Systemic Progesterone Concentrations Research Articles

PGF2α induces a pro-labour phenotypical switch in human myometrial cells that can be inhibited with PGF2α receptor antagonists.

Preterm birth is the leading cause of infant morbidity and mortality. There has been an interest in developing prostaglandin F2α (PGF2α) antagonists as a new treatment for preterm birth, although much of the rationale for their use is based on studies in rodents where PGF2α initiates labour by regressing the corpus luteum and reducing systemic progesterone concentrations. How PGF2α antagonism would act in humans who do not have a fall in systemic progesterone remains unclear. One possibility, in addition to an acute stimulation of contractions, is a direct alteration of the myometrial smooth muscle cell state towards a pro-labour phenotype. In this study, we developed an immortalised myometrial cell line, MYLA, derived from myometrial tissue obtained from a pregnant, non-labouring patient, as well as a novel class of PGF2α receptor (FP) antagonist. We verified the functionality of the cell line by stimulation with PGF2α, resulting in Gαq-specific coupling and Ca2+ release, which were inhibited by FP antagonism. Compared to four published FP receptor antagonists, the novel FP antagonist N582707 was the most potent compound [Fmax 7.67 ± 0.63 (IC50 21.26nM), AUC 7.30 ± 0.32 (IC50 50.43nM), and frequency of Ca2+ oscillations 7.66 ± 0.41 (IC50 22.15nM)]. RNA-sequencing of the MYLA cell line at 1, 3, 6, 12, 24, and 48h post PGF2α treatment revealed a transforming phenotype from a fibroblastic to smooth muscle mRNA profile. PGF2α treatment increased the expression of MYLK, CALD1, and CNN1 as well as the pro-labour genes OXTR, IL6, and IL11, which were inhibited by FP antagonism. Concomitant with the inhibition of a smooth muscle, pro-labour transition, FP antagonism increased the expression of the fibroblast marker genes DCN, FBLN1, and PDGFRA. Our findings suggest that in addition to the well-described acute contractile effect, PGF2α transforms myometrial smooth muscle cells from a myofibroblast to a smooth muscle, pro-labour-like state and that the novel compound N582707 has the potential for prophylactic use in preterm labour management beyond its use as an acute tocolytic drug.

Longitudinal patterns in progesterone metabolites in pregnant and non-pregnant Steller sea lions (Eumetopias jubatus)

Similar to the several pinniped and a few terrestrial carnivore species, the Steller sea lion has a seasonal synchronized mating scheme enabled by a female reproductive cycle that includes embryonic diapause, delayed implantation, and pseudopregnancy (a state in which the corpus luteum produces progesterone for approximately as long as in pregnant females). Due to this, circulating systemic progesterone concentrations cannot be used to differentiate pregnant and nonpregnant females during early gestation. With the use of advanced measurement technologies such as liquid chromatography tandem mass spectrometry (LC-MS/MS) additional steroid hormones are measurable which can provide additional information on the endocrine pathways throughout gestation. Our objectives were to further characterize endocrine patterns in female Steller sea lion pregnancy by 1) quantifying longitudinal profiles of hormone metabolites in pregnant and non-pregnant female sera, and 2) evaluating hormone profiles to identify pregnant animals within the early stage of gestation. Three gestation stages were delineated based on what is believed to be the period of implantation (September-October): EARLY (August– November), MID (December-February), and LATE (March to May). Five steroids, Progesterone (P4), 5α-dihydroprogesterone (DHP), 17αOH-progesterone (17OHP), 20αOH-progesterone (20OHP), and androstenedione (A4), were detected in both pregnant and non-pregnant animals. A significant difference in P4 concentrations was measured between EARLY and MID gestation (p ≤ 0.01) in both pregnant and non-pregnant animals. During MID gestation there was a significant difference (p ≤ 0.05) between pregnant and non-pregnant animals in all pregnanes measured. Significant patterns of correlation between P4 and 17OHP and between P4 and DHP were detected during EARLY and MID gestation in non-pregnant animals. While those significant correlations also exist in EARLY pregnant animals, this pattern was lost by MID gestation. This loss of correlation suggests a potential shift in progesterone metabolism from ovarian to alternative tissue (e.g. fetal gonads or adrenal glands) by MID gestation in Steller sea lions. We were unable to identifying a steroid hormone biomarker capable of differentiating pseudopregnancy from pregnant animals and conclude that such a biomarker likely falls outside of the traditional progesterone metabolic pathway.

