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For fertility preservation, preventing chemotherapy-induced premature ovarian insufficiency (POI) in patients with breast cancer is challenging. Our previous study suggested that caulerpin, a marine indole alkaloid, exerts antitumor effects on breast cancer cells. However, the potential effects of caulerpin on ovarian tissues remain unknown. In the present study, xenograft tumors derived from the MDA-MB-231 breast cancer cell line were established in a female BALB/c nude mouse model. Cyclophosphamide (CTX) alone caused remarkable ovarian damage, including irregular estrous cycles, follicle loss, and reduced expression of anti-Mullerian hormone (AMH) and follicle-stimulating hormone receptor (FSHR), whereas ovarian toxicity was largely reduced after caulerpin treatment in mice and in vitro. The gene signature of the ovaries of CTX-treated tumor-bearing mice revealed differentially expressed genes (DEGs) that regulate two important processes, namely, macrophage polarization and cellular senescence, as well as the activation of the p53/NF-κB signaling pathway. In vitro, CTX induced M1 macrophage polarization in THP-1 cells, which was accompanied by activation of the p53/NF-κB signaling pathway. Additionally, senescence was upregulated in the ovaries of CTX-treated tumor-bearing mice and in granulosa cells (GCs) cocultured with THP-1 cells exposed to LPS/IFN-γ, characterized by increased activity of senescence-associated β-galactosidase (SAβG), increased ROS levels and elevated levels of senescence-related markers (p53, p21 and p38MAPK). Furthermore, caulerpin or a p53 inhibitor (pifithrin-α) modulated CTX-induced M1 polarization in macrophages, thereby delaying GC senescence. These findings demonstrated that caulerpin contributes to alleviating CTX-induced ovarian toxicity by modulating M1 macrophage polarization through the p53/NF-κB signaling pathway, which promotes the senescence of GCs by inducing ROS production.Thus, caulerpin may be a potential therapeutic strategy for breast cancer patients., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Accumulating evidence suggests that exposure to ambient airborne PM2.5 increases the risk of primary ovarian insufficiency (POI). However, whether ferroptosis, a newly discovered type of cell death involved in PM2.5-induced lung injury and fibrosis, is involved in PM2.5-induced POI has not been determined. This study aimed to verify the involvement of PM2.5-induced ferroptosis in ovarian dysfunction and further demonstrate that melatonin inhibits ferroptosis by activating the Nrf2 signaling pathway to ameliorate POI in vivo and in vitro. In our study, PM2.5 promoted iron accumulation and induced lipid peroxidation, thus contributing to ferroptosis in KGN cells and ovaries. However, these effects were eliminated and enhanced in Nrf2-overexpressing and Nrf2-knockdown cells, respectively. In addition, melatonin and ferrostatin-1 (Fer-1) inhibited ferroptosis by activating the NRF2 signaling pathway, as evidenced by the silencing of Nrf2 in vivo and in vitro. Mechanistically, Nrf2-knockout mice were more susceptible to ferroptosis and PM2.5-induced POI than control mice. Moreover, melatonin suppressed changes in morphological and biochemical indicators related to ferroptosis, such as MDA and GSH depletion and GPX4 and XCT downregulation, by enhancing Nrf2 signaling. Here, we first reported that PM2.5 triggered ferroptosis by increasing ROS levels, lipid peroxidation and glutathione depletion. Notably, melatonin significantly decreased ferroptosis levels and improved ovarian function by activating the NRF2 signaling pathway in vivo and in vitro., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

When a naïve observer meets with a familiar conspecific in pain, mice may have a myriad of social (sniffing, allolicking, allogrooming, huddling) and non-social (self-grooming) behaviors under dyadic social interaction (DSI) paradigm. Unlike male, female observers express more allolicking behavior toward injury site of a familiar female in pain, but with less body allogrooming. In current study, we investigated roles of natural estrus cycle phases and ovarian estrogen in these behaviors and results showed that: (1) there was no changes in above behaviors in terms of latency, time and bouts across different natural estrus cycle phases in intact female. (2) however, ovariectomy (OVX) changed estrus cycle phases, lowered circulating level of ovarian estrogen, reduced time and bouts of allolicking behavior and increased time of self-grooming without affecting other behaviors. Moreover, OVX in observers decreased social buffering effect of DSI on spontaneous pain-related behavior in demonstrator relative to naïve and sham controls. (3) treatment of OVX-female with β-estradiol (E 2 ) or progesterone (PROG) as replacement therapies, only E 2 reversed impairment of allolicking behavior. (4) Additionally, socially transferred pain could be identified in intact female across all estrus cycle phases post-DSI, but disappeared in OVX-female, which could be reversed completely by E 2 but not by PROG. (5) Finally, serum levels of estrogen, PROG, oxytocin, arginine vasopressin (AVP), prolactin, norepinephrine and 5-HT were examined by ELISA after E 2 , results showed only AVP level was significantly increased. These results suggest both injury site-targeted caring behavior and socially transferred pain are selectively dependent on ovarian estrogen. This article is part of the Special Issue on "Empathic Pain"., Competing Interests: Declaration of competing interest The authors hereby declare that there are no conflicts of interest, financial or otherwise, that could have influenced the integrity and objectivity of the research conducted and presented in this paper. This includes, but is not limited to, the following aspects: 1. Financial Conflicts of Interest: All authors affirm that they have not received any direct or indirect financial support that could influence the outcomes of this research. Furthermore, no author is employed by, consults for, owns shares in, or receives funding from any company or organization that would benefit from the results of this study. 2. Personal and Academic Relationships: All authors declare that they have no personal relationships or academic competition related to the subject matter of this research that could interfere with the objectivity or fairness of the research. 3. Transparency in Data and Methods: To ensure transparency, the data and methodologies used in this study are openly available for peer review and public scrutiny. 4. Other Potential Conflicts: All authors confirm that there are no other potential conflicts of interest, beyond those disclosed, that could undermine the integrity of this research., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Sevoflurane is extensively employed as an inhalation anesthetic in medical practices, due to its promising pharmacokinetics. Conversely, the data regarding effects of prolonged exposure to trace amounts of sevoflurane on the female reproductive system is obscure. Therefore, this study aimed to investigate the reproductive toxicity and underlying mechanism of long-term sevoflurane inhalation in female rats. A total 60 SPF grade SD female rats were randomly alienated into four equal groups as control group (A), 50 ppm sevoflurane group (B), 150 ppm sevoflurane group (C), and 300 ppm sevoflurane groups (D). Ovaries and uterine organs were collected for gross as well as histopathological analysis, western blotting, and immuno-histochemistry evaluation. Results revealed that pregnancy rate, number of fetuses (fetal mice) and general body weight of group B, C, and D were substantially lower (P < 0.05), while were compared with control. On the contrary, estrous period in groups B, C, D was shortened noticeably (P < 0.05), and estrus interval and cycle were significantly longer (P < 0.05). In fact, the ovarian and uterine coefficients of group B, C and D were significantly reduced as compared with control. However, ovarian and uterine histomorphology remained normal in control group, while obvious pathological alterations were detected in groups B, C, and D. Although, the expression of SOD protein in the ovarian and uterine tissues of groups B, C, and D was significantly reduced (P < 0.05), in contrast to group A. However, the MDA protein expression increased significantly (P < 0.05) as compared with group A. While expression of apoptosis-related genes (Bcl2 and Bax) and humoral immunity related genes (IL-6, IL-10 and TNF-α) showed highest elevation in groups exposure with sevoflurane (p < 0.001) in comparison to control. Conclusively, long-term inhalation of trace amounts of sevoflurane is toxic to female reproductive system and can severely affect reproductive organs and fertility by induction of oxidative stress and apoptosis., Competing Interests: Declarations. Competing interest: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: Ecdysone-induced protein 93 F (E93, also known as Eip93F) plays a crucial role in the reproductive process of numerous insects. This study aims to delineate the function of E93 in Chilo suppressalis and elucidated the regulatory mechanism by which E93 influences the reproduction of C. suppressalis METHODS AND RESULTS: The results of the bioinformatics analysis indicate that C. suppressalis E93 shows the highest homology with E93 from Bombyx mori. We used qPCR to evaluate the expression profile of CsE93 from different developmental stages and tissues, revealed that CsE93 had the highest expression levels in the head, which peaked during the prepupal stage. Silencing CsE93 resulted in a significant reduction in yolk deposition and abnormal ovarian development. Moreover, the transcriptional levels of vitellogenin (Vg) and E74A, which are related to vitellogenesis and the 20E pathway, were significantly down-regulated in dsE93-treated female pupae. In addition, we identified Vitelline membrane protein 26Ab (VMP26Ab), a downstream gene associated with the integrity of the inner eggshell. The knockdown of VMP26Ab resulted in a significant reduction in the number of eggs and abnormal ovarian development, similar to RNAi E93. Finally, we identified an active promoter fragment (containing GAGA-containing motif) of CsVMP26Ab and demonstrated that CsE93 can bind to it., Results: Our results indicate that CsE93 plays an important role in C. suppressalis reproduction. CsE93 modulates the CsVMP26Ab expression by acting on its promoter involve in the reproduction of C. suppressalis finally., Competing Interests: Declarations. Ethical approval: This article does not contain any studies with human participants or animals performed by any of the authors. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Preeclampsia (PE) is a form of hypertension that manifests in the later stages of pregnancy. Since Kaempferol (Ka) has remedial potential hence this research was conducted to examine its therapeutic effect on Preeclampsia rats by regulating Wingless-related integration site/β-catenin (Wnt/B-catenin) pathway. To achieve this, thirty-two SD female rats were randomly allocated into four groups: control, preeclampsia (PE, LPS, 1 mg/kg), preeclampsia with kaempferol (PE + Ka), and preeclampsia with Dickkopf - 1 (DKK-1) and kaempferol (PE + DKK-1 + Ka). Rats in the PE + Ka and PE + DKK-1 + Ka groups received intraperitoneal injections at 50 mg/kg/d of kaempferol, whereas the PE + DKK-1 + Ka group was administered with 60 µg/kg/d of recombinant rat DKK-1 protein, an inhibitor of the Wnt/β-catenin signaling pathway. Our findings revealed that systolic blood pressure (SBP) in the PE + Ka group was significantly reduced in comparison to PE group (P < 0.05). The urine albumin levels in the PE + Ka group decreased noticeably (P < 0.05), whereas serum concentrations of Tumor Necrosis Factor Alpha (TNF-α), Interleukin-1β (IL-1β), and Interleukin-6 (IL-6) in the PE + Ka group were reduced (P < 0.05) in comparison to PE group. Although PE + Ka group exhibited elevated levels of superoxide dismutases (SOD), glutathione (GSH), and catalase (CAT) in placental tissue relative to the PE group, whilst levels of malondialdehyde (MDA), alkaline phosphatase (ALP), serum glutamic-pyruvic transaminase (SGPT), and serum glutamic-oxaloacetic transaminase (SGOT) considerably decreased (P < 0.05). Comparatively mRNA levels of Wnt1 and β-catenin in the PE + Ka group were elevated, whereas mRNA level of DKK-1 was diminished (P < 0.05). Administration of DKK-1 counteracted kaempferol effects on these parameters in Preeclampsia rats (P < 0.05). Devastatingly, ovarian and kidney histomorphology in the PE group exhibited significant degenerative alterations, whereas kaempferol groups demonstrated normal histomorphology in comparison to the PE group. Conclusively, Kaempferol can significantly lower systolic blood pressure and urine albumin in PE female rats while mitigating excessive oxidative stress. The therapeutic efficacy of kaempferol on Preeclampsia may be mediatated via Wnt/β-catenin signaling pathway., Competing Interests: Declarations. Ethical approval: The animal study was approved by Animal Care and Use Committee of laboratory animals, Department of Obstetrics, Yantaishan Hospital, Shandong China (Approval number 2023011). The study was conducted in accordance with the local legislation and institutional requirements. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The female reproductive system dysfunction considerably affects the overall health of women and children on a global scale. Over the decade, the incidence of reproductive disorders has become a significant source of suffering for women. Infertility in women may be caused by a range of acquired and congenital abnormalities. Ovaries play a central role in the female reproductive function. Any defect in the normal functioning of these endocrine organs causes health issues and reproductive challenges extending beyond infertility, as the hormones interact with other tissues and biological processes in the body. The complex pathophysiology of ovarian disorders makes it a multifactorial disease. The key etiological factors associated with the diseases include genetic factors, hormonal imbalance, environmental and lifestyle factors, inflammatory conditions, oxidative stress, autoimmune diseases, metabolic factors, and age. Oxidative stress is a major contributor to disease development and progression affecting the oocyte quality, fertilization, embryo development, and implantation. The choice of treatment for ovarian disorders varies among individuals and has associated complications. Reproductive tissue engineering holds great promise for overcoming the challenges associated with the current therapeutic approach to tissue regeneration. Furthermore, incorporating nanotechnology into tissue engineering could offer an efficient treatment strategy. This review provides an overview of incorporating antioxidant nanomaterials for engineering ovarian tissue to address the disease recurrence and associated pathophysiology. Cerium oxide nanoparticles (CeO 2 NPs) are prioritized for evaluation primarily due to their antioxidant properties. In conclusion, the review explores the potential applications of CeO 2 NPs for effective and clinically significant ovarian tissue regeneration., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Adult females of the primitively eusocial wasp Polistes jokahamae can change caste in response to photoperiod length. Short-day photoperiod females store more lipids but have less developed ovaries for hibernation (becoming gynes), whereas long-day photoperiod females have fewer lipid stores but more developed ovaries under queenless conditions (becoming egg-laying workers). To explore the physiological mechanisms underlying photoperiod-related caste determination, analyses of gene expression levels in the brains of short- or long-day females were performed. Results suggest that short-day females may exhibit higher expression of genes involved in tryptophan metabolism, insulin signaling, and nutrition, including the digestion of sugars and lipids, and production of royal jelly proteins. Oral administration of tryptophan resulted in a positive correlation between tryptophan levels in the brain and lipid stores in the abdomen, suggesting that tryptophan promotes lipid storage in gynes. Long-day females showed enhanced expression of genes involved in tyramine/dopamine syntheses, epidermal growth factor receptor, insulin degradation, and oogenesis. In these females, tyramine administration increased the brain levels of tyramine and dopamine, and activated initial ovarian development under queenless conditions. Together, these results indicate that photoperiod-related caste determination in P. jokahamae may involve gene expression pathways similar to those involved in caste determination during the preimaginal stage in other Polistes species., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

