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Simple Summary: AAs are crucial for various reproductive functions, including hormone synthesis and follicular development. However, their use in ruminants is limited by ruminal degradation. GAA, a precursor to creatine, offers a promising alternative as it bypasses ruminal breakdown to a significant extent. GAA has shown positive effects on growth performance and feed efficiency in cattle and sheep. While research in this area is still limited, preliminary findings suggest that GAA may improve placental vascularization in cows. Further research is needed to fully understand the effects of GAA on reproductive processes in ruminants. In this study, GAA supplementation was shown to influence ovarian function and follicular development but did not improve overall pregnancy outcomes in ewes. Guanidinoacetic acid (GAA), a precursor of creatine, has a recognized effect on ruminant performance when used as a dietary supplement. However, its impact on reproductive response remains to be elucidated. Therefore, this study aimed to contribute initially to this area by supplementing the diets of ewes with a high dose of GAA, evaluating its effects on reproductive response. Twenty adult sheep had their estrus synchronized using an MPA sponge, eCG, and PGF2α. After estrus detection ewes were mated. For 10 days until mating, ewes were grouped in groups of baseline diet (BSD; n = 10) and GAA diet (GAAD; n = 10), which was the BSD with daily 0.9 g/kg DM of GAA. After the eCG + PGF2α dose, the GAAD group exhibited an increase in the peak diastolic and pulsatility of the ovarian artery, a reduction in the systolic/diastolic peaks ratio, and a larger intraovarian blood perfusion area. A greater depletion of follicles with <3 mm was observed in the GAAD group and a higher number of follicles ≥3 mm. No differences were observed between the diets respect to pregnancy, and twin rates. Thus, a high GAA supply before mating significantly alters ovarian vasculature and improves follicular growth in ewes but does not affect the pregnancy rate. [ABSTRACT FROM AUTHOR]

Postpartum uterine disease in dairy cows affects ovarian function, but it is unclear how the type and timing of disease relates to ovarian function. To explore associations between uterine disease and contemporaneous ovarian function, postpartum uterine health was evaluated in 17 lactating primiparous Holstein cows that ovulated. Ovarian function was assessed by measuring daily hormone concentrations and the diameter of the largest follicle and subsequent corpus luteum. Metritis (≤14 d postpartum) was associated with reduced follicle diameter (P = 0.002) and tended to reduce plasma estradiol concentrations (P = 0.062) from d 7 to 16 postpartum compared to cows without metritis. Metritis was associated with reduced plasma progesterone (P = 0.026) but not corpus luteum diameter from d 17 to 26 postpartum. Endometritis (≥15 d postpartum) was associated with reduced corpus luteum diameter (P = 0.005) but did not alter plasma progesterone from d 17 to 26 postpartum compared to cows without endometritis. To explore longer term effects of uterine infection on luteal function, non-lactating primiparous Holstein cows received an intrauterine infusion of vehicle control (n = 11) or pathogenic Escherichia coli and Trueperella pyogenes to induce endometritis (n = 12) and corpora lutea were recovered on d 16 of the estrous cycle, 146 d after intrauterine infusion. Intrauterine infusion of bacteria had no effect on plasma progesterone or luteal diameter compared to control, and only altered the expression of 2 of 94 candidate genes (NCF1 and TLR9). Taken together, these studies imply that uterine diseases are principally associated with changes to ovarian function at a time contemporaneous with disease. • Follicle diameter, estradiol, and progesterone were reduced in cows with metritis. • Corpus luteum diameter was reduced in cows with endometritis. • Induced endometritis altered gene expression in the corpus luteum after infection. [ABSTRACT FROM AUTHOR]

BACKGROUND Successful ovulation is essential for natural conception and fertility. Defects in the ovulatory process are associated with various conditions of infertility or subfertility in women. However, our understanding of the intra-ovarian biochemical mechanisms underlying this process in women has lagged compared to our understanding of animal models. This has been largely due to the limited availability of human ovarian samples that can be used to examine changes across the ovulatory period and delineate the underlying cellular/molecular mechanisms in women. Despite this challenge, steady progress has been made to improve our knowledge of the ovulatory process in women by: (i) collecting granulosa cells across the IVF interval, (ii) creating a novel approach to collecting follicular cells and tissues across the periovulatory period from normally cycling women, and (iii) developing unique in vitro models to examine the LH surge or hCG administration-induced ovulatory changes in gene expression, the regulatory mechanisms underlying the ovulatory changes, and the specific functions of the ovulatory factors. OBJECTIVE AND RATIONALE The objective of this review is to summarize findings generated using in vivo and in vitro models of human ovulation, with the goal of providing new insights into the mechanisms underlying the ovulatory process in women. SEARCH METHODS This review is based on the authors' own studies and a search of the relevant literature on human ovulation to date using PubMed search terms such as 'human ovulation EGF-signaling', 'human ovulation steroidogenesis', 'human ovulation transcription factor', 'human ovulation prostaglandin', 'human ovulation proteinase', 'human ovulation angiogenesis' 'human ovulation chemokine', 'human ovulatory disorder', 'human granulosa cell culture'. Our approach includes comparing the data from the authors' studies with the existing microarray or RNA-seq datasets generated using ovarian cells obtained throughout the ovulatory period from humans, monkeys, and mice. OUTCOMES Current findings from studies using in vivo and in vitro models demonstrate that the LH surge or hCG administration increases the expression of ovulatory mediators, including EGF-like factors, steroids, transcription factors, prostaglandins, proteolytic systems, and other autocrine and paracrine factors, similar to those observed in other animal models such as rodents, ruminants, and monkeys. However, the specific ovulatory factors induced, their expression pattern, and their regulatory mechanisms vary among different species. These species-specific differences stress the necessity of utilizing human samples to delineate the mechanisms underlying the ovulatory process in women. WIDER IMPLICATIONS The data from human ovulation in vivo and in vitro models have begun to fill the gaps in our understanding of the ovulatory process in women. Further efforts are needed to discover novel ovulatory factors. One approach to address these gaps is to improve existing in vitro models to more closely mimic in vivo ovulatory conditions in humans. This is critically important as the knowledge obtained from these human studies can be translated directly to aid in the diagnosis of ovulation-associated pathological conditions, for the development of more effective treatment to help women with anovulatory infertility or, conversely, to better manage ovulation for contraceptive purposes. REGISTRATION NUMBER N/A. [ABSTRACT FROM AUTHOR]

Background: Polytrichia is a common clinical and trichoscopic feature in folliculitis decalvans (FD) that morphologically corresponds to compound follicular structures (CFS) of six or more follicles sharing a single infundibulum, surrounded by fibrosis. Objectives: To characterize the type of follicles in the CFS in scalp biopsy specimens from FD. Methods: We retrospectively reviewed 10 scalp biopsy specimens obtained by using the trichoscopy‐guided approach from the affected scalp of patients diagnosed with early‐stage FD between 2018 and 2023. We assessed the number and type of follicles within the CFS on horizontal sections. Results: The total number of assessed follicles was 205, out of which 159 were part of CFS. Of those, 156 were terminal follicles (146 anagen and ten telogen), three intermediate, and zero vellus follicles. Other common findings included dense mixed cell infiltrate consisting of histiocytes, plasma cells, lymphocytes, neutrophils, and eosinophils; psoriasiform hyperplasia; and fragmented hair shafts. Conclusions: We show that CFS in FD are composed largely of terminal anagen follicles. This may help with further understanding of the disease. [ABSTRACT FROM AUTHOR]

