Pub Date : 2025-12-31DOI: 10.1016/j.repbio.2025.101175
Marwah Suliman Maashi
By transmitting bioactive substances, including proteins, lipids, and a varied spectrum of non-coding RNAs, seminal exosomes—including epididymosomes and prostasomes—are shown to be crucial in sperm maturation, motility, and protection. The paper emphasizes how miRNAs, lncRNAs, and circRNAs within these exosomes affect gene expression during spermatogenesis and are starting to be interesting non-invasive biomarkers for several infertility abnormalities, including azoospermia and asthenozoospermia. It also addresses the therapeutic possibilities of exosome-based approaches in reducing inflammation, oxidative stress, and cellular damage in the male reproductive system. The paper identifies essential information gaps and suggests future research paths to clarify the molecular processes behind male infertility by combining knowledge from proteomic, lipidomic, and transcriptomic investigations. These results help to close the knowledge gap on seminal exosomes and open the door for creative therapeutic and diagnostic tools in reproductive medicine.
{"title":"Exosomal ncRNAs in seminal fluid: Unraveling their regulatory roles in male infertility","authors":"Marwah Suliman Maashi","doi":"10.1016/j.repbio.2025.101175","DOIUrl":"10.1016/j.repbio.2025.101175","url":null,"abstract":"<div><div>By transmitting bioactive substances, including proteins, lipids, and a varied spectrum of non-coding RNAs, seminal exosomes—including epididymosomes and prostasomes—are shown to be crucial in sperm maturation, motility, and protection. The paper emphasizes how miRNAs, lncRNAs, and circRNAs within these exosomes affect gene expression during spermatogenesis and are starting to be interesting non-invasive biomarkers for several infertility abnormalities, including azoospermia and asthenozoospermia. It also addresses the therapeutic possibilities of exosome-based approaches in reducing inflammation, oxidative stress, and cellular damage in the male reproductive system. The paper identifies essential information gaps and suggests future research paths to clarify the molecular processes behind male infertility by combining knowledge from proteomic, lipidomic, and transcriptomic investigations. These results help to close the knowledge gap on seminal exosomes and open the door for creative therapeutic and diagnostic tools in reproductive medicine.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101175"},"PeriodicalIF":2.5,"publicationDate":"2025-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145883682","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}
Bovine spermatozoa are highly vulnerable to oxidative stress (OS), which can impair motility, viability, membrane integrity, and DNA integrity. Human amniotic membrane-derived mesenchymal stem cells (AM-MSCs) secrete paracrine factors with antioxidant properties, making their secretome (AM-MSC-Sec) a potential tool for protecting sperm function. This study evaluated the protective effects of AM-MSC-Sec on bovine sperm under OS. Frozen Holstein bull semen was thawed, purified using a two-layer density gradient, and divided into four groups: control (no treatment), H₂O₂ (10 μM), H₂O₂ + 20 % AM-MSC-Sec, and H₂O₂ + 20 % DMEM/F12. After 2 h at 38.5°C and 5 % CO₂, sperm quality parameters—including motility, progressive motility, viability, plasma membrane integrity, acrosome reaction, DNA fragmentation, malondialdehyde (MDA), and total antioxidant capacity (TAC)—were assessed. AM-MSC-Sec significantly improved the percentage of total and progressive motility, enhanced viability and membrane integrity, and decreased DNA fragmentation compared with control and DMEM groups. The percentage of acrosome-reacted sperm increased, and TAC levels were elevated, indicating improved antioxidant defense. DMEM/F12 alone did not show a protective effect. MDA levels increased non-significantly under stress in the control group; however, the difference was not statistically significant. These findings suggest that AM-MSC-Sec could mitigate oxidative damage and preserve multiple functional parameters in bovine sperm. AM-MSC-Sec may serve as a promising therapeutic agent for maintaining sperm quality, with potential applications in bovine reproductive technologies.