This is the first study on how a substance with anti-progestogenic activity affects amphibian reproduction. Mifepristone, a synthetic anti-progestin used in abortion pills, was chosen as model compound. African clawed frog (Xenopus laevis) females were exposed to four mifepristone concentrations (0.7, 9, 120, and 1380 ng∙L -1 ) for 30 days. A control group was also included. Egg-laying during the experiment was significantly less at the highest concentration. At the experiment's end, mifepristone-exposed and control females were randomly mated with sexually mature males. Breeding rate for females exposed to 1380 ng∙L -1 mifepristone was only 50 %. Histology revealed no significant changes in gonads, thyroid, or liver. Females exposed to 1380 ng∙L -1 mifepristone had lower estradiol levels in plasma, lower mRNA expression of lh and fsh in brain-pituitary complex, and p450scc in ovaries. In liver, mRNA level of npr was significantly increased in females exposed to 120 ng∙L -1 mifepristone. mRNA expression of mpr, erβ, dio2, and dio3 were upregulated in animals exposed to 9 ng∙L -1 and 120 ng∙L -1 mifepristone, whereas vtg expression was significantly downregulated in females exposed to 1380 ng∙L -1 mifepristone. All these findings show that exposure to compounds with anti-progestogenic activity affects the hypothalamus-pituitary-gonad axis and decreases reproductive success., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Persistent organic pollutants (POPs) are easily accumulated in organisms due to its high lipophilicity. Bivalves are still one of the marine invertebrates with high species diversity despite experiencing with environmental pollution. However, studies investigating the adaptation mechanisms of bivalves towards POPs have been lacking. In this study, we chose benzo[a]pyrene (B[a]P), a typical POPs, to investigate the accumulation and excretion mechanism of clam Ruditapes philippinarum towards B[a]P. Laser confocal microscopy images of ovaries and testes confirmed significant colocalization between lipid droplets and B[a]P. However, B[a]P in the testes might mainly excrete with the excretory fluid. Furthermore, lipidomics results indicated that the upregulated TGs were TGs with 48-58 total carbon atoms and 6-12 double bonds in acyl side chains in ovaries, while the upregulated TGs mainly distributed in 48-58 total carbon atoms and 4-14 double bonds in acyl side chains in testes. In ovaries, gene expression of lipid metabolism and phospholipid metabolism basically presented an upward trend at Pro and Mat stages, while entirely performed a downregulation trend at Gro stage. Testes had the opposite regulation with that. Additionally, B[a]P prompted the excretion ability of female clams, but exhibited a complex effect in the male. This study provides a scientific basis for bivalve toxicology and healthy aquaculture, which is of great significance for marine ecological environment protection and aquatic product safety of China., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Anthropogenically environmental acidification impacts aquatic organisms, including teleosts, the largest group of vertebrates. Despite its significance, how teleosts allocate nutrient and energy among their organs to cope with acidic stress remains unclear. Our integrated analysis of physiological, metabolic, and gene expression data reveals that Japanese medaka (Oryzias latipes) mobilize energy resources among organs in response to acidic conditions. We found that the muscles lost carbohydrates and proteins and the liver accumulates all macronutrients in both sexes. Notably, female-specific energy mobilization between the liver and ovary were triggered by estrogen signaling, resulting in improved oocyte maturation and ovulation. Female produced more offspring under acidic stress. Furthermore, the offspring embryos exhibited smaller diameters and earlier hatching but demonstrated growth rates and acid tolerance. These metabolic changes suggest a trade-off in energy allocation by suppressing basal maintenance (33 % decrease in oxygen consumption) and growth (25 % decrease in muscle mass) but enhancing energy storage (159 % increase in liver mass in males and 127 % in females) and reproduction (165 % increase in ovary mass). This reallocation may improve medaka fitness and population sustainability in acidic environments. Further investigation into more species is needed to project the survival of aquatic animals in an acidified future., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Ompok bimaculatus is a commercially important food fish of Northeast India. For higher production of this species, the molecular mechanisms underlying its reproductive physiology is an important aspect to study. Vegetable oils are rich in ω3 and ω6 fatty acids and fat-soluble vitamins, that enhance reproductive performances. In the present study, O. bimaculatus were first fed with vegetable oil at 3, 5, 7 and 9% in the diet. The best treatment was determined by the analysis of relative fecundity, fertilisation rate, Gonadosomatic Index (GSI) and Hepatosomatic Index (HIS). Comparative transcriptomics analysis of gonadal tissues between control and best treatment was done using Illumina NextSeq500. The bioinformatics analysis by Trinity v2.11.0 and edgeR v3.32.1 identified 36 DEGs related to reproduction, those were later validated by qPCR analysis. The Differentially Expressed Genes (DEGs) revealed, regulation of Hypothalamic-pituitary-gonadal axis (HPG) axis through retinoid signalling pathway, IGF pathway and folliculogenesis pathway, including inhibitory effects of dietary vegetable oil on fish gonadal maturation., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Mandal et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

