 48 dairy cows (60 lactations) with average 305-day milk yield of 8149 kg per cow. Blood samples taken 1-14 days before calving and 1-14, 28-42 and 63-77 days after calving were analysed for aspartate aminotransferase, glucose, ketone bodies, triglycerides, non-esterified fatty acids and cholesterol. Milk progesterone (P(4)) profiles (samples collected twice a week, P(4) levels measured in whole milk by enzyme immunoassay) were used to evaluate the interval from calving to first luteal response, P(4) >5 ng/ml, and the interval from calving to first normal cycle. The MIXED procedure of the sas system was used to study the association of investigated parameters. A higher concentration of ketone bodies before calving was associated with shorter interval to recurrence of first normal cycle (P = 0.007) and tended to be related to shorter interval from calving to first luteal response (P = 0.071). A lower prepartum aminotransferase activity showed a tendency to be associated with shorter interval from calving to first luteal response (P = 0.084). Results suggest metabolic status up to 2 weeks prepartum to be related to the resumption of postpartum luteal activity in Estonian Holstein dairy cows.

Sympathetic skin response in premenstrual syndrome

Premenstrual syndrome is a term which includes a broad group of emotional, behavioral and physical symptoms that occur for several days before menses and subside following the menstrual period. Many women experience premenstrual syndrome symptoms, particularly physical ones such as breast tenderness and swelling. Approximately 5-10% women suffer from severe premenstrual syndrome and another 30-40% have moderate symptoms. Premenstrual syndrome continues to be an unsolved problem. In this study, we evaluated 24 premenstrual syndrome patients and 20 healthy women in the control group. The ages of the women were 22-34 years (mean +/- SD: 25+/-3) for the premenstrual syndrome group and 23-34 (25+/-3) for the control group. The sympathetic skin response was recorded from the palms, soles and genital regions by using electrical stimuli to the median nerve at the wrist. The sympathetic skin response was recorded twice, in the follicular and late luteal phases of menstruation. The follicular and late luteal phase sympathetic skin response of the two groups were compared. The amplitudes and latency values of the late luteal and follicular phase sympathetic skin response from the premenstrual syndrome group and control group women were statistically similar. We also did not find any latency or amplitude difference in the sympathetic skin response obtained from the three regions of the premenstrual syndrome patients and the control group. We checked sympathetic skin response in the symptomatic (late luteal phase) and asymptomatic (follicular phase) periods of patients with premenstrual syndrome, a disorder known to have many autonomic symptoms, to determine whether there was sudomotor sympathetic involvement. The results of our PMS patients indicate at the very least that there is no difference with the control subjects as regards peripheral sudomotor functions.

Overview of the response of anoestrous ewes to the ram effect.

The present review summarises knowledge of the reproductive response of anoestrous ewes to the introduction of rams - in other words, the ram effect. The ovarian and endocrine response, the factors that determine whether ewes will respond or not (associated with both the stimulus and the receptivity of the ewes) and some aspects of practical management are discussed. Information on the use of the ram effect to stimulate post-partum, prepubertal and cyclic ewes is also given. New insights are provided on ovarian response patterns, including recently collected information on luteal responses. The existence of delayed ovulation (5-7 days after the introduction of the rams) followed by luteal phases of normal or short length, luteal cysts and luteinised follicles is reported after scanning the ovaries with ultrasound. Endocrine parameters for depth of anoestrus, such as LH pulsatility and FSH concentrations, and how the concentrations of these hormones should be considered are discussed. Particular attention is paid to the observation of spontaneous, higher LH pulsatility and higher FSH concentrations in anoestrous ewes that respond to rams with luteal phases than in those that fail to respond. The use of progestogen priming and single progestogen administration and the possible advantages for synchronisation of oestrus are also discussed. Other factors that should be considered before the ram effect is applied, such as the strength of the stimulus and some practical considerations, are also included.

Overview of the response of anoestrous ewes to the ram effect

The present review summarises knowledge of the reproductive response of anoestrous ewes to the introduction of rams – in other words, the ram effect. The ovarian and endocrine response, the factors that determine whether ewes will respond or not (associated with both the stimulus and the receptivity of the ewes) and some aspects of practical management are discussed. Information on the use of the ram effect to stimulate post-partum, prepubertal and cyclic ewes is also given. New insights are provided on ovarian response patterns, including recently collected information on luteal responses. The existence of delayed ovulation (5–7 days after the introduction of the rams) followed by luteal phases of normal or short length, luteal cysts and luteinised follicles is reported after scanning the ovaries with ultrasound. Endocrine parameters for depth of anoestrus, such as LH pulsatility and FSH concentrations, and how the concentrations of these hormones should be considered are discussed. Particular attention is paid to the observation of spontaneous, higher LH pulsatility and higher FSH concentrations in anoestrous ewes that respond to rams with luteal phases than in those that fail to respond. The use of progestogen priming and single progestogen administration and the possible advantages for synchronisation of oestrus are also discussed. Other factors that should be considered before the ram effect is applied, such as the strength of the stimulus and some practical considerations, are also included.