Pub Date : 2025-12-20DOI: 10.1016/j.repbio.2025.101171
Douglas C. Selle , Raquel S. dos Santos , Thaiza R. de Freitas , Jhony L. Benato , Thales de S. França , Renata V. Dantas , Marco A. Rotta , Marcelle F.O. Barbosa , Vanessa Coimbra , Diógenes H. Siqueira-Silva , Danilo P. Streit Jr.
This study aimed to characterize the reproductive biology of Loricariichthys anus (violinha) from the Guaíba River (Brazil) to support conservation efforts and development of captive reproduction protocols. Monthly collections (November 2021-October 2022) of 144 specimens were analyzed for sexual dimorphism, gonadosomatic index (GSI), gonadal histology, and sperm parameters (motility, concentration, morphology). Sexual dimorphism was evidenced by elongated lower lip in males, predominantly during the reproductive period (November-March). GSI values peaked during November-March for both sexes (P < 0.05). Histological analysis revealed females reached maturity from November to February with evidence of multiple spawning, while males produced sperm year-round, albeit in low volumes. Sperm motility parameters differed significantly between individuals (P < 0.05) with mean concentration of 6.88 × 106 ± 8.18 × 105 spermatozoa/mL. L. anus reproductive period occurs from November to February, with males presenting low seminal volume and viable sperm concentration compared to other Loricariidae species, which has implications for both conservation and captive reproduction efforts.
{"title":"Reproductive biology characterization of violinha Loricariichthys anus","authors":"Douglas C. Selle , Raquel S. dos Santos , Thaiza R. de Freitas , Jhony L. Benato , Thales de S. França , Renata V. Dantas , Marco A. Rotta , Marcelle F.O. Barbosa , Vanessa Coimbra , Diógenes H. Siqueira-Silva , Danilo P. Streit Jr.","doi":"10.1016/j.repbio.2025.101171","DOIUrl":"10.1016/j.repbio.2025.101171","url":null,"abstract":"<div><div>This study aimed to characterize the reproductive biology of <em>Loricariichthys anus</em> (violinha) from the Guaíba River (Brazil) to support conservation efforts and development of captive reproduction protocols. Monthly collections (November 2021-October 2022) of 144 specimens were analyzed for sexual dimorphism, gonadosomatic index (GSI), gonadal histology, and sperm parameters (motility, concentration, morphology). Sexual dimorphism was evidenced by elongated lower lip in males, predominantly during the reproductive period (November-March). GSI values peaked during November-March for both sexes (P < 0.05). Histological analysis revealed females reached maturity from November to February with evidence of multiple spawning, while males produced sperm year-round, albeit in low volumes. Sperm motility parameters differed significantly between individuals (P < 0.05) with mean concentration of 6.88 × 106 ± 8.18 × 105 spermatozoa/mL. <em>L. anus</em> reproductive period occurs from November to February, with males presenting low seminal volume and viable sperm concentration compared to other Loricariidae species, which has implications for both conservation and captive reproduction efforts.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101171"},"PeriodicalIF":2.5,"publicationDate":"2025-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145786586","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 : 2025-12-20DOI: 10.1016/j.repbio.2025.101172
Joel Iván Pacheco , Hugo Wenceslao Deza , Víctor Manuel Vélez-Marroquín , Wilber García , Francisco Franco , Edward H. Cabezas-Garcia , Daniel Marcelo Lombardo
Pregnancy involves characteristics unique to each species, such as the development of fetal membranes and related fluids. In this descriptive exploratory study, a unique third fetal fluid located under the epidermal membrane, in addition to the allantoic and amniotic membranes, were described in alpacas (Vicugna pacos). The aim was to characterize the biochemical composition of fetal fluids as alpaca gestation progressed. Twenty-seven pregnant alpacas were examined; 21 of them were slaughtered at various pregnancy stages to both collect allantoic and amniotic fluids, and six were monitored until parturition to obtain postpartum allantoic fluid. In the last third of pregnancy, an additional fluid was observed between the fetal skin and the epidermal membrane, herein termed as ‘epidermal fluid’. Biochemical analyses were performed via spectrophotometry to measure glucose, triglycerides, cholesterol, total proteins, albumin, hemoglobin, calcium, phosphorus, uric acid, and creatinine. High variability was observed in all metabolites. Allantoic fluid showed increasing levels of uric acid, creatinine, and calcium throughout pregnancy, suggesting fetal renal function, while postpartum fluid showed reduced levels of several metabolites. The amniotic fluid displayed increased triglycerides and cholesterol, and decreased glucose levels at the end of pregnancy. Epidermal fluid exhibited the highest levels of glucose, calcium, and creatinine, and lowest uric acid compared to amniotic fluid, indicating a distinct biochemical composition. This study provides the first comprehensive insight into the biochemical characterization of fetal fluids in alpacas, updating current knowledge of fetal developmental physiology in South American camelids.