People are now exposed to higher levels of electromagnetic radiation (EMR) due to the widespread use of mobile phones in recent years. The possible effects of this exposure on human health are related to EMR. It has been suggested that exposure to EMR has serious effects on reproduction. The study aimed to investigate the impact of exposure to EMR (4.5 GB; 2600 MHz) emitted by mobile phones on rat ovaries. 18 adult female Wistar albino rats were used in the study, and the animals were divided into three groups (n = 6): control, stand-by, and dialing. For 8 weeks, the experimental groups were subjected to 4.5 GB EMR at 2600 MHz while on standby and making 10-min calls every hour. The rats in the control group received no exposure. Hematoxylin-eosin (H&E) staining of ovarian tissues was performed for histomorphological examinations. Additionally, immunoexpression of autophagy-related protein Beclin-1, apoptosis marker Caspase-3, ovarian reserve marker FSH, and oxidative stress marker iNOS were investigated in the rat ovaries. Microscopic examinations showed follicular degeneration in the ovaries of the rats in the stand-by and dialing groups. The immunoexpression of Beclin-1, Caspase-3, FSH, and iNOS was detected in granulosa cells and the corpus luteum in ovarian tissues obtained from the two EMR-exposed groups. There was a significant increase in the immunoexpression of Beclin-1 and Caspase-3 in the dialing group compared to the other two groups. Additionally, the iNOS and FSH expressions were increased in both EMR exposure groups compared to the control. Our results suggest that EMR exposure harms the ovaries, and autophagy and apoptosis are involved in this process., Competing Interests: Declarations. Competing interest: The authors declare no competing interests. Ethical approval: Animals were obtained from Kobay Animal Breeding and Experimental Research Laboratory in Ankara, Turkey. The animal ethics committee approved the experimental protocol (Approved number: 338) of Kobay Company (Turkey)., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The ovaries are key steroid hormone production sites in post-pubertal females. However, current research on steroidogenic enzymes, endogenous hormone concentrations and their effects on healthy ovarian function and malignant development is limited. Here, we discuss the importance of steroid enzymes in normal and malignant ovaries, alongside hormone concentrations, receptor expression and action. Key enzymes include STS, 3β-HSD2, HSD17B1, ARK1C3, and aromatase, which influence ovarian steroidal action. Both androgen and oestrogen action, via their facilitating enzyme, drives ovarian follicle activation, development and maturation in healthy ovarian tissue. In ovarian cancer, some data suggest STS and oestrogen receptor α may be linked to aggressive forms, while various oestrogen-responsive factors may be involved in ovarian cancer metastasis. In contrast, androgen receptor expression and action vary across ovarian cancer subtypes. For future studies investigating steroidogenesis and steroidal activity in ovarian cancer, it is necessary to differentiate between disease subtypes for a comprehensive understanding., (© 2024 The Author(s).)

Objective: The deleterious effects of caffeine consumption on reproductive functions of female Wistar rats were investigated in this study., Methods: In this experimental study, 35 female Wistar rats (180-200g) were divided into 7 groups: Control, II-IV received oral caffeine (10, 20, and 40mg/kg/day respectively) for 21 days. V-VII received similar caffeine doses for 21 days, followed by a 21-day withdrawal period. The ovaries, fallopian tubes, and uteri were assessed for levels of malondialdehyde (MDA), nitric oxide (NO), reduced glutathione (GSH), superoxide dismutase (SOD), and catalase activity using spectrophotometry. Serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), and estradiol levels were measured by ELISA. Organ histology was performed using microscopy. Statistical analysis employed ANOVA with significance at p<0.05., Results: Caffeine caused dose-dependent increases in MDA, NO, and catalase activity in the ovaries, fallopian tubes, and uteri which decreased upon withdrawal. GSH levels in the ovary and fallopian tubes decreased with caffeine intake but recovered during withdrawal. Caffeine reduced estradiol levels in a dose-dependent manner, its withdrawal led to reductions in serum LH at 20 and 40mg/kg/day and FSH at 40mg/kg/day. Histology revealed dose-dependent alterations in ovarian architecture with congested connective tissues. Caffeine caused sloughing of plicae in the muscularis of the fallopian tubes, degenerated epithelial layer in the uterus, and severe inflammation of the myometrial stroma cells that persisted during caffeine withdrawal., Conclusions: Caffeine consumption adversely impacted the female reproductive functions of rats, altering hormonal balance and organ structure which persisted even after caffeine withdrawal.

Background: N6-methyladenosine (m6A) modification is a widespread and reversible epigenetic alteration in eukaryotic mRNA, playing a pivotal role in various biological functions. Its significance in female reproductive development and associated diseases has recently become a focal point of research., Objective: This review aims to consolidate current knowledge of the role of m6A modification in female reproductive tissues, emphasizing its regulatory dynamics, functional significance, and implications in reproductive health and disease., Methods: A comprehensive analysis of recent studies focusing on m6A modification in ovarian development, oocyte maturation, embryo development, and the pathogenesis of reproductive diseases., Results: m6A modification exhibits dynamic regulation in female reproductive tissues, influencing key developmental stages and processes. It plays critical roles in ovarian development, oocyte maturation, and embryo development, underpinning essential aspects of reproductive health. m6A modification is intricately involved in the pathogenesis of several reproductive diseases, including polycystic ovary syndrome (PCOS), premature ovarian failure (POF), and endometriosis, offering insights into potential molecular mechanisms and therapeutic targets., Conclusion: The review highlights the crucial role of m6A modification in female reproductive development and related diseases. It underscores the need for further research to explore innovative diagnostic and therapeutic strategies for reproductive disorders, leveraging the insights gained from understanding m6A modification's impact on reproductive health., (© 2024 The Author(s). Immunity, Inflammation and Disease published by John Wiley & Sons Ltd.)

Epigenetic modifications regulate chromatin accessibility, gene expression, cell differentiation and tissue development. As epigenetic modifications can be inherited via mitotic and meiotic cell divisions, they enable a heritable memory of cell identity and function and can alter inherited characteristics in the next generation. Tight regulation of epigenetic information is critical for normal cell function and is often disrupted in diseases including cancer, metabolic, neurological and inherited congenital conditions. The ovary performs critical functions in female reproductive health and fertility, including oocyte and sex-hormone production. Oocytes undergo extensive epigenetic programming including the establishment of maternal genomic imprints, which are critical for offspring health and development. Epigenetic modifiers also regulate ovarian somatic cells, such as granulosa and theca cells which support oocytes and produce hormones. While ovarian dysfunction contributes to serious ovarian conditions such as primary ovarian insufficiency (POI), polycystic ovary syndrome (PCOS) and ovarian cancers, the roles of epigenetic modifications in the ovary and their contribution to ovarian dysfunction are not properly understood. Here we review recent advancements in understanding Polycomb proteins, important epigenetic modifiers that have emerging roles in ovarian development and maternal epigenetic inheritance. Polycomb group proteins (PcGs) contribute to the faithful establishment of epigenetic information in oocytes, a process essential for normal offspring development in mice. Emerging evidence also indicates that PcGs regulate ovarian function and female fertility. Understanding these and similar mechanisms will provide greater insight into the epigenetic regulation of ovarian and oocyte function, and how its disruption can impact reproductive health and maternal inheritance.

The role of anti-WNT ZNRF3 is central to determining gonadal fate: XY mice lacking functional ZNRF3 exhibit a highly variable gonadal sex reversal phenotype in the fetal period, characterised by appearance of ovarian tissue. To investigate this sex reversal further, we used single-cell RNA-seq to examine the transcriptomes of XY Znrf3-deficient gonads during the mouse sex-determining period. Analyses of cell trajectories in mutant gonads reveal the failure of pre-supporting cells to commit to the Sertoli cell fate, XY granulosa cell development, unstable commitment in those cells that reach the Sertoli path and enhanced contribution to a supporting-like cell fate. By developing a machine learning-based score for transcriptomic similarity to Sertoli and granulosa, we show pervasive disruption to acquisition of testicular cell fate in the mutant supporting cell lineage, with large numbers of cells co-expressing pro-Sertoli and pro-granulosa markers. These data reveal that loss of Znrf3 results in transcriptomic and cellular heterogeneity, with shifts in cellular sex identity that undermine a simple binary model in which mutant supporting cell precursors achieve either Sertoli or granulosa cell differentiation., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)

Lead (Pb) is a neurotoxicant with early life exposure linked to long-term health effects. Piwi-interacting RNAs (piRNAs) are small non-coding RNAs that associate with PIWIL proteins to induce DNA methylation. It remains unknown whether Pb exposure influences piRNA expression. This study evaluated how perinatal Pb exposure (32 ppm in drinking water) impacts piRNA expression in adult mice and assessed piRNA dysregulation as a potential mechanism for Pb-induced toxicity. Pb exposure effects on piRNA expression and associated gene repression in the germline (testis/ovary) and soma (liver and brain) were evaluated. Small RNA sequencing was used to determine differentially expressed piRNAs, RT-qPCR to examine piRNA target expression, and whole genome bisulfite sequencing to evaluate target DNA methylation status. Three piRNAs (mmpiR-1500602, mmpiR-0201406, and mmpiR-0200026) were significant after multiple testing correction (all downregulated in the male Pb-exposed brain in comparison to control; FDR < 0.05). Within piOxiDB, TAO Kinase 3 was identified as a downstream mRNA target for one of the three Pb-sensitive piRNA. The Pb-exposed male brain exhibited increased Taok3 expression ( p  < 0.05) and decreased DNA methylation (FDR < 0.01). The results demonstrate that perinatal Pb exposure stably influences longitudinal piRNA expression in a tissue- and sex-specific manner, potentially via DNA methylation-directed mechanisms.