Reintroducing serum FSH measurement during ovarian stimulation for ART

Research questionWhat is the impact of systemic FSH concentrations during ovarian stimulation for IVF/intracytoplasmic sperm injection on systemic progesterone concentrations in the late follicular phase? DesignPost-hoc analysis of a previously performed randomized controlled trial (RCT) performed between November 2017 and February 2020 in a tertiary IVF centre. The RCT included patients with infertility undergoing ovarian stimulation in a gonadotrophin-releasing hormone (GnRH) antagonist protocol. The GnRH antagonist was administered at 08:00 h and recombinant FSH at 20:00 h. Ultrasound and blood tests were performed 3–5 h after the GnRH antagonist. ResultsThe subgroup analysis comprised 105 patients. Systemic FSH concentrations increased from Day 2/3 until initiation of GnRH antagonist and remained constant until the day of trigger (DoT). The total group was split according to the median FSH DoT concentration (12.95 IU/l; Group A <12.95 IU/l; Group B ≥12.95 IU/l). Significant differences, with the higher concentrations in Group B, were found for: systemic FSH concentration on Day 2/3 (P = 0.04), total gonadotrophin dosage (P = 0.03), progesterone on DoT (P = 0.001) and progesterone per follicle (P = 0.004). In the total group, systemic DoT FSH concentration was statistically significantly positively correlated with the DoT progesterone concentration and the ratio of progesterone per follicle (ρ = 0.37 and 0.38, respectively, both P < 0.001). No significant correlations were seen between the systemic DoT FSH concentration and the number of retrieved oocytes. ConclusionWhile ovarian response seems to be independent from the systemic FSH concentrations on the DoT, high concentrations of circulatory FSH augment the production of progesterone.

Biochemical characterization of progesterone-induced alterations in bovine uterine fluid amino acid and carbohydrate composition during the conceptus elongation window†.

Pregnancy establishment in cattle is contingent on conceptus elongation-a fundamental developmental event coinciding with the time during which most pregnancies fail. Elongation in vivo is directly driven by uterine secretions, indirectly influenced by systemic progesterone concentrations, and has yet to be recapitulated in vitro. To better understand the microenvironment evolved to facilitate this phenomenon, the amino acid and carbohydrate composition of uterine fluid was interrogated using high-throughput metabolomics on days 12, 13, and 14 of the estrous cycle from heifers with normal and high circulating progesterone. A total of 99 biochemicals (79 amino acids and 20 carbohydrates) were consistently identified, of which 31 showed a day by progesterone interaction. Fructose and mannitol/sorbitol did not exhibit a day by progesterone interaction, but displayed the greatest individual fluctuations (P≤0.05) with respective fold increases of 18.39 and 28.53 in high vs normal progesterone heifers on day 12, and increases by 10.70-fold and 14.85-fold in the uterine fluid of normal progesterone animals on day 14 vs day 12. Moreover, enrichment analyses revealed that the phenylalanine, glutathione, polyamine, and arginine metabolic pathways were among the most affected by day and progesterone. In conclusion, progesterone had a largely stabilizing effect on amino acid flux, and identified biochemicals of likely importance to conceptus elongation initiation include arginine, fructose, glutamate, and mannitol/sorbitol.

Ovine placental steroid synthesis and metabolism in late gestation.

Steroid synthesis is required for pregnancy maintenance and for parturition, but comparatively little is known about the major metabolic routes that influence circulating concentrations. Dietary intake changes progesterone and estradiol concentrations in pregnant ewes but whether this reflects placental synthesis is unknown. Progesterone metabolism by 5alpha-reduction is a major metabolic route in other species and can influence the onset of parturition. Therefore, studies were conducted to (1) determine placental enzyme activity, progesterone, and estradiol measured by immunoassay in late gestation ewes on low-, moderate-, and high-nutritional planes, (2) to assess the significance of 5alpha-reduction of progesterone in determining progesterone concentrations in late gestation ewes (gestation day 145) given finasteride to inhibit 5alpha-reductase metabolism. In the second experiment, steroid profiles were examined comprehensively in blood and tissues by liquid chromatography tandem mass spectrometry for the first time in this species. Dietary intake altered progesterone and estradiol serum concentrations but without correlated changes in placental 3beta-hydroxysteroid dehydrogenase, 17alpha-hydroxylase/17,20-lyase cytochrome P450 or aromatase activity. 5alpha-reduced pregnane metabolites were identified in ewes at 145 days of gestation, but concentrations were lower than those of progesterone. Finasteride inhibited 5alpha-reduced progesterone metabolism but did not impact serum progesterone concentrations in these ewes. We conclude that (1) diet-induced changes in serum progesterone and estradiol concentrations are not likely a result of altered placental synthesis of sex steroid but most likely by their metabolism, and (2) metabolism by 5α-reduction is not a major determinant of systemic progesterone concentrations in late gestation ewes.

