Pub Date : 2025-09-15DOI: 10.1016/j.repbio.2025.101079
Marilú Cristofoli , Flavia Maria Pia Montenegro Donoso , Mayla Magalhães de Oliveira Alcobaça , Fabiana Morse Gosson Jorge , Danielle Cristina Calado de Brito , Moacir Franco de Oliveira , Antônio Chaves de Assis Neto
The rhea (Rhea americana) is a docile and highly adaptable ratite with notable zootechnical potential for the production of meat, eggs, feathers, and leather. Beyond its economic relevance, captive breeding plays an important role in conservation. However, information on its embryonic and gonadal development remains limited—knowledge that is essential for improving reproductive management. This study aimed to characterize gonadal differentiation during artificial incubation, focusing on developmental stages at days 9, 12, 15, 18, 21, and 24. We provide a detailed morphological and cytological description of germ and somatic cells, documenting the appearance of the gonadal crest at day 9, the formation of undifferentiated gonads by day 12, and the onset of sexual differentiation at day 15. Morphometric analyses of cortex thickness and gonad–mesonephros area relationships enabled comparisons between sexes. The gonads were positioned cranioventrally on the mesoneph, and testicular maturation occurred earlier than ovarian differentiation. Transmission electron microscopy revealed greater mitochondrial density in male germ cells than in female germ cells, indicating early functional divergence. These results contribute to the understanding of reproductive development in R. americana and support the development of conservation and breeding strategies for this and related species.
{"title":"Gonadal morphogenesis and sexual differentiation in rhea (Rhea americana)","authors":"Marilú Cristofoli , Flavia Maria Pia Montenegro Donoso , Mayla Magalhães de Oliveira Alcobaça , Fabiana Morse Gosson Jorge , Danielle Cristina Calado de Brito , Moacir Franco de Oliveira , Antônio Chaves de Assis Neto","doi":"10.1016/j.repbio.2025.101079","DOIUrl":"10.1016/j.repbio.2025.101079","url":null,"abstract":"<div><div>The rhea (<em>Rhea americana</em>) is a docile and highly adaptable ratite with notable zootechnical potential for the production of meat, eggs, feathers, and leather. Beyond its economic relevance, captive breeding plays an important role in conservation. However, information on its embryonic and gonadal development remains limited—knowledge that is essential for improving reproductive management. This study aimed to characterize gonadal differentiation during artificial incubation, focusing on developmental stages at days 9, 12, 15, 18, 21, and 24. We provide a detailed morphological and cytological description of germ and somatic cells, documenting the appearance of the gonadal crest at day 9, the formation of undifferentiated gonads by day 12, and the onset of sexual differentiation at day 15. Morphometric analyses of cortex thickness and gonad–mesonephros area relationships enabled comparisons between sexes. The gonads were positioned cranioventrally on the mesoneph, and testicular maturation occurred earlier than ovarian differentiation. Transmission electron microscopy revealed greater mitochondrial density in male germ cells than in female germ cells, indicating early functional divergence. These results contribute to the understanding of reproductive development in <em>R. americana</em> and support the development of conservation and breeding strategies for this and related species.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 4","pages":"Article 101079"},"PeriodicalIF":2.5,"publicationDate":"2025-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145060608","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-10DOI: 10.1016/j.repbio.2025.101074
Zheng Zhi , Haijun Yang , Zhangming Liu , Wei Pan , Yanzhi Hao , Xinming Zhang , Shuancheng Zhang , Huazhou Xu , Huirong Ma , Huilan Du , Cuimiao Song , Qiong Wu
Controlled ovarian hyperstimulation (COH) is a cornerstone of assisted reproductive technologies, yet its effects on endometrial function and embryo implantation remain poorly understood, particularly regarding the role of hypoxia-inducible factor 2α (HIF-2α) signaling. Therefore, the objective of this study was to investigate whether COH-induced endometrial dysfunction impairs mouse embryo implantation through the HIF-2α pathway. A COH mouse model was established using gonadotropin-releasing hormone agonist (GnRH-a)/human menopausal gonadotropin (hMG)/human chorionic gonadotropin (hCG) administration. Embryo implantation status was evaluated on gestational days 5, 6, and 20; endometrial tissues were analyzed for HIF-2α pathway activity via immunohistochemistry (IHC), immunofluorescence (IF), Western blot (WB) and quantitative real-time PCR (qRT-PCR); histological changes were assessed by Jones silver staining and transmission electron microscopy (TEM); serum estradiol (E2), progesterone (P4), and prolactin (PRL) levels were measured by radioimmunoassay (RIA). COH mice exhibited reduced total embryo implantation rates (on day 5, 6 and 20), together with decreased serum E2, P4, and PRL levels. COH mice exhibited preserved luminal epithelium integrity with increased microvillus density and continuous basement membrane structure. qRT-PCR and WB demonstrated significantly downregulated HIF-2α expression at both mRNA and protein levels, accompanied by reduced downstream RAB27B (member of RAS oncogene family)/matrix metalloproteinase 9 (MMP9) and lysyl oxidase (LOX)/adrenomedullin (ADM) signaling, which associated with impaired luminal epithelium detachment and compromised trophoblast invasion. Together, these findings identify HIF-2α as a potential key mediator of COH-induced endometrial microenvironment alterations, revealing molecular mechanisms underlying implantation failure. Importantly, the HIF-2α-RAB27B/MMP9 and HIF-2α-LOX/ADM axes are highlighted as promising therapeutic targets to optimize assisted reproductive outcomes.