Cadmium (Cd) is prevalent in aquatic ecosystems and accumulates in various tissues of aquatic organisms, leading to severe biological toxicity. Selenium (Se) is recognized for mitigating heavy metal toxicity, though its protective effects against Cd in aquatic crustaceans remain underexplored. This study, therefore, assessed the effects of dietary Cd (15 mg/kg) exposure and Se (6 mg/kg) supplementation on the hepatopancreas and ovaries of female crayfish to uncover the mechanisms of Cd toxicity and the protective role of Se. The results showed that Cd accumulation in the hepatopancreas caused a reduced hepatopancreas index (HPI), decreased protein content, histopathological damage, and oxidative stress, while Se supplementation reduced Cd levels, mitigated damage, and restored tissue integrity and antioxidant defenses. Transcriptomic analysis further revealed significant alterations in gene expression related to detoxification, lipid metabolism, and energy production in response to Cd exposure, which were partially or fully restored by Se supplementation. Additionally, Se alleviated Cd-induced inhibition of ovarian development, as evidenced by improved ovary index, enhanced oocyte development, and normalization of essential trace element levels. Mechanistically, Se restored the Cd-disrupted vitellogenin (Vtg) synthesis in the hepatopancreas via regulating the mRNA expression of hsp70 and genes related to the molt-inhibiting hormone (MIH) (mih, rxr, and ecr). Overall, these findings indicate that Se supplementation mitigated Cd-induced hepatopancreatic dysfunction, restored Vtg synthesis, and consequently counteracted the inhibition of ovarian development in adult female crayfish., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Perimenopausal syndrome is a significant issue that disturbs women's metabolism, mood and quality of life. Apigenin (4',5,7-trihydroxyflavone) is a natural flavonoid that exhibits antioxidant, anti-inflammatory and anticancer effects. The present study aims to investigate the effect of apigenin on perimenopausal syndrome by combining bioinformatics analysis with in vivo experiments. The mouse model with perimenopausal syndrome was established using 4-vinylcyclohexene diepoxide (VCD) treatment. Apigenin alleviated VCD-induced disorder of estrous cycle and shrinkage of ovarian tissue. The reduction of anti-Muller hormone and the increase of follicle stimulation hormone and luteinizing hormone triggered by VCD were reversed by apigenin in a dose-dependent manner. Apigenin suppressed the VCD-induced decrease of primordial, primary, secondary and antral follicle number in ovarian tissue. Oxidative stress in ovarian tissue was activated by VCD treatment through increasing the reactive oxygen species production. High concentration of apigenin significantly reversed the alteration induced by VCD. Apigenin alleviated VCD-induced cell apoptosis through regulating Bax, Bcl-2, cleaved PARP1 and caspase-3. Furthermore, the phosphorylation of AKT and FOXO3a was inhibited by VCD and activated by apigenin in a dose-dependent manner. Collectively, apigenin effectively mitigates the ovarian dysfunction through suppressing oxidative stress and apoptosis via the AKT/FOXO3a signaling pathway., (© 2024 John Wiley & Sons Ltd.)

Aims: Present study demonstrates dose and time dependent effects of NiONPs (<30 nm) on the ovaries of Wistar rat., Methods: Female rats were gavaged NiONPs or NiOMPs (5 mg/kg b.w.) for 24 h, 15 days and 30 days, euthanized and ovaries thus removed were analyzed for nickel bioaconcentration and processed for scanning electron microscopy. Serum samples were analyzed to compare the effects of nickel nano & microparticles on progesterone and estradiol values., Results: Results confirmed the bioaccumulation of Ni in ovarian tissue. Its concentration was higher in NiONPs treated rats than NiOMPs treated rats. Progesterone level increased whereas estradiol values decreased in NiONPs and NiOMPs treated rats. SEM results also exhibited dose dependent effects on the morphology of corpoluteal complex. The structural changes varied from formation of blebs to distorted microvilli and germinal epithelium., Conclusion: It is hypothesized that NiONPs/NiOMPs are biodegraded into smaller fragments that conjugate with amino acids and or alter downstream signaling pathways, generate ROS and modulate protein structure activity relationships. Finally, these processes manifest into morphological alterations in the ovary. Biopersistence of nickel in female reproductive system may compromise with fertility and reproductive performance of exposed population., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Microtubule (MT) regulation is essential for oocyte development. In Drosophila , MT stability, polarity, abundance, and orientation undergo dynamic changes across developmental stages. In our effort to identify novel microtubule-associated proteins that regulate MTs in the Drosophila ovary, we identified a previously uncharacterized gene, CG18190, which encodes a novel MT end-binding (EB) protein, which we propose to name EB-SUN. We show that EB-SUN colocalizes with EB1 at growing MT plus-ends in Drosophila S2 cells. Tissue-specific and developmental expression profiles from Paralog Explorer reveal that EB-SUN is predominantly expressed in the ovary and early embryos, while EB1 is ubiquitously expressed. Furthermore, as early as oocyte determination, EB-SUN comets are highly concentrated in oocytes during oogenesis. EB-SUN knockout (KO) results in decreased MT density at the onset of mid-oogenesis (stage 7) and delays oocyte growth during late mid-oogenesis (stage 9). Combining EB-SUN KO with EB1 knockdown (KD) in germ cells significantly further reduces MT density at stage 7. Hatching assays of single protein depletion reveal distinct roles for EB-SUN and EB1 in early embryogenesis, likely due to differences in their expression and binding partners. Notably, all eggs from EB-SUN KO/EB1 KD females fail to hatch, suggesting partial redundancy between these proteins., Competing Interests: Conflicts of interest: The authors declare no financial conflict of interest.

The present investigation aimed to evaluate the therapeutic potential of exopolysaccharides (EPSs) derived from endophytic fungi against arsenic [As(III)]-mediated metabolic and reproductive ailments. Two endophytic fungi, Diaporthe arengae (CleR1) and Fusarium proliferatum (CleR3), were isolated from Clerodendrum infortunatum (Cle), and used for the extraction of two types of EPSs. GC-MS analysis confirmed the presence of hydroxymethyl furfural (HMF) and α-d-glucopyranose in the EPS1 (CleR1) and EPS2 (CleR3), respectively. FTIR analysis revealed the potential As(III)-chelation properties of both EPSs. EPS1 and EPS2 significantly mitigated As(III)-induced oxidative stress and lipid peroxidation by restoring the activities of antioxidative enzymes. EPSs successfully corrected the gonadotropin imbalance and steroidogenic alterations. The downregulation of proinflammatory (NF-κB and TNF-α) and proapoptotic (BAX) mediators and the upregulation of antiapoptotic (Bcl-2) markers were also detected following the treatment with EPSs. Histomorphological restoration of reproductive and metabolic organs was also observed in both the EPS groups. Moreover, the As(III)-induced increase in the immunoreactivity of the androgen receptor (AR) was successfully reversed by these EPSs. Molecular docking predicted that HMF and α-d-glucopyranose of EPS1 and EPS2 interact with the active site of AR by limiting its activity. Hence, EPS could be effective for developing new therapeutic strategies for managing As(III) toxicity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The kidding traits of goats are an important index of production. However, the molecular regulatory mechanisms of kidding traits in goats have not been fully elucidated. This study aimed to investigate the molecular regulatory network of kidding traits in goats. Multi-omics revealed the enrichment of 10 signaling pathways, with fatty acid biosynthesis, biosynthesis of unsaturated fatty acids, and steroid hormone biosynthesis pathways being closely related to reproduction. Interestingly, the key rate-limiting enzymes, fatty acid synthase (FASN), stearoyl-CoA desaturase 5 (SCD5), fatty acid desaturase 1 (FADS1), 3β-hydroxysteroid dehydrogenase/isomerase (3BHSD), and steroidogenic acute regulatory protein (STAR) enriched in these pathways regulate changes in reproduction-related metabolites. In interference experiments, it was observed that suppressing these key rate-limiting enzymes inhibited the expression of CYP19A1, ESR2, and FSHR. Furthermore, interference inhibited granulosa cell proliferation, caused cell cycle arrest, and promoted apoptosis. Thus, these results suggest that the specific markers of nanny goats with multiple kids are the key rate-limiting enzymes FASN, SCD5, FADS1, 3BHSD, and STAR. These findings may greatly enhance the understanding of regulatory mechanisms that govern goat parturition., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Molybdenum (Mo) plays a crucial role in regulating normal physiological function. However, its potential effect on female infertility has received little attention., Methods: In this study, we explored the potential molecular mechanisms of Mo's action on mouse ovaries and oocytes by establishing a busulfan-induced infertility model. Adult female Kunming mice were randomly divided into three groups: control, +busulfan, and +busulfan+Mo. After 30 days of busulfan treatment [Myleran, 20 mg/kg body weight ip], mice in the busulfan+Mo group were provided with 7.5 mg/L Mo per day in drinking water for an additional 42 days. On day 72, we examined the morphology of the oocytes and ovarian tissue after H&E staining, measured the concentrations of serum hormones by ELISA, and detected Bax, Bcl-2, caspase-3 and caspase-9 by immunohistochemical staining and western immunoblotting. We also assessed the oxidative stress in cells by measuring the activity of the antioxidant enzyme, SOD, the concentrations of MDA and LDH, and the percentage of apoptotic cells using kits. The number of litters born was counted after mating with male mice, and the organ coefficients were calculated after weighing on an analytic balance., Results: Results showed that Mo treatment restored female reproductive hormone levels to near normal. Mo also significantly inhibited the mitochondrial stress-induced expression of apoptotic proteins., Conclusion: Our findings demonstrate that Mo treatment at a dose of 7.5 mg/L can ameliorate busulfan-induced infertility in female mice. These data may provide a reference for the development of treatments for female infertility., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier GmbH.)

