Monosodium glutamate (MSG) has been implicated in female reproductive toxicity through endocrine disruption, oxidative stress, inflammation, and apoptosis. This study investigated the protective role of Sorghum bicolor extract (SBE), a polyphenol-rich nutraceutical, against MSG-induced reproductive dysfunction in female Wistar rats, integrating in vivo outcomes with in silico docking analysis. Rats were exposed to MSG (4 g/kg) with or without SBE supplementation (100 mg/kg) orally for 28 days. Estrous cycle, sexual behaviors, hormonal profiles, oxidative and nitrosative stress markers, inflammatory mediators, and apoptotic regulators were assessed. Molecular docking evaluated the interactions of SBE phytochemicals with LHB and LHCGR. MSG disrupted the pituitary–gonadal axis, shortened estrous phases, suppressed reproductive behaviors, increased lipid peroxidation, activated NF-κB, and triggered mitochondrial apoptosis. SBE restored gonadotropins and steroid hormones, normalized estrous cycle, enhanced antioxidant defenses, suppressed inflammation, and prevented apoptosis. Docking analysis revealed strong ligand–protein interactions, supporting endocrine and cellular protection. Conclusively, SBE confers robust gonadoprotective effects against MSG toxicity through endocrine restoration, antioxidant, anti-inflammatory, and anti-apoptotic mechanisms validated in vivo and in silico.
{"title":"Mechanisms underlying Sorghum bicolor extract protection against monosodium glutamate-induced reproductive toxicity in female rats","authors":"Dumebi Anita Konwea , Jerome Ndudi Asiwe , Julian Enwerim Nwangwa , Blessing Zeinab Odili-Ovili , Saviour God’swealth Usin , Eze Kingsley Nwangwa","doi":"10.1016/j.repbio.2025.101168","DOIUrl":"10.1016/j.repbio.2025.101168","url":null,"abstract":"<div><div>Monosodium glutamate (MSG) has been implicated in female reproductive toxicity through endocrine disruption, oxidative stress, inflammation, and apoptosis. This study investigated the protective role of <em>Sorghum bicolor</em> extract (SBE), a polyphenol-rich nutraceutical, against MSG-induced reproductive dysfunction in female Wistar rats, integrating in vivo outcomes with in silico docking analysis. Rats were exposed to MSG (4 g/kg) with or without SBE supplementation (100 mg/kg) orally for 28 days. Estrous cycle, sexual behaviors, hormonal profiles, oxidative and nitrosative stress markers, inflammatory mediators, and apoptotic regulators were assessed. Molecular docking evaluated the interactions of SBE phytochemicals with LHB and LHCGR. MSG disrupted the pituitary–gonadal axis, shortened estrous phases, suppressed reproductive behaviors, increased lipid peroxidation, activated NF-κB, and triggered mitochondrial apoptosis. SBE restored gonadotropins and steroid hormones, normalized estrous cycle, enhanced antioxidant defenses, suppressed inflammation, and prevented apoptosis. Docking analysis revealed strong ligand–protein interactions, supporting endocrine and cellular protection. Conclusively, SBE confers robust gonadoprotective effects against MSG toxicity through endocrine restoration, antioxidant, anti-inflammatory, and anti-apoptotic mechanisms validated <em>in vivo</em> and <em>in silico</em>.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101168"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145679900","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 : 2026-03-01Epub Date: 2025-11-20DOI: 10.1016/j.repbio.2025.101100
Rossella Fucci , Irene Oliva , Arianna Marcellini , Francesca Rizzello , Laura Badolato , Denise De Angelis , Paolo Evangelisti , Elisabetta Baldi , Asia Iserani , Patrizia Falcone , Maria Elisabetta Coccia
To date, the Gardner Grading System remains the gold standard for the conventional evaluation of blastocysts. However, the use of Time-lapse technology in Assisted Reproductive Technologies (ART) cycles provides a dynamic, morphokinetic assessment that complements traditional morphological evaluation, as vitrification and subsequent warming process may induce morphological and biochemical variations in blastocysts. After warming, 203 blastocysts, obtained from homologous ART cycles, were cultured in EmbryoScope to evaluate their morphokinetic modifications using Time-Lapse Technology (TLT). (I) The degree of trophectoderm expansion at insertion into the incubator (t0 exp), (II) the first signal of trophectoderm re-expansion (t first exp), and (III) the degree of expansion and morphology at two hours (t2h exp, t2h grad) were recorded. Statistical analysis was performed by SPSS version 29.0 and relations between I, II, III parameters and beta-human Chorionic Gonadotropin (beta-hCG) outcome, heartbeat, and live birth were assessed.
Results
Showed that t first exp was significantly associated with beta-hCG levels, heartbeat, and live birth outcomes. Additionally, blastocysts achieving complete re-expansion within two hours and presented optimal morphology exhibited a higher likelihood of successful implantation. Early biomarkers were identified through morphokinetic parameters derived from Time-Lapse technology, and pregnancy outcome prediction was improved by the combination of morphological evaluation with post-warming kinetic assessments.