Tannic acid, as a polyphenol, is widely present in dicotyledonous plants and angiosperms. The study found that tannins could affect the ovarian development of Brandt's voles. However, the specific effects of tannic acid on reproduction and its underlying mechanisms are still poorly understood. Therefore, this study explored the effects of tannic acid on the reproduction of Brandt's voles. We treated female Brandt's voles from four weeks postpartum with varying tannic acid doses (0 (control), 0.3% (low dose) or 0.6% (high dose) tannic acids) until 8 weeks (puberty) and 13 weeks (sexual maturity)). Ovarian specimens were collected for immunohistochemical analysis and RT-PCR. The treated Brandt's voles were mated with male Brandt's voles of the same age and we recorded the number of embryos, litters and the male to female ratio of the offspring. We found that the female Brandt's voles in the low-dose tannic acid group had enhanced reproductive abilities, characterized by reduced follicular atresia and increased numbers of embryos and litters of offspring. The different doses of tannic acid altered the female-male ratio of the Brandt's voles' offspring. While the low-dose of tannic acid enhanced the level of ovarian autophagy by significantly increasing the expression of Beclin1 and LC3 proteins in the ovaries of the Brandt's voles during sexual maturity. In short, tannic acid can promote the reproduction of Brandt's voles by affecting the level of ovarian autophagy and improving the development of follicles. Tannins can boost the reproduction of Brandt's voles. [ABSTRACT FROM AUTHOR]

The transcriptional regulation of gene expression in the latter stages of follicular development in laying hen ovarian follicles is not well understood. Although differentially expressed genes (DEGs) have been identified in pre-recruitment and pre-ovulatory stages, the master regulators driving these DEGs remain unknown. This study addresses this knowledge gap by utilizing Master Regulator Analysis (MRA) combined with the Algorithm for the Reconstruction of Accurate Cellular Networks (ARACNe) for the first time in laying hen research to identify master regulators that are controlling DEGs in pre-recruitment and pre-ovulatory phases. The constructed ARACNe network included 10,466 nodes and 292,391 edges. The ARACNe network was then used in conjunction with the Virtual Inference of Protein-activity by Enriched Regulon (VIPER) for the MRA to identify top up- and down-regulated master regulators. VIPER analysis revealed FOXO1 as a master regulator, influencing 275 DEGs and impacting pathways related to apoptosis, proliferation, and hormonal regulation. Additionally, CLOCK, known as a crucial regulator of circadian rhythm, emerged as an upregulated master regulator in the pre-ovulatory stage. These findings provide new insights into the transcriptional landscape of laying hen ovarian follicles, offering a foundation for further exploration of follicle development and enhancing reproductive efficiency in avian species. [ABSTRACT FROM AUTHOR]

Objective To observe the therapeutic effects of endometrial stem cells (EnSCs) and their exosomes on rats with autoimmune premature ovarian failure (POF). Methods EnSCs were isolated and cultured from menstrual cycle decidua in female health volunteers, and were separately cultured in exosome-free medium or exosome-free medium supplemented with 50 µmol/L metformin. Exosomes were extracted from the culture supernatant and identified, named EnSC-Exo and EnSCs-Met-Exo, respectively. Fifteen 4-week-old SD female rats with normal estrous cycles were randomly divided into the control group, POF group, POF-EnSCs group, POF-Exo group, and POF-Met-Exo group, with three rats in each group; the autoimmune POF rat models were established by intraperitoneal subcutaneous double-point injection of immunological reagents in all groups except the control group, and then intraperitoneal injections of normal saline, EnSCs, EnSCs-Exo, and EnSCs-Met-Exo were given, respectively. Serum estradiol (E2), follicle-stimulating hormone (FSH) levels in each group were detected by ELISA. The ovarian tissues of each group were collected, and we observed the appearance and calculated the ovarian volume. Pathological conditions were observed and the number of primordial, primary, secondary, and atretic follicles were recorded after HE staining. Results Compared with the control group, the serum E2 level, ovarian volume, and the numbers of primary and secondary follicles in the POF group significantly decreased, while the serum FSH level and the number of atretic follicles significantly increased (all P<0. 05). Compared with the POF model group, the serum FSH levels and the numbers of atretic follicles in the POF-EnSCs group, POF-Exo group, and POF-Met-Exo group significantly decreased, and the serum E2 level and the number of primary and secondary follicles in the POF-Met-Exo group increased (all P<0. 05). There was no statistically significant difference in the number of primordial follicles among all groups (all P>0. 05). The ovarian appearance and pathological results of the control group were normal; the ovaries of the POF group were congested, the surface was collapsed, the granular sensation was not obvious, and they appeared atrophic, with ovarian tissue showing stromal fibrosis, twisted zona pellucida, and infiltration of granulosa cells and inflammatory cells; the ovarian appearance of the POF-EnSCs group, POF-Exo group, and POF-MetExo group rats was close to that of the control group, and the ovarian tissue inflammation and stromal fibrosis were alleviated in comparison with those of the POF group. Conclusion Intraperitoneal injection of human EnSCs and their exosomes EnSCs-Exo and EnSCs-Met-Exo can alleviate pathological changes such as ovarian fibrosis and inflammation and promote follicular development in autoimmune POF rats, with EnSCs-Met-Exo having a better therapeutic effect. [ABSTRACT FROM AUTHOR]

Objective: To assess the impacts of Platelet-Rich Plasma(PRP) and Granulocyte Colony-Stimulating Factor(G-CSF) on a rat model with induced ovarian follicular damage caused by cyclophosphamide(Cy). Materials and Methods: Forty-two Sprague–Dawley rats were randomly allocated into seven distinct groups as; Group 1(control): NaCl intraperitoneal (IP) injection was administered on days D1, D7, and D14. Group 2(Cy):Cy IP injection on D1 + NaCl IP injection on D7 and D14 were administered. Group 3(PRP): PRP IP injection on D1,D7 and D14 were administered. Group 4(Cy + PRP):Cy IP injection on D1 and PRP IP injection on D1, D7 and D14 were administered. Group 5(G-CSF): G-CSF IP injection on D1, D7 and D14 were administered. Group 6(Cy + G-CSF):Cy IP injection on D1+ G-CSF IP injection on D1, D7 and D14 were administered. Group 7(Cy + PRP + G-CSF):Cy IP injection on D1+ PRP IP injection on D1,D7 and D14+ G-CSF IP injection on D1,D7 and D14 were administered. Follicular number, histological scores of AMH and INSL3 stained follicles at different stages of follicular development, and serum Anti-Müllerian hormone(AMH) were evaluated. Results: The primary, secondary, and antral follicle intensity scores for AMH-positive staining were most prominent in Groups 3 and 5. There was no significant difference between groups 4, 6 and 7 compared to group 1 in terms of follicule counts and AMH staining. The intensity scores of AMH-positive staining follicles were notably reduced in group 2 compared to groups 4, 6, and 7, with a significant difference (p <.01). Among the groups, group 2 exhibited the least intense antral follicle staining for INSL3, displaying a significant difference(p <.01) compared to the remaining groups. Conclusions: Autologous PRP and G-CSF might protect ovarian function in the face of ovarian damage caused by Cy-induced effects. [ABSTRACT FROM AUTHOR]

