Preovulatory Ovarian Follicles Research Articles

Evolutionary-developmental (evo-devo) dynamics of hominin brain size.

Brain size tripled in the human lineage over four million years, but why this occurred remains uncertain. Here, to study what caused this brain expansion, I mathematically model the evolutionary and developmental (evo-devo) dynamics of hominin brain size. The model recovers (1) the evolution of brain and body sizes of seven hominin species starting from brain and body sizes of the australopithecine scale, (2) the evolution of the hominin brain-body allometry and (3) major patterns of human development and evolution. I show that the brain expansion recovered is not caused by direct selection for brain size but by its genetic correlation with developmentally late preovulatory ovarian follicles. This correlation is generated over development if individuals experience a challenging ecology and seemingly cumulative culture, among other conditions. These findings show that the evolution of exceptionally adaptive traits may not be primarily caused by selection for them but by developmental constraints that divert selection.

The effects of MEK and PKA inhibition on Spire proteins during oocyte maturation.

Asymmetric division of oocytes driven by chromosome migration is a crucial step of oocyte maturation. Actin filaments take key roles in chromosome migration in oocytesThe aim of this study was to determine the effects of MEK and PKA inhibition on the levels of Spire-1 and Spire-2 proteins that are known to be related to actin nucleation.MEK inhibitor PD98059 and PKA inhibitor H89 were applied during IVM to the oocytes retrieved from preovulatory ovarian follicles of PMSG induced 3-5 weeks old female BalbC mice. GVBD and PBE rates were determined. Spire-1 and Spire-2 proteins were detected by immunofluorescence and western blot in oocytes at different maturation stages.Though GVBD rates were similar in different groups, PBE rates were lower in the MEK inhibition group. Through immunofluorescence, cortical localizations of Spire-1 and Spire-2 were determined. MEK inhibition resulted in a decrease in cortical Spire-1 and Spire-2 levels in PBE oocytes. PKA inhibition led to an increase in cortical Spire-1 levels in spindle migration stage oocytes, and an increase in cortical and total Spire-2 levels in PBE oocytes. Application of both MEK and PKA inhibition resulted in compensation of the decrease in Spire-1, while Spire-2 levels remained low with no compensation of PKA inhibition.According to the results of this study, chemical inhibition of MEK and PKA during oocyte maturation alters Spire-1 and Spire-2 protein levels.

Association of glucocorticoid receptor expression with key members of the insulin signaling pathway and heat shock proteins in the bovine ovary

Glucocorticoids (GCs) act through their receptor (GR) as regulators in different biological processes such as reproduction. In the absence of GCs, the GR remains inactive in the cytoplasm by associating with heat shock proteins (HSPs), which act as molecular chaperones, among which the most relevant are HSP90 and HSP70. Cytoplasmic GC-activated GR mediates non-genomic effects, interacting with members of signaling pathways such as PI3K/Akt, which participates in several metabolic processes, including the insulin signaling pathway. The aim of the present study was to evaluate possible associations between the cytoplasmic GR and the main intermediates of the insulin signaling pathway and HSP90 and HSP70 in ovaries of dairy cows. To this end, the protein expression of cytoplasmic GR, key members of the insulin signaling pathway, and HSPs was evaluated in ovarian preovulatory follicles of non-lactating Holstein cows in proestrus. Positive associations were observed between protein expression of GR and HSP90, IRS1, pIRS1, PI3K and pAkt (p < 0.05; β > 0) in granulosa cells of dominant follicles of dairy cows. Instead, in theca cells, no associations were observed between protein expression of GR and members of the insulin signaling pathway or HSPs. These data provide evidence of the possible association between the non-genomic mechanisms of action of the GR and the insulin signaling pathway in the bovine ovary.

Single-cell RNA-seq analysis and cell-cluster deconvolution of the human preovulatory follicular fluid cells provide insights into the pathophysiology of ovarian hyporesponse.