Pre-ovulatory follicle affects corpus luteum diameter, blood flow, and progesterone production in mares

Color Doppler ultrasonography was used to study the temporal relationships between pre-ovulatory follicle (POF) and corpus luteum (CL) diameter and blood flow, with systemic progesterone (P4) concentration during two transitional ovulatory seasons in mares. Variables of POF and CL/P4 were evaluated for 6days before and 17days after ovulation, respectively. Evaluations were performed during two consecutive estrous cycles in spring and fall seasons, and during the last estrous cycle of the season. There were significant correlations among POF and CL variables, and P4 concentration that ranged from 0.24 to 0.95, and among the ratios of different variables that ranged from 0.39 to 0.92. There were linear regressions (P<0.01−0.001) for all comparisons among different variables. The POF diameter before the first ovulation of the season was larger (P<0.05), and POF vascularity was less (P<0.05), than in the last estrous cycle during the season. The CL blood flow was less (P<0.01) during the last compared with first pre-ovulatory period of the season. The POF diameters were positively correlated (r=0.67) during the two pre-ovulatory periods of spring and fall. Results provide evidence that the POF affects CL diameter and blood flow, and subsequently P4 production, and that POF diameter is repeatable within the same individual during different seasons.

Human Chorionic Gonadotropin Has Anti-Inflammatory Effects at the Maternal-Fetal Interface and Prevents Endotoxin-Induced Preterm Birth, but Causes Dystocia and Fetal Compromise in Mice.

Human chorionic gonadotropin (hCG) is implicated in the maintenance of uterine quiescence by down-regulating myometrial gap junctions during pregnancy, and it was considered as a strategy to prevent preterm birth after the occurrence of preterm labor. However, the effect of hCG on innate and adaptive immune cells implicated in parturition is poorly understood. Herein, we investigated the immune effects of hCG at the maternal-fetal interface during late gestation, and whether this hormone can safely prevent endotoxin-induced preterm birth. Using immunophenotyping, we demonstrated that hCG has immune effects at the maternal-fetal interface (decidual tissues) by: 1) increasing the proportion of regulatory T cells; 2) reducing the proportion of macrophages and neutrophils; 3) inducing an M1 → M2 macrophage polarization; and 4) increasing the proportion of T helper 17 cells. Next, ELISAs were used to determine whether the local immune changes were associated with systemic concentrations of progesterone, estradiol, and/or cytokines (IFNgamma, IL1beta, IL2, IL4, IL5, IL6, IL10, IL12p70, KC/GRO, and TNFalpha). Plasma concentrations of IL1beta, but not progesterone, estradiol, or any other cytokine, were increased following hCG administration. Pretreatment with hCG prevented endotoxin-induced preterm birth by 44%, proving the effectiveness of this hormone as an anti-inflammatory agent. However, hCG administration alone caused dystocia and fetal compromise, as proven by Doppler ultrasound. These results provide insight into the mechanisms whereby hCG induces an anti-inflammatory microenvironment at the maternal-fetal interface during late gestation, and demonstrate its effectiveness in preventing preterm labor/birth. However, the deleterious effects of this hormone on mothers and fetuses warrant caution.

Pregnancy losses in cattle: potential for improvement.

For heifers, beef and moderate-yielding dairy cows, it appears that the fertilisation rate generally lies between 90% and 100%. For high-producing dairy cows, there is a less substantive body of literature, but it would appear that the fertilisation rate is somewhat lower and possibly more variable. In cattle, the major component of embryo loss occurs in the first 16 days following breeding (Day 0), with emerging evidence of greater losses before Day 8 in high-producing dairy cows. In cattle, late embryo mortality causes serious economic losses because it is often recognised too late to rebreed females. Systemic concentrations of progesterone during both the cycle preceding and following insemination affect embryo survival, with evidence of either excessive or insufficient concentrations being negatively associated with survival rate. The application of direct progesterone supplementation or treatments to increase endogenous output of progesterone to increase embryo survival cannot be recommended at this time. Energy balance and dry matter intake during the first 4 weeks after calving are critically important in determining pregnancies per AI when cows are inseminated at 70-100 days after calving. Level of concentrate supplementation of cows at pasture during the breeding period has minimal effects on conception rates, although sudden reductions in dietary intake should be avoided. For all systems of milk production, more balanced breeding strategies with greater emphasis on fertility and feed intake and/or energy must be developed. There is genetic variability within the Holstein breed for fertility traits, which can be exploited. Genomic technology will not only provide scientists with an improved understanding of the underlying biological processes involved in fertilisation and the establishment of pregnancy, but also, in the future, could identify genes responsible for improved embryo survival. Such information could be incorporated into breeding objectives in order to increase the rate of genetic progress for embryo survival. In addition, there is a range of easily adoptable management factors, under producer control, that can either directly increase embryo survival or ameliorate the consequences of low embryo survival rates. The correction of minor deficits in several areas can have a substantial cumulative positive effect on herd reproductive performance.

