The effects of progesterone on oocyte maturation and embryo development (original) (raw)
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Biology of Reproduction, 2011
Progesterone (P4) exerts its effects by binding to specific genomic (nPR-A/B) and non-genomic (mPRα/β, PGRMC1/2) receptors. P4 has a role in the regulation of the ovulatory cycle, but its participation in oocyte maturation in mammals has not yet been clarified. Therefore, the aim of the present study was to characterize the protein expression of P4 receptors in bovine oocytes and cumulus cells during in vitro maturation (IVM) and to study the effect of P4 and its receptors on oocyte developmental competence. Cumulus-oocyte complexes (COCs) were subjected to IVM, fertilization (IVF) and culture (IVC). IVM was performed for 24 h in the presence or absence of P4, LH, FSH, Trilostane, promegestone (R5020), mifepristone (RU 486) or antibodies against mPRα or mPRβ. Protein expression of PRs was studied by western blotting and immunofluorescence. The results demonstrate the presence of both genomic and non genomic P4 receptors in bovine COCs. The dynamic changes observed in the protein expression of PRs following IVM or in response to supplementation with LH, FSH or P4 suggest an important role during bovine oocyte maturation. Inhibition of P4 synthesis by cumulus cells or blocking of nPR and mPRα activity produced a decrease in bovine embryo development, indicating that P4 intracellular signaling is mediated by its interaction with nuclear and membrane progesterone receptors and is important for oocyte developmental competence.
Journal of Dairy Science, 2011
Two experiments evaluated the influence of altering the concentrations of progesterone during the development of the ovulatory follicle on the composition of the follicular fluid, circulating LH and PGF 2α metabolite (PGFM), and expression of endometrial progesterone receptor and estrogen receptor-α. In both experiments, the estrous cycles were presynchronized (GnRH and progesterone insert followed by insert removal and PGF 2α 7 d later, and GnRH after 48 h) and cows were then enrolled in 1 of 2 treatments 7 d later (study d −16): high progesterone (HP) or low progesterone (LP). In experiment 1 (n = 19), cows had their estrous cycle synchronized starting on study d −9 (GnRH and progesterone insert on d −9, and insert removal and PGF 2α on d −2). In experiment 2 (n = 25), cows were submitted to the same synchronization protocol as in experiment 1, but had ovulation induced with GnRH on study d 0. In experiment 1, plasma was sampled on d −4 and analyzed for concentrations of LH; the dominant follicle was aspirated on d 0 and the fluid analyzed for concentrations of progesterone, estradiol, and free and total IGF-1. In experiment 2, follicular development and concentrations of progesterone and estradiol in plasma were evaluated until study d 16. Uterine biopsies were collected on d 12 and 16 for progesterone receptor and estrogen receptor-α protein abundance. An estradiol/oxytocin challenge for PGFM measurements in plasma was performed on d 16. In experiments 1 and 2, LP cows had lower plasma concentrations of progesterone and greater concentrations of estradiol, and had larger ovulatory follicle diameter (20.4 vs. 17.2 mm) at the end of the synchronization protocol than HP cows. Concentration of LH tended to be greater for LP than HP cows (0.98 vs. 0.84 ng/ mL). The dominant follicle of LP cows had greater concentration of estradiol (387.5 vs. 330.9 ng/mL) and a lower concentration of total IGF-1 (40.9 vs. 51.7 ng/ mL) than that of HP cows. In experiment 2, estradiol and progesterone concentrations did not differ between treatments from d 0 to 16; however, the proportion of cows with a short luteal phase tended to increase in LP than HP (25 vs. 0%). Concentrations of PGFM were greater for LP than HP. Uterine biopsies had a greater abundance of progesterone receptor, and tended to have less estrogen receptor-α abundance on d 12 compared with d 16. An interaction between treatment and day of collection was detected for estrogen receptor-α because of an earlier increase in protein abundance on d 12. Reduced concentrations of progesterone during the development of the ovulatory follicle altered follicular dynamics and follicular fluid composition, increased basal LH concentrations, and prematurely increased estrogen receptor-α abundance and exacerbated PGF 2α release in the subsequent estrous cycle.