Rbpms2, an RNA-binding protein with multiple splicing (Rbpms), can interact with RNAs to involve oocyte development, thereby influencing female sex differentiation in vertebrates. Here, two splicing variants of the Rbpms2 gene from Japanese flounder (Paralichthys olivaceus) were identified, namely Rbpms2.1 and Rbpms2.2. The two variants exhibited 98.22 % amino acid homology, both featuring an RNA recognition motif (RRM) domain spanning positions 98-170 amino acids. They were relatively conserved throughout phylogenetic evolution. Differently, the C-terminal region of the Rbpms2.1 contains five additional sequential amino acids (-VRDQP-) compared to Rbpms2.2. The real-time qPCR results demonstrated that Rbpms2.1 and Rbpms2.2 had relatively abundant expression in the gonads of adult Japanese flounder, with higher expression levels in the ovary compared to the testis (P < 0.05). In situ hybridization results showed strong positive expression of Rbpms2 mRNA in oocytes at stages I-III during the V stage of ovarian development. In the testis atstage IV, the expression of Rbpms2 mRNA was mainly concentrated on primary spermatocytes. Importantly, Rbpms2 binding sites were found in the 3'UTR, 5'UTR, and ORF regions of the sex-related genes including dmrt1, sox9, amh, foxl2, and wnt4. siRNA interference and overexpression analysis of Rbpms2.1 and Rbpms2.2 in primary cells of the ovary and testis showed that Rbpms2 can repress the expression of male-related genes (dmrt1, sox9, and amh) and significantly promote the expression of female-related genes (foxl2 and wnt4). Our results revealed that Rbpms2 may play a critical role by targeting the sex-related genes in the gonad development of Japanese flounder., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

Prior to the pubertal molt and mating, the ovarian development of the mud crab Scylla paramamosain was primarily at stage II. However, immediately after mating, female crabs initiate vitellogenesis, and their ovaries quickly develop. The aim of this study was to identify differentially expressed genes associated with ovarian development in the mud crab before and after mating, in order to elucidate the influence of mating on ovarian development using comparative transcriptomics. The KEGG pathway analysis results indicated that ribosome and ribosome-related pathways were highly associated with ovarian development at stage II across both transcriptomes, likely to support the subsequent vitellogenesis by providing the necessary materials. Additionally, the neurodegeneration, MAPK, cAMP and PLD pathways were active in regulating oogonia differentiation, oocyte proliferation and vitellogenesis after mating. Meanwhile, certain intra-ovarian factors, such as the cell cycle-related genes cyclin B and APC, the forkhead box family genes Foxl2 and slp1, the SOX family gene SOX5-like, the hormone-related genes SULT1E1 and Eip74EF-like, the growth factor-related genes VEGFD-like and CUBE1-like, as well as HPS10 and tra1-like, have essential functions in regulating ovarian development after mating. Furthermore, the receptors of extra-ovarian hormones, such as RPCHR, HR4, and ILR1, as well as the neurotransmitter receptor 5-HTR4, were involved in ovarian development. It is believed that ovarian development is controlled by the coordinated action of both intrinsic and extrinsic endocrine factors, and these factors are influenced by mating. Finally, the analysis of epigenic modification-related genes, transcription factors, and target genes revealed the regulation of gene expression. Our study indicated that, those genes work in a coordinated manner to regulate the complex processes of follicle cell development, oogonia differentiation, oocyte proliferation, and vitellogenesis during ovarian development., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Gonadal sex determination (GSD) is a complex but poorly understood process in the early stages of embryonic development. This process determines whether the bipotential gonadal primordium (BGP) will differentiate into testes or ovaries through the activation of genetic factors related to Sertoli or Granulosa cells, respectively. The study of this developmental process remains challenging due to experimental limitations and the complexity of the underlying genetic interactions. Boolean Networks (BNs) are binary networks that simulate genetic behavior and are commonly used for modeling gene regulatory networks (GRNs) due to their simplicity when dealing with a high number of gene interactions. Reported BNs usually use a synchronous (parallel) update scheme, which means that all the nodes (representing genes) update their values simultaneously. However, the use of this update scheme has been criticized because it cannot represent biological systems that are highly regulated at a temporal scale. Asynchronous and block-sequential updating schemes appear as an alternative to tackle this issue. In the first case, the updating scheme follows a random behavior while, in the second case, the set of network nodes is partitioned into blocks such that the nodes within a block are updated simultaneously, and the blocks are considered in a specific order sequence. To assess the impact of different updating approaches in a GRN associated to GSD we first made a node reduction without losing the main dynamics of the original network which are related to the formation of testes and ovaries. Then, we tested the effect of perturbations given by the inactivation of genes on the network attractors, specifically the SRY and WNT4 genes, since the former is only present in the Y chromosome and the latter is of importance in early embryo development. We found that both genes were crucial, but WNT4 alone showed a higher percentage of attractors towards a phenotype than the SRY alone. Finally, we found that using asynchronous and block-sequential updating schemes, the attraction basins - i.e., the set of configurations that reach an attractor - remain with similar percentages to those of the original network, which supports the robustness of the model., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Infertility is a worldwide reproductive health problem influenced by the embryo implantation efficiency. We previously revealed that dietary chenodeoxycholic acid (CDCA) positively influence the early embryo implantation. But how CDCA regulate embryo implantation is largely unexplored. Herein, we investigated the mechanism behind CDCA's regulation on embryo implantation in rats. Results showed that CDCA promoted uterine receptivity, leading to increased number of implantation sites. Mechanistically, CDCA reshaped maternal amino acid metabolism and enhanced serum progesterone levels. CDCA enhanced ovarian progesterone synthesis by improving steroidogenesis-related protein (StAR and CYP11A1) expression via Takeda G-protein-coupled receptor 5. Elevated progesterone exaggerated uterine progesterone but weakened the estradiol signaling in the CDCA group, contributing to better uterine receptive for embryo implantation. Additionally, elevated transcription repressor Stat5b induced the down-regulation of progesterone-metabolizing enzyme 20-hydroxysteroid dehydrogenase 20α-HSD, complementally explained uterine progesterone signaling enhancement. Overall, our data revealed that CDCA drove ovarian steroidogenesis to improve embryo implantation through enhancing uterine receptivity via progesterone receptor pathway in rats. Therefore, CDCA diet may be a potential favorable nutritional strategy for infertility and pregnancy management., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

This study aimed to elucidate the role of TP53-induced glycolysis and apoptosis regulator (TIGAR) in polycystic ovary syndrome (PCOS). A rat model PCOS was constructed by subcutaneous injection with dehydroepiandrosterone (DHEA). Follicular atresia and reduced granular cells (GCs) in ovaries suggested successful modeling. The low expression of TIGAR was observed in ovarian tissue of PCOS rat. To explore the role of TIGAR in PCOS, lentivirus carrying the TIGAR were used to up-regulate TIGAR expression. TIGAR overexpression reduced the DHEA-induced increase of ovarian weight, the levels of estradiol (E2), and the ratio of luteinizing hormone/follicle-stimulating hormone (LH/FSH) in the serum, as well as improved the morphology of the follicle, especially increased the thickness of the GC layer, which attributed to the inhibition of apoptosis by TIGAR. In addition, high expression of TIGAR inhibited oxidative stress in ovaries of PCOS rat, as evidenced by decreased level of malondialdehyde (MDA), and reactive oxygen species (ROS), and enhanced activity of glutathione peroxidase (GPX) and superoxide dismutase (SOD). Mechanically, Nrf2/OH-1 signal pathway was activated by TIGAR. The effect of TIGAR on PCOS were verified in the primary rat GCs treated with dihydrotestosterone, but also the rescue experiment was performed. Downregulation of Nrf2 reversed the effects of TIGAR, indicating that TIGAR suppressed oxidative stress and GC apoptosis by activating Nrf2/OH-1 pathway in PCOS. Finally, non-targeted metabolomics revealed that TIGAR might affect the energy metabolic pathway, thereby altering the metabolic profile of primary rat GCs. This study provided new insights into the prevention and treatment of PCOS., Competing Interests: Declaration of competing interest The authors disclosed no relevant financial or non-financial interests., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Bisphenol A (BPA) belongs to the endocrine disruptor chemicals (EDCs) causing various reproductive disorders in females. We analysed the toxic effects of BPA in the uterus and ovaries. The BPA was administered orally with the repeated low dose (LD, 1 mg/kg) and high dose (HD, 5 mg/kg) of body weight on alternate days for 4 months via oral gavage to Swiss mice. BPA administration decreases body weight, ovarian weight and size at LD, but increases ovarian weight and size at HD. The uterus weight, length, and diameter were increased in both the treated groups. The histopathological data show decreased ovarian follicle size, epithelial hyperplasia, and lymphocytic infiltration in the ovary. The BPA-treated uterus shows increased vascularization, atrophied endometrium and myometrium, and endometrial hyperplasia (EH) with aberrant glandular growth. The cancer stem cells (CSCs) in the ovaries were identified based on staining with anti-mouse CD44 and anti-mouse CD133 antibodies and analysed by flow cytometry. Three different populations of ovarian CSCs: CD44 + CD133 - , CD44 + CD133 + , and CD44-CD133+, can be recognised based on the intensity of these receptors. CD44 + CD133 - and CD44 + CD133 + cell percentages were increased in BPA-treated groups. CD44 - CD133 + were increased in LD but decreased in HD. The BPA administration also induces ROS production, which decreases the expression of antioxidant genes Superoxide dismutase 1 (SOD1), Superoxide dismutase 2 (SOD2), Catalase (CAT), Glutathione peroxidase 1 (GPX1), and Forkhead box O3 (FOXO3) in ovarian cells. In conclusion, BPA exposure induced an inflammatory response, increased CSC proportions, induced ROS, and decreased antioxidant responses in the ovaries., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Nitin Bhardwaj reports financial support was provided by Indian Council of Medical Research. Nitin Bhardwaj reports financial support was provided by University Grant commission. Mohammad Z. Ahmed reports financial support was provided by King Soud University. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024 Elsevier Inc. All rights reserved.)