Objective: Today, there is only limited knowledge of the spatial organization of hair chemistry. Infrared microspectroscopy is a well‐established tool to provide such information and has significantly contributed to this field. In this study, we present new results combining multiple infrared microspectroscopy methods at different length scales to create a better chemical histology of human hair, including the hair follicle, hair shaft, hair medulla and hair cuticle. Methods: We used hyperspectral IR imaging & spectroscopy (HIRIS) and synchrotron‐radiation FTIR microspectroscopy (SR‐μFTIR) to measure transversal hair sections and SR‐μFTIR to obtain high‐resolution maps of longitudinal sections from the hair shaft and from the hair follicle. We used optical photothermal IR microspectroscopy (OPTIR) to analyse the cuticle surface of intact hairs. Results: By mapping longitudinal sections of the human hair follicle with confocal SR‐μFTIR, we report the first demonstration of glycogen presence in the outer root sheath of the hair follicle by spectroscopy, and its quantification at the micron scale. Spectral maps, combined with machine learning‐based analysis, enabled us to differentiate the various layers of the hair follicle and provided insights into the chemical changes that occur during hair formation in the follicle. Using HIRIS and SR‐μFTIR to analyse the hair medulla in transversal sections of human hairs, we report here, for the first time by vibrational spectroscopy methods, the detection of unsaturated lipids at very low concentrations in the medulla. By analysing longitudinal sections of the hair shaft with SR‐μFTIR, we found that calcium carboxylates are present in large regions of the hair cuticle, and not just in small focal areas as previously thought. We then use OPTIR to analyse the hair cuticle of intact hairs at submicron resolution without sectioning and report the distribution of calcium carboxylates at the surface of intact hair for the first time. Conclusion: These new findings illustrate the potential of infrared microspectroscopy for imaging the chemical composition of human hair and may have implications for biomedical research or cosmetology. [ABSTRACT FROM AUTHOR]

Abnormal interaction between granulosa cells and oocytes causes disordered development of ovarian follicles. However, the interactions between oocytes and cumulus granulosa cells (CGs), oocytes and mural granulosa cells (MGs), and CGs and MGs remain to be fully explored. Using single-cell RNA-sequencing (scRNA-seq), we determined the transcriptional profiles of oocytes, CGs and MGs in antral follicles. Analysis of scRNA-seq data revealed that CGs may regulate follicular development through the BMP15-KITL-KIT-PI3K-ARF6 pathway with elevated expression of luteinizing hormone receptor (LHR). Because internalization of the LHR is regulated by Arf6, we constructed LHRN316S mice by CRISPR/Cas9 to further explore mechanisms of follicular development and novel treatment strategies for female infertility. Ovaries of LHRN316S mice exhibited reduced numbers of corpora lutea and ovulation. The LHRN316S mice had a reduced rate of oocyte maturation in vitro and decreased serum progesterone levels. Mating LHRN316S female mice with ICR wild type male mice revealed that the infertility rate of LHRN316S mice was 21.4% (3/14). Litter sizes from LHRN316S mice were smaller than those from control wild type female mice. The oocytes from LHRN316S mice had an increased rate of maturation in vitro after progesterone administration in vitro. Furthermore, progesterone treated LHRN316S mice produced offspring numbers per litter equivalent to WT mice. These findings provide key insights into cellular interactions in ovarian follicles and provide important clues for infertility treatment. [ABSTRACT FROM AUTHOR]

The system forming ovarian follicles is developed to investigate in vitro folliculogenesis in a confined environment to obtain functional oocytes. Several studies have reported the successful generation of fully functional oocytes using mouse-induced pluripotent stem cells (iPSCs) and mouse female germline stem cells (fGSCs) as sources of stem cells for in vitro gametogenesis models. In addition, human oogonia have been generated through heterologous co-culture of differentiated human primordial germ cell-like cells (hPGCLCs) with mouse germline somatic cells, although oocyte formation remains challenging. Thus, studies on in vitro ovarian formation in other species are utilized as an introductory approach for in vitro mammalian gametogenesis by understanding the differences in culture systems between species and underlying mechanisms. In this study, we optimized the method of the entire oogenesis process from rat embryonic gonads. We identified well-maturated MII oocytes from rat gonads using our constructed method. Moreover, we generated the first successful in vitro reconstitution of xenogeneic follicles from mouse primordial germ cells (PGCs) and rat somatic cells. We also established an appropriate culture medium and incubation period for xenogeneic follicles. This method will be helpful in studies of xenogeneic follicular development and oocyte generation.

The purpose of this research was to determine the correlation between the weight of the ovary, the number of follicles, and the quality of the oocytes from the culling female bovine obtained from the slaughterhouse. The variation may affect the potential of the ovary to produce follicles. Paired ovaries were obtained from female bovines. Follicle numbers were recorded and divided into three size categories (small: 3 mm, medium: < 3 mm-8 mm, and large: > 8 mm). Oocytes were aspirated and the number of oocytes was recorded and graded into four categories (grades A, B, C, and D). There was a positive correlation between the weight of the ovary and the number of follicles, which is 0.560 with the regression equation y = 3.52 + 0.501 x. There was a positive correlation between the number of follicles and the number of oocytes, which is 0.546 with the regression equation y =2.48 + 1.204 x. There was a positive correlation between the number of oocytes and the grade of oocytes, which is 0.520, with the regression equation y = 0.93 + 0.800 x. There was no correlation between the weight of the ovary and oocyte grade A, which is 0.013.