{"title":"Controlled ovarian hyperstimulation reduces mouse embryo implantation capacity through uterine HIF-2α mediated pathway","authors":"Zheng Zhi , Haijun Yang , Zhangming Liu , Wei Pan , Yanzhi Hao , Xinming Zhang , Shuancheng Zhang , Huazhou Xu , Huirong Ma , Huilan Du , Cuimiao Song , Qiong Wu","doi":"10.1016/j.repbio.2025.101074","DOIUrl":"10.1016/j.repbio.2025.101074","url":null,"abstract":"<div><div>Controlled ovarian hyperstimulation (COH) is a cornerstone of assisted reproductive technologies, yet its effects on endometrial function and embryo implantation remain poorly understood, particularly regarding the role of hypoxia-inducible factor 2α (HIF-2α) signaling. Therefore, the objective of this study was to investigate whether COH-induced endometrial dysfunction impairs mouse embryo implantation through the HIF-2α pathway. A COH mouse model was established using gonadotropin-releasing hormone agonist (GnRH-a)/human menopausal gonadotropin (hMG)/human chorionic gonadotropin (hCG) administration. Embryo implantation status was evaluated on gestational days 5, 6, and 20; endometrial tissues were analyzed for HIF-2α pathway activity via immunohistochemistry (IHC), immunofluorescence (IF), Western blot (WB) and quantitative real-time PCR (qRT-PCR); histological changes were assessed by Jones silver staining and transmission electron microscopy (TEM); serum estradiol (E<sub>2</sub>), progesterone (P<sub>4</sub>), and prolactin (PRL) levels were measured by radioimmunoassay (RIA). COH mice exhibited reduced total embryo implantation rates (on day 5, 6 and 20), together with decreased serum E<sub>2</sub>, P<sub>4</sub>, and PRL levels. COH mice exhibited preserved luminal epithelium integrity with increased microvillus density and continuous basement membrane structure. qRT-PCR and WB demonstrated significantly downregulated HIF-2α expression at both mRNA and protein levels, accompanied by reduced downstream RAB27B (member of RAS oncogene family)/matrix metalloproteinase 9 (MMP9) and lysyl oxidase (LOX)/adrenomedullin (ADM) signaling, which associated with impaired luminal epithelium detachment and compromised trophoblast invasion. Together, these findings identify HIF-2α as a potential key mediator of COH-induced endometrial microenvironment alterations, revealing molecular mechanisms underlying implantation failure. Importantly, the HIF-2α-RAB27B/MMP9 and HIF-2α-LOX/ADM axes are highlighted as promising therapeutic targets to optimize assisted reproductive outcomes.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 4","pages":"Article 101074"},"PeriodicalIF":2.5,"publicationDate":"2025-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145026550","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-09DOI: 10.1016/j.repbio.2025.101076
Yang Yang , Linfeng Li , Ning Zhang , Zheng Zhang , Zhengyang Wu , Dongmei Ji
Current research indicates that polyethylene terephthalate microplastics (PET-MPs) may significantly impair male reproductive function. This study aimed to investigate the potential molecular mechanisms underlying this impairment. Potential gene targets of PET-MPs were predicted via the SwissTargetPrediction database. GWAS summary statistics for male infertility were obtained from the GWAS Catalog, and eQTL data were acquired from the eQTLGen database. SMR analysis was subsequently performed to identify genes exhibiting significant causal associations with male infertility. The potential targets of PET-MPs contributing to male infertility were ultimately identified by intersecting the two gene sets. Additionally, bulk RNA-seq and scRNA-seq analyses were employed to elucidate the potential molecular mechanisms underlying PET-MPs-induced male infertility. Our findings suggest that CLK4 may serve as a functional target through which PET-MPs contribute to male infertility, with elevated CLK4 expression representing a significant risk factor. Molecular docking and molecular dynamics simulations demonstrated that PET-MPs can form stable binding conformations with the CLK4 protein. We further identified terminally differentiated CD4+ T cells as a prominent risk factor for male infertility, revealing a significant positive correlation between CLK4 expression levels and Th1 cell infiltration. Additionally, CLK4 exhibited a biphasic expression pattern during spermatocyte-to-sperm differentiation, initially increasing before subsequent downregulation. These observations indicate that PET-MPs may participate in the pathogenesis of male infertility by targeting CLK4 to modulate Th1 cell infiltration and disrupt normal spermatogenic processes. In conclusion, our study demonstrates that CLK4 may serve as a potential target for PET-MP-induced male infertility, and we further elucidate the underlying molecular mechanisms.