Procambarus clarkii (Girard, 1852) has important economic value in China and internationally. In this research, the comparative transcriptome analysis was used to reveal molecular mechanisms of influences of photoperiod and light intensity on ovarian development in P. clarkii for the first time. Some genes (such as laminin, collagen, integrin beta, catenin) and pathways (including TGF-beta signaling pathway, focal adhesion, ECM-receptor interaction) associated with ovarian development and oocyte maturation were significantly upregulated. Some genes related to circadian clock (such as CLK, PER) were identified in this research. The results indicated that when light intensity or photoperiod increased, P. clarkii could up-regulate the expression levels of the laminin and collagen, thereby synthesizing related proteins, promoting meiosis of the oocytes, thus increasing the number of oocytes in the ovary. At the same time, P. clarkii could up-regulate the expression levels of integrin beta, integrin alpha 6, and diacylglycerol to synthesize related proteins, thereby promoting the formation of proteins and fats such as triglycerides, these proteins and fats can provide material basis for maturation and development of oocytes, resulting in oocyte maturation and ovarian development. P. clarkii could synthesize related proteins by upregulating expression levels of genes (such as catenin), these proteins or hormones can adhere to other actins (such as integrins), thereby stabilizing the morphology of the oocytes and ensuring normal development. Meantime, the increase in light intensity or photoperiod could cause release GSH and VTG, resulting in oocytes development and maturation. The data in this research can reveal molecular mechanisms of impacts of photoperiod and light intensity on oocyte maturation and ovarian development in P. clarkii, can offer crucial genomic data for studying developmental mechanisms of ovary and oocyte in crustacean., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

Ovarian tissue autografting is a valuable clinical option to help restore fertility in women with cancer. However, many follicles are lost due to ischemia-reperfusion (IR) injury, which depletes follicles after grafting. We aimed to investigate the effect of vitamin C, an antioxidant with anti-apoptotic and anti-inflammatory properties, on improving the structure and function of autografted ovaries in mice. Thirty-six female NMRI mice (4-5 weeks old) were divided into three groups of 12: control (no grafting), autograft + vitamin C (50 mg/kg/day intraperitoneally), and autograft + saline (100 µl/day/animal, intraperitoneally). After the ovarian autografting and before the start of the experiment, each group was further divided into 7-day and 28-day subgroups. Seven days after ovary autografting, serum levels of malondialdehyde (MDA), total antioxidant capacity (TAC), and inflammatory factors were measured. On day 28, ovarian histology, DNA fragmentation, and estradiol and progesterone levels were assessed. Results were analyzed using one-way ANOVA and Tukey's test, with significance set at p<0.05. In the autograft + vitamin C group, there were significant increases in the mean total volume of the ovary, cortex (p<0.05), medulla, number of follicles, and levels of IL-10, progesterone, estradiol, and TAC (p<0.001), compared to the autograft group. Conversely, the rate of apoptosis and serum levels of MDA, IL-6, and TNF-α were notably reduced in the autograft + vitamin C group (p<0.001). These results suggest that vitamin C can significantly enhance the recovery of autografted ovaries through its antioxidant, anti-inflammatory, and anti-apoptotic effects., Competing Interests: Declaration of Competing Interest The authors have no conflict of interest to disclose., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Despite the negative environmental and biologic effects, organophosphates have currently been widely used. We aimed to examine the possible negative effects of diazinon, a type of organophosphate, on rat ovarian tissue. Wistar Albino rats were divided into four groups. No treatment was given to control, olive oil was applied to sham group. Experimental groups were injected intraperitoneally with 30 and 60 mg/kg/day diazinon, respectively. 24 h later, ovarian tissues were extracted, preparated, examined via light and electron microscope. In the experimental groups granulosa and corpus luteum showed degenerative changes. Dilatation of endoplasmic reticulum cisterns and morphological alterations of mitochondria in granulosa cells were detected utrastructurally. Also, accumulation of lipid droplets and autophagic vacuoles was observed in cells of corpus luteum. A statistically significant dose-dependent decrease in superoxide dismutase and catalase reactivity and a statistically significant increase in caspase-3 expression in cells of atretic follicles and corpus luteum were observed. Results show that exposure to a single dose of diazinon may disrupt antioxidant system, trigger atresia in follicles and negatively effect corpus luteum functions. It was concluded that studies applying possible antioxidant treatments should be carried out to reduce and prevent the negative effects of diazinon on the reproductive system., Competing Interests: Declarations Competing interests The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Globally, approximately 6-20 % of women who are of reproductive age suffer from polycystic ovary syndrome (PCOS), with environmental factors believed to be significant contributors. Di-2-ethylhexyl phthalate (DEHP) is known to be an endocrine disruptor, and is also suspected of being associated with the occurrence of PCOS, but in vivo studies to verify this association are lacking. In this study, female SD rats were exposed to DEHP at levels of 0.1, 1.0, and 10 mg/kg/d, which are comparable to daily human exposure, to explore its potential role in the development of PCOS. The findings indicated that DEHP exposure reduced ovarian and uterine coefficients, decreased accumulation of primordial follicles, increased the prevalence of atretic and cystic follicles and fibrosis in ovarian tissues, altered serum hormone levels, elevated blood glucose levels and insulin resistance, disrupted the endocrine system and resulted in significant oxidative damage in the ovarian tissues. These results imply that DEHP exposure may cause lesions resembling PCOS to develop. By analyzing the differential expression of the proteome, and using GO and KEGG enrichment analyses, we found they were mainly enriched in the metabolic pathway and in the PPAR signaling pathway. We confirmed that activation of the PPARγ signaling pathway caused by DEHP exposure, is related to the emergence of PCOS-like lesions. This research provides direct in vivo experimental evidence for the association between DEHP exposure and PCOS., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024 Elsevier B.V. All rights reserved.)

Methyltransferase-like 3 (METTL3) is extensively reported to be involved in organ fibrosis. Ovarian fibrosis is a main characteristic of polycystic ovary syndrome (PCOS). However, the reaction mechanism of METTL3 in PCOS is poorly investigated. This paper was intended to reveal the role and the mechanism of METTL3 in PCOS. Animal and cell models of PCOS were induced by dehydroepiandrosterone (DHEA). H&E staining was performed to detect the pathological alterations in ovary tissues. Masson staining, immunofluorescence, along with western blot measured fibrosis both in vitro and in vivo. To evaluate estrous cycle, vaginal smear was performed. Lipid peroxidation and ferroptosis were evaluated by MDA assay kits, GSH assay kits, immunohistochemistry, Prussian blue staining and western blot. qRT-PCR and western blot were adopted to estimate METTL3 and GPX4 expression. The m6A and hormone secretion levels were respectively assessed by m6A RNA Methylation Quantitative Kit and corresponding kits. The interaction between METTL3 and GPX4 was testified by immunoprecipitation. The fibrosis and ferroptosis were aggravated and m6A and METTL3 expression were increased in ovarian tissues of DHEA-induced PCOS mice. METTL3 silencing alleviated pathological changes, affected hormone secretion level, and repressed fibrosis, lipid peroxidation and ferroptosis in the ovarian tissues of PCOS mice. In vitro, DHEA stimulation increased m6A and METTL3 expression and induced ferroptosis and fibrosis. METTL3 knockdown promoted GPX4 expression in DHEA-induced granulosa cells by m6A modification and restrained DHEA-induced fibrosis, lipid peroxidation and ferroptosis in granulosa cells via elevating GPX4. METTL3 silence inhibited ovarian fibrosis in PCOS, which was mediated through suppressing ferroptosis by upregulating GPX4 in m6A-dependent manner., Competing Interests: Declarations Competing interests The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Bisphenol-A (BPA) is a widely used chemical that can harm the human body, including the reproductive system. BPA accumulates in the body and is found in 95 % of individuals due to everyday exposure through food, water, and skin absorption. BPA can impair female fertility by interfering with ovarian folliculogenesis, inhibiting follicular growth, and inducing atresia, leading to polycystic ovary syndrome (PCOS). PCOS is a prevalent endocrine disorder that affects many reproductive-aged women. While current treatments can help manage symptoms, they do not entirely prevent complications. Luteolin, a natural flavonoid with medicinal properties, is commonly used to treat metabolic and inflammatory disorders. Therefore, we evaluated Luteolin's properties against PCOS in Network pharmacology and molecular docking studies; further, the antioxidant and anti-inflammatory properties in protecting the Chinese Hamster ovarian (CHO) cells from Reactive Oxygen Species, cellular damage, and negative mitochondrial membrane potential were evaluated. Additionally, an in-vivo PCOS-like model was developed using zebrafish, and the localization of Luteolin was identified using fluorescein isothiocyanate (FITC). Luteolin protected the CHO cells from cellular damage, ROS, and negative mitochondrial membrane potential. Luteolin alleviated the total SOD levels in the Zebrafish ovary, induced follicular maturation, and altered the key genes in ovarian proliferation and pro-inflammatory cytokines TNF-α and IL-1β expression. Natural Phyto-oxidants such as Luteolin may protect follicular development and early PCOS in adult zebrafish to prevent oxidative stress and inflammation. This study suggests using Luteolin as a phytomedicine to alleviate ovarian function decline., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Polycystic ovary syndrome (PCOS) is a hormonal disorder and significantly affects reproductive and metabolic function. Bromodomain-containing protein 4 (BRD4) is reported to promote ovarian fibrosis in PCOS. The present work was conducted to investigate the detailed role of BRD4 and the corresponding functional mechanism in PCOS. Functional experiments including CCK-8 method, EDU staining and TUNEL staining were used to detect the key cellular processes. Western blot examined the expression of BRD4, apoptosis- and endoplasmic reticulum stress (ERS)-associated proteins. HDOCK server predicted the binding of BRD4 with Glucose-Regulated Protein 78 (GRP78), which was validated by Co-IP assay. BRD4 expression was increased and ERS was activated in dehydroepiandrosterone (DHEA)-induced KGN cells. Inhibition of BRD4 improved the viability whereas it inhibited the apoptosis and ERS of KGN cells induced by DHEA. In addition, BRD4 bound to GRP78. GRP78 elevation or ERS activator tunicamycin (TM) partly abolished the impacts of BRD4 silencing on the ERS, proliferation and apoptosis in DHEA-treated KGN cells. Anyway, knockdown of BRD4 may reduce DHEA-induced ovarian granular cell damage in PCOS via inactivating GRP78-mediated ERS., (Copyright © 2024. Published by Elsevier Ltd.)