Form of supplemental selenium fed to cycling cows affects systemic concentrations of progesterone but not those of estradiol

In areas where soils are deficient in selenium (Se), dietary supplementation of this trace mineral directly to cattle is recommended. Selenium status affects fertility, and the form of Se supplemented to cows affects tissue-specific gene expression profiles. The objective of this study was to determine whether the form of Se consumed by cows would affect follicular growth and the production of steroids. Thirty-three Angus-cross cows that had ad libitum access of a mineral mix containing 35 ppm of Se in free-choice vitamin–mineral mixes as either inorganic (ISe), organic (OSe), or a 50/50 mix of ISe and OSe (MIX) for 180 days were used. After 170 days of supplementation, all cows were injected with 25-mg PGF2α to induce regression of the CL and then monitored for behavioral estrus (Day 0). From Day 4 to Day 8 after estrus, follicular growth was determined by transrectal ultrasonography. On Day 6, cows were injected with PGF2α (20 then 15 mg, 8–12 hours apart) to induce regression of the developing CL and differentiation of the dominant follicle of the first follicular wave into a preovulatory follicle. On Day 8, 36 hours after PGF2α (20 mg), the contents of the preovulatory follicle were aspirated by ultrasound-guided follicular puncture. Blood collected on Days 6 and 8 and follicular fluid collected on Day 8 was analyzed for concentrations of progesterone and estradiol. Form of Se supplemented to cows affected (P = 0.04) the systemic concentration of progesterone on Day 6, but not on Day 8. Form of Se did not affect the systemic concentration of estradiol on Day 6 or Day 8. Form of Se tended to affect (P = 0.07) the concentration of progesterone, but not that of estradiol, in the follicular fluid. Form of Se did not affect diameter of the dominant ovarian follicle on Days 4 to 6, but tended to affect (P = 0.08) the diameter of the preovulatory follicle on Day 8. Our results suggest that form of Se fed to cows affects the production of progesterone but not that of estradiol. Further investigation of organic Se–induced increases in progesterone and potentially the effects of increased progesterone on the establishment of pregnancy, especially in cows of lower fertility, is warranted.

Bilateral ovarian leiomyoma treated by standing laparoscopic ovariectomy

SummaryAn 11‐year‐old Warmblood mare presented to a referral facility due to behavioural problems. A hormonal profile performed prior to referral included increased systemic concentrations of testosterone and progesterone but a low concentration of inhibin. Transrectal ultrasound revealed ovaries consistent with a normal cycling mare. At the request of the owner, standing bilateral laparoscopic ovariectomy was performed without complication. Both ovaries were submitted for histopathological examination which revealed bilateral ovarian leiomyoma.

Ultrasound monitoring of blood flow and echotexture of the corpus luteum and uterus during early pregnancy of beef heifers

The aim was to characterize changes in the ultrasound characteristics of the CL and uterus in pregnant, inseminated nonpregnant, and cyclic beef heifers and to correlate findings with systemic progesterone (P4) concentrations with the intention of identifying possible markers for early identification of pregnancy. Heifers were randomly selected for artificial insemination after estrus synchronization. Ultrasound examinations of the CL and uterus were carried out by transrectal ultrasonography using a high-resolution ultrasound scanner equipped with a 12 MHz linear array probe on Days 7, 11, 14, 16, and 18 after artificial insemination (Day 0; i.e., estrus). Cross-sectional B-mode images of the CL were captured for calculation of CL tissue area and echotexture. Images of the CL and associated blood flow were captured and stored for analysis of luteal blood flow area and ratio. Longitudinal B-mode images of the uterine horns were captured just beyond the bifurcation of the uterine horns and stored for analysis of contrast and homogeneity (MaZda v4.6; Technical University of Lodz, Institute of Electronics, Poland). A total of three images were captured for each structure of interest. Serum concentrations of P4 were determined from blood samples collected at each ultrasound examination. After pregnancy diagnosis by ultrasound, heifers were retrospectively allocated as being pregnant (embryonic heartbeat on Day 28; n = 14) or nonpregnant (interestrous interval 18–21 days; n = 8) and their data were compared with noninseminated cyclic heifers (n = 10). Corpus luteum tissue area did not appear to change between pregnant, nonpregnant, or cyclic control groups between Days 7 and 18 (P > 0.05). No significant differences in CL echotexture characteristics were found between groups at any time point. There were no significant differences between pregnant, nonpregnant, and cyclic control groups for CL blood flow area (P > 0.05). However, CL blood flow ratio decreased significantly (P < 0.05) in both inseminated nonpregnant and cyclic heifers between Days 14 and 18, whereas it remained unchanged in pregnant heifers (P > 0.05). Uterine homogeneity was not significantly different between groups at any time point (P > 0.05). However, uterine contrast was significantly greater (P < 0.05) in pregnant compared with cyclic control heifers on Days 16 and 18. Concentrations of P4 were lower (P < 0.05) in nonpregnant and control heifers than in pregnant heifers from Days 16 to 18. In conclusion, there were differences between nonpregnant and cyclic heifers compared with pregnant heifers in P4 concentrations from Day 16. On Day 18, the CL and uterine characteristics were different between the nonpregnant and pregnant heifers. Ultrasound measures of CL blood flow and uterine echotexture may be useful to establish pregnancy status. Further investigation is required to identify if pregnancy diagnosis can be made on Day 18 or at a later day postpartum.