Effect of circulating progesterone on in vitro developmental competence of bovine oocytes
Animal reproduction, 2009
This study evaluated the effects of systemic progesterone concentration on oocyte quality and in vitro embryo production. Oocytes were retrieved from 15 crossbred cows (Bos taurus x Bos indicus). These cows were randomly allocated into three groups to provide low; high, or very low (LP4, HP4 and VLP4, respectively) plasma progesterone concentrations and received either a previously used CIDR, two new CIDR devices, or no progesterone treatment (Day 0). The CIDR devices were replaced every 8 days along with 150 µg of D-cloprostenol injections. The ovum pickup (OPU) procedure was performed every 4 days from Day 4 to 24. Simultaneous to OPU procedure, plasma was collected to measure progesterone and on Day 18, serial blood samples were collected to assess the pattern of LH release. Hormone concentrations were analyzed by ANOVA and the binomial variables were analyzed by Chi-square. Plasma progesterone concentration was higher in the HP4, intermediate in the LP4, and lower in the VLP4 group (3.6, 1.6, and 0.5 ng/ml; P < 0.05). Plasma LH was higher in the LP4, intermediary in the VLP4, and lower in the HP4 group (1.6, 1.0, and 0.8 ng/ml). A greater percentage of viable oocytes (grades I to III) was retrieved from LP4 (79.4%; 131/165) than from the HP4 (68.4%; 119/174) group (P = 0.07); the VLP4 group did not differ from the others (72.3%; 60/83). Furthermore, the blastocyst production and blastocyst rate was higher in LP4 (1.3 ± 0.4; 28.2%), than in HP4 (0.8 ± 0.4; 16.0%) or the VLP4 (0.4 ± 0.4; 15.0%) group (P = 0.06 and 0.03 for blastocyst production and rate, respectively). In conclusion, intermediate plasma P4 concentration that results in higher circulating LH in cows may improve in vitro embryo production.
In Vitro Cellular & Developmental Biology - Animal, 2013
The role of progesterone (P 4 ) and prostaglandins (PGs) in bovine early embryonic development and embryomaternal crosstalk is almost unknown. Here, the in vitro steroidogenic (P 4 ) and prostanoid (PGE 2 and PGF 2α) interactions between bovine embryos and luteal cells (LC) were evaluated. In two experiments, embryos (n=1.900) were either co-cultured with LC or cultured alone, from days 2 to 7 (day 0=in vitro insemination). LC were also cultured alone, and medium was used as a control, all groups being cultured either with or without oil overlay of culture medium. Oil overlay of culture medium significantly decreased the amount of P 4 , but not of PGE 2 and PGF 2α measured in culture medium. Embryos and LC had transcripts of genes coding for enzymes of the PGs (PTGS2, PGES, and PGFS) and P 4 (StAR, P450scc, and 3β-HSD) synthesis pathways, and produced P 4 , PGF 2α , and PGE 2 into culture medium. Co-culture with LC exerted an embryotrophic effect, significantly increasing blastocyst yield and quality. This indicates a possible direct effect of LC in early embryo development. Embryos did not exert a luteotrophic effect upon LC. This may indicate that early embryos (until day 7) probably do not exert influence in LC main function. It is suggested that production of P 4 , PGE 2 , and PGF 2α by early embryos may be associated to autocrine signaling leading to events in development and to paracrine signaling in the endometrium leading to local uterine receptivity.
