Molecular Reproduction and Development最新文献

{"title":"From Mesenchymal Stromal Cells to Extracellular Vesicles: Scalable Strategies for Ovarian Stroma Regeneration","authors":"Stefhani Martins Barcelos,&nbsp;Amandda Évelin Silva-Carvalho,&nbsp;Felipe Perecin,&nbsp;Juliana Lott Carvalho","doi":"10.1002/mrd.70113","DOIUrl":"10.1002/mrd.70113","url":null,"abstract":"<p>Ovarian aging is characterized by the reduction of the follicular pool and a progressive loss in oocyte quality. Although these changes have initially been attributed to oocyte alterations, increasing evidence indicates that stromal deterioration plays a central role in this process. Stromal remodeling involves complex interactions among structural, molecular, and cell communication disruptions. This review discusses the main ovarian stroma shifts related to aging, emphasizing ECM remodeling, biomechanical changes, and cell signaling dysfunction. In light of the multifactorial nature of ovarian aging, no single therapeutic strategy is likely to fully address stromal dysfunction. In this context, Mesenchymal Stromal Cells (MSCs) and their secreted Extracellular Vesicles (EVs) have emerged as promising approaches, given their ability to deliver a diverse and functionally complementary set of bioactive molecules that collectively modulate inflammation, support extracellular matrix remodeling, and restore cell communication. Notably, EVs retain key paracrine effects of MSCs while offering advantages in safety and manufacturability, supporting the transition from cell-based to cell-free therapies. Accordingly, this review also examines this therapeutic shift, highlighting the role of MSCs and EVs in stromal regeneration. By integrating ovarian biology with biotechnological bioprocesses, we discuss key translational challenges—including scalability, quality control, and regulatory considerations—that are critical for the development of effective stroma-targeted interventions aimed at preserving or restoring ovarian function.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mrd.70113","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148032826","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Single-Embryo RNA Sequencing Reveals Transcriptomic Alterations in Blastocysts Derived From IVM or Vitrified Oocytes","authors":"Hubdar Ali Kolachi,&nbsp;Pei-Pei Zhang,&nbsp;Jesse Oluwaseun Ayantoye,&nbsp;Muhammad Shahzad,&nbsp;Baigao Yang,&nbsp;Jianhua Dong,&nbsp;Yang Liu,&nbsp;Xiaoyi Feng,&nbsp;Muhammad Ibrahim Panhwar,&nbsp;Pengcheng Wan,&nbsp;Xueming Zhao","doi":"10.1002/mrd.70116","DOIUrl":"10.1002/mrd.70116","url":null,"abstract":"<div>\u0000 \u0000 <p>In vitro production (IVP) of bovine embryos often results in lower developmental competence compared to in vivo-derived embryos. This disparity is further exacerbated by oocyte cryopreservation (vitrification). This study aimed to investigate the molecular basis of reduced embryo quality by profiling the transcriptomic impact of in vitro maturation (IVM) and vitrification on bovine blastocysts.Day 7 blastocysts from three groups were analyzed: (1) in vivo: control embryos produced entirely in vivo, (2) IVF: embryos from fresh oocytes subjected to IVM and in vitro fertilization, and (3) V_IVF: embryos from vitrified-thawed oocytes subjected to IVM and fertilization. Whole-blastocyst transcriptome profiling was performed using single-embryo RNA sequencing (SUPeR-seq; <i>n</i> = 3 blastocysts per group) and analyzed without cellular dissociation. Developmental competence was significantly reduced in the V_IVF group (~ 11.8% blastocyst rate) compared to the IVF group (~ 41.5%) and in vivo controls. Transcriptome analysis revealed 362 differentially expressed mRNAs in V_IVF embryos and 78 in IVF embryos compared to in vivo controls. Genes such as <i>ATP1A1, GOT1L1</i>, and <i>QSOX1</i> were significantly altered, influencing pathways related to oxidative phosphorylation, lysosome activity, and chromatin organization. In addition, 694 DE-lncRNAs in the V_IVF group and 187 in the IVF group were identified. Exonic circRNAs (608) were detected with differential expression and enrichment in signaling and epigenetic pathways, including Hedgehog signaling and stem cell pluripotency regulation. Integrated network analysis revealed coordinated dysregulation across mRNA, lncRNA, and circRNA layers, suggesting disruption of multi-layered regulatory circuits. Overall, this study presents a detailed transcriptomic landscape of bovine blastocysts affected by IVM and vitrification.</p></div>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148032887","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}

