Molecular Reproduction and Development最新文献

{"title":"Characteristics of Exosomes and Microbiota in the Decidua of Patients With Early Pregnancy Loss and Their Impact on Trophoblast Cells.","authors":"Fengping He, Lujing Chen, Jiwu Lou, Yanle Guo, Shushu Fan, Tizhen Yan, Yongmei Zhang","doi":"10.1002/mrd.70104","DOIUrl":"10.1002/mrd.70104","url":null,"abstract":"<p><p>Early pregnancy loss (EPL) is a common complication with incompletely understood pathogenesis. Recent studies highlight the roles of exosomes and their miRNAs in intercellular communication, and the influence of uterine decidual microbiota on pregnancy outcomes. This study aimed to characterize decidual exosomes and microbiota in EPL patients and explore their mechanistic roles in EPL development. Exosomes were extracted from decidual tissues of women undergoing normal pregnancy termination (NC group) and those with early pregnancy embryonic loss (EPL group). These exosomes were applied to treat trophoblast cells (HTR8/SVneo), and cell functions were examined using CCK-8, Transwell assays, and Annexin V-PI staining. Inflammation-related gene expression was quantified by qRT-PCR. Microarray analysis identified differentially expressed exosomal miRNAs, validated by qRT-PCR. 16S rRNA sequencing analyzed microbial community differences between groups, with key taxa identified using LEfSe. The roles of miR-483-5p mimics on Lactobacillales and trophoblast cells were also examined. Exosomes from both groups expressed CD63, HSP70, and Histone-H3, with typical membrane structures observed and particle sizes of 100-120 nm. Exosomes from the EPL group remarkably repressed trophoblast cell proliferation, migration, and invasion, promoted apoptosis, and upregulated TLR4, NF-κB, IL-6, TNF-α, and IL-8 mRNA levels. Microarray analysis revealed significant downregulation of miR-483-5p in exosomes from the EPL group. 16S rRNA sequencing showed distinct microbial communities in decidual tissues of the EPL group, particularly in Lactobacillales and Erysipelotrichaceae abundance. Hsa-miR-483-5p mimics enhanced Lactobacillales viability and colocalized with them. Combined treatment with miR-483-5p mimics and Lactobacillales reversed the inhibitory effects of exosomes from the EPL group on trophoblast cells and downregulated inflammatory factor expression. This study characterized exosomes in the decidual tissues of the EPL group and their inhibitory effects on trophoblast cells, identifying key miRNAs (miR-483-5p) and microbial taxa (Lactobacillales). These results indicate that exosomes and microbiota could play a role in EPL development by regulating trophoblast functions and inflammatory responses.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 5","pages":"e70104"},"PeriodicalIF":3.0,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13137401/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147817803","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":"Kisspeptin-10 Promotes Hormone Secretion, Ovarian Follicles Development and Fecundity via PI3K/AKT/ERK Signal Pathway in Mice","authors":"Wei Suocheng,&nbsp;Xu Linglong,&nbsp;Gao Jingshuang,&nbsp;Guo Zhenya,&nbsp;Cai Yuan","doi":"10.1002/mrd.70093","DOIUrl":"10.1002/mrd.70093","url":null,"abstract":"&lt;div&gt;\u0000 \u0000 \u0000 &lt;section&gt;\u0000 \u0000 &lt;h3&gt; Objectives&lt;/h3&gt;\u0000 \u0000 &lt;p&gt;The present study aimed to comprehensively elucidate how KP-10 modulates hormone secretion, the proliferation and autophagy of ovarian follicles by regulating in-vivo and in-vitro levels of associated genes and proteins.