Reduction in responsiveness to gonadotropins or hyporesponsiveness may lead to the failure of in vitro fertilization (IVF), due to a low number of retrieved oocytes. The ovarian sensitivity index (OSI) is used to reflect the ovarian responsiveness to gonadotropin stimulation before IVF. Although introduced to clinical practice already years ago, its usefulness to predict clinical outcomes requires further research. Nevertheless, pathophysiological mechanisms of ovarian hyporesponse, along with advanced maternal age and in younger women, have not been fully elucidated. Follicles consist of multiple cell types responsible for a repertoire of biological processes including responding to pituitary gonadotropins necessary for follicle growth and oocyte maturation as well as ovulation. Encouraging evidence suggests that hyporesponse could be influenced by many contributing factors, therefore, investigating the variability of ovarian follicular cell types and their gene expression in hyporesponders is highly informative for increasing their prognosis for IVF live birth. Due to advancements in single-cell analysis technologies, the role of somatic cell populations in the development of infertility of ovarian etiology can be clarified. Here, somatic cells were collected from the fluid of preovulatory ovarian follicles of patients undergoing IVF, and RNA-seq was performed to study the associations between OSI and gene expression. We identified 12 molecular pathways differentially regulated between hypo- and normoresponder patient groups (FDR<0.05) from which extracellular matrix organization, post-translational protein phosphorylation, and regulation of Insulin-like Growth Factor (IGF) transport and uptake by IGF Binding Proteins were regulated age-independently. We then generated single-cell RNA-seq data from matching follicles revealing 14 distinct cell clusters. Using cell cluster-specific deconvolution from the bulk RNA-seq data of 18 IVF patients we integrated the datasets as a novel approach and discovered that the abundance of three cell clusters significantly varied between hypo- and normoresponder groups suggesting their role in contributing to the deviations from normal ovarian response to gonadotropin stimulation. Our work uncovers new information regarding the differences in the follicular gene expression between hypo- and normoresponders. In addition, the current study fills the gap in understanding the inter-patient variability of cell types in human preovulatory follicles, as revealed by single-cell analysis of follicular fluid cells.

Medullary bone in male budgerigars (Melopsittacus undulatus) with testicular neoplasms

Medullary bone is a calcium-rich, labile bone normally occurring in female birds with each egg-laying cycle. The stimulus for formation of medullary bone is, in part, the cyclic increase in serum estrogens produced by preovulatory ovarian follicles. Increased bone density due to formation of medullary bone, particularly in pneumatic bones, has been termed polyostotic hyperostosis, even if physiologic. This study investigated the formation of medullary bone in nonpneumatic (femur) and pneumatic (humerus) bones in sexually mature male budgerigars submitted for autopsy. Of the 21 sexually mature male budgerigars submitted for autopsy, 7 (33%) had medullary bone in 1 or more bones examined. All 7 male budgerigars with medullary bone had a testicular neoplasm, which was morphologically consistent with a testicular sustentacular cell tumor, seminoma, or interstitial cell tumor. Medullary bone was not present in the 14 cases with other diseases. Medullary bone formation in pneumatic and nonpneumatic bones can occur in male budgerigars with testicular neoplasms. Radiographic increases in medullary bone density, particularly in the humerus, could provide antemortem indication of testicular neoplasia in male budgerigars.

Characterization of XR_311113.2 as a MicroRNA Sponge for Pre-ovulatory Ovarian Follicles of Goats via Long Noncoding RNA Profile and Bioinformatics Analysis.

Long noncoding RNAs (lncRNAs) were identified recently as a large class of noncoding RNAs (ncRNAs) with a length ≥200 base pairs (bp). The function and mechanism of lncRNAs have been reported in a growing number of species and tissues. In contrast, the regulatory mechanism of lncRNAs in the goat reproductive system has rarely been reported. In the present study, we sequenced and analyzed the lncRNAs using bioinformatics to identify their expression profiles. As a result, 895 lncRNAs were predicted in the pre-ovulatory ovarian follicles of goats. Eighty-eight lncRNAs were differentially expressed in the Macheng black goat when compared with Boer goat. In addition, the lncRNA XR_311113.2 acted as a sponge of chi-miR-424-5p, as assessed via a luciferase activity assay. Taken together, our findings demonstrate that lncRNAs have potential effects in the ovarian follicles of goats and may represent a promising new research field to understand follicular development.