{"title":"First Report of CRISPR-Cas9 Ribonucleoprotein Delivery Into Teleost Spermatozoa With Preserved Membrane and Genomic Integrity","authors":"Laís dos Santos Gonçalves,&nbsp;William Borges Domingues,&nbsp;Leandro Silva Nunes,&nbsp;Eduardo Bierhals Blödorn,&nbsp;Eduardo Nunes Dellagostin,&nbsp;Amanda Weege da Silveira Martins,&nbsp;Izani Bonel Acosta,&nbsp;Carine Dahl Corcini,&nbsp;Antônio Sergio Varela Júnior,&nbsp;Charles Nunes Fróes,&nbsp;Vinicius Farias Campos","doi":"10.1002/mrd.70114","DOIUrl":"10.1002/mrd.70114","url":null,"abstract":"<p>Delivery of large molecular complexes into spermatozoa remains a challenge in reproductive biotechnology. Teleost sperm possess highly compacted chromatin and minimal cytoplasmic volume, limiting intracellular access of ribonucleoprotein assemblies. Here, we evaluated whether CRISPR-Cas9 ribonucleoprotein (RNP) complexes can be introduced into Nile tilapia spermatozoa using capillary electroporation while preserving cellular integrity. RNP uptake was quantified via EGFP fluorescence, and sperm performance was assessed through motility, membrane integrity, mitochondrial activity, and DNA fragmentation analyses. Approximately 10%–13% of spermatozoa exhibited detectable EGFP signal following electroporation. Increased pulse numbers reduced motility and mitochondrial activity, whereas membrane integrity was preserved and no increase in DNA fragmentation was detected. A single pulse (1250 V, 40 ms) provided the best balance between RNP uptake and functional preservation. These findings support capillary electroporation as a minimally disruptive strategy for CRISPR-RNP delivery in teleost reproductive biology.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mrd.70114","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147974031","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"","authors":"","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148085564","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}

{"title":"","authors":"","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148073031","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}

{"title":"","authors":"","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148073030","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}

{"title":"METTL3-Mediated m6A Modification Promotes the Stability of PLCζ mRNA in Sperm","authors":"Chao Yang,&nbsp;Longling Han,&nbsp;Wenting Xu,&nbsp;Donghong Li","doi":"10.1002/mrd.70106","DOIUrl":"10.1002/mrd.70106","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 <p>Male infertility is a global health concern, with sperm dysfunction being a major contributing factor. Phospholipase C zeta (PLCζ), a key sperm-specific factor essential for oocyte activation, is closely linked to fertilization failure. This study investigated the role of METTL3-mediated N6-methyladenosine (m6A) modification in regulating PLCζ expression and sperm function. The mRNA and protein expression levels of PLCζ, METTL3, METTL14, and WTAP in sperm were measured using qRT–PCR and western blotting, respectively. The morphology of the sperm was evaluated by Giemsa staining. The zinc ion content of the semen was measured using the 5-Br-PAPS colorimetric method, and the citrate concentration and the total m6A level were measured using commercial kits. The m6A modification level of PLCζ mRNA in mouse sperm and the binding interaction between METTL3 and PLCζ mRNA were analyzed by RNA immunoprecipitation (RIP). Finally, the stability of PLCζ mRNA was examined using an actinomycin d-mediated transcription inhibition assay. The results showed that PLCζ expression was downregulated in the semen of patients with low fertilization rates (LFR), and its level correlated positively with reduced zinc/citrate content. These patients also exhibited a marked decrease in global m6A modification levels. Inhibition of METTL3 downregulated PLCζ expression in mouse sperm and was accompanied by impaired prostate secretory function. Mechanistically, METTL3 inhibition reduced the stability of PLCζ mRNA by decreasing its m6A modification. In conclusion, METTL3 enhances the stability of PLCζ mRNA through m6A methylation, thereby maintaining its normal expression level in sperm and ultimately supporting fertilization capacity and secretory homeostasis of the prostate.</p>\u0000 </section>\u0000 </div>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147869030","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}