{"title":"Biochemical profile of fetal fluids in alpacas (Vicugna pacos) during pregnancy and immediate postpartum","authors":"Joel Iván Pacheco , Hugo Wenceslao Deza , Víctor Manuel Vélez-Marroquín , Wilber García , Francisco Franco , Edward H. Cabezas-Garcia , Daniel Marcelo Lombardo","doi":"10.1016/j.repbio.2025.101172","DOIUrl":"10.1016/j.repbio.2025.101172","url":null,"abstract":"<div><div>Pregnancy involves characteristics unique to each species, such as the development of fetal membranes and related fluids. In this descriptive exploratory study, a unique third fetal fluid located under the epidermal membrane, in addition to the allantoic and amniotic membranes, were described in alpacas (<em>Vicugna pacos</em>). The aim was to characterize the biochemical composition of fetal fluids as alpaca gestation progressed. Twenty-seven pregnant alpacas were examined; 21 of them were slaughtered at various pregnancy stages to both collect allantoic and amniotic fluids, and six were monitored until parturition to obtain postpartum allantoic fluid. In the last third of pregnancy, an additional fluid was observed between the fetal skin and the epidermal membrane, herein termed as ‘epidermal fluid’. Biochemical analyses were performed via spectrophotometry to measure glucose, triglycerides, cholesterol, total proteins, albumin, hemoglobin, calcium, phosphorus, uric acid, and creatinine. High variability was observed in all metabolites. Allantoic fluid showed increasing levels of uric acid, creatinine, and calcium throughout pregnancy, suggesting fetal renal function, while postpartum fluid showed reduced levels of several metabolites. The amniotic fluid displayed increased triglycerides and cholesterol, and decreased glucose levels at the end of pregnancy. Epidermal fluid exhibited the highest levels of glucose, calcium, and creatinine, and lowest uric acid compared to amniotic fluid, indicating a distinct biochemical composition. This study provides the first comprehensive insight into the biochemical characterization of fetal fluids in alpacas, updating current knowledge of fetal developmental physiology in South American camelids.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101172"},"PeriodicalIF":2.5,"publicationDate":"2025-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145786494","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":"2025-12-19","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 : 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":"2025-12-17","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}
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":"2025-12-03","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 : 2025-11-29DOI: 10.1016/j.repbio.2025.101169
Lea Bejstrup Jensen , Laura Victoria Højer , Lisette Schönhage , Cristina Subiran Adrados , Jane Alrø Bøtkjær , Tanni Borgbo , Jesús Cadenas , Kirsten Tryde Macklon , Stine Ringholm , Anette Tønnes Pedersen , Stine Gry Kristensen
Luteinization is a vital process in female reproduction, where granulosa cells differentiate into progesterone-secreting luteal cells. While luteinizing hormone (LH) and human chorionic gonadotropin (hCG) are established promoters of luteinization, the regulatory role of activin A remains incompletely understood. This study aimed to investigate whether activin A can prevent or reverse luteinization in granulosa cells from small antral and preovulatory follicles using an in vitro model. Granulosa cells were collected from women undergoing either IVF treatment or ovarian tissue cryopreservation. Cells were cultured for 48 h with FSH, hCG, or activin A. Hormone secretion (progesterone and estradiol) and content of steroidogenic markers (CYP19A1, STAR, 17βHSD1, 3βHSD2, FSHR, and LHCGR) were analyzed via ELISA, RT-qPCR, and Western blot.
Results
demonstrated that activin A suppressed luteinization markers and progesterone production while promoting estrogen synthesis and maintaining granulosa cell features. Conversely, hCG increased luteinization marker expression and progesterone secretion, consistent with its known role in luteal transformation. Notably, granulosa cells from small antral follicles showed a distinct, stage-specific response to activin A compared to those from preovulatory follicles, indicating a temporal regulation of luteinization competence.
These findings support a regulatory role for activin A in maintaining granulosa cell identity and delaying luteinization. Understanding the differential responsiveness of granulosa cells across follicular development stages may inform new therapeutic approaches for luteal phase deficiencies and fertility preservation strategies.