Context: The number of developmentally competent cumulus–oocyte complexes (COCs) retrieved during Ovum Pick-Up (OPU) determines success in both bovine and human assisted reproduction. Follicular flushing for COC retrieval is practicsed widely in humans but not in cattle. Aims: To determine the benefits of follicular flushing in cattle and assess the merits of a novel 16G double-lumen needle ('OxIVF') that flushes laterally to the needle shaft. Methods: Experiment 1 flushed 655 antral follicles (≥7 mm) from 255 abattoir-derived cattle ovaries. In Experiment 2, 12 Holstein heifers underwent two cycles of OPU in a cross-over design comparing both needle types. In Experiment 3, 11 Holstein heifers underwent two cycles of OPU using the OxIVF needle in a cross-over design: flushing (≥7 mm follicles) vs a 'Hybrid' approach of flushing (≥7 mm follicles) and aspiration (5–7 mm follicles); followed by two cycles of standard follicle aspiration (>5 mm follicles). Key results: In Experiment 1, COC recovery was greater (P = 0.034) for the OxIVF vs Standard needle (mean ± s.e.; 74.1 ± 2.10% vs 67.0 ± 2.23%); yield of Grade 1 COCs was also greater (20.1 ± 1.97% vs 8.2 ± 1.38%; P < 0.001). In Experiment 2, recovery of COCs was greater (P = 0.045) for the OxIVF vs Standard needle (89.1 ± 2.98% vs 79.6 ± 3.47%). Day 6 embryo yield was also greater (P = 0.017) for the OxIVF vs Standard needle (87.2 ± 4.38% vs 67.6 ± 6.73%). In Experiment 3, recovery of COCs was greater (P = 0.033) for 'Flush' vs 'Aspirate' groups (82.1 ± 5.06% vs 66.2 ± 3.48%). However, number of Day 8 blastocysts for the 'Hybrid' vs 'Flush' approach (9.2 ± 1.39 vs 6.5 ± 1.05 per cycle) did not reach statistical significance. Conclusions: Follicular flushing using the OxIVF needle, embracing the 'Hybrid' approach, has the potential to increase oocyte retrieval and blastocyst number per donor cycle in cattle but requires further validation. Implications: Larger scale studies will seek to confirm benefits of follicular flushing using the OxIVF needle in cattle. Future studies should consider applications in both equine and human assisted reproduction. The number of oocytes retrieved during assisted reproduction is a key determinant of success in both cattle and humans. Ovarian follicular flushing is not practised in cattle but offers the prospect of increasing oocyte yield and number of transferrable embryos. We determined that a 'hybrid' approach of flushing larger follicles and aspirating smaller follicles has potential in this regard. These findings serve as an impetus for follow-up studies in human and equine assisted reproduction. Image provided by authors. This article belongs to the collection: The biology of the ovary – Honouring the contributions of Ken P McNatty and Rex J Scaramuzzi. [ABSTRACT FROM AUTHOR]

Premature ovarian failure (POF) is intricately linked to cellular fates such as senescence, apoptosis, and impaired granulosa cell (GC) differentiation, each of which contributes to ovarian dysfunction and follicular depletion. Autophagy is essential in preventing POF by maintaining cellular homeostasis through the degradation and recycling of damaged organelles and proteins, thereby preserving ovarian function and preventing follicular depletion. Recent studies have revealed that the targeted regulation and disruption of autophagy through various molecular mechanisms ultimately lead to the pathogenesis of POF. In this review, we provide a comprehensive analysis of the disruption in regulatory mechanisms of autophagy contributing to POF. Specifically, we elucidate the molecular mechanisms that can be targeted to restore autophagy homeostasis, offering therapeutic potential for the treatment of POF. Significance Statement: This review investigates the critical role of autophagy in preventing POF, a condition causing infertility and other health issues in women. The purpose is to understand how disruptions in autophagy contribute to POF by examining its impact on several types of cells during follicle development. Autophagy dysregulation leads to compromised germ cell viability, disrupted oogonia‐to‐oocyte transformation, and impaired differentiation, which are central to POF pathogenesis. This work highlights the potential of targeting autophagy for developing new treatments, offering valuable insights for future research aimed at preserving ovarian function and fertility. [ABSTRACT FROM AUTHOR]

In this study, the fruit and seed morphology of all the Periplocoideae (Periploca gracilis and P. graeca) and the majority of Asclepiadoideae (Araujia sericifera, Cionura erecta, Cynanchum acutum and Gomphocarpus fruticosus) taxa in Türkiye was evaluated from a taxonomic point of view using, for the first time, both light and scanning electron microscopy. Macro- and micro-morphological investigations were carried out on herbarium samples. An identification key for the examined Periplocoideae and Asclepiadoideae taxa, and expanded fruit and seed descriptions for each taxon were prepared. Numerical analysis showed that shape and size of follicle, size, shape and surface texture of seed were found to be effective in delimiting the studied Periplocoideae and Asclepiadoideae taxa at subfamilial and generic levels. [ABSTRACT FROM AUTHOR]

Objectives: To determine the effect of short-term metformin administration on follicular fluid (FF) total oxidant status (TOS), total antioxidant status (TAS), oxidative stress index (OSI) and nuclear factor kappa B (NF-kB) in women with clomiphene citrate-resistant polycystic ovary syndrome (PCOS). Methods: Fifty-eight patients aged 23–34 who were planned to have intracytoplasmic sperm injection due to clomiphene citrate-resistant PCOS were included in the study. Participants were divided into two groups according to whether they used metformin or not. While 30 of 58 PCOS patients were using short-term metformin in combination with controlled ovarian stimulation, 28 PCOS patients were not using metformin. Metformin was started in the mid-luteal period and continued until the day before oocyte retrieval at 850 mg twice daily. To determine FF-NF-kB, TAS, TOS and OSI values, a dominant follicle ≥17–18 mm in diameter was selected for aspiration. Results: The number of mature follicles and fertilization rates of the metformin group were significantly higher than those not taking metformin. FF-TOS and OSI of the metformin group were significantly lower than those of the group not receiving metformin. Patients receiving metformin had higher FF-TAS than the group not receiving metformin. FF-NF-kB levels of the metformin group were significantly lower than the group not receiving metformin. Insulin resistance, FF-NF-kB and FF-TOS were negatively correlated with the number of mature oocytes. FF-TAS was positively correlated with the number of oocytes. Conclusions: Short-term metformin treatment in clomiphene-resistant PCOS improves the number of mature follicles and fertilization rates by regulating the intra-follicle redox balance. [ABSTRACT FROM AUTHOR]

In mammalian ovaries, most follicles do not ovulate and are eliminated by atresia, which primarily depends on granulosa cell (GC) apoptosis. Autophagy is an alternative mechanism involved in follicle depletion in mammals through independent or tandem action with apoptosis. However, follicular autophagy has not yet been investigated in sheep; therefore, the present study aimed to investigate the involvement of autophagy in atresia among a pool of growing antral follicles in ewe ovaries. The abundance of the autophagic marker LC3B-II was determined using western blotting in GCs collected from ewe antral follicles. The antral follicles were classified as healthy or atretic based on morphological criteria and steroid measurements in follicular fluid (FF). Immunofluorescence and confocal microscopy analyses were performed on GCs to evaluate the presence of autophagic proteins and their subcellular localisation. Caspase-3 and DNA fragmentation were assessed using western blotting and TUNEL assays, respectively, in the same GC population to investigate the simultaneous apoptosis. The novel results of this study demonstrated enhanced LC3B-II protein expression in GCs of atretic follicles compared to that of healthy ones (1.3-fold increase; P = 0.0001, ANOVA), indicating a correlation between autophagy enhancement in GCs and antral follicular atresia. Autophagy, either functioning independently or in tandem with apoptosis, may be involved in the atresia of growing antral follicles in ewe ovaries because atretic GCs also showed high levels of apoptotic markers. The findings of this study might have important implication on scientific understanding of ovarian follicle dynamics. • Ovine granulosa cells undergo autophagy during follicular atresia at antral stage • The autophagic marker level is higher in atretic than in healthy granulosa cells • Granulosa cells of atretic follicles undergo apoptosis as well • Autophagy and apoptosis could regulate granulosa cell death • Both autophagy and apoptosis could be involved in antral follicle atresia in sheep [ABSTRACT FROM AUTHOR]