{"title":"Analyzing the toxicological effects of PET-MPs on male infertility: Insights from network toxicology, mendelian randomization, and transcriptomics","authors":"Yang Yang , Linfeng Li , Ning Zhang , Zheng Zhang , Zhengyang Wu , Dongmei Ji","doi":"10.1016/j.repbio.2025.101076","DOIUrl":"10.1016/j.repbio.2025.101076","url":null,"abstract":"<div><div>Current research indicates that polyethylene terephthalate microplastics (PET-MPs) may significantly impair male reproductive function. This study aimed to investigate the potential molecular mechanisms underlying this impairment. Potential gene targets of PET-MPs were predicted via the SwissTargetPrediction database. GWAS summary statistics for male infertility were obtained from the GWAS Catalog, and eQTL data were acquired from the eQTLGen database. SMR analysis was subsequently performed to identify genes exhibiting significant causal associations with male infertility. The potential targets of PET-MPs contributing to male infertility were ultimately identified by intersecting the two gene sets. Additionally, bulk RNA-seq and scRNA-seq analyses were employed to elucidate the potential molecular mechanisms underlying PET-MPs-induced male infertility. Our findings suggest that <em>CLK4</em> may serve as a functional target through which PET-MPs contribute to male infertility, with elevated <em>CLK4</em> expression representing a significant risk factor. Molecular docking and molecular dynamics simulations demonstrated that PET-MPs can form stable binding conformations with the <em>CLK4</em> protein. We further identified terminally differentiated CD4<sup>+</sup> T cells as a prominent risk factor for male infertility, revealing a significant positive correlation between <em>CLK4</em> expression levels and Th1 cell infiltration. Additionally, <em>CLK4</em> exhibited a biphasic expression pattern during spermatocyte-to-sperm differentiation, initially increasing before subsequent downregulation. These observations indicate that PET-MPs may participate in the pathogenesis of male infertility by targeting <em>CLK4</em> to modulate Th1 cell infiltration and disrupt normal spermatogenic processes. In conclusion, our study demonstrates that <em>CLK4</em> may serve as a potential target for PET-MP-induced male infertility, and we further elucidate the underlying molecular mechanisms.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 4","pages":"Article 101076"},"PeriodicalIF":2.5,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145019726","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-04DOI: 10.1016/j.repbio.2025.101073
Yi Wang , Yuting Zhu , Yan Cui , Jiali Fang , Hong Zhong , Yujie Shi , Lan Liu , Xianwei Cui
Gestational diabetes mellitus (GDM) is a common and serious complication during pregnancy. Depleted next-generation probiotic, Akkermansia muciniphila (AKK) in GDM women indicates its potential on GDM prevention. However, the functions and mechanisms of AKK on GDM remain unclear. Due to the limited strategies for GDM therapy, combined with the anaerobic properties of AKK, herein, we reported pasteurized AKK functions as novel postbiotics which ameliorated glucose intolerance and insulin resistance in the GDM mouse model induced by high-fat diet (HFD) feeding combined with STZ. The oral administration of pasteurized AKK enhanced glucose homeostasis and alleviated placental inflammation in the GDM mouse model. Specifically, placental macrophage polarization was transferred by AKK treatment. In addition, the outer membrane protein of AKK, Amuc_1100, mimicked anti-inflammatory properties and improvement of GDM, which served as an effector protein. These findings demonstrate that oral AKK supplementation alleviated placental inflammatory responses through modulating macrophage polarization. Mechanically, we uncovered that a heat-stable outer membrane protein of AKK, Amuc_1100, mimics the anti-diabetic properties of pasteurized AKK through oral administration. Taken together, our findings demonstrated an effective treatment of GDM from the perspective of potential probiotic agents.