迄今为止，加德纳分级系统仍然是传统囊胚评估的金标准。然而，在辅助生殖技术（ART）周期中使用延时技术提供了一种动态的形态动力学评估，补充了传统的形态学评估，因为玻璃化和随后的升温过程可能会诱导囊胚的形态和生化变化。加热后，从同源ART周期中获得203个囊胚，在EmbryoScope中培养，使用延时技术（Time-Lapse Technology， TLT）评估其形态动力学改变。(I)记录滋养外胚层插入培养箱时的膨胀程度（第0 exp）， （II）滋养外胚层再次膨胀的第一次信号（第t first exp）， （III） 2h时的膨胀程度和形态（第2h exp，第2h grad）。采用SPSS 29.0进行统计分析，评估I、II、III参数与β -人绒毛膜促性腺激素（β - hcg）结局、心跳、活产之间的关系。结果表明，t first exp与β - hcg水平、心跳和活产结局显著相关。此外，囊胚在2小时内完全再膨胀并呈现最佳形态，表明植入成功的可能性更高。通过Time-Lapse技术获得的形态动力学参数来识别早期生物标志物，并通过形态评估和升温后动力学评估相结合来改善妊娠结局预测。
{"title":"Morphokinetic assessment of the blastocyst’s trophectoderm re-expansion post-warming: predictive markers for clinical pregnancy in ART","authors":"Rossella Fucci , Irene Oliva , Arianna Marcellini , Francesca Rizzello , Laura Badolato , Denise De Angelis , Paolo Evangelisti , Elisabetta Baldi , Asia Iserani , Patrizia Falcone , Maria Elisabetta Coccia","doi":"10.1016/j.repbio.2025.101100","DOIUrl":"10.1016/j.repbio.2025.101100","url":null,"abstract":"<div><div>To date, the Gardner Grading System remains the gold standard for the conventional evaluation of blastocysts. However, the use of Time-lapse technology in Assisted Reproductive Technologies (ART) cycles provides a dynamic, morphokinetic assessment that complements traditional morphological evaluation, as vitrification and subsequent warming process may induce morphological and biochemical variations in blastocysts. After warming, 203 blastocysts, obtained from homologous ART cycles, were cultured in EmbryoScope to evaluate their morphokinetic modifications using Time-Lapse Technology (TLT). (I) The degree of trophectoderm expansion at insertion into the incubator (t0 exp), (II) the first signal of trophectoderm re-expansion (t first exp), and (III) the degree of expansion and morphology at two hours (t2h exp, t2h grad) were recorded. Statistical analysis was performed by SPSS version 29.0 and relations between I, II, III parameters and beta-human Chorionic Gonadotropin (beta-hCG) outcome, heartbeat, and live birth were assessed.</div></div><div><h3>Results</h3><div>Showed that t first exp was significantly associated with beta-hCG levels, heartbeat, and live birth outcomes. Additionally, blastocysts achieving complete re-expansion within two hours and presented optimal morphology exhibited a higher likelihood of successful implantation. Early biomarkers were identified through morphokinetic parameters derived from Time-Lapse technology, and pregnancy outcome prediction was improved by the combination of morphological evaluation with post-warming kinetic assessments.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101100"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145568963","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 : 2026-03-01Epub Date: 2025-11-21DOI: 10.1016/j.repbio.2025.101103
José Anderson da Silva Gomes , Renan Gabriel da Silva Ferreira , Jennyfer Martins de Carvalho , Maria Luísa Figueira de Oliveira , Rubem Carlos Araújo Guedes , Elba Verônica Matoso Maciel de Carvalho , Leucio Duarte Vieira Filho , Bruno Mendes Tenorio , Fernanda das Chagas Angelo Mendes Tenorio
The aim of the present study was to investigate the activity of exogenous melatonin as a potential agent in reversing testicular structural damage induced by early weaning. For this purpose, Wistar rats were used and divided into four experimental groups: control; early weaning; early weaning treated with melatonin in a daily dose 10 mg/kg of body weight; and early weaning treated with a vehicle composed of ethanol and saline solution. Except for the control group, all animals were weaned on the 16th day after birth. Body weight was measured weekly, and euthanasia was performed on day 51. The testes were weighed and collected for histopathological, morphometric, immunohistochemical, and oxidative stress analyses, while serum was collected for biochemical and hormonal analyses. At the end of the experiment, the early weaning group exhibited an increase in body mass, as well as structural alterations, including a reduction in the diameter and height of the seminiferous tubule epithelium, along with atrophy, vacuolization, and degeneration of germ cells. This group also showed lower production of pachytene spermatocytes, Sertoli cells, and Leydig cells, increased oxidative stress, decreased testosterone concentration, an adverse lipid profile, and reduced PCNA labeling. In contrast, the group treated with melatonin demonstrated improvement in these parameters when compared to early weaning group. These findings suggest that melatonin exerts a restorative role in testicular tissue.