Introduction: Survival rates of the childhood cancer patients are improving, however cancer treatments such as chemotherapy may lead to infertility due to loss of the primordial follicle (PMF) reserve. Doxorubicin (DXR) is a gonadotoxic chemotherapy agent commonly used in childhood cancers. Anti-Müllerian Hormone (AMH) has been reported to have a protective effect on the mouse ovarian reserve against DXR in vivo. However, whether AMH can prevent PMF loss in conjunction with DXR in human ovarian tissue in vivo has not been determined. Methods: In order to investigate this, we first established an optimum dose of DXR that induced PMF loss in cultured mouse ovaries and investigated the efficacy of AMH on reducing DXR-induced PMF loss in mice in vitro. Second, we investigated the effects of DXR on pre-pubertal human ovarian tissue and the ability of AMH to prevent DXR-induced damage comparing using a mouse xenograft model with different transplantation sites. Results: Mouse ovaries treated with DXR in vitro and in vivo had reduced PMF populations and damaged follicle health. We did not observe effect of DXR-induced PMF loss or damage to follicle/stromal health in human ovarian cortex, this might have been due to an insufficient dose or duration of DXR. Although AMH does not prevent DXR-induced PMF loss in pre-pubertal and adult mouse ovaries, in mouse ovaries treated with higher concentration of AMH in vitro, DXR did not cause a significant loss in PMFs. This is the first study to illustrate an effect of AMH on DXR-induced PMF loss on pre-pubertal mouse ovaries. However, more experiments with higher doses of AMH and larger sample size are needed to confirm this finding. Discussion: We did not observe that AMH could prevent DXR-induced PMF loss in mouse ovaries in vivo. Further studies are warranted to investigate whether AMH has a protective effect against DXR in xenotransplanted human ovarian tissue. Thus, to obtain robust evidence about the potential of AMH in fertility preservation during chemotherapy treatment, alternative AMH administration strategies need to be explored alongside DXR administration to fully interrogate the effect of DXR and AMH on human xenografted tissues. [ABSTRACT FROM AUTHOR]

Ovulation is vital for successful reproduction. Following ovulation, cumulus cells and oocyte are released, while mural granulosa cells (mGCs) remain sequestered within the post‐ovulatory follicle to form the corpus luteum. However, the mechanism underlying the confinement of mGCs has been a longstanding mystery. Here, in vitro and in vivo evidence is provided demonstrating that the stiffening of mGC‐layer serves as an evolutionarily conserved mechanism that prevents mGCs from escaping the post‐ovulatory follicles. The results from spatial transcriptome analysis and experiments reveal that focal adhesion assembly, triggered by the LH (hCG)‐cAMP‐PKA‐CREB signaling cascade, is necessary for mGC‐layer stiffening. Disrupting focal adhesion assembly through RNA interference results in stiffening failure, mGC escape, and the subsequent development of an abnormal corpus luteum characterized by decreased cell density or cavities. These findings introduce a novel concept of "mGC‐layer stiffening", shedding light on the mechanism that prevents mGC escape from the post‐ovulatory follicle. [ABSTRACT FROM AUTHOR]

This study aims to compare the concentration of the hormone PNX-14 in obese women with polycystic ovary syndrome (PCOS) with that in women without PCOS, and to investigate its relationship with certain sexual hormones. Materials and Methods: This study was conducted at Al-Salam Teaching Hospital in Mosul from July 1, 2022, to the end of September 2022. The study included 100 samples from women aged between 16 and 47 years, of whom 50 were obese and thin women diagnosed with polycystic ovary syndrome, and 50 were healthy women. Samples were collected from women during the early follicular phase (the second or fifth day of the menstrual cycle). The group of obese women was divided into three categories based on the degree of obesity (morbid obesity, class I obesity, class II obesity). Both the patient group and the control group were divided into four age categories: (>40), (31-40), (21-30), and (<20) years. Serum samples were separated and stored in sealed tubes, and we then measured the hormone concentrations (PNX-14, progesterone, luteinizing hormone, follicle-stimulating hormone, testosterone). An increase was observed in the concentrations of both luteinizing hormone, estrogen, prolactin, and testosterone in the serum of women with polycystic ovary syndrome compared to the healthy women. Meanwhile, the hormones (follicle-stimulating hormone and progesterone) showed a significant decrease in their concentrations in women with PCOS compared to the healthy women [ABSTRACT FROM AUTHOR]

The objective of the study was to characterise the expression patterns of the two key components of cortisol action namely HSD11B1 (11‐beta‐hydroxysteroid dehydrogenase type 1) and NR3C1 (nuclear receptor subfamily 3, group C, member 1, also known as the glucocorticoid receptor) in superovulation induced bovine follicles during the periovulation and subsequent corpus luteum (CL) formation. Bovine ovaries containing preovulatory follicles or CL were timely defined during induced ovulation as follows: 0 h before GnRH (Gonadotropin‐releasing hormone) application, and 4, 10, 20, 25 (follicles) and 60 h (early CL) after GnRH. The low mRNA expression of HSD11B1 and NR3C1 in the follicle group before the GnRH application increased significantly in the follicle group 20 h after GnRH and remained high afterward also in the early CL group. In contrast, the high NR3C1 mRNA decreased in follicles 25 h after GnRH (close to ovulation) and significantly increased again after ovulation (early CL). Our results indicated the involvement of HSD11B1 and NR3C1 as the two key components of cortisol action in the local mechanisms coordinating final follicle maturation, ovulation, follicular‐luteal transition and CL development in the cow. [ABSTRACT FROM AUTHOR]

The purpose of this research was to determine the correlation between the weight of the ovary, the number of follicles, and the quality of the oocytes from the culling female bovine obtained from the slaughterhouse. The variation may affect the potential of the ovary to produce follicles. Paired ovaries were obtained from female bovines. Follicle numbers were recorded and divided into three size categories (small: 3 mm, medium: < 3 mm-8 mm, and large: > 8 mm). Oocytes were aspirated and the number of oocytes was recorded and graded into four categories (grades A, B, C, and D). There was a positive correlation between the weight of the ovary and the number of follicles, which is 0.560 with the regression equation y = 3.52 + 0.501 x. There was a positive correlation between the number of follicles and the number of oocytes, which is 0.546 with the regression equation y =2.48 + 1.204 x. There was a positive correlation between the number of oocytes and the grade of oocytes, which is 0.520, with the regression equation y = 0.93 + 0.800 x. There was no correlation between the weight of the ovary and oocyte grade A, which is 0.013. [ABSTRACT FROM AUTHOR]

Abstract Premature ovarian insufficiency (POI) is defined as onset of menopause characterized by amenorrhea, hypergonadotropism, and hypoestrogenism, before the age of 40 years. The POI is increasing, which seriously affects the quality of patients’ life. Due to its diversity of pathogenic factors, complex pathogenesis and limited treatment methods, the search for finding effective treatment of POI has become a hotspot. Stem cells are characterized by the ability of self-renewal and differentiation and play an important role in the regeneration of injured tissues, which is therapy is expected to be used in the treatment of POI. The aim of this review is to summarize the pathogenic mechanisms and the research progress of POI treatment with stem cells from different sources.