Composition of the bovine uterine proteome is associated with stage of cycle and concentration of systemic progesterone

Early embryonic loss accounts for over 70% of total embryonic and foetal loss in dairy cattle. Early embryonic development and survival is associated with the concentration of systemic progesterone. To determine if the uterine proteome is influenced by stage of cycle or systemic progesterone concentrations, uterine flushings were collected from the ipsi- and contralateral uterine horns of beef heifers on Days 7 (n = 10) and 15 (n = 10) of the oestrous cycle. Animals were separated into low or high progesterone groups based on plasma progesterone concentrations on Day 5 of the cycle. Samples were albumin depleted before iTRAQ R labeling and subsequent strong cation exchange-LC-MS/MS analyses. A total of 20 proteins were up to 5.9-fold higher (p<0.05) and 20 were up to 2.3-fold lower on Day 15 compared toDay 7. In addition, the expression of a number of proteins on Day 7 and/or 15 of the cycle was correlated with progesterone concentrations during Days 3–7 or the rate of change in progesterone between Days 3 and 7. This study highlights the dynamic changes occurring in the microenvironment surrounding the embryo during this period. The findings here also support the hypothesis that progesterone supports embryonic development by altering the maternal uterine environment.

Insulin-like growth factor and insulin-like growth factor-binding proteins in the bovine uterus throughout the oestrous cycle

The aims of the present study were to assess several components of the insulin-like growth factor (IGF) system in bovine uterine flushings across different days of the oestrous cycle and to examine the relationship between the IGF system and systemic progesterone concentrations. Uterine flushings and plasma were collected from cows on Days 3, 7, 11 and 15 of the oestrous cycle. The IGF-1 concentration was more than 5-fold higher in the uterus compared with plasma on Days 7 and 11 of the cycle, with values similar on Days 3 and 15. Similarly, uterine concentrations of IGF-binding protein (IGFBP)-2 and IGFBP-3 were up to 10- and 4-fold higher than in plasma, respectively, suggesting synthesis and/or transportation of the IGFBPs into the uterus. In addition, concentrations of IGFBP-2 and IGFBP-3 were higher in the uterine horns, ipsilateral to the corpus luteum, on Day 15. This difference could indicate a local controlling mechanism with progesterone possibly playing a role in regulating the concentration of IGFBPs between the uterine horns. There was no significant relationship between systemic progesterone concentrations and IGFBP concentrations on Day 7 of the oestrous cycle. The present study shows that uterine concentrations of IGFBPs are cycle stage specific and also suggests IGF-dependent and -independent functions for IGFBPs during a time of major change in the developing embryo.

Evidence for independent evolution of functional progesterone withdrawal in primates and guinea pigs

Cervix remodeling (CRM) is a critical process in preparation for parturition. Early cervix shortening is a powerful clinical predictor of preterm birth, and thus understanding how CRM is regulated is important for the prevention of prematurity. Humans and other primates differ from most other mammals by the maintenance of high levels of systemic progesterone concentrations. Humans have been hypothesized to perform functional progesterone withdrawal (FPW). Guinea pigs are similar to humans in maintaining high-progesterone concentrations through parturition, thus making them a prime model for studying CRM. Here, we analyze the phylogenetic history of FPW and document gene expression in the guinea pig uterine cervix. Data on progesterone withdrawal were collected from the literature, and character evolution was analyzed. Uterine cervix samples were collected from non-pregnant, mid-pregnant and late pregnant guinea pigs. RNA was extracted and sequenced. Relative transcript levels were estimated and compared among sample groups. The phylogenetic analysis shows that FPW evolved independently in primates and guinea pigs. The transcriptome data confirms that guinea pigs down-regulate progesterone receptor toward parturition, in contrast to humans. Some of the similarities between human and guinea pig are: down-regulation of estrogen receptor, up-regulation of VCAN and IGFBP4 as well as likely involvement of prostaglandins. (i) FPW in guinea pigs evolved independently from that in primates. (ii) A small set of conserved gene regulatory changes has been detected.