Fisetin, a polyphenolic flavonoid, exhibits numerous pharmacological activities against metabolic syndromes. The present research aims to explore the therapeutic efficacy of fisetin in experimental polycystic ovary syndrome (PCOS). Female Sprague-Dawley rats were administered mifepristone (20 mg/kg/day) to induce PCOS. PCOS rats were treated with fisetin (20 mg/kg and 40 mg/kg) and further compared with metformin HCl, the conventional drug for PCOS. The mechanism of fisetin was explored using dorsomorphin (an AMPK inhibitor). Then, rats were sacrificed for further analysis of biochemical and histological parameters. PCOS rats exhibited irregular estrous cycles, increased serum testosterone (4.72 ± 0.139 ng/ml), estradiol (750.2 ± 16.56 pg/ml), LH (30.33 ± 1.563 mIU/ml), HOMA-IR (1.115 ± 0.049), TNF-α (86.59 ± 3.93 pg/ml), IL-6 (55.34 ± 4.432 pg/ml), and TBARS (3.867 ± 0.193 µmol/mg) along with declined progesterone (11.67 ± 1.54 ng/ml), FSH (13.33 ± 1.256 mIU/ml), GSH (33.47 ± 1.348 µmol/mg) levels, and SOD (2.163 ± 0.298 U/mg) activity as compared to normal control group. Fisetin high dose significantly lowers testosterone (3.014 ± 0.234 ng/ml), estradiol (533.7 ± 15.39 pg/ml), LH (16.67 ± 1.62 mIU/ml), HOMA-IR (0.339 ± 0.20), TNF-α (46.02 ± 2.66 pg/ml), IL-6 (31.77 ± 3.47 pg/ml), and TBARS (1.747 ± 0.185 µmol/mg) and enhances progesterone (33.17 ± 1.447 ng/ml), FSH (27.17 ± 1.42 mIU/ml), GSH (60.35 ± 1.1.102 µmol/mg) levels, and SOD (4.513 ± 0.607 U/mg) activity. The histology of ovarian tissues shows a significant increase in cystic follicles in PCOS rats compared with the normal control group. These alterations were attenuated with fisetin treatment. Administration of dorsomorphin with fisetin can reverse the beneficial effects of fisetin in PCOS rats. Altogether, these present findings highlight the potential of fisetin as a promising therapeutic intervention for the management of PCOS by modulating AMPK/SIRT1 signaling in rats., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Premature ovarian insufficiency (POI) critically affects female reproductive health, with obesity being a significant and recognized risk factor. Interleukin-27 (IL-27), known for its role in immune modulation and inflammation, has garnered attention in metabolic syndrome research. Nonetheless, the role of these immunometabolic factors on the initiation of POI remains to be unraveled. Our investigation delves into the influence of impaired IL-27 signaling on POI induction, particularly under the challenge of a high-fat diet (HFD). We analyzed patients' serum profiles and established a correlation of increased serum triglycerides with decreased IL-27 levels in POI cases. Experiments on C57BL/6 mice lacking the IL-27 receptor alpha (Il27ra -/- ) revealed that when subjected to HFD, these mice developed hallmark POI symptoms. This includes escalated lipid deposition in both liver and ovarian tissues, increased ovarian macrophages cellular aging, and diminished follicle count, all pointing to compromised ovarian function. These findings unveil a novel pathway wherein impaired IL-27 signaling potentiates the onset of POI in the presence of HFD. Understanding the intricate interplay between IL-27, metabolic alterations, and immune dysregulation sheds light on potential therapeutic avenues for managing POI, offering hope for improved reproductive health outcomes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Objectives: Polycystic ovary syndrome (PCOS) and subclinical hypothyroidism (SCH) are prevalent gynecological conditions. However, the interrelationship between the two remains elusive. This study aims to elucidate the association between these conditions and determine the potential impact of SCH on the physiological and metabolic characteristics of patients with PCOS., Methods: This cross-sectional study enrolled 133 patients with PCOS from our Hospital. Participants were categorized into two groups: those with PCOS + SCH ( n  = 58) and those with PCOS ( n  = 75). Serum hormonal levels, metabolic markers, ovarian volume, and follicle count were compared between the groups., Results: There was a significant difference in BMI between the two groups, with a higher prevalence of obesity in the PCOS + SCH group ( p  = .014). Compared to the PCOS group, patients with PCOS + SCH had significantly higher levels of TSH ( p  < .001), triglycerides ( p  = .025), and HOMA-IR ( p  < .001), while LH levels were significantly lower ( p  = .048). However, multivariate linear regression analysis revealed that TSH, triglycerides, LH, and HOMA-IR were not determinants for the occurrence of SCH in patients with PCOS. Additionally, there was a notable reduction in follicle count in the left ovary for the PCOS + SCH group compared to the PCOS group ( p  = .003), and the overall follicle diameter of the PCOS + SCH group was also smaller ( p  = .010)., Conclusion: SCH may exert effects on the physiological and metabolic profiles of patients with PCOS. Further investigation into the relationship between these disorders is warranted to delineate their clinical implications.

The ovary in mammals has developed specialized mechanisms for protection against pathogen infections; however, the understanding of the innate immune system in the ovary of crustaceans is still limited. To elucidate the ovary's defense mechanisms in response to viral challenges, we subjected oriental river prawns (Macrobrachium nipponense) to poly I:C, a double-stranded RNA analog that emulates viral dsRNA, and analyzed the ovary's transcriptome profiles. Concurrently, RNA-seq analysis was performed on the hepatopancreas, a well-recognized immune-related tissue, following poly I:C challenge to investigate the distinct response mechanisms of the ovary and hepatopancreas and to gain a comprehensive understanding of the immune responses in both tissues. The results indicate that 1368 genes are differentially expressed in the ovary, with 903 genes upregulated and 465 genes downregulated. Subsequent analysis reveals that these differentially expressed genes (DEGs) include numerous genes associated with innate immunity, such as members of the C-type lectin, fibrinogen-related protein (Frep), Toll-like receptor, and NOD-like receptor (NLR) gene families, as well as acid phosphatase, scavenger receptor, crustin, Down syndrome cell adhesion molecule (Dscam), hemocyanin, and lipopolysaccharide and beta-1,3-glucan binding protein (LGBP). Furthermore, the DEGs include several genes related to ovary development, such as sox8, vitellogenin, progranulin, cyclin-dependent kinase, ecdysone receptor, frizzled, and members of the Fox gene family. In the hepatopancreas, a total of 729 DEGs were identified. Comparison of the DEGs in both tissues indicates that only 91 genes are common to both groups, highlighting significant tissue-specific responses to poly I:C stimulation. This study aims to enhance our understanding of the immune protective mechanisms employed by the ovary in response to pathogen exposure and establishes a foundation for investigating ovarian reproductive immunity in crustaceans., Competing Interests: Declaration of competing interest We declare that we have no competing interests., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Objectives: This study aimed to explore the effect and mechanism of Yunkang oral liquid (YK) on polycystic ovary syndrome (PCOS). Methods: PCOS model rats were prepared by injecting exogenous androgen dehydroepiandrosterone, and YK was administered simultaneously for 28 days during modeling. The morphology of ovaries and uterus was observed using H&E staining, and serum levels of testosterone (T), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were determined by radioimmunoassay. Additionally, serum lipids (TG, HDL-c), blood glucose (GLU), and aminotransferase (AST, ALT) levels were detected. The expression of androgen receptor (AR) protein was determined by Western blotting. Results: YK treatment resulted in reduced serum levels of T, LH and FSH, ameliorated ovarian polycystic-like pathological changes and uterine morphology in PCOS rats, and decreased serum TG, GLU, AST and ALT levels, elevated serum HDL-c levels, and improved abnormalities of glycolipid metabolism accompanying PCOS. Moreover, YK decreased the expression of ovarian AR in PCOS rats. Conclusions: This study indicates that YK may protect the ovaries by inhibiting the expression of AR, which could be a potential treatment for PCOS.

