Conception rate with or without hCG trigger in clomiphene induced cycles

Background: Clomiphene citrate (CC) is still the most common drug for ovulation induction. Most physicians use hCG trigger routinely for follicle rupture. Ideally hCG is recommended only where there is no spontaneous LH surge. Hence this study was conducted to see the role of hCG for follicle rupture in CC induced cycle. Aims and objectives of the study were to compare the ovulation rate in CC induced cycle with or without hCG trigger and finally the conception rate.Methods: Study was conducted in the department of OBG, LLRM medical college Meerut on women with anovulatory infertility. All women were given 50-100 mg CC. Follicular study from D-9 was done till follicle rupture. Women in group A were observed without any trigger and women in group B were given inj. hCG trigger 10,000 IU when follicle size reached 20-22 mm.Results: Conception rates were 25% Vs 31% in group A and group B. Follicle rupture was seen in 84% cases in group A and 71% in group B. Results were comparable in both the groups.Conclusions: By adding inj. hCG for ovulation trigger does not increase the conception rate. LH surge is already there in CC induced cycles. It is having role only in cases where no LH surge is there.

A platform utilizing Drosophila ovulation for nonhormonal contraceptive screening

A significant unmet need for new contraceptive options for both women and men remains due to side-effect profiles, medical concerns, and the inconvenience of many currently available contraceptive products. Unfortunately, the development of novel nonsteroidal female contraceptive medicine has been stalled in the last couple of decades due to the lack of effective screening platforms. Drosophila utilizes conserved signaling pathways for follicle rupture, a final step in ovulation that is essential for female reproduction. Therefore, we explored the potential to use Drosophila as a model to screen compounds that could inhibit follicle rupture and be nonsteroidal contraceptive candidates. Using our ex vivo follicle rupture assay, we screened 1,172 Food and Drug Administration (FDA)–approved drugs and identified six drugs that could inhibit Drosophila follicle rupture in a dose-dependent manner. In addition, we characterized the molecular actions of these drugs in the inhibition of adrenergic signaling and follicle rupture. Furthermore, we validated that three of the four drugs consistently inhibited mouse follicle rupture in vitro and that two of them did not affect progesterone production. Finally, we showed that chlorpromazine, one of the candidate drugs, can significantly inhibit mouse follicle rupture in vivo. Our work suggests that Drosophila ovulation is a valuable platform for identifying lead compounds for nonsteroidal contraceptive development and highlights the potential of these FDA-approved drugs as novel nonsteroidal contraceptive agents.

Progesterone receptor-A isoform interaction with RUNX transcription factors controls chromatin remodelling at promoters during ovulation

Progesterone receptor (PGR) plays diverse roles in reproductive tissues and thus coordinates mammalian fertility. In the ovary, acutely induced PGR is the key determinant of ovulation through transcriptional control of a unique set of genes that culminates in follicle rupture. However, the molecular mechanisms for PGR specialised function in ovulation is poorly understood. To address this, we assembled a detailed genomic profile of PGR action through combined ATAC-seq, RNA-seq and ChIP-seq analysis in wildtype and isoform-specific PGR null mice. We demonstrated the unique action of PGR-A isoform in the ovary through a transcriptional complex involving physical interaction with RUNX and JUN/FOS transcription factors. The assembly of this unique complex directs targeting of PGR binding to proximal promoter regions and enables chromatin accessibility, leading to ovulatory gene induction. This PGR signalling mechanism is specific to ovulation and provides potential targets for infertility treatments as well as new contraceptives that block ovulation.

A central role for cAMP/EPAC/RAP/PI3K/AKT/CREB signaling in LH-induced follicular Pgr expression at medaka ovulation

Nuclear progestin receptor (PGR) is a ligand-activated transcription factor that has been identified as a pivotal mediator of many processes associated with ovarian and uterine function, and aberrant control of PGR activity causes infertility and disease including cancer. The essential role of PGR in vertebrate ovulation is well recognized, but the mechanisms by which PGR is rapidly and transiently induced in preovulatory follicles after the ovulatory LH surge are not known in lower vertebrates. To address this issue, we utilized the small freshwater teleost medaka Oryzias latipes, which serves as a good model system for studying vertebrate ovulation. In the in vitro ovulation system using preovulatory follicles dissected from the fish ovaries, we found that inhibitors of EPAC (brefeldin A), RAP (GGTI298), PI3K (Wortmannin), AKT (AKT inhibitor IV), and CREB (KG-501) inhibited LH-induced follicle ovulation, while the PKA inhibitor H-89 had no effect on follicle ovulation. The inhibitors capable of inhibiting follicle ovulation also inhibited follicular expression of Pgr and matrix metalloproteinase-15 (Mmp15), the latter of which was previously shown to not only be a downstream effector of Pgr but also a proteolytic enzyme indispensable for follicle rupture in medaka ovulation. Further detailed analysis revealed for the first time that the cAMP/EPAC/RAP/PI3K/AKT/CREB signaling pathway mediates the LH signal to induce Pgr expression in preovulatory follicles. Our data also showed that phosphorylated Creb1 is a transcription factor essential for pgr expression and that Creb1 phosphorylated by Akt1, rather than PKA, may be preferably used to induce pgr expression.