{"title":"The secretome from human amniotic membrane-derived mesenchymal stem cells alleviate oxidative damage in bovine sperm","authors":"Negin Amirjannati , Mohammadmehdi Barfar , Mostafa Pournourali , Niloofar Khalili , Parviz Tajik , Razie Shams , Alireza Jahandideh , Leila Balaei Goli , Seyed Jafar Hashemian , Ablofazl Shirazi , Hannaneh Golshahi","doi":"10.1016/j.repbio.2025.101178","DOIUrl":"10.1016/j.repbio.2025.101178","url":null,"abstract":"<div><div>Bovine spermatozoa are highly vulnerable to oxidative stress (OS), which can impair motility, viability, membrane integrity, and DNA integrity. Human amniotic membrane-derived mesenchymal stem cells (AM-MSCs) secrete paracrine factors with antioxidant properties, making their secretome (AM-MSC-Sec) a potential tool for protecting sperm function. This study evaluated the protective effects of AM-MSC-Sec on bovine sperm under OS. Frozen Holstein bull semen was thawed, purified using a two-layer density gradient, and divided into four groups: control (no treatment), H₂O₂ (10 μM), H₂O₂ + 20 % AM-MSC-Sec, and H₂O₂ + 20 % DMEM/F12. After 2 h at 38.5°C and 5 % CO₂, sperm quality parameters—including motility, progressive motility, viability, plasma membrane integrity, acrosome reaction, DNA fragmentation, malondialdehyde (MDA), and total antioxidant capacity (TAC)—were assessed. AM-MSC-Sec significantly improved the percentage of total and progressive motility, enhanced viability and membrane integrity, and decreased DNA fragmentation compared with control and DMEM groups. The percentage of acrosome-reacted sperm increased, and TAC levels were elevated, indicating improved antioxidant defense. DMEM/F12 alone did not show a protective effect. MDA levels increased non-significantly under stress in the control group; however, the difference was not statistically significant. These findings suggest that AM-MSC-Sec could mitigate oxidative damage and preserve multiple functional parameters in bovine sperm. AM-MSC-Sec may serve as a promising therapeutic agent for maintaining sperm quality, with potential applications in bovine reproductive technologies.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101178"},"PeriodicalIF":2.5,"publicationDate":"2025-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145883680","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 : 2025-12-30DOI: 10.1016/j.repbio.2025.101176
Paulina Surmacz, Monika Petrajtis-Gołobów, Paula Kiełbik, Katarzyna Siewruk, Ewa Kautz-Wasilewska, Sławomir Giziński
In recent years, special attention has been paid to proAKAP4, the precursor of A-kinase anchoring protein 4 (AKAP4), which plays a crucial role in the structure and function of the sperm flagellum. This testis-specific protein forms the fibrous sheath and local signal transduction required for proper sperm capacitation and motility. Highly conserved across species, proAKAP4 levels are strongly correlated with fertility and sperm survivability post-thaw. This review summarizes the biological roles of proAKAP4 and its potential as a semen quality biomarker in various mammalian species, including humans, horses, bulls, boars, dogs, rodents and other species. We discuss structural and molecular aspects of proAKAP4 (e.g., prodomain, PKA interactions, phosphorylation sites), as well as its involvement in key signaling pathways controlling sperm movement, such as the cAMP-PKA cascade and calcium signaling. The review also outlines various methods for measuring proAKAP4, such as ELISA, Western blotting and immunocytochemistry. Particular attention is given to commercial assays, which allow rapid and species-specific quantification of proAKAP4, independent of seminal plasma or cryoprotectant presence. Although the data are promising, researchers highlight the need for further validation in larger populations under varying environmental conditions. Nonetheless, current findings suggest that proAKAP4 may emerge as a standard marker in andrological diagnostics and a valuable tool for selecting breeding males and optimizing semen cryopreservation procedures.
{"title":"ProAKAP4 as a potential biomarker of fertility and sperm freezability in males of different species - A review","authors":"Paulina Surmacz, Monika Petrajtis-Gołobów, Paula Kiełbik, Katarzyna Siewruk, Ewa Kautz-Wasilewska, Sławomir Giziński","doi":"10.1016/j.repbio.2025.101176","DOIUrl":"10.1016/j.repbio.2025.101176","url":null,"abstract":"<div><div>In recent years, special attention has been paid to proAKAP4, the precursor of A-kinase anchoring protein 4 (AKAP4), which plays a crucial role in the structure and function of the sperm flagellum. This testis-specific protein forms the fibrous sheath and local signal transduction required for proper sperm capacitation and motility. Highly conserved across species, proAKAP4 levels are strongly correlated with fertility and sperm survivability post-thaw. This review summarizes the biological roles of proAKAP4 and its potential as a semen quality biomarker in various mammalian species, including humans, horses, bulls, boars, dogs, rodents and other species. We discuss structural and molecular aspects of proAKAP4 (e.g., prodomain, PKA interactions, phosphorylation sites), as well as its involvement in key signaling pathways controlling sperm movement, such as the cAMP-PKA cascade and calcium signaling. The review also outlines various methods for measuring proAKAP4, such as ELISA, Western blotting and immunocytochemistry. Particular attention is given to commercial assays, which allow rapid and species-specific quantification of proAKAP4, independent of seminal plasma or cryoprotectant presence. Although the data are promising, researchers highlight the need for further validation in larger populations under varying environmental conditions. Nonetheless, current findings suggest that proAKAP4 may emerge as a standard marker in andrological diagnostics and a valuable tool for selecting breeding males and optimizing semen cryopreservation procedures.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"26 1","pages":"Article 101176"},"PeriodicalIF":2.5,"publicationDate":"2025-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145879631","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.2025.101171
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}
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