MiR-135a Suppresses Granulosa Cell Growth by Targeting Tgfbr1 and Ccnd2 during Folliculogenesis in Mice.

The success of female reproduction relies on high quality oocytes, which is determined by well-organized cooperation between granulosa cells (GCs) and oocytes during folliculogenesis. GC growth plays a crucial role in maintaining follicle development. Herein, miR-135a was identified as a differentially expressed microRNA in pre-ovulatory ovarian follicles between Large White and Chinese Taihu sows detected by Solexa deep sequencing. We found that miR-135a could significantly facilitate the accumulation of cells arrested at the G1/S phase boundary and increase apoptosis. Mechanically, miR-135a suppressed transforming growth factor, beta receptor I (Tgfbr1) and cyclin D2 (Ccnd2) expression by targeting their 3′UTR in GCs. Furthermore, subcellular localization analysis and a chromatin immunoprecipitation-quantitative real-time PCR (ChIP-qPCR) assay demonstrated that the TGFBR1-SMAD3 pathway could enhance Ccnd2 promoter activity and thus upregulate Ccnd2 expression. Finally, estrogen receptor 2 (ESR2) functioned as a transcription factor by directly binding to the miR-135a promoter region and decreasing the transcriptional activity of miR-135a. Taken together, our study reveals a pro-survival mechanism of ESR2/miR-135a/Tgfbr1/Ccnd2 axis for GC growth, and also provides a novel target for the improvement of female fertility.

Endocrine and molecular milieus of ovarian follicles are diversely affected by human chorionic gonadotropin and gonadotropin-releasing hormone in prepubertal and mature gilts

Different strategies are used to meet optimal reproductive performance or manage reproductive health. Although exogenous human chorionic gonadotropin (hCG) and gonadotropin-releasing hormone (GnRH) agonists (A) are commonly used to trigger ovulation in estrous cycle synchronization, little is known about their effect on the ovarian follicle. Here, we explored whether hCG- and GnRH-A-induced native luteinizing hormone (LH) can affect the endocrine and molecular milieus of ovarian preovulatory follicles in pigs at different stages of sexual development. We collected ovaries 30 h after hCG/GnRH-A administration from altrenogest and pregnant mare serum gonadotropin (eCG)-primed prepubertal and sexually mature gilts. Several endocrine and molecular alternations were indicated, including broad hormonal trigger-induced changes in follicular fluid steroid hormones and prostaglandin levels. However, sexual maturity affected only estradiol levels. Trigger- and/or maturity-dependent changes in the abundance of hormone receptors (FSHR and LHCGR) and proteins associated with lipid metabolism and steroidogenesis (e.g., STAR, HSD3B1, and CYP11A1), prostaglandin synthesis (PTGS2 and PTGFS), extracellular matrix remodeling (MMP1 and TIMP1), protein folding (HSPs), molecular transport (TF), and cell function and survival (e.g., VIM) were observed. These data revealed different endocrine properties of exogenous and endogenous gonadotropins, with a potent progestational/androgenic role of hCG and estrogenic/pro-developmental function of LH.

The mRNA-destabilizing protein Tristetraprolin targets “meiosis arrester” Nppc mRNA in mammalian preovulatory follicles

C-natriuretic peptide (CNP) and its receptor guanylyl cyclase, natriuretic peptide receptor 2 (NPR2), are key regulators of cyclic guanosine monophosphate (cGMP) homeostasis. The CNP-NPR2-cGMP signaling cascade plays an important role in the progression of oocyte meiosis, which is essential for fertility in female mammals. In preovulatory ovarian follicles, the luteinizing hormone (LH)-induced decrease in CNP and its encoding messenger RNA (mRNA) natriuretic peptide precursor C (Nppc) are a prerequisite for oocyte meiotic resumption. However, it has never been determined how LH decreases CNP/Nppc In the present study, we identified that tristetraprolin (TTP), also known as zinc finger protein 36 (ZFP36), a ubiquitously expressed mRNA-destabilizing protein, is the critical mechanism that underlies the LH-induced decrease in Nppc mRNA. Zfp36 mRNA was transiently up-regulated in mural granulosa cells (MGCs) in response to the LH surge. Loss- and gain-of-function analyses indicated that TTP is required for Nppc mRNA degradation in preovulatory MGCs by targeting the rare noncanonical AU-rich element harbored in the Nppc 3' UTR. Moreover, MGC-specific knockout of Zfp36, as well as lentivirus-mediated knockdown in vivo, impaired the LH/hCG-induced Nppc mRNA decline and oocyte meiotic resumption. Furthermore, we found that LH/hCG activates Zfp36/TTP expression through the EGFR-ERK1/2-dependent pathway. Our findings reveal a functional role of TTP-induced mRNA degradation, a global posttranscriptional regulation mechanism, in orchestrating the progression of oocyte meiosis. We also provided a mechanism for understanding CNP-dependent cGMP homeostasis in diverse cellular processes.