{"title":"","authors":"","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148073032","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}

{"title":"High-Resolution Imaging Reveals Mitochondrial Protein Imbalance in Sperm of Oligoasthenospermic Men","authors":"Pauline Teixeira,&nbsp;Magalie Boguenet,&nbsp;Florence Pascaretti-Grizon,&nbsp;Florence Manero,&nbsp;Solenn Plouzennec,&nbsp;Pierre-Emmanuel Bouet,&nbsp;Pascal Reynier,&nbsp;Arnaud Chevrollier,&nbsp;Pascale May-Panloup","doi":"10.1002/mrd.70110","DOIUrl":"10.1002/mrd.70110","url":null,"abstract":"<p>Male infertility accounts for nearly half of global infertility cases and is often linked to poor sperm quality, such as oligospermia, asthenospermia, and teratospermia. Although the exact mechanisms remain unclear, mitochondria, which are critical for ATP production and reactive oxygen species generation during fertilization, are increasingly recognized as key players in male infertility but remain underexplored. ATP synthesis relies on respiratory chain complexes that establish a proton gradient across the inner mitochondrial membrane, powering the F0F1-ATP synthase. Proper stoichiometry of these complexes, encoded by both nuclear and mitochondrial DNA, is essential for efficient energy production. However, no previous studies have directly correlated mitochondrial protein expression with sperm quality. Due to the small size and helical structure of sperm mitochondria, imaging them is challenging, yet crucial for understanding sperm function. This study investigates the role of mitochondrial oxidative phosphorylation proteins in sperm quality using single-cell immunolabeling and high-resolution microscopy. It compares spermatozoa from oligoasthenospermic (OA) patients to normospermic controls. OA sperm show increased levels of mitochondrial genome encoded cytochrome <i>c</i> oxidase subunit 1 and significantly reduced F0F1-ATP synthase subunits. This imbalance may contribute to mitochondrial dysfunction and underscores the need for deeper exploration of the pathophysiological mechanisms involved, which could affect sperm function and fertility. These initial results suggest a potential pathway toward the identification of novel biomarkers of male infertility.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13159058/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147869011","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Assessment of Survival and Expansion of Mouse Pre-Antral Follicles in 3D System Within Hyaluronic Acid-Alginate Hydrogel Cocultured With Human Theca Stem and Progenitor Cells","authors":"Leila Badban,&nbsp;Reihaneh Pirjani,&nbsp;Mojtaba Rezazadeh Valojerdi","doi":"10.1002/mrd.70108","DOIUrl":"10.1002/mrd.70108","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 <p>Cancer treatments can impair fertility in young women, and existing fertility preservation methods are often unsuitable for prepubertal patients. Replicating the natural follicular microenvironment in a three-dimensional (3D) in vitro culture of ovarian follicles presents a promising alternative, and co-culturing follicles with ovarian cells may enhance oocyte development. This study used a 3D culture system of hyaluronic acid and alginate to grow mouse preantral follicles and human theca stem and progenitor cells (hTSCs and hTPCs). Ovarian tissue was obtained from five transsexual women; hTSCs were isolated, expanded, confirmed, and differentiated into hTPCs. Preantral mouse follicles were collected and co-cultured with hTSCs and hTPCs in 0.5% hyaluronic acid-alginate (HA-Alg) hydrogel for 14 days. hTSCs were identified by spindle-shaped fibroblast morphology, positive alkaline phosphatase activity, expressing mesenchymal markers like CD29, CD44, and CD90, and differentiation potential to adipocytes and osteocytes-like cells. hTSCs differentiation into hTPCs causes an increase in steroidogenic gene expression, hormone release, and follicular recruitment. Coculturing of mouse preantral follicles with hTSCs and hTPCs in HA-Alg hydrogel induced follicle survival, hormone release, and oocyte maturation. Human theca cells in the stem and progenitor cells stage may support mouse follicle growth, survival, and steroidogenesis.</p>\u0000 </section>\u0000 </div>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147869022","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}