{"title":"Pasteurized Akkermansia muciniphila ameliorates insulin resistance by reducing placental inflammation in GDM mouse model","authors":"Yi Wang , Yuting Zhu , Yan Cui , Jiali Fang , Hong Zhong , Yujie Shi , Lan Liu , Xianwei Cui","doi":"10.1016/j.repbio.2025.101073","DOIUrl":"10.1016/j.repbio.2025.101073","url":null,"abstract":"<div><div>Gestational diabetes mellitus (GDM) is a common and serious complication during pregnancy. Depleted next-generation probiotic, Akkermansia muciniphila (AKK) in GDM women indicates its potential on GDM prevention. However, the functions and mechanisms of AKK on GDM remain unclear. Due to the limited strategies for GDM therapy, combined with the anaerobic properties of AKK, herein, we reported pasteurized AKK functions as novel postbiotics which ameliorated glucose intolerance and insulin resistance in the GDM mouse model induced by high-fat diet (HFD) feeding combined with STZ. The oral administration of pasteurized AKK enhanced glucose homeostasis and alleviated placental inflammation in the GDM mouse model. Specifically, placental macrophage polarization was transferred by AKK treatment. In addition, the outer membrane protein of AKK, Amuc_1100, mimicked anti-inflammatory properties and improvement of GDM, which served as an effector protein. These findings demonstrate that oral AKK supplementation alleviated placental inflammatory responses through modulating macrophage polarization. Mechanically, we uncovered that a heat-stable outer membrane protein of AKK, Amuc_1100, mimics the anti-diabetic properties of pasteurized AKK through oral administration. Taken together, our findings demonstrated an effective treatment of GDM from the perspective of potential probiotic agents.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 4","pages":"Article 101073"},"PeriodicalIF":2.5,"publicationDate":"2025-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144988998","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Radiation and doxorubicin (Dox) exert destructive effects on testicular tissue. Resveratrol, a natural antioxidant, may protect the spermatogenic system against the toxic effects of these agents. This study evaluated the protective and antioxidant properties of resveratrol-loaded solid lipid nanoparticles (RES-SLNs) against Dox- and radiation-induced testicular injury in mice. Following the synthesis of RES-SLNs and characterization of their physicochemical properties, 60 adult male mice were randomly assigned to six groups (n = 10 per group): control, RES-SLNs, Dox, radiation (Rad), RES-SLNs + Dox, and RES-SLNs + Rad. Five mice from each group underwent anesthesia and bilateral orchiectomy for determination of malondialdehyde (MDA) and superoxide dismutase (SOD) levels. The remaining mice were sacrificed for histopathological evaluation of testicular tissue. Radiation and Dox administration significantly increased MDA levels, reduced SOD activity, and altered histological parameters. In contrast, intraperitoneal RES-SLN administration significantly lowered MDA levels, enhanced SOD activity, and mitigated morphological alterations induced by radiation or Dox. These findings indicate that RES-SLNs attenuate oxidative stress in the testes caused by radiation or Dox, thereby preventing severe structural damage.