121 RELATIONSHIP BETWEEN PROSTAGLANDIN E2 METABOLITE, UTERINE ARTERIAL VASODILATORY CHANGES, AND PROGESTERONE DURING THE OESTROUS CYCLE AND EARLY PREGNANCY IN BEEF COWS

Vasodilation of the female reproductive vasculature is essential to supply the increasing demands associated with reproductive processes; including maintenance of a healthy functional luteal structure, and the establishment and continuance of pregnancy. Due to its vasodilatory effects, prostaglandin E2 has been identified as a potential candidate sustaining such vascular effects. Here, the relationship between systemic concentrations of prostaglandin E2 metabolite (PGEM) and progesterone (P4), and uterine arterial vasodilatory changes were evaluated during the oestrous cycle and early pregnancy. Synchronised Angus cows (n = 19) were artificially inseminated (AI) and examined for pregnancy 45 d post-AI. Jugular blood samples were collected on Days 0, 3, 6, 10, 16, 20, 25, and 32 (Day 0 = AI) for serum PGEM and P4 evaluation. Vasodilatory changes of the uterine arteries were also characterised using the B-mode ultrasound luminal circumference (UAC). Retrospective comparisons were carried out between pregnant (n = 10) and nonpregnant (n = 9) animals. The GLM Procedure of SAS (SAS Institute Inc., Cary, NC, USA) was used to evaluate the effects of pregnancy status and sampling day over PGEM, UAC, and P4. Pearson correlations between PGEM, UAC, and P4 were obtained by the CORR Procedure of SAS. All variables were measured in triplicate and thresholds for significance and tendency were fixed at P ≤ 0.05 and 0.06 to 0.10, respectively. From Day 0 to Day 20, the overall PGEM concentration tended to be higher in nonpregnant cows compared to pregnant ones (P = 0.08; 90.97 ± 4.38 v. 78.28 ± 3.01 pg mL–1, respectively), while a shift in favor of pregnant cows was observed at Day 32 (P = 0.29; 89.88 ± 14.23 v. 106.3 ± 7.17 pg mL–1, respectively). Both uterine arteries showed similar UAC (P ≥ 0.05); therefore, they were combined in one average value per animal each sampling day. There was an overall increase of the UAC over time, from 10.49 ± 0.01 mm at Day 0 to 12.17 ± 0.02 mm at Day 20 (P ≤ 0.0001), but no significant differences were found between pregnant and nonpregnant cows during this period (P ≥ 0.05). An overall sharp increase in P4 concentrations was observed from Day 3 to Day 16 (0.60 ± 0.12 to 9.66 ± 0.45 ng mL–1, P ≤ 0.0001); while from Day 16 to Day 32, pregnant cows maintained steady and higher P4 concentrations than nonpregnant ones (9.87 ± 0.31 v. 4.40 ± 1.09 ng mL–1, respectively; P = 0.002). As expected, nonpregnant cows showed a sharp decrease in P4 concentrations from Day 16 to Day 25, followed by a second increase at Day 32 (9.27 ± 0.20, 0.85 ± 0.33, and 6.04 ± 1.67 ng mL–1, respectively; P = 0.0007). The UAC and PGEM concentrations were positively correlated in pregnant (r = 0.48; P = 0.002), but not in nonpregnant animals (r = 0.29; P = 0.21). Similarly, UAC and P4 concentrations were also positively associated in pregnant (r = 0.47; P = 0.002), but not in nonpregnant cows (r = 0.01; P = 0.96). Our findings support the role of PGE2 as a potential mediator of the vascular function in the female reproductive tract and, subsequently, as an essential factor to ensure adequate P4 production able to sustain early pregnancy in the bovine. USDA-ARS Grant#58-6402-3-0120.