Mitochondria-targeted therapeutics, MitoQ and BGP-15, reverse aging-associated meiotic spindle defects in mouse and human oocytes.

STUDY QUESTIONDo mitochondria-targeted therapies reverse ageing- and oxidative stress-induced spindle defects in oocytes from mice and humans?SUMMARY ANSWERExposure to MitoQ or BGP-15 during IVM protected against spindle and chromosomal defects in mouse oocytes exposed to oxidative stress or derived from reproductively aged mice whilst MitoQ promoted nuclear maturation and protected against chromosomal misalignments in human oocytes.WHAT IS KNOWN ALREADYSpindle and chromosomal abnormalities in oocytes are more prevalent with maternal aging, increasing the risk of aneuploidy, miscarriage and genetic disorders such as Down’s syndrome. The origin of compromised oocyte function may be founded in mitochondrial dysfunction and increased reactive oxygen species (ROS).STUDY DESIGN, SIZE, DURATIONOocytes from young and old mice were treated with MitoQ and/or BGP-15 during IVM. To directly induce mitochondrial dysfunction, oocytes were treated with H2O2, and then treated the MitoQ and/or BGP-15. Immature human oocytes were cultured with or without MitoQ. Each experiment was repeated at least three times, and data were analyzed by unpaired-sample t-test or chi-square test.PARTICIPANTS/MATERIALS, SETTING, METHODSImmature germinal vesicle (GV) stage oocytes from 1-, 12- and 18-month-old mice were obtained from preovulatory ovarian follicles. Oocytes were treated with MitoQ and/or BGP-15 during IVM. GV-stage human oocytes were cultured with or without MitoQ. Mitochondrial membrane potential and mitochondrial ROS were measured by live-cell imaging. Meiotic spindle and chromosome alignments were visualized by immunofluorescent labeling of fixed oocytes and the 3-dimensional images were analyzed by Imaris.MAIN RESULTS AND THE ROLE OF CHANCEMitoQ or BGP-15 during IVM protects against spindle and chromosomal defects in oocytes exposed to oxidative stress and in oocytes from aged mice (P < 0.001). In human oocytes, the presence of MitoQ during IVM promoted nuclear maturation and had a similar positive effect in protecting against chromosomal misalignments (P < 0.001).LIMITATIONS, REASONS FOR CAUTIONOur study identifies two excellent candidates that may help to improve fertility in older women. However, these potential therapies must be tested for efficacy in clinical IVM systems, and undergo thorough examination of resultant offspring in preclinical models before utilization.WIDER IMPLICATIONS OF THE FINDINGSOur results using in-vitro systems for oocyte maturation in both mouse and human provide proof of principle that mitochondrially targeted molecules such as MitoQ and BGP-15 may represent a novel therapeutic approach against maternal aging-related spindle and chromosomal abnormalities.STUDY FUNDING/COMPETING INTEREST(S)The project was financially supported by the National Health and Medical Research Council and Australian Research Council, Australia. U.A.-Z. was supported by the Iraqi Higher Education and Scientific Research Ministry PhD scholarship and O.C. was supported by TUBITAK-1059B191601275. M.P.M. consults for MitoQ Inc. and holds patents in mitochondria-targeted therapies. R.L.R. is an inventor on patents relating to the use of BGP-15 to improve gamete quality.TRIAL REGISTRATION NUMBERN/A

Reproductive biology and placentotrophic embryonic development of the smalleye smooth-hound shark, Mustelus higmani, from the south-eastern Caribbean