{"title":"Evaluation of the protective effect of resveratrol-loaded solid lipid nanoparticles on radiation- or doxorubicin-induced spermatogenic damage in mice","authors":"Mohammad Farhadi Rad , Faranak Aghaz , Khodabakhsh Rashidi , Sousan Gharibnejad , Elham Arkan , Kamran Mansouri , Elahe Motevaseli , Masoud Najafi","doi":"10.1016/j.repbio.2025.101058","DOIUrl":"10.1016/j.repbio.2025.101058","url":null,"abstract":"<div><div>Radiation and doxorubicin (Dox) exert destructive effects on testicular tissue. <em>Resveratrol</em>, a natural antioxidant, may protect the spermatogenic system against the toxic effects of these agents. This study evaluated the protective and antioxidant properties of resveratrol-loaded solid lipid nanoparticles (RES-SLNs) against Dox- and radiation-induced testicular injury in mice. Following the synthesis of RES-SLNs and characterization of their physicochemical properties, 60 adult male mice were randomly assigned to six groups (n = 10 per group): control, RES-SLNs, Dox, radiation (Rad), RES-SLNs + Dox, and RES-SLNs + Rad. Five mice from each group underwent anesthesia and bilateral orchiectomy for determination of malondialdehyde (MDA) and superoxide dismutase (SOD) levels. The remaining mice were sacrificed for histopathological evaluation of testicular tissue. Radiation and Dox administration significantly increased MDA levels, reduced SOD activity, and altered histological parameters. In contrast, intraperitoneal RES-SLN administration significantly lowered MDA levels, enhanced SOD activity, and mitigated morphological alterations induced by radiation or Dox. These findings indicate that RES-SLNs attenuate oxidative stress in the testes caused by radiation or Dox, thereby preventing severe structural damage.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 4","pages":"Article 101058"},"PeriodicalIF":2.5,"publicationDate":"2025-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144988997","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cadmium (Cd) is a toxic heavy metal known to cause DNA damage, genetic alterations, and reproductive issues in males through free radical production. This study investigates L-arginine, an amino acid with antioxidant and metal-chelating properties, in mitigating Cd-induced genetic and reproductive toxicity in male mice. Thirty-six male mice were divided into six groups: a control group, a Cd group (2 mg/kg), three groups receiving Cd (2 mg/kg) combined with L-arginine (50, 100, 200 mg/kg), and a positive control group receiving vitamin C (500 mg/kg) with Cd. After 42 days, oxidative stress markers, including ROS, protein carbonyl, lipid peroxidation, and glutathione levels, were assessed in bone marrow and testicular tissue. Sperm parameters and histopathological evaluations of testes were also conducted. Chronic Cd exposure significantly reduced glutathione levels and increased oxidative stress markers in both tissues. Sperm count and motility decreased, while abnormal sperm increased. Conversely, L-arginine treatment notably reduced oxidative stress markers, improved antioxidant levels, sperm quality, and testicular histopathology compared to the Cd group. Additionally, Cd exposure increased the number of micronuclei in polychromatic erythrocytes (PCEs), which decreased with L-arginine treatment, enhancing the PCE/(PCE+normochromatic erythrocytes) ratio. These findings suggest that L-arginine effectively mitigates Cd-induced toxicity. The results indicate that L-arginine has significant protective effects against Cd-induced reproductive and bone marrow toxicity.
{"title":"The ameliorative effects of L-arginine on testicular and genotoxic toxicity induced by chronic exposure to cadmium chloride in male murine models","authors":"Nasibeh Hemati , Milad Chahardori , Mohammad Hosseinzadeh , Fereshteh Taleb-Pour , Fatemeh Shaki","doi":"10.1016/j.repbio.2025.101072","DOIUrl":"10.1016/j.repbio.2025.101072","url":null,"abstract":"<div><div>Cadmium (Cd) is a toxic heavy metal known to cause DNA damage, genetic alterations, and reproductive issues in males through free radical production. This study investigates L-arginine, an amino acid with antioxidant and metal-chelating properties, in mitigating Cd-induced genetic and reproductive toxicity in male mice. Thirty-six male mice were divided into six groups: a control group, a Cd group (2 mg/kg), three groups receiving Cd (2 mg/kg) combined with L-arginine (50, 100, 200 mg/kg), and a positive control group receiving vitamin C (500 mg/kg) with Cd. After 42 days, oxidative stress markers, including ROS, protein carbonyl, lipid peroxidation, and glutathione levels, were assessed in bone marrow and testicular tissue. Sperm parameters and histopathological evaluations of testes were also conducted. Chronic Cd exposure significantly reduced glutathione levels and increased oxidative stress markers in both tissues. Sperm count and motility decreased, while abnormal sperm increased. Conversely, L-arginine treatment notably reduced oxidative stress markers, improved antioxidant levels, sperm quality, and testicular histopathology compared to the Cd group. Additionally, Cd exposure increased the number of micronuclei in polychromatic erythrocytes (PCEs), which decreased with L-arginine treatment, enhancing the PCE/(PCE+normochromatic erythrocytes) ratio. These findings suggest that L-arginine effectively mitigates Cd-induced toxicity. The results indicate that L-arginine has significant protective effects against Cd-induced reproductive and bone marrow toxicity.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 4","pages":"Article 101072"},"PeriodicalIF":2.5,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144916422","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-27DOI: 10.1016/j.repbio.2025.101060