{"title":"Effect of exogenous melatonin on the testes of Wistar rats undergoing early weaning","authors":"José Anderson da Silva Gomes , Renan Gabriel da Silva Ferreira , Jennyfer Martins de Carvalho , Maria Luísa Figueira de Oliveira , Rubem Carlos Araújo Guedes , Elba Verônica Matoso Maciel de Carvalho , Leucio Duarte Vieira Filho , Bruno Mendes Tenorio , Fernanda das Chagas Angelo Mendes Tenorio","doi":"10.1016/j.repbio.2025.101103","DOIUrl":"10.1016/j.repbio.2025.101103","url":null,"abstract":"<div><div>The aim of the present study was to investigate the activity of exogenous melatonin as a potential agent in reversing testicular structural damage induced by early weaning. For this purpose, Wistar rats were used and divided into four experimental groups: control; early weaning; early weaning treated with melatonin in a daily dose 10 mg/kg of body weight; and early weaning treated with a vehicle composed of ethanol and saline solution. Except for the control group, all animals were weaned on the 16th day after birth. Body weight was measured weekly, and euthanasia was performed on day 51. The testes were weighed and collected for histopathological, morphometric, immunohistochemical, and oxidative stress analyses, while serum was collected for biochemical and hormonal analyses. At the end of the experiment, the early weaning group exhibited an increase in body mass, as well as structural alterations, including a reduction in the diameter and height of the seminiferous tubule epithelium, along with atrophy, vacuolization, and degeneration of germ cells. This group also showed lower production of pachytene spermatocytes, Sertoli cells, and Leydig cells, increased oxidative stress, decreased testosterone concentration, an adverse lipid profile, and reduced PCNA labeling. In contrast, the group treated with melatonin demonstrated improvement in these parameters when compared to early weaning group. These findings suggest that melatonin exerts a restorative role in testicular tissue.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101103"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145568961","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 : 2026-03-01Epub Date: 2025-11-20DOI: 10.1016/j.repbio.2025.101096
Chenming Zhang , Xiaofei Han , Yifei Wang , Ruimin Ma , Sicheng Ma , Wenbang Liu , Zhe Chang , Zixue Sun
Polystyrene microplastics (PS-MPs) are extensively utilized in plastic goods worldwide. The ingestion of PS-MPs has resulted in a high rate of DNA fragmentation index (DFI), which can potentially result in infertility and recurrent spontaneous abortion. This study established and characterized a mouse model of polystyrene microplastic (PS-MP)-induced sperm DNA damage (DnaD), and concurrently analyzed the associated transcriptomic and proteomic profiles. Over a period of 60 days, male mice assigned to the PS group were given PS-MPs at a dose of 1 mg/kg/d while the control group was administered an equivalent volume of normal saline. Sperm DNA Fragmentation Index (DFI) was then assessed using the Sperm Chromatin Structure Assay (SCSA).The testis was examined using RNA-seq and data-independent acquisition (DIA) to detect the patterns of mRNA and protein expression. The PS group exhibited an significant increase in the sperm DFI. Compared with the control group, 874 differentially expressed genes (DEGs) and 164 differentially expressed proteins (DEPs) were identified in the PS group. These included Agt, Gstt1, Fetub, Akr1c12, Eln, Gaa, Ppic and Ltbp2. The PI3K/Akt and metabolic pathways exhibited significant enrichment of these genes. After a 60-day period of intragastric injection, our findings indicated that the administration of PS-MPs at a 1 mg/kg/d dosage can lead to DnaD in the sperm of male mice. The metabolic and PI3K/Akt signaling pathways could be associated with the reproductive toxicity of PS-MPs.
Summary sentence
The intake of PS-MPs mainly reduces DFI in mice via the metabolic and PI3K/Akt signaling pathways.
{"title":"The mouse model of induced sperm DNA damage caused by polystyrene microplastics exhibited distinct transcriptomic and proteomic features","authors":"Chenming Zhang , Xiaofei Han , Yifei Wang , Ruimin Ma , Sicheng Ma , Wenbang Liu , Zhe Chang , Zixue Sun","doi":"10.1016/j.repbio.2025.101096","DOIUrl":"10.1016/j.repbio.2025.101096","url":null,"abstract":"<div><div>Polystyrene microplastics (PS-MPs) are extensively utilized in plastic goods worldwide. The ingestion of PS-MPs has resulted in a high rate of DNA fragmentation index (DFI), which can potentially result in infertility and recurrent spontaneous abortion. This study established and characterized a mouse model of polystyrene microplastic (PS-MP)-induced sperm DNA damage (DnaD), and concurrently analyzed the associated transcriptomic and proteomic profiles. Over a period of 60 days, male mice assigned to the PS group were given PS-MPs at a dose of 1 mg/kg/d while the control group was administered an equivalent volume of normal saline. Sperm DNA Fragmentation Index (DFI) was then assessed using the Sperm Chromatin Structure Assay (SCSA).The testis was examined using RNA-seq and data-independent acquisition (DIA) to detect the patterns of mRNA and protein expression. The PS group exhibited an significant increase in the sperm DFI. Compared with the control group, 874 differentially expressed genes (DEGs) and 164 differentially expressed proteins (DEPs) were identified in the PS group. These included Agt, Gstt1, Fetub, Akr1c12, Eln, Gaa, Ppic and Ltbp2. The PI3K/Akt and metabolic pathways exhibited significant enrichment of these genes. After a 60-day period of intragastric injection, our findings indicated that the administration of PS-MPs at a 1 mg/kg/d dosage can lead to DnaD in the sperm of male mice. The metabolic and PI3K/Akt signaling pathways could be associated with the reproductive toxicity of PS-MPs.</div></div><div><h3>Summary sentence</h3><div>The intake of PS-MPs mainly reduces DFI in mice via the metabolic and PI3K/Akt signaling pathways.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101096"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145568960","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}