AbstractThe reproductive biology and embryonic development of Mustelus higmani were examined between January 2015 and December 2016 in the south-eastern Caribbean. Captures comprised 813 females (23.2–72.5 cm TL), and 960 males (22.6–62.5 cm TL). The total length at 50% maturity was estimated as 47.8 and 47.5 cm for females and males, respectively. Uterine fecundity ranged from 1 to 8 embryos and ovarian fecundity between 1 and 9 vitellogenic follicles. The time of parturition and mating season of M. higmani may occur throughout the year, peaking between November and February. The presence of pre-ovulatory ovarian follicles along with advanced embryos indicates an annual reproductive cycle for female M. higmani. The main embryonic development stages were recorded in the samples, from uterine eggs (1 to 6 per female) to term embryos (23.0–26.0 cm TL). The transition between placental pre-implantation and post-implantation occurs when embryos have attained a TL of 5.0–6.0 cm. The observation of abundant uterine histotrophic secretions in late pregnant and post-partum females demonstrates that histotrophy may intensify close to birth in this species. The local population of M. higmani appears to have relatively high productivity; nonetheless, this species is heavily harvested and lacks management measures in the study area.

Effect of eCG treatment on gene expression of selected matrix metalloproteinases (MMP-2, MMP-7, MMP-9, MMP-10, and MMP-13) and the tissue inhibitors of metalloproteinases (TIMP-2 and TIMP-3) in the chicken ovary

Several metalloproteinases (MMPs) are present and functional in the chicken ovary and regulate the extracellular matrix (ECM) during follicle development, ovulation, atresia, and regression. The regulation of the abundance of MMPs in avian ovarian follicles, however, is largely unknown. The aim of the present study was to examine effects of equine chorionic gonadotropin (eCG) on abundance of selected MMPs and relevant tissue inhibitors of MMPs (TIMPs) in the hen ovary. The MMP-2 and MMP-9 activity was also determined. Results indicated there were effects of eCG on abundances of MMP-2, MMP-7, MMP-9, MMP-10, MMP-13, TIMP-2, and TIMP-3 mRNA transcript and/or protein relative abundances in white, yellowish, small yellow, and the largest yellow preovulatory (F3-F1) ovarian follicles. The response to eCG depended on the stage of follicle development, layer of follicular wall, and the type of MMPs or TIMPs affected by eCG. Furthermore, there was a pause in egg laying when eCG was administered and there were morphological changes in the ovary following eCG treatment that were associated with alterations in MMP-2 and MMP-9 activity. In general, the results indicate that eCG, which has primarily follicle stimulating hormone (FSH)-like bioactivities, is a negative regulator of MMP abundance and activity in the largest yellow preovulatory follicles. Results from the present study indicate the gonadotropins, especially FSH, by the regulation of transcription, translation, and/or activity of proteins of the MMP system have effects on the mechanisms that underlie ECM remodeling and cell function throughout ovarian follicle development in the chicken ovary.

Neutrophil granulocytes: participation in homeostatic and reparative processes. Part I

After exiting from the bone marrow (BM) into the circulation, mature neutrophil granulocytes undergo a set of sequential phenotypic and physiological changes collectively called “aging” in the absence of inflammation, by constitutively sensing prime signals from commensal microbiota and acquiring higher functional alertness in case of activation upon tissue damage or pathogen invasion. Physiological aging of blood neutrophils and their subsequent return to the BM result in signals modulating size and function of the hematopoietic niche. Circadian physiological infiltration of BM by neutrophils contributes to maintaining baseline level of circulating hematopoietic progenitor cells capable of regeneration and immune surveillance. Apart from the BM, neutrophils actively enter other healthy tissues, probably exerting some effects on their baseline physiologic state. Using lung tissue, it has been shown that neutrophils can “govern” action of gene set regulating cell growth, migration, proliferation, differentiation, and carcinogenesis. Neutrophils participate in destruction of endometrial tissues during desquamation phase as well as subsequent repair and physiological angiogenesis during proliferative phase of the menstrual cycle; promote wall rupture of the preovulatory ovarian follicle and oocyte exit; contribute to degradation and resorption of the corpus luteum in pregnancy failure; play an important physiological role in vascular remodeling in pregnant uterus and developing maternal immune tolerance to semi-allogeneic fetus. Neutrophils actively migrating to the surface of intestinal epithelium during local infection and/or damage stimulate epithelial restitution and recovery of its barrier function. On the other hand, neutrophils recruited into the oral cavity regulate quantitative and qualitative composition of microbial communities in oral biofilms, and ensure healthy state of periodontal structures. Being a major player and regulator in healing of skin wounds at early stage, inflammation, neutrophils not only destroy potential pathogens, but also participate in cleansing wounds from cell debris, produce cytokines, enzymes, and growth factors affecting further stages in repair process. Both apoptosis and NETosis underlying neutrophil death greatly contribute to wound healing. However, dysregulation and imbalance in both apoptosis and NETosis may lead to unfavorable consequences as well as developing chronic non-healing wounds.