Daniyal Ezati , Reyhane Vardiyan , Ali Reza Talebi , Fatemeh Alipour , Hasan Pahang , Shabnam Mohammadi
Sperm cryopreservation is a conventional method to preserve sperm cells for a long time. This technique may cause various effects on sperm parameters. Therefore, applying beneficial antioxidants to decrease the adverse effects of freezing is noteworthy. Fisetin is a compound with antioxidant and anti-inflammatory properties. The main aim of the present study is to investigate the protective and beneficial effects of fisetin against cryo-damage on sperm functional parameters. In this experimental study, we analyzed 20 semen samples from asthenoteratozoospermic (AT) patients. Each sample was divided into three treatment groups: (1) fresh control (non-frozen), (2) cryopreservation medium (without fisetin), and (3) cryomedium supplemented with 50 μM fisetin. Freezing and thawing procedures were performed via the conventional method. Post-thaw analyses revealed that cryopreservation significantly reduced sperm motility, chromatin integrity, and mitochondrial membrane potential while increasing DNA fragmentation, malondialdehyde (MDA) levels, and apoptosis (p < 0.05). Fisetin supplementation markedly improved progressive motility (p < 0.05), reduced non-motile sperm percentage (p < 0.05), and decreased DNA fragmentation and MDA levels (p < 0.05). Additionally, it enhanced chromatin condensation and reduced apoptosis rates (p < 0.05). Fisetin attenuates cryo-damage through its antioxidant and anti-apoptotic properties, improving post-thaw sperm quality. Thus, incorporating fisetin into cryopreservation media could enhance sperm viability for assisted reproductive technologies (ART).
{"title":"Fisetin attenuates the adverse effects of freezing and thawing procedures on the biological characteristics of human asthenoteratozoospermia samples","authors":"Daniyal Ezati , Reyhane Vardiyan , Ali Reza Talebi , Fatemeh Alipour , Hasan Pahang , Shabnam Mohammadi","doi":"10.1016/j.repbio.2025.101060","DOIUrl":"10.1016/j.repbio.2025.101060","url":null,"abstract":"<div><div>Sperm cryopreservation is a conventional method to preserve sperm cells for a long time. This technique may cause various effects on sperm parameters. Therefore, applying beneficial antioxidants to decrease the adverse effects of freezing is noteworthy. Fisetin is a compound with antioxidant and anti-inflammatory properties. The main aim of the present study is to investigate the protective and beneficial effects of fisetin against cryo-damage on sperm functional parameters. In this experimental study, we analyzed 20 semen samples from asthenoteratozoospermic (AT) patients. Each sample was divided into three treatment groups: (1) fresh control (non-frozen), (2) cryopreservation medium (without fisetin), and (3) cryomedium supplemented with 50 μM fisetin. Freezing and thawing procedures were performed via the conventional method. Post-thaw analyses revealed that cryopreservation significantly reduced sperm motility, chromatin integrity, and mitochondrial membrane potential while increasing DNA fragmentation, malondialdehyde (MDA) levels, and apoptosis (p < 0.05). Fisetin supplementation markedly improved progressive motility (p < 0.05), reduced non-motile sperm percentage (p < 0.05), and decreased DNA fragmentation and MDA levels (p < 0.05). Additionally, it enhanced chromatin condensation and reduced apoptosis rates (p < 0.05). Fisetin attenuates cryo-damage through its antioxidant and anti-apoptotic properties, improving post-thaw sperm quality. Thus, incorporating fisetin into cryopreservation media could enhance sperm viability for assisted reproductive technologies (ART).</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 4","pages":"Article 101060"},"PeriodicalIF":2.5,"publicationDate":"2025-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144903737","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-26DOI: 10.1016/j.repbio.2025.101071
Abdel-Khalek Kh A E. , Saima Qadeer , Sh.A. Gabr , A.M. Shehabeldin , A.E.B. Ghodaia , Sara Latif , Asma Ul Husna
This study aimed to investigate the cryoprotective mechanism of nano-curcumin (CRNPs) on rabbit sperm via molecular docking and to compare the effects of two extenders, Tris-egg yolk (TEYE) and Tris-soybean lecithin (TSBLE) supplemented with CRNPs on post thaw sperm quality. Molecular docking revealed strong binding of nano-curcumin to Caspase-3 and Caspase-9, suggesting its potential to inhibit pro-apoptotic activity. In silico toxicological profiling predicted a low acute toxicity for nano-curcumin (LD50 = 2000mg/kg; class4). Semen from ten white New Zealand rabbits was collected and pooled, then divided between TEYE and TSBLE. Each extender was further supplemented with CRNPs (0, 1.5, 2 and 2.5 µg/mL). After equilibration, samples were cryopreserved and evaluated post-thaw using CASA and flow cytometry. Supplementation with 1.5 µg/mL CRNPs significantly (p < 0.05) improved total, progressive, and rapid motility, normal morphology, motion velocities, and viability compared to the control. Although reductions in immotility, abnormal forms, apoptosis, and necrosis were observed, these differences were not significant (p > 0.05). Both extenders significantly (p < 0.05) enhanced antioxidant defense by increasing TAC, GPx, and SOD levels while reducing MDA and H2O2, with TSBLE showing slightly better performance. In conclusion, CRNPs at 1.5 µg/mL improved sperm motility, viability, morphology, and antioxidant status, likely by mitigating oxidative stress and apoptosis via caspase inhibition. Both extenders were effective, and further in vivo studies are needed to assess fertility outcomes.