Dissection of the Ovulatory Process Using ex vivo Approaches

Ovulation is a unique physiological phenomenon that is essential for sexual reproduction. It refers to the entire process of ovarian follicle responses to hormonal stimulation resulting in the release of mature fertilization-competent oocytes from the follicles and ovaries. Remarkably, ovulation in different species can be reproduced out-of-body with high fidelity. Moreover, most of the molecular mechanisms and signaling pathways engaged in this process have been delineated using in vitro ovulation models. Here, we provide an overview of the major molecular and cytological events of ovulation observed in frogs, primarily in the African clawed frog Xenopus laevis, using mainly ex vivo approaches, with the focus on meiotic oocyte maturation and follicle rupture. For the purpose of comparison and generalization, we also refer extensively to ovulation in other biological species, most notoriously, in mammals.

Diclofenac Administered After the Initiation of Luteinizing Hormone Surge Increases Live Births in Natural-Cycle IVF-ET: A Cohort Study

Background: Diclofenac inhibits follicle rupture and its use in natural-cycle in vitro fertilization and embryo transfer (IVF-ET) has been reported to increase oocyte retrieval chances but has not been reported to improve the therapeutic outcome (live birth). The question is whether the therapeutic utility of diclofenac is demonstrable when administered to a subgroup of women with an imminent LH surge, a higher risk group for premature ovulation.Methods: Infertile women indicated for the natural-cycle IVF-ET between September 2014 and February 2015 (n=183) were recruited in a private infertility clinic and diclofenac use (50 mg suppositories, thrice every 8 h before oocyte retrieval) was offered when their serum LH level was ≥14.0 IU/L on an LH-triggering day (n=137). Of the 137 women, 108 electively used diclofenac and 29 did not. Oocytes were retrieved from both dominant and subordinate nondominant follicles and were fertilized. The resulting blastocysts were frozen, thawed, and transferred one by one in the following spontaneous ovulatory or hormone replacement cycles. Results: Cumulative live birth rate (after the single oocyte retrieval) was calculated from the dominant and nondominant follicles. The live birth rate from dominant follicles was higher in the diclofenac group (21/108, 19%) than in the no diclofenac group (1/29, 3%) (P < .05). Conversely, the live birth rate from nondominant follicles, which had no potential for ovulation, was not different between the diclofenac group (13/108, 12%) and the no diclofenac group (3/29, 10%). Conclusion: Diclofenac improved the live birth rate from dominant follicles when it was administered to women with an imminent LH surge. However, diclofenac did not affect the live birth rate from non-dominant follicles which were not at risk of follicle rupture.

Prematurely ruptured dominant follicles often retain competent oocytes in infertile women

Ovulation consists of a follicle’s rupture and subsequent oocyte extrusion, although there is a paucity of evidence regarding whether every follicle’s rupture is associated with extrusion of its oocyte. We examined this issue in a large-scale window-of-opportunity study by attempting aspiration of single dominant follicles that were found to have ruptured before a scheduled oocyte retrieval during in vitro fertilisation and embryo transfer treatment of infertile women. We were able to aspirate 587 of 1,071 ultrasonographically confirmed post-rupture dominant follicles from 1,071 women (i.e. one dominant follicle per woman) and retrieved 225 oocytes (oocyte recovery ratio: 43.4% of aspirated follicles), which yielded 28 live births (live birth ratio: 11.0% of retrieved oocytes). Interestingly, the live birth ratio for post-rupture dominant follicles was not statistically different from that achieved using regular pre-rupture aspiration of dominant follicles (1,085/8,977, 12.1%). These findings suggest that oocyte extrusion frequently does not occur after follicle rupture in infertile women undergoing in vitro fertilisation treatment, although the oocyte retained in the follicle can remain competent for use during that treatment.

The regulation of oocyte maturation and ovulation in the closest sister group of vertebrates

Ascidians are the closest living relatives of vertebrates, and their study is important for understanding the evolutionary processes of oocyte maturation and ovulation. In this study, we first examined the ovulation of Ciona intestinalis Type A by monitoring follicle rupture in vitro, identifying a novel mechanism of neuropeptidergic regulation of oocyte maturation and ovulation. Ciona vasopressin family peptide (CiVP) directly upregulated the phosphorylation of extracellular signal–regulated kinase (CiErk1/2) via its receptor. CiVP ultimately activated a maturation-promoting factor, leading to oocyte maturation via germinal vesicle breakdown. CiErk1/2 also induced expression of matrix metalloproteinase (CiMMP2/9/13) in the oocyte, resulting in collagen degradation in the outer follicular cell layer and liberation of fertile oocytes from the ovary. This is the first demonstration of essential pathways regulating oocyte maturation and ovulation in ascidians and will facilitate investigations of the evolutionary process of peptidergic regulation of oocyte maturation and ovulation throughout the phylum Chordata.

In Vitro Reconstruction of Xenopus Oocyte Ovulation

Progesterone is widely used to induce maturation of isolated fully grown oocytes of the African clawed frog, Xenopus laevis. However, the hormone fails to release oocytes from the layer of surrounding follicle cells. Here, we report that maturation and follicle rupture can be recapitulated in vitro by treating isolated follicular oocytes with progesterone and low doses of the matrix metalloproteinase (MMP), collagenase, which are ineffective in the absence of the steroid. Using this in vitro ovulation model, we demonstrate that germinal vesicle breakdown (GVBD) and oocyte liberation from ovarian follicles occur synchronously during ovulation. Inhibition of the MAPK pathway in these experimental settings suppresses both GVBD and follicular rupture, whereas inhibition of MMP activity delays follicular rupture without affecting GVBD. These results highlight importance of MAPK and MMP activities in the ovulation process and provide the first evidence for their involvement in the release of oocytes from ovarian follicles in frogs. The in vitro ovulation model developed in our study can be employed for further dissection of ovulation.