Endometriosis is a condition in which functional endometrial glands and stroma are found to grow outside the uterine cavity that can lead to symptoms like dysmenorrhea, dyspareunia, adhesions, and infertility. Current treatment strategies can provide only symptomatic relief as its etiology remains unclear. Several studies have linked the cellular process of epithelial–mesenchymal transition (EMT) to endometriosis development; however, what triggers EMT states in endometriosis is unknown. Polycomb group (PcG) proteins are histone modifiers that control gene expression by catalyzing repressive histone modifications such as H3K27me3/2 and H2AK119ub1. The misexpression of PcGs and/or genes controlled by them reportedly causes several types of cancers, such as breast, colon, pancreatic, and liver. We investigated whether dysregulation of PcG proteins such as RING1B, BMI1, and EZH2 in ectopic endometriotic tissue as compared to eutopic correlates with genes required for EMT in endometriotic tissue from the same patient with laparoscopically confirmed endometriosis. We quantified the expression of Polycomb repressive complex 1 (PRC1) genes (RING1B and BMI1), EMT-associated genes (TWIST, SNAI1, SNAI2, ZEB1, CDH1, CDH2, and VIM) in paired eutopic and ectopic (endometriotic) tissue samples obtained from 12 women who underwent laparoscopy for severe endometriosis identified as per #ENZIAN classification. Our results showed that the endometriotic lesions had higher gene expression of PRC1 proteins – RING1B and BMI1 as well as higher expression of EMT associated genes than the endometrial tissue. Thus, our results suggest that PRC1 proteins may have a role in the pathogenesis of endometriosis.
子宫内膜异位症是一种功能正常的子宫内膜腺体和间质生长在子宫腔外的情况，可导致痛经、性交困难、粘连和不孕等症状。目前的治疗策略只能提供症状缓解，因为其病因尚不清楚。一些研究将上皮-间质转化（EMT）的细胞过程与子宫内膜异位症的发展联系起来；然而，是什么触发了子宫内膜异位症的EMT状态尚不清楚。Polycomb group （PcG）蛋白是组蛋白修饰因子，通过催化抑制组蛋白修饰如H3K27me3/2和H2AK119ub1来控制基因表达。据报道，PcGs和/或由PcGs控制的基因的错误表达会导致几种类型的癌症，如乳腺癌、结肠癌、胰腺癌和肝癌。我们研究了与异位子宫内膜异位症患者相比，异位子宫内膜异位症组织中PcG蛋白如RING1B、BMI1和EZH2的失调是否与子宫内膜异位症组织中EMT所需的基因相关。我们量化了从12名因严重子宫内膜异位症接受腹腔镜检查的妇女中获得的配对异位和异位（子宫内膜异位症）组织样本中Polycomb抑制复合体1 （PRC1）基因（RING1B和BMI1）、emt相关基因（TWIST、SNAI1、SNAI2、ZEB1、CDH1、CDH2和VIM）的表达。我们的研究结果表明，与子宫内膜组织相比，子宫内膜异位症病变中PRC1蛋白- RING1B和BMI1基因表达较高，EMT相关基因表达较高。因此，我们的研究结果表明PRC1蛋白可能在子宫内膜异位症的发病机制中起作用。
{"title":"Correlation between Polycomb repressive complex proteins and epithelial–mesenchymal transition-associated genes in endometriotic tissues","authors":"Anushree Nandan , Abhishek Mangeshikar , Vaijayanti Kale , Anuradha Vaidya , Prasad Pethe","doi":"10.1016/j.repbio.2025.101173","DOIUrl":"10.1016/j.repbio.2025.101173","url":null,"abstract":"<div><div>Endometriosis is a condition in which functional endometrial glands and stroma are found to grow outside the uterine cavity that can lead to symptoms like dysmenorrhea, dyspareunia, adhesions, and infertility. Current treatment strategies can provide only symptomatic relief as its etiology remains unclear. Several studies have linked the cellular process of epithelial–mesenchymal transition (EMT) to endometriosis development; however, what triggers EMT states in endometriosis is unknown. Polycomb group (PcG) proteins are histone modifiers that control gene expression by catalyzing repressive histone modifications such as H3K27me3/2 and H2AK119ub1. The misexpression of PcGs and/or genes controlled by them reportedly causes several types of cancers, such as breast, colon, pancreatic, and liver. We investigated whether dysregulation of PcG proteins such as RING1B, BMI1, and EZH2 in ectopic endometriotic tissue as compared to eutopic correlates with genes required for EMT in endometriotic tissue from the same patient with laparoscopically confirmed endometriosis. We quantified the expression of Polycomb repressive complex 1 (PRC1) genes (<em>RING1B</em> and <em>BMI1</em>), EMT-associated genes (<em>TWIST</em>, <em>SNAI1</em>, <em>SNAI2</em>, <em>ZEB1</em>, <em>CDH1</em>, <em>CDH2,</em> and <em>VIM</em>) in paired eutopic and ectopic (endometriotic) tissue samples obtained from 12 women who underwent laparoscopy for severe endometriosis identified as per #ENZIAN classification. Our results showed that the endometriotic lesions had higher gene expression of PRC1 proteins – <em>RING1B</em> and <em>BMI1</em> as well as higher expression of EMT associated genes than the endometrial tissue. Thus, our results suggest that PRC1 proteins may have a role in the pathogenesis of endometriosis.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101173"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145786492","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 : 2026-03-01Epub Date: 2025-10-27DOI: 10.1016/j.repbio.2025.101091
Huijia Jin , Yujiao Guo , Yanshe Xie , Xin Xiang , Zhengguang Wang , Jizhong Xiang