&lt;/p&gt;\u0000 &lt;/section&gt;\u0000 \u0000 &lt;section&gt;\u0000 \u0000 &lt;h3&gt; Methods&lt;/h3&gt;\u0000 \u0000 &lt;p&gt;Follicular granulosa cells (FGCs) were cultured in EMDM/F12 medium. The proliferation viability, apoptosis, and autophagy rates of FGCs were determined, respectively. For in-vivo tests, 0, 5, 10, 20, 40 μg/g of KP-10 were injected into 75 mice allocated into control group (CG) and four KP-10 treated groups (including KP-1, KP-2, KP-3, and KP-4 groups). Maximum transverse diameter (MTD), maximum longitudinal diameter (MLD) and follicle-wall thickness (FWT) of secondary follicles were measured in days 0, 5, 10, 15, and 25. RT-qPCR and Western blot were employed to determine the contents of genes and proteins.&lt;/p&gt;\u0000 &lt;/section&gt;\u0000 \u0000 &lt;section&gt;\u0000 \u0000 &lt;h3&gt; Results&lt;/h3&gt;\u0000 \u0000 &lt;p&gt;The KP-10-treated groups exhibited increased FGCs proliferation, accompanied by decreased apoptosis and autophagy rates as compared with the control group (CG). The expression levels of KISS1R, Bcl-2, LC3-II, mTOR, p62/Sqstm1, PI3K, AKT, and ERK genes and proteins in FGCs were significantly enhanced (&lt;i&gt;p&lt;/i&gt; &lt; 0.05). Additionally, the number of secondary follicles, MTD, MLD, FWT, and the serum concentrations of progesterone and estradiol were notably elevated (&lt;i&gt;p&lt;/i&gt; &lt; 0.05). Moreover, the in-vivo levels of PI3K, AKT, ERK mRNAs, as well as StAR and CYP11A1 genes and proteins, were significantly higher than those in CG. FGCs proliferation in KP-10-treated groups was increased with reduction of apoptosis and autophagy rates. Levels of KISS1R, Bcl-2, LC3-Ⅱ, mTOR, p62/Sqstm1, PI3K, AKT and ERK genes and proteins in FGCs were enhanced (&lt;i&gt;p&lt;/i&gt; &lt; 0.05). Numbers of the secondary follicles, MTD, MLD, FWT and serum concentrations of progesterone and estradiol were significantly promoted (&lt;i&gt;p&lt;/i&gt; &lt; 0.05). In-vivo levels of PI3K, AKT, ERK mRNAs, StAR and CYP11A1 genes and proteins were accelerated as compared to CG.&lt;/p&gt;\u0000 &lt;/section&gt;\u0000 \u0000 &lt;section&gt;\u0000 \u0000 &lt;h3&gt; Conclusions&lt;/h3&gt;\u0000 \u0000 &lt;p&gt;KP-10 accelerated the proliferation and suppressed the apoptosis and autophagy of FGCs, facilitated the development of the secondary follicles, and nd enhanced the synthesis and secretion of P4 and E2. Ten microgram per gram KP-10 had the best efficacy. Our study first time explored comprehensively the effects and mechanisms KP-10 in-vitro and in-vivo regulating reproductive hormone secretion, ovarian follicles development and fecundity in mice. These effects of KP-10 were achieved probably by activation of PI3K/AK","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 4","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147639323","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":"Growth Differentiation Factor 9 Supplementation Restores the Lipopolysaccharide-Induced Negative Effects on Bovine Oocyte Developmental Competence","authors":"Dragos Scarlet,&nbsp;Idil Serbetci,&nbsp;Gerhard Schuler,&nbsp;Heinrich Bollwein,&nbsp;Mariusz P. Kowalewski","doi":"10.1002/mrd.70099","DOIUrl":"10.1002/mrd.70099","url":null,"abstract":"<p>Growth differentiation factor 9 (GDF9) plays a key role in enhancing developmental competence and blastocyst yield during in vitro maturation (IVM). This study investigated whether GDF9 could counteract the harmful effects of lipopolysaccharide (LPS), a gram-negative bacterial component, on bovine in vitro embryo production, while also examining related gene expression in cumulus cells and oocytes. We hypothesized that GDF9 may compensate for LPS through the NF-κB pathway. Ovaries were collected from a slaughterhouse, and oocytes (≥ 50 per group per replicate) were matured under four conditions: control, GDF9, LPS, and GDF9 + LPS. After IVM, fertilization and culture were performed, with blastocyst development evaluated on Day 7. Cumulus cells and oocytes were analyzed for gene expression (RT-qPCR), while IVM media were tested for progesterone and estradiol. Results showed that co-treatment with GDF9 and LPS restored cumulus expansion, cleavage, blastocyst rate, and embryo quality compared with LPS alone (<i>p</i> &lt; 0.05), with no differences from controls. LPS increased mRNA levels of <i>CXCL8</i>, <i>TNFα</i>, and <i>TLR2</i> in cumulus cells (<i>p</i> &lt; 0.05), but candidate gene expression in oocytes and cumulus cells remained unaffected. Steroid concentrations and estradiol:progesterone ratios were similar across groups. In summary, GDF9 supplementation alleviates LPS-induced impairment in bovine oocytes during IVM, though the underlying mechanisms remain unclear.