Jing Lin,1 Fenglu Wu,2 Yanwen Zhu,2 Qianqian Zhu,2 Tong Du,2 Jiaying Lin2 1Center for Reproductive Medicine, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China; 2Department of Assisted Reproduction, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of ChinaCorrespondence: Jiaying Lin; Tong Du, Department of Assisted Reproduction, Shanghai Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Zhizaoju Road No. 639, Shanghai, People’s Republic of China, Email lemon_1114@126.com; ocyte@qq.comObjective: Our objective was to investigate the efficacy of letrozole co-treatment in an antagonist protocol for infertile women with polycystic ovary syndrome (PCOS).Patients and Methods: This retrospective cohort study included infertile women with PCOS undergoing IVF/ICSI with and without letrozole co-treatment in an antagonist protocol from 2007– 2021 at Shanghai Ninth People’s Hospital (Shanghai, China). A total of 1559 participants were enrolled, with 1227 women in the antagonist group and 332 women in the letrozole co-treatment group. Propensity score-based patient-matching model was conducted to balance covariates between the groups. The primary outcome was the number of retrieved oocytes, with secondary outcomes including endocrine parameters, ovarian stimulation outcomes, pregnancy outcomes, and obstetrical and neonatal complications.Results: Letrozole co-treatment induced significant changes in hormonal regulation, increased the percentage of large follicles, and resulted in fewer retrieved oocytes (P < 0.05). However, there was no negative impact on the number of usable embryos or good-quality embryos (P > 0.05). The live birth rates following fresh embryo transfer were comparable between the letrozole and control groups (single embryo transfer: 28.9% vs 29.7%, P > 0.05; double embryo transfer: 37.3% vs 45.6%, P > 0.05). Additionally, there were no significant differences between the two groups in the live birth rate per patient after frozen embryo transfer and the cumulative live birth rate (P > 0.05). No significant differences in obstetrical and neonatal complications were observed between the groups (P > 0.05).Conclusion: The addition of letrozole to the antagonist protocol for women with PCOS undergoing IVF induces a higher percentage of large follicles during oocyte retrieval, while reducing the overall number of retrieved oocytes. Moreover, the use of letrozole demonstrates comparable clinical outcomes following embryo transfers. These findings highlight the potential application of letrozole in an antagonist protocol for women with PCOS.Keywords: letrozole, antagonist protocol, polycystic ovary syndrome, follicle, in vitro fertilization

Abstract Genetic variations in the ovine ovulation rate, which are associated with the FecB mutation, provide useful models by which to explore the mechanisms regulating the development of mammalian antral follicles. In order to study the effects of the FecB mutation on cumulus cell differentiation, preovulatory follicles were aspirated and cumulus cells were isolated from three FecB genotypes (homozygous, heterozygous and wild type) of Small Tail Han (STH) sheep superstimulated with FSH. Transcriptome information from tens of thousands of cumulus cells was determined with the 10 × Genomics single-cell RNA-seq technology. Under the superovulation treatment, the observed number of preovulatory follicles in the ovaries of FecB carriers was still significantly higher than that in the wild-type (P

Premature ovarian insufficiency (POI) is defined as onset of menopause characterized by amenorrhea, hypergonadotropism, and hypoestrogenism, before the age of 40 years. The POI is increasing, which seriously affects the quality of patients' life. Due to its diversity of pathogenic factors, complex pathogenesis and limited treatment methods, the search for finding effective treatment of POI has become a hotspot. Stem cells are characterized by the ability of self-renewal and differentiation and play an important role in the regeneration of injured tissues, which is therapy is expected to be used in the treatment of POI. The aim of this review is to summarize the pathogenic mechanisms and the research progress of POI treatment with stem cells from different sources. [ABSTRACT FROM AUTHOR]

Simple Summary: Microencapsulated hot chili pepper (MHCP) has properties that affect the metabolism of bovines, so the objective of this research was to study the effects of MHCP on milk production and the reproduction of dairy cows. Twenty-four animals were separated into two groups. In the first group, each cow consumed 1 g of microencapsulated hot chili per day. The second group served as a control group for 42 days. Daily milk production and weekly milk composition were monitored for 42 days. These animals were subjected to a hormonal synchronization protocol, tests to monitor the size and vascularization of the ovarian structures, as well as the evaluation of serum progesterone. Ultimately, the supplemented group presented differences in milk production and milk components. On the other hand, a trend was found in which the ovarian follicle was mostly irrigated in the group that consumed the MHCP. We conclude that MHCP provides benefits in dairy production. The present study evaluates the effect of including microencapsulated hot chili pepper (MHCP) in the diet of crossbred dairy cows on the volume and quality of milk and on ovarian morphofunctionality. Twenty-four crossbred females in their lactating period were used. The cows were divided into two experimental groups, a control (CT) and an MHCP -supplemented group (CP) given 1 g a day per animal of microencapsulated hot chili in concentrate for 42 days. Over seven weeks of daily milk production was measured, and sample milk was collected weekly for composition analysis. Animals were subject to an ovulation synchronization protocol on day 0 (D0), and an intravaginal progesterone (P4) implant, estradiol benzoate, and prostaglandin (PGF2α) were administered. On D8, the P4 implant was removed and PGF2α, equine chorionic gonadotropin, and estradiol cypionate were administered to the animals. The ovarian dynamics were evaluated in B mode and color Doppler. There were significant differences (p < 0.05) in the group X time interaction, the volume of milk produced, and the amount in kg/day of milk components. There was a higher percentage of vascularization in the preovulatory follicle in the CP group (p ≥ 0.10). The findings show that the inclusion of MHCP in the diet of dairy cows does influence their milk production and reproduction. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to assess differences in various scalp parameters between patients with androgenetic alopecia (AGA) and healthy volunteers using 22 MHz ultrasound. Methods: Thirty patients with AGA (AGA group) and 30 healthy volunteers (control group) who visited the Department of Dermatology at the Second Affiliated Hospital of Soochow University from September 2021 to June 2022 were randomly selected. The patients with AGA met the diagnostic criteria outlined in the Chinese Guidelines for the Diagnosis and Treatment of Androgenetic Alopecia. The severity of alopecia was assessed for males between grades 2 and 4 on the Norwood–Hamilton scale, and for females between stages 2 and 3 on the Ludwig scale. No artificial interventions were conducted at the vertex, and all examination conditions remained consistent. Ultrasound examinations at 22 MHz were performed on the scalp at the vertex in both the AGA and control groups. Seven parameters were measured, namely, epidermis + dermis thickness, entire scalp thickness, subcutaneous tissue thickness, average follicle width, average follicle length, follicle count, and the presence of color flow signals in the subcutaneous tissue. The differences in these parameters were then compared. Results: The AGA group showed reduced thickness of the entire scalp and subcutaneous tissue, narrower average follicle width, shorter average follicle length, lower hair follicle count, and fewer instances of color flow signals in the subcutaneous tissue at the vertex area (p < 0.05). Conclusion: High‐frequency (22 MHz) ultrasonography can be employed to visualize the entrance echo, dermis, subcutaneous tissue, and hair follicles of the scalp, thereby providing imaging for the clinical assessment of hair loss. [ABSTRACT FROM AUTHOR]