The establishment of endometrial receptivity is required for successful embryo implantation during early pregnancy in many mammals. Extracellular vesicles (EVs) microRNAs (miRNAs) play an important role during embryo implantation. While their roles have been characterized in other species, the specific functions of trophoblast-derived EVs miRNAs in ovine endometrial receptivity remain undefined. In this study, we systematically investigated the effects of ovine placental trophoblast (OTR) cells-derived EVs on ovine endometrial epithelial cells (EECs) by cck-8 assay, EdU assay, cell migration assay, RT-qPCR and ultrastructural examination of apical plasma membranes. Subsequently, miRNA expression profiles of EV-treated EECs were identified and analyzed by miRNA-Seq. The results showed that OTR cells-derived EVs were taken up by EECs, enhancing the migration of EECs. EVs treatment reduced microvilli on the apical plasma membranes of EECs. The expression of genes involved in endometrial receptivity increased. OTR cells-derived EVs induced changes consistent with a receptive phenotype through coordinated cellular remodeling and gene expression changes. The miRNA-Seq results revealed 287 detectable miRNAs, including 34 with significant differential expression (20 upregulated and 14 downregulated) in EV-treated versus control EECs. The predicted target genes of these differentially expressed miRNAs were enriched in signaling pathways regulating embryo implantation and endometrial receptivity, such as MAPK, Toll-like receptor, adherens junction and focal adhesion. Our in vitro findings suggested that OTR cells-derived EVs may promote endometrial receptivity by facilitating the transformation of EECs, as indicated by receptivity-associated morphological and molecular changes. It provided novel insights for improving successful pregnancy rate in sheep.
{"title":"Extracellular vesicles derived from placental trophoblast regulate ovine endometrial receptivity by promoting the transformation of endometrial epithelial cells","authors":"Huijia Jin , Yujiao Guo , Yanshe Xie , Xin Xiang , Zhengguang Wang , Jizhong Xiang","doi":"10.1016/j.repbio.2025.101091","DOIUrl":"10.1016/j.repbio.2025.101091","url":null,"abstract":"<div><div>The establishment of endometrial receptivity is required for successful embryo implantation during early pregnancy in many mammals. Extracellular vesicles (EVs) microRNAs (miRNAs) play an important role during embryo implantation. While their roles have been characterized in other species, the specific functions of trophoblast-derived EVs miRNAs in ovine endometrial receptivity remain undefined. In this study, we systematically investigated the effects of ovine placental trophoblast (OTR) cells-derived EVs on ovine endometrial epithelial cells (EECs) by cck-8 assay, EdU assay, cell migration assay, RT-qPCR and ultrastructural examination of apical plasma membranes. Subsequently, miRNA expression profiles of EV-treated EECs were identified and analyzed by miRNA-Seq. The results showed that OTR cells-derived EVs were taken up by EECs, enhancing the migration of EECs. EVs treatment reduced microvilli on the apical plasma membranes of EECs. The expression of genes involved in endometrial receptivity increased. OTR cells-derived EVs induced changes consistent with a receptive phenotype through coordinated cellular remodeling and gene expression changes. The miRNA-Seq results revealed 287 detectable miRNAs, including 34 with significant differential expression (20 upregulated and 14 downregulated) in EV-treated versus control EECs. The predicted target genes of these differentially expressed miRNAs were enriched in signaling pathways regulating embryo implantation and endometrial receptivity, such as MAPK, Toll-like receptor, adherens junction and focal adhesion. Our in vitro findings suggested that OTR cells-derived EVs may promote endometrial receptivity by facilitating the transformation of EECs, as indicated by receptivity-associated morphological and molecular changes. It provided novel insights for improving successful pregnancy rate in sheep.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101091"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145395682","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 : 2026-03-01Epub Date: 2025-12-17DOI: 10.1016/j.repbio.2025.101170
Higor da Silva Ferreira , Gabriella Borba de Oliveira , Eduardo de Oliveira Sanguinet , Ana Laura da Silva Feijó , César Augusto Pinzón-Osorio , Verônica Rafaela Benvenutti , Louise Fontoura Köhler , Marianna Bertolini , Fernando Almeida-Souza , Felipe de Jesus Moraes Junior , David Driemeier , Ana Lúcia Abreu-Silva , José Luiz Rigo Rodrigues , Marcelo Bertolini
Seminal somatic cells (SSCs) represent promising donor nuclei for cloning by somatic cell nuclear transfer (SCNT), particularly for genetic conservation. This study aimed to identify and characterize SSCs present in bovine semen, determine their possible tissue origins, and evaluate their suitability for cloning. SSCs were isolated using sucrose or mini-Percoll® density gradients and analyzed for morphology, viability, and cell cycle status, as well as their potential use in SCNT cloning. Cytological and histological assessments of the bovine genitourinary tract were performed side-by-side to identify the anatomical origins of each SSC type, and epithelial marker expression was examined by RT-qPCR in SSCs, urethral, and bladder tissues. The mini-Percoll® gradient efficiently separated SSCs from sperm cells, concentrating viable somatic cells in the 20 % fraction. Three SSC types were identified (polygonal, round, and elongated), with polygonal cells being the most abundant and highly viable (>90 %), although they failed to adhere or proliferate in culture. Flow cytometry revealed that most polygonal SSCs (89 %) were arrested at the G0/G1 phase, indicating cell-cycle compatibility for cloning, although with low membrane fusion capacity in SCNT assays. Cytological, histological, and molecular findings collectively indicate that polygonal SSCs most likely originate from the non-keratinized stratified epithelium of the urethral navicular fossa, whereas round and elongated SSCs may derive from the proximal urethra and the ampullary-vesicular-ductal complex, respectively. These results provide a comprehensive anatomical and molecular characterization of SSC subtypes in bovine semen, offering a foundation for improving SCNT cloning standardization and applications in genetic preservation.