Serum Anti-Mullerian Hormone and Estradiol Concentrations in Gilts and Their Age at Puberty.

Simple SummaryIt has been documented in several species that blood levels of the hormone, anti-Mullerian hormone (AMH), are an indirect measure of the number of ovarian follicles and that the number of ovarian follicles may be an indication of potential female fertility. In the present study, we determined AMH levels at various ages in immature female pigs and related these to potential measures of future fertility including age at puberty and numbers of ovarian follicles at puberty, and the gilt response to ovarian stimulation after puberty. Our results support the suggestion that blood levels of AMH are reflective of future fertility in gilts.For experiment one, blood samples were obtained from 200 gilts at 90, 120, 150, 180, and 200 days of age. Serum samples from the 30 youngest (166.1 ± 0.7 days) and 30 oldest (198.8 ± 0.6 days) gilts exhibiting estrus by 200 days, and a further 18 gilts that remained anestrus at 200 days, were assayed for serum concentrations of anti-Mullerian hormone (AMH) and estradiol (E2). Gilts younger at puberty had higher (p < 0.05) AMH levels than those older at puberty, and both groups had higher AMH levels than anestrus gilts (p < 0.05). Regardless of age, serum E2 was higher (p < 0.05) in gilts that achieved puberty than in gilts remaining anestrus. At spontaneous pubertal estrus detection, there was no effect of pubertal age on the number of preovulatory ovarian follicles. For experiment two, 152 prepubertal gilts received an intramuscular (IM) injection of 400 IU eCG plus 200 IU hCG and then received fence-line boar contact to detect estrus onset. Serum AMH concentrations were higher (p < 0.05) in the first 25 gilts to exhibit puberty than the last 28 gilts, with the first gilts also having more preovulatory follicles (p < 0.0001). Taken together, these data support an association between serum AMH concentrations and degree of physiological maturity and ovarian follicular development in gilts.

Estimation of dietary manganese requirement for laying duck breeders: effects on productive and reproductive performance, egg quality, tibial characteristics, and serum biochemical and antioxidant indices