Progesterone enhances in vitro development of bovine embryos
Theriogenology, 2012
Increased pregnancy rates in cattle given progesterone (P 4) prior to 5 d after breeding have recently been reported. The objective was to determine if this increase in pregnancy rate could be attributed to a direct positive effect of P 4 on the developing embryo. In Experiment 1, 280 bovine oocytes were inseminated in vitro and at Day 3 (insemination ϭ Day 0), good quality 8 cell embryos (n ϭ 206) were randomly allocated to be cultured in either CR1aaϩserum with 0 or ϳ15 ng/mL (n ϭ 102 and n ϭ 104, respectively). In Experiment 2, 881 bovine oocytes were used; on Day 3, good quality 8 cell embryos (n ϭ 511) were randomly allocated to either the control (CR1aaϩFCS, n ϭ 168), vehicle (CR1aa ϩ FCS ϩ ethanol, n ϭ 170), or P 4 treatment (CR1aa ϩ FCS ϩ ϳ15 ng/mL P 4 in ethanol, n ϭ 173). On Day 7, in both experiments, there were increased numbers of blastocysts developing in the P 4 group (Experiment 1, 59% and Experiment 2, 71%) compared to the vehicle (Experiment 2, 53%) or control (40 and 62% in Experiments 1 and 2, respectively). The addition of P 4 (8%) stimulated the rate of embryo development (early blastocysts or more advanced stages on Day 6) compared to vehicle (3%) and control (0%) and the P 4 group had more hatched or hatching blastocysts (33%) on Day 9 compared to the control or vehicle group (21 or 22%). Additionally, the P 4 group had greater embryo diameter and significantly more Grade 1 blastocysts on Day 7. In conclusion, P 4 had a direct, positive effect on developing bovine embryos cultured in vitro.
Animal Reproduction Science, 2018
This study evaluated the effect of progesterone priming during follicular growth on oocyte competence to undergo oocyte cleavage and embryo development in sheep. Two experiments were performed on a total of 195 females that either received or did not receive a progesterone treatment (CIDR-type device) during the first follicular wave, Day 0. After follicular aspiration on Day 3, the cleavage rate and the embryo development rate following in vitro fertilization and culture were greater in those females that received the progesterone treatment (P< 0.05). In conclusion, these studies provide evidence that progesterone treatment during follicular growth affects oocyte competence, with the greater progesterone concentrations enhancing the oocyte's capacity to undergo cleavage and embryo development.
Progesterone regulation of primordial follicle assembly in bovine fetal ovaries
Molecular and cellular endocrinology, 2009
Fertility in mammals is dependant on females having an adequate primordial follicle pool to supply oocytes for fertilization. The formation of primordial follicles is called ovarian follicular assembly. In rats and mice progesterone and estradiol have been shown to inhibit follicle assembly with assembly occurring after birth when the pups are removed from the high-steroid maternal environment. In contrast, primordial follicle assembly in other species, such as cattle and humans, occurs during fetal development before birth. The objective of the current study is to determine if progesterone levels regulate primordial follicle assembly in fetal bovine ovaries. Ovaries and blood were collected from bovine fetuses. Interestingly, ovarian progesterone and estradiol concentrations were found to decrease with increasing fetal age and correlated to increased primordial follicle assembly. Microarray analysis of fetal ovary RNA suggests that progesterone membrane receptor and estrogen nuclea...
International Journal of Fertility and …, 2009
The aim of the present study was to investigate the role of progesterone on the developmental competence of cumulus-oocyte complexes (COCs) and cumulus-denuded oocytes (CDOs) at germinal vesicle (GV) stage. Materials and Methods: GV oocytes of pregnant mare's serum gonadotropin (PMSG)-primed prepubertal mice were divided into two groups: CDOs and COCs. The oocytes were cultured in TCM199 with different concentrations of progesterone (10, 38, 50 and 100 μM) and without progesterone (controls). The number of oocytes at the GV, germinal vesicle breakdown (GVBD) and metaphase II (MII) stages were counted. In vitro fertilization (IVF) of MII oocytes and their development to the blastocyst stage were evaluated. Results: Significantly different MII rates were observed between the COCs (85%) and CDOs (68%) control groups. The MII rates of 83%, 48%, 14% and 0% for COCs and 65%, 53%, 20% and 0% for CDOs were obtained in TCM199 that contained 10, 38, 50 and 100μM progesterone concentrations, respectively. These MII rates were lower (p<0.05) in both COCs and CDOs as compared to their respective control groups, except for 10 μM. The fertilization and blastocyst rates of COCs (83% and 35%, respectively) were higher (p <0.05) than those of the CDOs (51% and 5%, respectively) control groups. The fertilization and blastocyst rates in the presence of 10 μM (81% and 36%, respectively) and 38 μM (85% and 30%, respectively) progesterone in COCs and CDOs (52% and 4% for 10 μM; 56% and 4% for 38 μM) were similar to their respective control groups. Conclusion: Adding progesterone to the medium could not improve maturation of mouse GV oocytes and their development to the blastocyst stage.