In recent years, blood anti-Müllerian hormone (AMH) levels have been investigated in female animals to diagnose many conditions, such as the presence of ovarian tissue, follicle reserve, and granulosa cell tumors. Since blood collection is an invasive method, diagnosis with a non-invasive method is important in terms of practicality and animal welfare. This study aimed to investigate the presence of AMH in cat urine and determine whether a correlation exists between blood and urine AMH levels. In addition, it was aimed at revealing whether there was a change in blood and urine AMH levels according to ovarian follicle distribution. Twenty-seven healthy, fertile female cats in the follicular phase were included. Following blood and urine sample collection, a routine ovariohysterectomy was performed. Histological analysis of the removed ovarian tissue was used to determine ovarian follicle types. While both AMH and estrogen levels were determined in blood samples, only AMH levels were investigated in urine samples. Blood AMH levels averaged 10.61 ± 0.75 ng/mL (range: 5–16 ng/mL), while urine AMH levels averaged 5.67 ± 0.91 ng/mL (range: 0.2–13 ng/mL). While urinary AMH level was <1 ng/mL in 7 cats, urinary AMH was >1 ng/mL in all remaining cats. While the study demonstrated AMH excretion in urine, no correlation was found between blood and urine AMH values. However, a significant positive correlation was observed between blood AMH levels and serum estrogen levels (P < 0.001). These findings suggest that urinary AMH may be a product of proteolytic degradation, potentially leading to inaccurate estimations of ovarian activity based solely on urine AMH levels. • In cats, AMH is excreted in the urine. • There is no correlation between blood and urine AMH levels in cats. • Urinary AMH levels in intact cats may be < 1 ng/mL. [ABSTRACT FROM AUTHOR]

During the development of animal organs, various adverse stimuli or toxic environments can induce oxidative stress and delay ovarian development. Paeoniflorin (PF), the main active ingredient of the traditional Chinese herb Paeonia lactiflora Pall., has protective effects on various diseases by preventing oxidative stress. However, the mechanism by which PF attenuates oxidative damage in mouse ovaries remains unclear. We evaluated the protective effects of PF on ovaries in an H2O2-induced mouse oxidative stress model. The H2O2-induced mouse ovarian oxidative stress model was used to explore the protective effect of PF on ovarian development. Histology and follicular development were observed. We then detected related indicators of cell apoptosis, oxidative stress, and autophagy in mouse ovaries. We found that PF inhibited H2O2-induced ovarian cell apoptosis and ferroptosis and promoted granulosa cell proliferation. PF prevented oxidative stress by increasing nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) expression levels. In addition, the autophagic flux of ovarian cells was activated and was accompanied by increased lysosomal biogenesis. Moreover, PF-mediated autophagy was involved in clearing mitochondria damaged by H2O2. Importantly, PF administration significantly increased the number of primordial follicles, primary follicles, secondary follicles, and antral follicles. PF administration improved ovarian sizes compared with the H2O2 group. The present study suggested that PF administration reversed H2O2-induced ovarian developmental delay and promoted follicle development. PF-activated mitophagy is crucial for preventing oxidative stress and improving mitochondrial quality. [ABSTRACT FROM AUTHOR]

Cell death is an important process in the body, as it occurs throughout every tissue during development, disease, and tissue regeneration. Phagocytes are responsible for clearing away dying cells and are typically characterized as either professional or nonprofessional phagocytes. Professional phagocytes, such as macrophages, are found in nearly every part of the body while nonprofessional phagocytes, such as epithelial cells, are found in every tissue type. However, there are organs that are considered “immune-privileged” as they have little to no immune surveillance and rely on nonprofessional phagocytes to engulf dying cells. These organs are surrounded by barriers to protect the tissue from viruses, bacteria, and perhaps even immune cells. The Drosophila ovary is considered immune-privileged, however the presence of hemocytes, the macrophages of Drosophila, around the ovary suggests they may have a potential function. Here we analyze hemocyte localization and potential functions in response to starvation-induced cell death in the ovary. Hemocytes were found to accumulate in the oviduct in the vicinity of mature eggs and follicle cell debris. Genetic ablation of hemocytes revealed that the presence of hemocytes affects oogenesis and that they phagocytose ovarian cell debris and in their absence fecundity decreases. Unpaired3, an IL-6 like cytokine, was found to be required for the recruitment of hemocytes to the oviduct to clear away obsolete follicle cells. These findings demonstrate a role for hemocytes in the ovary, providing a more thorough understanding of phagocyte communication and cell clearance in a previously thought immune-privileged organ. [ABSTRACT FROM AUTHOR]

Objective: Chronic inflammation is considered to be of key importance in the pathogenesis of polycystic ovarian syndrome (PCOS). Ganoderma lucidum polysaccharide (GLP) and Hypericum perforatum (HP) have been reported to have anti-inflammatory and antioxidant activities. We studied the effects of these agents on ovarian tissue in a rat model of experimental PCOS. Materials and Methods: Forty-two Sprague-Dawley female rats were divided into 6 groups with 7 animals in each group as listed below: Group 1: Control, Group 2: PCOS, Group 3: PCOS + HP, Group 4: HP only, Group 5: PCOS+ GLP, Group 6: GLP only. At the end of the experimental procedures, all the animals underwent bilateral oophorectomy and blood samples were collected. Ovarian tissue and blood samples were used for biochemical and histopathological analysis. Results: Follicle degeneration in the PCOS group showed a statistically significant increase compared to the other groups (p < 0.05). Cystic follicles were significantly reduced in the PCOS+GLP and PCOS+HP groups as compared to the PCOS group. Levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) were elevated in PCOS rats (p < 0.01). Levels of sex hormone binding globulin (SHBG) levels were diminished (p < 0.01). Levels of malondialdehyde (MDA) and insulin-like growth factor 1 (IGF-1) were increased in PCOS rats as compared to the other groups (p < 0.02, p < 0.02, respectively). GLP supplementation diminished the levels of IGF-1 and MDA. GLP or HP supplementation increased reduced glutathione (GSH). Conclusion: GLP and HP treatment normalizes SHBG levels while correcting PCOS-induced hyperandrogenemia. Both herbs regulate the redox balance by decreasing the levels of MDA and increasing the level of GSH. [ABSTRACT FROM AUTHOR]