This study was aimed at estimating the dietary manganese (Mn) requirement for laying duck breeders. A total of 504 Longyan duck breeders (body weight: 1.20 ± 0.02 kg) aged 17 wk were randomly allocated to 6 treatments. The birds were fed with a basal diet (Mn, 17.5 mg/kg) or diets supplemented with 20, 40, 80, 120, or 160 mg/kg of Mn (as MnSO4·H2O) for 18 wk. Each treatment had 6 replicates of 14 ducks each. As a result of this study, dietary Mn supplementation did not affect the productive performance of laying duck breeders in the early laying period (17–18 wk), but affected egg production, egg mass, and feed conversion ratio (FCR) from 19 to 34 wk (P < 0.05), and there was a linear and quadratic effect of supplement level (P < 0.05). The proportion of preovulatory ovarian follicles increased (P < 0.01) linearly and quadratically, and atretic follicles (weight and percentage) decreased (P < 0.05) quadratically with dietary Mn supplementation. The density and breaking strength of tibias increased (quadratic; P < 0.05), the calcium content of tibias decreased (linear, quadratic; P < 0.01), and Mn content increased (linear, quadratic; P < 0.001) with increase in Mn. The addition of Mn had a quadratic effect on serum contents of estradiol, prolactin, progesterone, luteinizing hormone, and follicle-stimulating hormone (P < 0.001). Dietary Mn supplementation decreased serum contents of total protein (linear, P < 0.05), glucose (quadratic, P < 0.05), total bilirubin, triglycerides, total cholesterol, low-density lipoprotein cholesterol, and calcium (linear, quadratic; P < 0.05). The serum total antioxidant capacity and total and Mn-containing superoxide dismutase activities increased (linear, quadratic; P < 0.001), and malondialdehyde content decreased (linear, quadratic; P < 0.001) in response to Mn supplemental levels. The dietary Mn requirements, in milligram per kilogram for a basal diet containing 17.5 mg/kg of Mn, for Longyan duck breeders from 19 to 34 wk of age were estimated to be 84.2 for optimizing egg production, 85.8 for egg mass, and 95.0 for FCR. Overall, dietary Mn supplementation, up to 160 mg/kg of feed, affected productive performance, tibial characteristics, and serum biochemical and antioxidant status of layer duck breeders. Supplementing this basal diet (17.5 mg/kg of Mn) with 85 to 95 mg/kg of additional Mn was adequate for laying duck breeders during the laying period.

Predictive value of ovarian hormone-producing function assessment in poor responders undergoing assisted reproductive technologies

Hypothesis/aims of study. The management of poor responders to ovarian stimulation for in vitro fertilization (IVF) has always been a challenge. It is difficult for both doctors and patients to make decision to proceed to oocyte donation or abandon fertility treatment. More predictors of successful IVF treatment in poor responders are needed. The aim of this study was to assess hormonal ovarian function in poor responders undergoing ovarian stimulation and to identify predictors of the chance of clinical pregnancy after IVF cycle.&#x0D; Study design, materials and methods. The study included 45 infertile patients undergoing ovarian stimulation with poor ovarian response according to the Bologna criteria. All patients underwent standard IVF or IVF/ICSI protocol using gonadotropin releasing hormone antagonists. Letrozole (5 mg/day) was administered during the first 5 days of stimulation in standard antagonist FSH/hMG protocol to 13 of the patients included in the study, with standard antagonist FSH/hMG protocol being only administered to other 32 patients. Serum and follicular fluid were collected at the time of follicle aspiration, and the concentrations of total testosterone, estradiol and androstenedione were determined.&#x0D; Results. Follicular fluid concentrations of testosterone and androstenedione were higher and serum estradiol level was lower in the letrozole group. The serum concentrations of testosterone and androstenedione were comparable in both groups, while the serum estradiol/testosterone ratio was lower in the letrozole group. The threshold level of estradiol/testosterone ratio 1000 on the day of oocyte retrieval above which pregnancy and implantation rates were increased was 1532.68 (odds ratio 7.0 (95% CI 1.3935.35), р = 0.02).&#x0D; Conclusion. Evaluating of the serum estradiol / testosterone ratio has been shown to determine aromatase activity of ovarian preovulatory follicles and to predict IVF outcome in poor responders undergoing assisted reproductive technology.

MiR-17-5p affects porcine granulosa cell growth and oestradiol synthesis by targeting E2F1 gene.

Female fertility potential is based on the development and growth of ovarian follicles. Our previous study showed that miR-17-5p was significantly differently expressed in pre-ovulatory ovarian follicles of Large White (LW) and Chinese Taihu (CT) sows. In the present study, we investigated the role of miR-17-5p in ovarian follicle development. We demonstrated that miR-17-5p overexpression significantly decreased the luciferase reporter activity containing the E2F transcription factor 1 (E2F1) 3'untranslated region (3'UTR) and suppressed the E2F1 expression, whereas the miR-17-5p inhibition increased the E2F1 expression in porcine granulosa cells (pGCs). Meanwhile, miR-17-5p overexpression or E2F1 knockdown promoted cell growth, follicular development marker genes (LHR, CYP19A1 and AREG) expression and oestradiol synthesis, and miR-17-5p inhibition suppressed cell growth, follicular development marker genes (LHR, CYP19A1 and AREG) expression and oestradiol synthesis in pGCs. Furthermore, E2F1 knockdown increased CYP19A1 promoter activity. This study suggests that miR-17-5p regulates pGC growth and oestradiol synthesis by targeting E2F1 gene.