{"title":"Origin, viability, and cell cycle status of seminal somatic cells for potential application in cattle cloning via nuclear transfer","authors":"Higor da Silva Ferreira , Gabriella Borba de Oliveira , Eduardo de Oliveira Sanguinet , Ana Laura da Silva Feijó , César Augusto Pinzón-Osorio , Verônica Rafaela Benvenutti , Louise Fontoura Köhler , Marianna Bertolini , Fernando Almeida-Souza , Felipe de Jesus Moraes Junior , David Driemeier , Ana Lúcia Abreu-Silva , José Luiz Rigo Rodrigues , Marcelo Bertolini","doi":"10.1016/j.repbio.2025.101170","DOIUrl":"10.1016/j.repbio.2025.101170","url":null,"abstract":"<div><div>Seminal somatic cells (SSCs) represent promising donor nuclei for cloning by somatic cell nuclear transfer (SCNT), particularly for genetic conservation. This study aimed to identify and characterize SSCs present in bovine semen, determine their possible tissue origins, and evaluate their suitability for cloning. SSCs were isolated using sucrose or mini-Percoll® density gradients and analyzed for morphology, viability, and cell cycle status, as well as their potential use in SCNT cloning. Cytological and histological assessments of the bovine genitourinary tract were performed side-by-side to identify the anatomical origins of each SSC type, and epithelial marker expression was examined by RT-qPCR in SSCs, urethral, and bladder tissues. The mini-Percoll® gradient efficiently separated SSCs from sperm cells, concentrating viable somatic cells in the 20 % fraction. Three SSC types were identified (polygonal, round, and elongated), with polygonal cells being the most abundant and highly viable (>90 %), although they failed to adhere or proliferate in culture. Flow cytometry revealed that most polygonal SSCs (89 %) were arrested at the G0/G1 phase, indicating cell-cycle compatibility for cloning, although with low membrane fusion capacity in SCNT assays. Cytological, histological, and molecular findings collectively indicate that polygonal SSCs most likely originate from the non-keratinized stratified epithelium of the urethral navicular fossa, whereas round and elongated SSCs may derive from the proximal urethra and the ampullary-vesicular-ductal complex, respectively. These results provide a comprehensive anatomical and molecular characterization of SSC subtypes in bovine semen, offering a foundation for improving SCNT cloning standardization and applications in genetic preservation.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101170"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145783948","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 : 2026-03-01Epub Date: 2025-11-25DOI: 10.1016/j.repbio.2025.101167
Huamei Ju , Ziliang Geng , Binyan Chen , Yuwei Shang , Xia Chen , Danni Wang , Wenbin Wang , Huiting Sun , Yichao Shi , Jiajun Yu
The heterotrimeric complex serine-threonine protein phosphatase 2 A (PP2A) is integral to the regulation of essential cellular processes. It is particularly crucial in spermatogenesis, where it is indispensable for meiosis, mitosis, sperm capacitation, and apoptosis. Previous research has concentrated on the knockdown of the catalytic subunit of PP2A, PPP2CA, in germ cells using Ddx4-Cre, resulting in male mouse sterility, disrupted meiotic recombination, and meiotic arrest of spermatocytes. To further elucidate the role of PP2A in spermatogenesis, we performed transcriptomic and proteomic sequencing analyses on the testes of knockout and control mice. A nine-quadrant map was developed to depict the differential expression of genes and proteins. Our analyses identified 1732 differentially expressed genes, which exhibited a strong positive correlation with the trends in differential protein expression. Gene Ontology (GO) enrichment analysis indicated a significant downregulation of genes involved in spermatogenesis, sperm cell development, and sperm cell differentiation. Furthermore, KEGG enrichment analysis revealed a notable enrichment of differentially expressed genes within the ubiquitin-mediated proteolysis pathway. In knockout mouse testicular tissue, testicular expression of the ubiquitin-related gene, UBE2K, was markedly downregulated, which was associated with the accumulation of histone H3, upregulation of the methyltransferase SETDB1, and increased levels of H3K9me3. Similarly, knockdown of Ppp2ca in GC2 cells resulted in decreased UBE2K expression, histone H3 accumulation, SETDB1 upregulation, and elevated H3K9me3 levels, consistent with mirroring the phenotype observed in the knockout mice. Notably, the ubiquitin-related gene UBE2K was identified as a significant outlier in the nine-quadrant map, and real-time quantitative PCR confirmed that UBE2K transcript levels were significantly reduced in knockout mice compared to wild-type controls. These findings suggest that PP2A may regulate histon.