Feeding level and dietary energy source have no effect on embryo survival in gilts, despite changes in systemic progesterone levels

This study was designed to assess the effect of feeding level and dietary energy source on luteal function, systemic progesterone concentration and embryo survival in gilts during early gestation. At Day 0 of pregnancy, 104 gilts were allocated to one of four experimental diets (LStarch: 1.2 × maintenance requirement (M) Starch diet (43.3% starch), n = 31; HStarch: 2.4 × M Starch diet (43.3% starch), n = 21; HFat: 2.4 × M Fat diet (13.5% fat), n = 23; and HFibre: 2.4 × M Fibre diet (7.2% fibre), n = 23). On Day 5 of gestation, no significant difference in circulating concentration of systemic progesterone was seen among the treatments. However, on Day 15 of pregnancy, gilts on the HStarch diet had a significantly lower concentration of systemic progesterone than did gilts on both the LStarch and HFat diets (P &lt; 0.05; 24.8 ± 2.4 v. 32.7 ± 2.4 and 36.1 ± 2.1 ng/mL, respectively). At Day 35 of gestation, there was also a tendency for gilts on the HStarch and HFat diets to have a higher total luteal weight than for gilts on the LStarch diets (7.2 ± 0.2 and 7.1 ± 0.2 v. 6.7 ± 0.2 g (P &lt; 0.05)). No difference in embryo survival was seen among the treatments. From the present study, we can conclude that altering feeding level and dietary energy source did not affect embryo survival, despite the fact that systemic progesterone concentrations were affected on Day 15 of gestation. Also, luteal weight was greater for those gilts on the high feeding level than for those on the low feeding level when fed the same energy source.

Correlations between ultrasonographic characteristics of corpora lutea and systemic concentrations of progesterone during the discrete stages of corpora lutea lifespan and secretory activity in cyclic ewes

Associations between physical characteristics and functionality of corpora lutea (CL) have previously been reported in monovulatory species, albeit several studies in cattle and humans have refuted the existence of temporal relationships between CL size, echotexture and serum progesterone (P(4)) concentrations. The main objective of the present study was to examine whether or not there were correlations between ultrasonographic image attributes of CL and systemic concentrations of P(4) during the discrete stages of the luteal phase in two breeds of sheep differing in ovulation rates (non-prolific Western White Face [WWF] ewes and prolific Finn [F] sheep). Transrectal ovarian ultrasonography utilized a 7.5-MHz linear-array transducer connected to a portable scanner (Aloka SSD-500) and the images were analyzed using commercially available image analytical software (Image ProPlus(®)) validated for the present application in sheep. The correlations were assessed using the Pearson's Product Moment (PPM) analysis and also, to increase the accuracy of statistical tests, the analysis of covariance (ANCOVA), with the number of CL as a co-factor. In WWF ewes, serum concentrations of P(4) correlated significantly with the total luteal area (TLA) during the CL growth phase (days 3-6; day 0 = ovulation) and functional luteolysis (days 12-15), and with numerical pixel values (NPVs--pixel intensity) during luteolysis; the results obtained by using two different statistical methods were generally similar. In prolific F ewes, serum P(4) concentrations were directly correlated with TLA during CL growth (days 3-6; ANCOVA), functional luteolysis (days 13-14; PPM), and structural CL regression (days 11-14; PPM and ANCOVA), and with NPVs during functional luteolysis (PPM and ANCOVA). We concluded that systemic P(4) concentrations could only be accurately predicted from the changes in luteal area during CL growth and regression, and from NPVs during luteolysis, in both prolific and non-prolific ewes, but the changes in size and echotexture of the luteal glands at mid-cycle were not indicative of serum P(4) concentrations in sheep.

Alterations in systemic concentrations of progesterone during the early luteal phase affect RBP4 expression in the bovine uterus

Systemic progesterone affects the timing and duration of uterine endometrial gene and protein expression and has significant effects on conceptus development. The objective of the present study was to examine how changes in progesterone concentrations during the early luteal phase affect retinol-binding protein (RBP4) mRNA and protein concentrations in the uterus. Endometrial tissue and uterine flushings were recovered on Days 7 and 13 of the oestrous cycle in heifers with high, normal and low progesterone concentrations. RBP4 mRNA and protein concentrations were higher (P<0.05) on Day 13 compared with Day 7 in heifers with high and control progesterone concentrations. However, there was no difference in RBP4 protein concentrations between Days 7 and 13 in heifers with low progesterone (P>0.05). On Day 7, although heifers with low progesterone had lower RBP4 mRNA expression compared with controls (P<0.05) there was no difference in protein concentrations between treatment groups. On Day 13, RBP4 mRNA was 2-fold higher (P<0.001) in heifers with high and control progesterone compared with their low-progesterone counterparts and RBP4 protein concentrations were over 2-fold higher (P<0.001) in heifers with high compared to low progesterone. In conclusion, progesterone modulates uterine RBP4 mRNA and protein abundance in a time- and concentration-dependent manner.