Chinese herbs contain substances that regulate female hormones. Our study confirmed that Zingiberis rhizoma carbonisata contains Zingiberis rhizoma -based carbon dots (ZR-CDs), which exert regulatory effects on serum oestradiol and FSH in mice and show impacts on endometrial growth and follicular development that potentially affect the ability of female fertility. ZR-CDs were characterized to clarify the microstructure, optical features, and functional group characteristics. It shows that ZR-CDs are spherical carbon nanostructures ranging from 0.97 to 2.3 nm in diameter, with fluorescent properties and a surface rich in functional groups. We further investigated the impact of ZR-CDs on oestradiol and FSH in serum, growth, and the development of ovarian and uterine using normal female mice and exogenous oestradiol intervention model. It was observed that ZR-CDs accelerated oestrogen metabolism and attenuated oestradiol-induced endometrial hyperplasia. Simultaneously, ZR-CDs triggered an increase in FSH, even in the presence of high-serum oestradiol that inhibits FSH secretion. Our findings suggest that ZR-CDs could be a potential therapeutic treatment for anovulatory menstruation.

Loss of ovarian homeostasis is associated with ovary dysfunction and female diseases; however, the underlying mechanisms responsible for the establishment of homeostasis and its function in the ovary have not been fully elucidated. Here, we showed that conditional knockout of Rab37 in oocytes impaired macroautophagy/autophagy proficiency in the ovary and interfered with follicular homeostasis and ovary development in mice. Flunarizine treatment upregulated autophagy, thus rescuing the impairment of follicular homeostasis and ovarian dysfunction in rab37 knockout mice by reprogramming of homeostasis. Notably, both the E2F1 and EGR2 transcription factors synergistically activated Rab37 transcription and promoted autophagy. Thus, RAB37-mediated autophagy ensures ovary function by maintaining ovarian homeostasis. Abbreviations: AMH: anti-Mullerian hormone; ATG: autophagy related; BECN1: beclin 1; cKO: conditional knockout; Cre: cyclization recombination enzyme; dpp: days postpartum; E2: estradiol; E2F1: E2F transcription factor 1; EBF1: EBF transcription factor 1; EGR2: early growth response 2; FSH: follicle stimulating hormone; LH: luteinizing hormone; mpp: months postpartum; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; RAB37: RAB37, member RAS oncogene family; SQSTM1: sequestosome 1; TFEB: transcription factor EB; Zp3 : zona pellucida glycoprotein 3.

Background: Puberty onset signifies the beginning of sexual maturation and reproductive phase in poultry indeed, and plays an essential role in genetics and breeding. Studying gonadal development is one of the important approaches to exploring the genetic mechanism of puberty onset., Result: In our study, the phenotype data of the testes and ovaries of the 120-day-old Nonghua duck showed a large coefficient of variation, indicating that their gonads were in different developmental states. The CNV-based GWAS results for 358 Nonghua ducks showed two deleted-type CNVRs were associated with testicular weight (TW) and testicular percentage (TP), namely CNVR492 (Chr2: 59473501-59478500 bp) and CNVR494 (Chr2: 59514001-59517000 bp). Additionally, two both-type CNVRs were associated with ovarian weight (OW) and ovarian percentage (OP), namely CNVR557 (Chr2: 99951001-99956500 bp) and CNVR891 (Chr7: 39115001-39122500 bp). RNA-seq analysis showed 6228 and 1070 differentially expressed genes (DEGs) related to the TW and OW. These DEGs were mainly enriched in the MAPK signaling pathway, cytokine-cytokine receptor interaction, and focal adhesion, which were reported to affect gonadal development. Further, by joint analysis of CNV-based GWAS and RNA-seq data, 3 genes, including LOC106019197, CDH19 (LOC101793040), and TYW5 were identified as potential candidate genes for TW and OW. LOC106019197 and CDH19 were down-regulated in the heavier-testes group (> 5 g), while TYW5 was also down-regulated in the heavier-ovaries group (> 3 g). The qRT-PCR revealed that LOC106019197 and CDH19 exhibited higher expression levels in the wild/CN0 and CN0/CN0 genotypes compared to the wild/wild genotype. TYW5 showed the highest expression level in the wild/CN0 genotype and the lowest in the CN2/CN2 genotype. In addition, the expression levels of LOC106019197 and CDH19 were significantly higher at 0w than at 8w and 24w., Conclusion: Our results revealed that LOC106019197 and CDH19 may act as inhibitors of duck testicular development. TYW5 may play a role in delaying ovarian development. These findings provide new insights into the mechanism of puberty onset in ducks., Competing Interests: Declarations. Ethics approval and consent to participate: All methods were carried out following relevant guidelines and regulations. All duck work was conducted following a protocol approved by Sichuan Agricultural University China’s animal ethics and welfare committee (AEWC). Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Aim: The interaction of MerTK, which negatively regulates immune responses, with its ligand Pros1 contributes to the resolution of apoptosis and inflammation, participating in the healing process of tissues. The levels of MerTK and Pros1, intensely expressed in macrophages (Mϕs), are affected by sex hormones. The expression levels of these proteins in Mϕs, which have a role in corpus luteum (CL) development or regression and folliculogenesis, were investigated in this study since their expressions have not been evaluated in pregnant mouse ovaries., Method: We analyzed mouse ovaries from non-pregnant mice at estrus and gestation days 5, 8, and 15 (each n:10). We used qPCR to evaluate Mertk and Pros1 mRNA levels and assessed their protein expression and localization using immunohistochemistry and double immunofluorescence staining for co-localization., Results: Mertk and Pros1 mRNA and protein levels significantly increased in GD15. MerTK and Pros1 protein levels in mouse CL on GD15 were significantly higher than all other groups. MerTK and Pros1 positive Mϕs were observed in CL of GD15 by double immunofluorescence. MerTK protein levels were increased in granulosa cells GD15 of primary and growing follicles., Conclusion: Our study revealed for the first time that the expression of MerTK and Pros1 was significantly increased in CL at GD15 in mice. These results suggest that increased levels of MerTK andPros1 may enhance their interaction as receptor-ligand binding partners in CL potentially contributing to the balance of apoptosis and inflammation., Competing Interests: Declarations. Institutional review board statement: The Animal Research Ethics Local Committee, Akdeniz University, approved the research by B.30.2.AKD.0.05.07.00/2 number. The Akdeniz University Institutional Animal Care and Use Committee standards carried out all experimental protocols on mice. Conflict of interest: The authors report no declarations of interest., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Obesity is one of the major risk factor for infertility since it causes decreased quality and quantity of gametes and a disrupted uterine environment which might result in miscarriage, stillbirth, and fetal abnormal growth. Obesity induces oxidative stress which is strongly associated with infertility. The clearing of oxidative stress by autophagy is maintained through the p62/ Keap1/Nrf2 pathway. In this pathway, oxidative stress induces p62 for binding to Keap1, thereby Keap1 cannot bind to the Nrf2 transcription factor. Then, Nrf2 translocates into the nucleus and initiates antioxidant-related gene expression. While p62, bound to Keap1, acts as an adaptor protein between autophagosome and damaged substrates which needs to be degraded for homeostasis. Up to date, obesity is strongly linked to abnormal autophagy activity. However, p62 protein expression has not been investigated in the obese ovary, testis, and uterus in detail. Thus, in the present study, we aimed to evaluate the effects of a high-fat diet (HFD)-induced obesity on p62 protein levels of the ovary, testis, and uterus in mice. Our results demonstrated that the p62 expression level was significantly altered by HFD in uterine glands, epithelium, myometrium, and stroma, and in the ovarian corpus luteum, testicular spermatogonium and spermatocytes., Competing Interests: Declarations. Ethical approval: The experimental protocol was approved by the Animal Care and Usage Committee of Ankara University (Protocol no: 2020-17-145). Competing interests: The authors declare no competing interests. Conflict of interest: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: Obesity significantly impacts female reproductive health and increases the risk of gynecological tumors. However, the specific transcriptional changes that occur in the ovarian microenvironment during obesity-induced stress and the relationship between obesity and ovarian cancer remain unclear., Methods: Our study investigated the single-cell landscape of the ovarian cortex in individuals with varying BMI levels by snRNA-seq, revealing weight-stage related cellular composition deviations and expression profile irregularities., Results: Using single-cell high-dimensional Weighted Gene Co-expression Network Analysis (hdWGCNA), we identified distinct obesity-related gene modules within various subpopulations of stroma cells and blood vascular endothelial cells. Notably, we observed a negative correlation between BMI and heat shock protein (HSP) family genes. Specifically, we found that HSPD1 might function as a potential regulator of ovarian carcinogenesis and progression under conditions of obesity, as supported by our co-analysis with data from three bulk RNA-seq ovarian cancer databases. Our findings suggested that lower expression of HSPD1 indicated a poorer prognosis for ovarian cancer., Conclusions: Our study identified a cluster of genes in ovarian cells that are suppressed by obesity, including those belonging to HSP family genes. These findings provide valuable insights for investigating the link between obesity and ovarian diseases., Competing Interests: Declarations. Ethics approval and consent to participate: This study has proved by the Medical Ethics Committee of the Second Hospital of Shandong University (KYLL-2023-314). Consent for publication: Not applicable. Competing interests: The authors have declared that no conflict of interest exists., (© 2024. The Author(s).)