异三聚体复合体丝氨酸-苏氨酸蛋白磷酸酶2 A （PP2A）是必不可少的细胞过程的调节。它在精子发生中尤其重要，在那里它对减数分裂、有丝分裂、精子获能和细胞凋亡是必不可少的。先前的研究主要集中在使用Ddx4-Cre敲除生殖细胞中PP2A的催化亚基PPP2CA，导致雄性小鼠不育、减数分裂重组中断和精母细胞减数分裂停滞。为了进一步阐明PP2A在精子发生中的作用，我们对敲除小鼠和对照小鼠的睾丸进行了转录组学和蛋白质组学测序分析。一个九象限的图谱被用来描绘基因和蛋白质的差异表达。我们的分析确定了1732个差异表达基因，这些基因与差异蛋白表达趋势表现出很强的正相关。基因本体（GO）富集分析表明，参与精子发生、精子细胞发育和精子细胞分化的基因显著下调。此外，KEGG富集分析显示，在泛素介导的蛋白水解途径中，差异表达基因显著富集。在敲除小鼠睾丸组织中，睾丸中泛素相关基因UBE2K的表达明显下调，这与组蛋白H3的积累、甲基转移酶SETDB1的上调以及H3K9me3水平的升高有关。同样，在GC2细胞中敲低Ppp2ca导致UBE2K表达降低，组蛋白H3积累，SETDB1上调，H3K9me3水平升高，与敲除小鼠的表型一致。值得注意的是，泛素相关基因UBE2K在九象限图谱中被鉴定为显著异常值，实时定量PCR证实，与野生型对照相比，敲除小鼠的UBE2K转录水平显著降低。这些发现表明PP2A可能调节组蛋白。
{"title":"Mechanism of PP2A affecting ubiquitination pathway in spermatogenesis","authors":"Huamei Ju , Ziliang Geng , Binyan Chen , Yuwei Shang , Xia Chen , Danni Wang , Wenbin Wang , Huiting Sun , Yichao Shi , Jiajun Yu","doi":"10.1016/j.repbio.2025.101167","DOIUrl":"10.1016/j.repbio.2025.101167","url":null,"abstract":"<div><div>The heterotrimeric complex serine-threonine protein phosphatase 2 A (PP2A) is integral to the regulation of essential cellular processes. It is particularly crucial in spermatogenesis, where it is indispensable for meiosis, mitosis, sperm capacitation, and apoptosis. Previous research has concentrated on the knockdown of the catalytic subunit of PP2A, PPP2CA, in germ cells using Ddx4-Cre, resulting in male mouse sterility, disrupted meiotic recombination, and meiotic arrest of spermatocytes. To further elucidate the role of PP2A in spermatogenesis, we performed transcriptomic and proteomic sequencing analyses on the testes of knockout and control mice. A nine-quadrant map was developed to depict the differential expression of genes and proteins. Our analyses identified 1732 differentially expressed genes, which exhibited a strong positive correlation with the trends in differential protein expression. Gene Ontology (GO) enrichment analysis indicated a significant downregulation of genes involved in spermatogenesis, sperm cell development, and sperm cell differentiation. Furthermore, KEGG enrichment analysis revealed a notable enrichment of differentially expressed genes within the ubiquitin-mediated proteolysis pathway. In knockout mouse testicular tissue, testicular expression of the ubiquitin-related gene, UBE2K, was markedly downregulated, which was associated with the accumulation of histone H3, upregulation of the methyltransferase SETDB1, and increased levels of H3K9me3. Similarly, knockdown of Ppp2ca in GC2 cells resulted in decreased UBE2K expression, histone H3 accumulation, SETDB1 upregulation, and elevated H3K9me3 levels, consistent with mirroring the phenotype observed in the knockout mice. Notably, the ubiquitin-related gene UBE2K was identified as a significant outlier in the nine-quadrant map, and real-time quantitative PCR confirmed that UBE2K transcript levels were significantly reduced in knockout mice compared to wild-type controls. These findings suggest that PP2A may regulate histon.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101167"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145614450","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 : 2026-03-01Epub Date: 2025-11-26DOI: 10.1016/j.repbio.2025.101166
Mehmet Can Nacar , Recai Aci̇ , Şengül Tural , Serbülent Yi̇ği̇t
This study aimed to investigate the association between the SOD1 gene I>D polymorphism, a key component of the antioxidant defense system, and polycystic ovary syndrome (PCOS), as well as to evaluate the potential effects of this genetic variation on genotypic distribution and metabolic parameters. A total of 100 PCOS patients and 135 healthy controls were included, and genomic DNA extracted from peripheral blood was analyzed for the SOD1 I>D polymorphism using conventional PCR, while clinical and biochemical parameters such as body mass index (BMI), insulin levels, HOMA-IR, FSH, LH, and other hormone levels were assessed. The ID genotype was found to be less frequent in the PCOS group compared to controls, with borderline statistical significance (p = 0.050; OR = 0.57), and was significantly less common in PCOS patients with a positive family history (p = 0.013), suggesting a potential protective effect, while correlations were observed between the ID genotype and markers of insulin resistance, though no significant differences were detected in allele frequencies. These findings indicate that the SOD1 gene I>D polymorphism may influence both susceptibility and metabolic characteristics of PCOS, supporting the role of oxidative stress in its pathogenesis and emphasizing the need for further functional and large-scale studies to confirm these associations.