{"title":"Inhibitory role of activin A during luteinization of follicular and lutein granulosa cells from small antral and preovulatory human follicles","authors":"Lea Bejstrup Jensen , Laura Victoria Højer , Lisette Schönhage , Cristina Subiran Adrados , Jane Alrø Bøtkjær , Tanni Borgbo , Jesús Cadenas , Kirsten Tryde Macklon , Stine Ringholm , Anette Tønnes Pedersen , Stine Gry Kristensen","doi":"10.1016/j.repbio.2025.101169","DOIUrl":"10.1016/j.repbio.2025.101169","url":null,"abstract":"<div><div>Luteinization is a vital process in female reproduction, where granulosa cells differentiate into progesterone-secreting luteal cells. While luteinizing hormone (LH) and human chorionic gonadotropin (hCG) are established promoters of luteinization, the regulatory role of activin A remains incompletely understood. This study aimed to investigate whether activin A can prevent or reverse luteinization in granulosa cells from small antral and preovulatory follicles using an in vitro model. Granulosa cells were collected from women undergoing either IVF treatment or ovarian tissue cryopreservation. Cells were cultured for 48 h with FSH, hCG, or activin A. Hormone secretion (progesterone and estradiol) and content of steroidogenic markers (CYP19A1, STAR, 17βHSD1, 3βHSD2, FSHR, and LHCGR) were analyzed via ELISA, RT-qPCR, and Western blot.</div></div><div><h3>Results</h3><div>demonstrated that activin A suppressed luteinization markers and progesterone production while promoting estrogen synthesis and maintaining granulosa cell features. Conversely, hCG increased luteinization marker expression and progesterone secretion, consistent with its known role in luteal transformation. Notably, granulosa cells from small antral follicles showed a distinct, stage-specific response to activin A compared to those from preovulatory follicles, indicating a temporal regulation of luteinization competence.</div><div>These findings support a regulatory role for activin A in maintaining granulosa cell identity and delaying luteinization. Understanding the differential responsiveness of granulosa cells across follicular development stages may inform new therapeutic approaches for luteal phase deficiencies and fertility preservation strategies.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101169"},"PeriodicalIF":2.5,"publicationDate":"2025-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145650690","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 : 2025-11-29DOI: 10.1016/j.repbio.2025.101105
Zhiqin Zhang , Xuechen Sun , Xin Li , Peipei Liu , Liyun Cao , Shenggen Long , Jun Tan
This study examined pannexin 1 (PANX1) expression in ovarian granulosa cells of women with advanced maternal age and its role in cell proliferation and apoptosis, aiming to clarify mechanisms of age-related follicular dysplasia. Ninety women undergoing assisted reproductive technology were divided into reproductive-age (<35 years), advanced-age (35–41 years), and very advanced-age (≥42 years) groups. Clinical data and granulosa cell function were analyzed. PANX1 expression was detected in primary cells, while KGN cells were transfected with PANX1 plasmid or siRNA. Cell proliferation, apoptosis, and extracellular ATP levels were evaluated. With increasing age, oocyte yield, blastocyst formation, and pregnancy rates declined, granulosa cell proliferation decreased, apoptosis increased, and PANX1 expression was elevated. PANX1 overexpression inhibited proliferation and increased extracellular ATP, whereas knockdown enhanced proliferation without affecting apoptosis. PANX1 upregulation in aging granulosa cells mediates ATP efflux, depletes intracellular ATP, and suppresses proliferation, contributing to abnormal follicular development and reduced fertility.
{"title":"Downregulation of intracellular ATP levels by PANX1 inhibits ovarian granulosa cell proliferation and mediates follicular dysplasia in elderly women","authors":"Zhiqin Zhang , Xuechen Sun , Xin Li , Peipei Liu , Liyun Cao , Shenggen Long , Jun Tan","doi":"10.1016/j.repbio.2025.101105","DOIUrl":"10.1016/j.repbio.2025.101105","url":null,"abstract":"<div><div>This study examined pannexin 1 (PANX1) expression in ovarian granulosa cells of women with advanced maternal age and its role in cell proliferation and apoptosis, aiming to clarify mechanisms of age-related follicular dysplasia. Ninety women undergoing assisted reproductive technology were divided into reproductive-age (<35 years), advanced-age (35–41 years), and very advanced-age (≥42 years) groups. Clinical data and granulosa cell function were analyzed. PANX1 expression was detected in primary cells, while KGN cells were transfected with PANX1 plasmid or siRNA. Cell proliferation, apoptosis, and extracellular ATP levels were evaluated. With increasing age, oocyte yield, blastocyst formation, and pregnancy rates declined, granulosa cell proliferation decreased, apoptosis increased, and PANX1 expression was elevated. PANX1 overexpression inhibited proliferation and increased extracellular ATP, whereas knockdown enhanced proliferation without affecting apoptosis. PANX1 upregulation in aging granulosa cells mediates ATP efflux, depletes intracellular ATP, and suppresses proliferation, contributing to abnormal follicular development and reduced fertility.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101105"},"PeriodicalIF":2.5,"publicationDate":"2025-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145650664","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 : 2025-11-26DOI: 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":"2025-11-26","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 : 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":"2025-11-25","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 : 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":"2025-11-21","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}
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