Biology of Reproduction, 2012
Uterine gland development (adenogenesis) in mice begins on Postnatal Day (PND) 5 and is completed in adulthood. Adenogenesis depends on estrogen receptor 1, and progesterone (P4) inhibits mitogenic effects of estrogen on uterine epithelium. This progestin-induced effect has been used to inhibit uterine gland development; progestin treatment of ewes for 8 wk from birth has produced infertile adults lacking uterine glands. The goals of the present study were to determine if a window of susceptibility to P4-mediated inhibition of uterine gland development exists in mice and whether early P4 treatment abolishes adenogenesis and fertility. Mice were injected daily with P4 (40 lg/g) or vehicle during various postnatal windows. Adenogenesis, cell proliferation, and expression of key morphoregulatory transcripts and proteins were examined in uteri at PNDs 10 and 20. Additionally, adenogenesis was assessed in isolated uterine epithelium. Treatment during PNDs 3-9, 5-9, or 3-7 abolished adenogenesis at PND 10, whereas treatments during PNDs 3-5 and 7-9 did not. Critically, mice treated during PNDs 3-9 lacked glands in adulthood, indicating that adenogenesis did not resume after this treatment. However, glands were present by PND 20 and later following treatment during PNDs 5-9 or 3-7, whereas treatment during PNDs 10-16 produced partial inhibition of adenogenesis at PND 20 and later. Epithelial proliferation at PND 10 was low following P4 treatment (PNDs 3-9) but exceeded that in controls at PND 20, indicating a rebound of epithelial proliferation following treatment. Messenger RNA for Wnt, Fzd, and Hox genes was altered by neonatal P4 treatment. All groups cycled during adulthood. Mice treated with P4 during PNDs 3-9, but not during other developmental windows, showed minimal fertility in adulthood. In summary, brief P4 treatment (7 days) during a critical neonatal window (PNDs 3-9) transiently inhibited epithelial proliferation but totally and permanently blocked adenogenesis and adult fertility. This resulted in permanent loss of uterine glands and, essentially, total infertility during adulthood. The narrow window for inhibition of adenogenesis identified here may have implications for development of this methodology as a contraceptive strategy for animals.
136 Evidence of a Direct Effect of P4 on Ivf-Derived Bovine 8-CELL Embryos
Reproduction, Fertility and Development
There has been much debate over a direct role for progesterone (P4) in early bovine embryo development. While previous attempts to supplement bovine embryos in vitro with P4 produced results that vary and are often contradictory, this may be a response of administering P4 at inappropriate times. Therefore, the objective of these experiments was to determine if P4 could exert a direct effect on developing IVF-derived bovine embryos when administered at an appropriate time of embryo development. In Exp. I, IVF-derived bovine 8-cell embryos were randomly allotted to treatments: (1) control, CR1aa medium (n = 168); (2) vehicle, CR1aa + ETOH (0.01%) (n = 170); and (3) P4, CR1aa + ETOH + P4 (20 ng/mL in 50-μL droplet) (n = 173). In Exp. II, IVF-derived bovine 8-cell embryos were randomly allotted to treatments: (1) control, CR1aa medium (n = 160); (2) vehicle, CR1aa + DMSO (0.01%) (n = 180); and (3) P4, CR1aa + DMSO (0.01%) + P4 (20 ng/mL in 50-μL droplet) (n = 170). All embryos were eval...