The role of EGF-like factor signaling pathway in granulosa cells in regulation of oocyte maturation and development

The surge of luteinizing hormone (LH) in preovulatory ovarian follicles triggers the resumption of meiosis in oocytes and induces the proliferation of surrounding cumulus granulosa cells. It is believed that LH receptors are expressed in the mural granulosa cells, but not the oocytes and the surrounding cumulus cells, suggesting that the LH signaling is mediated by factors produced by the granulosa cells. However, the mechanism underlying oocyte maturation induced by LH before ovulation has been controversial. Current studies suggest that LH binds on to its receptor on granulosa cells of the follicular wall to promote the production of EGF-like factors, which activate various signaling cascades and induce oocyte maturation and development. Since the in vitro maturation system is difficult to simulate the in vivo physiological environment, in vitro cultured follicles are likely to be de?cient in the EGF-like factors, which could result in the poor developmental competency of in vitro cultured oocytes and restrict their efficient utilization. In this review, we summarize the EGF-like factor signaling system in granulosa cells and its regulation of oocyte maturation and development. It aims to optimize the in vitro maturation culture system of oocytes and increase the EGF-like factor signaling system in cumulus granulosa cells, thereby providing a framework for improving the efficiency on in vitro maturation of oocytes.

Growth hormone during in vitro fertilization in older women modulates the density of receptors in granulosa cells, with improved pregnancy outcomes

To study the effect of aging and granulosa cell growth hormone receptor (GHR) expression, and the effect of growth hormone (GH) co-treatment during IVF on receptor expression. Laboratory study. University. A total of 445 follicles were collected from 62 women undergoing standard infertility treatment. Preovulatory ovarian follicle biopsies of granulosa cells and follicular fluid. Older women with a poor ovarian reserve were co-treated with GH to determine the effect of the adjuvant during IVF on the granulosal expression density of FSH receptor (FSHR), LH receptor (LHR), bone morphogenetic hormone receptor (BMPR1B), and GHR. Ovarian reserve, granulosa cell receptor density, oocyte quality, and pregnancy and live birth rates were determined. Growth hormone co-treatment increased the receptor density for granulosal FSHR, BMPR1B, LHR, and GHR compared with the non-GH-treated patients of the same age and ovarian reserve. Growth hormone co-treatment increased GHR density, which may increase GHR activity. The GH co-treatment was associated with a significant increase in pregnancy rate. Growth hormone co-treatment restored the preovulatory down-regulation of FSHR, BMPR1B, and LHR density of the largest follicles, which may improve the maturation process of luteinization in older patients with reduced ovarian reserve. The fertility of the GH-treated patients improved.

SLA-11 mutations are associated with litter size traits in Large White and Chinese DIV pigs

Litter size is an important economic traits in pigs. SLA-11 gene is a member of SLA (swine leukocyte antigen) complex. In our previous study, the SLA-11 gene was differentially expressed in PMSG-hCG stimulated preovulatory ovarian follicles of Chinese Taihu and Large White sows. Here, we identified two mutations (c.754-132 T > C and c.1421 + 38 T > C) in SLA-11 gene and analyzed the associations of two SNPs with litter size traits in Large White (n = 263) and DIV (n = 117) sows. The results showed that in Large White pigs, SLA-11 c.754-132 CC sows produced 0.74 and 0.87 more pigs per litter for TNB and NBA of all parities than did TT sows (p < .05); In DIV pigs, SLA-11 c.754-132 CC sows produced 1.17 more pigs per litter for TNB of all parities than did TC sows (p < .05). In Large White pigs, SLA-11 c.1421 + 38 CC sows produced 0.9 more pigs per litter for TNB of all parities than did TT sows (p < .05), while in DIV pigs SLA-11 c.1421 + 38 CC sows produced 0.84 and 0.7 less pigs per litter for TNB and NBA of all parities than did TT sows (p < .05). Our research indicated that SLA-11 mutations were potential molecular markers for improving the litter size traits in pigs.