{"title":"Genotypic and metabolic impact of the SOD1 I>D polymorphism in polycystic ovary syndrome","authors":"Mehmet Can Nacar , Recai Aci̇ , Şengül Tural , Serbülent Yi̇ği̇t","doi":"10.1016/j.repbio.2025.101166","DOIUrl":"10.1016/j.repbio.2025.101166","url":null,"abstract":"<div><div>This study aimed to investigate the association between the SOD1 gene I>D polymorphism, a key component of the antioxidant defense system, and polycystic ovary syndrome (PCOS), as well as to evaluate the potential effects of this genetic variation on genotypic distribution and metabolic parameters. A total of 100 PCOS patients and 135 healthy controls were included, and genomic DNA extracted from peripheral blood was analyzed for the SOD1 I>D polymorphism using conventional PCR, while clinical and biochemical parameters such as body mass index (BMI), insulin levels, HOMA-IR, FSH, LH, and other hormone levels were assessed. The ID genotype was found to be less frequent in the PCOS group compared to controls, with borderline statistical significance (p = 0.050; OR = 0.57), and was significantly less common in PCOS patients with a positive family history (p = 0.013), suggesting a potential protective effect, while correlations were observed between the ID genotype and markers of insulin resistance, though no significant differences were detected in allele frequencies. These findings indicate that the SOD1 gene I>D polymorphism may influence both susceptibility and metabolic characteristics of PCOS, supporting the role of oxidative stress in its pathogenesis and emphasizing the need for further functional and large-scale studies to confirm these associations.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101166"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145614579","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 : 2026-03-01Epub Date: 2025-11-01DOI: 10.1016/j.repbio.2025.101097
Haris Setiawan , Agung Irawan
Animal cloning remains inefficient, with live birth rates below 5 % in most mammalian species, slightly higher in bovines (20–25 %). Among various factors, nuclear donor cells play crucial roles, and their passage numbers may influence cloning efficiency. However, a definitive association between passage numbers and outcomes in cloning remains inconclusive due to insufficient data and inconsistent results. In the present meta-analytical study, we compared research using high (>6) and low (≤6) cell passage numbers across various types, breeds, and sexes of donor cells to assess their impact on embryo development and associated gene expressions in bovine clones. Our findings revealed that cell passaging influences developmental competence at the cleavage and blastocyst rates, with lower passage numbers yielding better results. Higher acetylation of H3K9 in low-passage cells is consistently associated with improved developmental competence through the blastocyst stage, although the difference was not statistically significant. Donor cells with higher histone acetylation may undergo reprogramming more easily and completely, thus improving cloning efficiency. Further analysis elucidated that the types and breeds of donor cells also affected the blastocyst outcome. Nevertheless, high heterogeneity and meta-bias were identified in the meta-analytical outcomes, particularly in cleavage, 2-cell, 8-cell, blastocyst, the total cell number (TCN), the ratio of inner cell mass and total cell number (ICM/TCN), NANOG, and birth rate, which may contribute to inconsistencies in embryo quality results and hinder comparisons between development-related gene expressions and the embryo transfer outcome.
{"title":"Influence of different cell passage numbers on bovine cloned embryo: A systematic review and meta-analysis","authors":"Haris Setiawan , Agung Irawan","doi":"10.1016/j.repbio.2025.101097","DOIUrl":"10.1016/j.repbio.2025.101097","url":null,"abstract":"<div><div>Animal cloning remains inefficient, with live birth rates below 5 % in most mammalian species, slightly higher in bovines (20–25 %). Among various factors, nuclear donor cells play crucial roles, and their passage numbers may influence cloning efficiency. However, a definitive association between passage numbers and outcomes in cloning remains inconclusive due to insufficient data and inconsistent results. In the present meta-analytical study, we compared research using high (>6) and low (≤6) cell passage numbers across various types, breeds, and sexes of donor cells to assess their impact on embryo development and associated gene expressions in bovine clones. Our findings revealed that cell passaging influences developmental competence at the cleavage and blastocyst rates, with lower passage numbers yielding better results. Higher acetylation of H3K9 in low-passage cells is consistently associated with improved developmental competence through the blastocyst stage, although the difference was not statistically significant. Donor cells with higher histone acetylation may undergo reprogramming more easily and completely, thus improving cloning efficiency. Further analysis elucidated that the types and breeds of donor cells also affected the blastocyst outcome. Nevertheless, high heterogeneity and meta-bias were identified in the meta-analytical outcomes, particularly in cleavage, 2-cell, 8-cell, blastocyst, the total cell number (TCN), the ratio of inner cell mass and total cell number (ICM/TCN), NANOG, and birth rate, which may contribute to inconsistencies in embryo quality results and hinder comparisons between development-related gene expressions and the embryo transfer outcome.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101097"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145418810","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}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号