{"title":"Molecular docking of nano-curcumin to caspase-3 and caspase-9: Unraveling its cryoprotective mechanism in rabbit sperm extenders","authors":"Abdel-Khalek Kh A E. , Saima Qadeer , Sh.A. Gabr , A.M. Shehabeldin , A.E.B. Ghodaia , Sara Latif , Asma Ul Husna","doi":"10.1016/j.repbio.2025.101071","DOIUrl":"10.1016/j.repbio.2025.101071","url":null,"abstract":"<div><div>This study aimed to investigate the cryoprotective mechanism of nano-curcumin (CRNPs) on rabbit sperm via molecular docking and to compare the effects of two extenders, Tris-egg yolk (TEYE) and Tris-soybean lecithin (TSBLE) supplemented with CRNPs on post thaw sperm quality. Molecular docking revealed strong binding of nano-curcumin to Caspase-3 and Caspase-9, suggesting its potential to inhibit pro-apoptotic activity. In silico toxicological profiling predicted a low acute toxicity for nano-curcumin (LD<sub>50</sub> = 2000mg/kg; class4). Semen from ten white New Zealand rabbits was collected and pooled, then divided between TEYE and TSBLE. Each extender was further supplemented with CRNPs (0, 1.5, 2 and 2.5 µg/mL). After equilibration, samples were cryopreserved and evaluated post-thaw using CASA and flow cytometry. Supplementation with 1.5 µg/mL CRNPs significantly (p < 0.05) improved total, progressive, and rapid motility, normal morphology, motion velocities, and viability compared to the control. Although reductions in immotility, abnormal forms, apoptosis, and necrosis were observed, these differences were not significant (p > 0.05). Both extenders significantly (p < 0.05) enhanced antioxidant defense by increasing TAC, GPx, and SOD levels while reducing MDA and H<sub>2</sub>O<sub>2</sub>, with TSBLE showing slightly better performance. In conclusion, CRNPs at 1.5 µg/mL improved sperm motility, viability, morphology, and antioxidant status, likely by mitigating oxidative stress and apoptosis via caspase inhibition. Both extenders were effective, and further in vivo studies are needed to assess fertility outcomes.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 4","pages":"Article 101071"},"PeriodicalIF":2.5,"publicationDate":"2025-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144896699","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-26DOI: 10.1016/j.repbio.2025.101059
Rabail Azhar Iqbal , Sophia Nazir
Gestational diabetes mellitus (GDM) is one of the commonly encountered medical disorders of pregnancy affecting women of childbearing age globally at an alarming rate and causing significant perinatal morbidity and adverse fetal outcomes. The intricate interchanges between genetic susceptibility, metabolic changes and environmental factors contribute to the altered β-cell function, insulin sensitivity and glucose metabolism leading to hyperglycaemia in pregnancy. However, the precise etiology of GDM remains incompletely understood. There is a compelling need for the identification of GDM in at-risk pregnant females at an early stage to improve maternal and fetal outcomes. Ongoing research has revealed numerous biomarkers that can be both predictive and diagnostic for GDM. This review aims to explore the various biomarkers that have increasing evidence for their use as predictors of GDM in the first and early second trimesters. The sensitivity, specificity, accuracy, and predictive power of these biomarkers have been compared to identify the ideal candidate(s), despite the effect of confounding factors. Throughout this review, research gaps were highlighted and future research directions were suggested to accurately diagnose GDM through a panel of efficacious biomarkers well before the disease manifests in later pregnancy.