Embryo death in cattle: an update

For heifers, beef and moderate-yielding dairy cows, fertilisation generally exceeds 90%. In high-producing dairy cows, it may be lower and possibly more variable. The major component of embryo loss occurs before Day 16 following breeding, with emerging evidence of greater losses before Day 8 in high-producing dairy cows. Late embryo loss causes serious economic losses because it is often recognised too late to rebreed females. Systemic concentrations of progesterone during the cycles both preceding and following insemination affect embryo survival; too-high or too-low a concentration has been shown to be negatively associated with survival rate. Energy balance and dry matter intake during the 4 weeks after calving are critically important in determining conception rate when cows are inseminated 70 to 100 days after calving. More balanced breeding strategies with greater emphasis on fertility, feed intake and energy must be developed. Genetic variability for fertility traits can be exploited; genomic technology will not only provide scientists with an improved understanding of the underlying biological processes involved in fertilisation and the establishment of pregnancy, but could identify genes responsible for improved embryo survival. Their incorporation into breeding objectives would increase the rate of genetic progress for embryo survival. There is a range of easily adoptable management factors, under producer control, that can either directly increase embryo survival or ameliorate the consequences of low embryo survival rates. The correction of minor deficits in several areas can have a substantial overall effect on herd reproductive performance.

Maternal Obesity and Overnutrition Is Associated with Reduced Systemic Progesterone During the Estrous Cycles of Adult Female Offspring and a Marked Increase in Insulin Resistance at Midpregnancy.

Previously published studies from our laboratory and others have demonstrated that maternal undernutrition (MU) and overnutrition (MO) alter fetal growth and development during intrauterine life, predisposing offspring to similar metabolic disturbances in postnatal life. We have recently demonstrated that MO predisposes nonpregnant female offspring to hyperphagia, increased obesity, and insulin resistance as adults. Further, we have reported that MU of ewes is associated with increased oxidative base lesions within DNA of mid-gestational (Day 78) fetal oogonia in sheep. More recently we have reported that female offspring born to MU ewes exhibited consistently reduced circulating progesterone concentrations during their estrous cycles, and reduced fertility when compared to female offspring born to ewes fed to NRC requirements. This study evaluated the impact of MO on systemic progesterone concentrations during an estrous cycle, fertility, and insulin resistance of adult female offspring at midgestation. Adult nonpregnant female offspring (2-3 years old) born to obese (OB) overnourished ewes fed 150% of NRC recommendations (n = 10), or 100% of NRC (Controls; CON, n=7) from 60 days prior to conception to term were utilized in this study. During an estrous cycle, ewes were bled daily from one estrus (Day 0) to the next, and serum collected and frozen at -80°C until analyzed for progesterone (P4) via validated RIA. At the second estrus, ewes were hand mated to an intact ram at 12hr intervals until the end of estrus. Conception rates and fetal numbers (singles and twins) were determined via real time ultrasonography, and pregnant ewes were subjected to an intravenous glucose tolerance test (IVGTT) at midgestation (75 ± 1 day). Plasma glucose and insulin was measured in triplicate by colorimetric assay and validated RIA, respectively. Estrous cycle length was reduced (P < 0.05) in the offspring of OB vs. CON ewes (17.2 ± 0.2 days vs. 18.0 ± 0.1 days). Further, serum progesterone concentrations were decreased (area under the curve, P < 0.05) during the estrous cycle of offspring from OB versus CON ewes. Baseline glucose and insulin concentrations were greater (P < 0.05) in offspring from OB vs. CON ewes, (64.03 ± 3.97 vs. 54.68 ± 1.30 mg/dL and 7.01 ± 2.18 vs. 3.43 ± 0.15 mIU/L, respectively). During the IVGTT, glucose concentrations remained greater (area under the curve; P < 0.01) than those of offspring from CON ewes during the post infusion period. Plasma insulin concentrations of OB offspring during IVGTT were also markedly increased (P < 0.03) when compared to offspring from CON ewes. These data suggest that the decrease in systemic progesterone exhibited by the OB offspring during their estrous cycles may contribute to the shortened estrous cycle lengths observed in this group. In spite of the decrease in circulating progesterone of OB offspring, however, conception rate and fecundity remained unaffected. Maternal overnutrition also resulted in a marked insulin resistance at midgestation in offspring of OB vs. CON ewes, suggesting that the F2 generation could exhibit the same metabolic alterations as their mothers resulting in a transgenerational programming effect. Supported in part by NIH INBRE #P20RR016474. (platform)