This study aimed to compare the effects of a single dose of GnRH or hCG administered at the beginning of the short-term oestrus synchronization protocol on ≥4 mm diameter follicles in Saanen goats during the breeding season. The goats estrus cycles were synchronized with intravaginal sponges containing 60 mg of medroxyprogesterone acetate for five days. Additionally, 1 ml of physiological saline solution (control-group; n=30), 0.004 mg of buserelin acetate (GnRH-group; n=31) or 150 IU of hCG (hCG-group; n=31) were injected intramuscularly to the goats during the sponges insertion. Transrectal ultrasonographic examination was performed immediately before and 24 h after intravaginal sponge application into all goats and follicles with a diameter of ≥4 mm in the ovaries were counted. Blood samples were collected on the same days to determine serum progesterone (P4) and estradiol (E2) concentrations. At the first ultrasonographic examination, the percentages of ≥4 mm diameter follicles were 56.66% (17/30), 54.83% (17/31) and 70.96% (22/31) in the control, GnRH and hCG groups, respectively. The percentage of goats with reduced follicle diameters 24 h later was 29.41% (5/17), 52.94 (9/17) and 59.09% (13/22) in the same groups, respectively. The mean regression rates of follicle diameters between days 0 and 1 in each group were significantly different (P<0.05). Serum E2 concentrations were significantly different (P<0.05) between days 0 and 1 in hCG group. There were no differences in serum E2, P4 concentrations and mean regression rates in follicle diameters between days 0 and 1 for all groups. As a result, a significant relationship between the administration of hCG or GnRH and the reduction of large follicle diameters could not be established. [ABSTRACT FROM AUTHOR]

Simple Summary: Ovarian translocation and intravital window implantation surgery contribute to high-resolution imaging of the ovarian structure of female rabbits in vivo using UBM. High-resolution UBM could effectively detect the ovarian structure and measure the size of follicles. The combination of UBM and intravital window implantation provides a chance to achieve real-time dynamic and continuous imaging of the ovary. This study offers a relatively novel method to visualize ovarian dynamics, ovulation induction, and the cumulus–oocyte complex in vivo instead of conventional histological examination. Continuous ovarian imaging has been proven to be a method for monitoring the development of follicles in vivo. The aim of this study was to evaluate the efficacy of combining ultrasound bio-microscopy (UBM) with an intravital window for follicle imaging in rabbits and to monitor the ovarian dynamic processes. New Zealand White female rabbits (n = 10) received ovarian translocation to a subcutaneous position. The ovarian tissue was sutured onto the abdominal muscles and covered with an intravital window for the continuous monitoring of the follicles using UBM. Results show that physiological changes (red blood cell and white blood cell counts, feed intake, and body weight change) in rabbits induced by surgery returned to normal physiological levels in one week. Furthermore, UBM could provide high-resolution imaging of follicles through the intravital window. Daily monitoring of ovarian dynamic processes for 6 days displayed variabilities in follicle counts and size. Collectively, these results provide a relatively new method to monitor ovarian dynamic processes and to understand the reproductive physiology of female rabbits. [ABSTRACT FROM AUTHOR]

Genetic variations in the ovine ovulation rate, which are associated with the FecB mutation, provide useful models by which to explore the mechanisms regulating the development of mammalian antral follicles. In order to study the effects of the FecB mutation on cumulus cell differentiation, preovulatory follicles were aspirated and cumulus cells were isolated from three FecB genotypes (homozygous, heterozygous and wild type) of Small Tail Han (STH) sheep superstimulated with FSH. Transcriptome information from tens of thousands of cumulus cells was determined with the 10 × Genomics single-cell RNA-seq technology. Under the superovulation treatment, the observed number of preovulatory follicles in the ovaries of FecB carriers was still significantly higher than that in the wild-type (P < 0.05). The expression patterns of cumulus cells differed between FecB carriers and wild-type ewes. The screened cumulus cells could also be further divided into different cell clusters, and the differentiation states and fates of each group of cumulus cells also remained different, which supports the notion that heterogeneity in gene expression is prevalent in single cells. The oxidative phosphorylation pathway was significantly enriched in differentially expressed genes among the cell differentiation branch nodes of cumulus cells and among the differentially expressed genes of cumulus cells from the three genotypes. Combined with the important role of oxidative phosphorylation in the maturation of COCs, we suggest that the oxidative phosphorylation pathway of cumulus cells plays a crucial role in the differentiation process of cumulus cells and the mutation effect of the FecB gene. [ABSTRACT FROM AUTHOR]

Shaping of developing organs requires dynamic regulation of force and resistance to achieve precise outcomes, but how organs monitor tissue mechanical properties is poorly understood. We show that in developing Drosophila follicles (egg chambers), a single pair of cells performs such monitoring to drive organ shaping. These anterior polar cells secrete a matrix metalloproteinase (MMP) that specifies the appropriate degree of tissue elongation, rather than hyper- or hypo-elongated organs. MMP production is negatively regulated by basement membrane (BM) mechanical properties, which are sensed through focal adhesion signaling and autonomous contractile activity; MMP then reciprocally regulates BM remodeling, particularly at the anterior region. Changing BM properties at remote locations alone is sufficient to induce a remodeling response in polar cells. We propose that this small group of cells senses both local and distant stiffness cues to produce factors that pattern the organs BM mechanics, ensuring proper tissue shape and reproductive success.

Abstract Background Two-dimensional ultrathin Ti3C2 (MXene) nanosheets have gained significant attention in various biomedical applications. Although previous studies have described the accumulation and associated damage of Ti3C2 nanosheets in the testes and placenta. However, it is currently unclear whether Ti3C2 nanosheets can be translocated to the ovaries and cause ovarian damage, thereby impairing ovarian functions. Results We established a mouse model with different doses (1.25, 2.5, and 5 mg/kg bw/d) of Ti3C2 nanosheets injected intravenously for three days. We demonstrated that Ti3C2 nanosheets can enter the ovaries and were internalized by granulosa cells, leading to a decrease in the number of primary, secondary and antral follicles. Furthermore, the decrease in follicles is closely associated with higher levels of FSH and LH, as well as increased level of E2 and P4, and decreased level of T in mouse ovary. In further studies, we found that exposure toTi3C2 nanosheets increased the levels of Beclin1, ATG5, and the ratio of LC3II/Ι, leading to autophagy activation. Additionally, the level of P62 increased, resulting in autophagic flux blockade. Ti3C2 nanosheets can activate autophagy through the PI3K/AKT/mTOR signaling pathway, with oxidative stress playing an important role in this process. Therefore, we chose the ovarian granulosa cell line (KGN cells) for in vitro validation of the impact of autophagy on the hormone secretion capability. The inhibition of autophagy initiation by 3-Methyladenine (3-MA) promoted smooth autophagic flow, thereby partially reduced the secretion of estradiol and progesterone by KGN cells; Whereas blocking autophagic flux by Rapamycin (RAPA) further exacerbated the secretion of estradiol and progesterone in cells. Conclusion Ti3C2 nanosheet-induced increased secretion of hormones in the ovary is mediated through the activation of autophagy and impairment of autophagic flux, which disrupts normal follicular development. These results imply that autophagy dysfunction may be one of the underlying mechanisms of Ti3C2-induced damage to ovarian granulosa cells. Our findings further reveal the mechanism of female reproductive toxicity induced by Ti3C2 nanosheets.