{"title":"Potential novel biomarkers for gestational diabetes mellitus in first and early second trimesters: A comprehensive review","authors":"Rabail Azhar Iqbal , Sophia Nazir","doi":"10.1016/j.repbio.2025.101059","DOIUrl":"10.1016/j.repbio.2025.101059","url":null,"abstract":"<div><div>Gestational diabetes mellitus (GDM) is one of the commonly encountered medical disorders of pregnancy affecting women of childbearing age globally at an alarming rate and causing significant perinatal morbidity and adverse fetal outcomes. The intricate interchanges between genetic susceptibility, metabolic changes and environmental factors contribute to the altered β-cell function, insulin sensitivity and glucose metabolism leading to hyperglycaemia in pregnancy. However, the precise etiology of GDM remains incompletely understood. There is a compelling need for the identification of GDM in at-risk pregnant females at an early stage to improve maternal and fetal outcomes. Ongoing research has revealed numerous biomarkers that can be both predictive and diagnostic for GDM. This review aims to explore the various biomarkers that have increasing evidence for their use as predictors of GDM in the first and early second trimesters. The sensitivity, specificity, accuracy, and predictive power of these biomarkers have been compared to identify the ideal candidate(s), despite the effect of confounding factors. Throughout this review, research gaps were highlighted and future research directions were suggested to accurately diagnose GDM through a panel of efficacious biomarkers well before the disease manifests in later pregnancy.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 4","pages":"Article 101059"},"PeriodicalIF":2.5,"publicationDate":"2025-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144903738","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The circadian clock is constructed by a transcription–translation feedback loop system of clock genes and regulates various reproductive physiological functions. Understanding the rhythms of clock gene expression is important for understanding biological rhythms. However, the in vivo clock gene expression rhythm in Japanese Black breeding cows has not yet been clarified. Herein, we investigated the circadian expression rhythm of clock genes in hair follicle cells in the hair roots of Japanese Black breeding cows. We found that the clock genes of Japanese Black breeding cows exhibit an expression rhythm with a period of approximately 24 h under natural light, similar to the pattern observed in mice and humans, and that this expression rhythm disappears under constant light conditions. Furthermore, since the expression levels of clock genes are significantly reduced in pregnant cows exposed to constant light conditions, we analysed the diurnal variation in the plasma concentration of progesterone, which plays an important role in maintaining pregnancy. We found that the progesterone secretion rhythm observed under natural light in pregnant cows also disappeared under constant light conditions. These results indicate that in Japanese Black breeding cows, clock genes exhibit a diurnal expression rhythm in response to light; this rhythm disappears under constant light conditions. These clock genes play important roles in the diurnal variation in hormone secretion by interacting with progesterone in the blood.
{"title":"Diurnal variation in clock gene expression and progesterone secretion is inhibited under constant light conditions in Japanese Black cows","authors":"Tsuyoshi Otsuka , Hiroki Mitsuishi , Hiroki Onishi , Masato Yayota","doi":"10.1016/j.repbio.2025.101056","DOIUrl":"10.1016/j.repbio.2025.101056","url":null,"abstract":"<div><div>The circadian clock is constructed by a transcription–translation feedback loop system of clock genes and regulates various reproductive physiological functions. Understanding the rhythms of clock gene expression is important for understanding biological rhythms. However, the <em>in vivo</em> clock gene expression rhythm in Japanese Black breeding cows has not yet been clarified. Herein, we investigated the circadian expression rhythm of clock genes in hair follicle cells in the hair roots of Japanese Black breeding cows. We found that the clock genes of Japanese Black breeding cows exhibit an expression rhythm with a period of approximately 24 h under natural light, similar to the pattern observed in mice and humans, and that this expression rhythm disappears under constant light conditions. Furthermore, since the expression levels of clock genes are significantly reduced in pregnant cows exposed to constant light conditions, we analysed the diurnal variation in the plasma concentration of progesterone, which plays an important role in maintaining pregnancy. We found that the progesterone secretion rhythm observed under natural light in pregnant cows also disappeared under constant light conditions. These results indicate that in Japanese Black breeding cows, clock genes exhibit a diurnal expression rhythm in response to light; this rhythm disappears under constant light conditions. These clock genes play important roles in the diurnal variation in hormone secretion by interacting with progesterone in the blood.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 4","pages":"Article 101056"},"PeriodicalIF":2.5,"publicationDate":"2025-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144867304","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}