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 4","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13062881/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147639316","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":"Sex-Specific Gene Expression Ontogeny During Gonadal Development in Post-Metamorphic Xenopus tropicalis","authors":"Daniele Marini,&nbsp;Mauricio Roza,&nbsp;Cecilia Berg,&nbsp;Vanessa Brouard","doi":"10.1002/mrd.70098","DOIUrl":"10.1002/mrd.70098","url":null,"abstract":"<p>Sexual differentiation in amphibians involves dynamic coordination between germ cells, somatic tissues, and gene expression. However, post-metamorphic gonadal maturation and sex-specific transcriptional programs underlying it remain poorly characterized. In this study, pre-pubertal gonadal development was analyzed in <i>Xenopus tropicalis</i> by integrating morphological, histological, and gene expression analyses. Froglets were sampled weekly from one until 8 weeks post-metamorphosis. Morphometric and histological assessments revealed progressive gonadal maturation in both sexes. In males, testicular growth correlated with body size, whereas in females, ovarian development occurred independently of somatic parameters. Expression profiling of 12 genes showed sex-specific mRNA patterns: <i>cyp17</i>, <i>amh</i>, and <i>amhr2</i> displayed a male-biased expression correlated with testicular development, while <i>aldh1a2</i> and <i>ddx4</i> were preferentially upregulated in females and associated with ovarian growth and follicular oocyte number. <i>Id4</i> declined in females but remained stable in males, reflecting, together with <i>ddx4</i> expression, distinct pregametogenesis and germline maintenance strategies. Opposite regulation of <i>aldh1a2</i> and <i>cyp26b1</i> supported a transient meiotic wave in females versus its basal maintenance in males. Multivariate and network analyses highlighted <i>dmrt1</i>, <i>amhr2</i>, <i>cyp17</i>, <i>esr1</i>, and <i>ddx4</i> as potential nodes of sex-specific regulatory networks. These findings reveal post-metamorphic, sex-specific transcriptional programs, and provide integrated molecular and histological endpoints for studies on vertebrate sex differentiation.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 4","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-04-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13051529/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147623229","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":"The Impact of Melatonin on Cellular Dynamics and Gene Expression of Bovine Embryos Cultured Under Low and High Oxygen Tension","authors":"Isabella Rodrigues dos Santos Oliveira,&nbsp;Carlos Frederico Martins,&nbsp;Fabiana Lima Rodrigues,&nbsp;Victor Carlos Mello,&nbsp;Maria Tereza de Oliveira Rodrigues,&nbsp;Lucas Costa de Faria,&nbsp;Hallya Beatriz Sousa Amaral,&nbsp;Rosângela Vieira de Andrade,&nbsp;Marcio José Poças Fonseca,&nbsp;Margot Alves Nunes Dode,&nbsp;Sônia Nair Báo","doi":"10.1002/mrd.70102","DOIUrl":"10.1002/mrd.70102","url":null,"abstract":"<p>This study investigated the effects of melatonin supplementation in the culture medium on the development, cellular dynamics and gene expression of bovine embryos produced <i>in vitro</i> under low (5%) or high (20%) oxygen tension. Zygotes were cultured without melatonin or with (10⁻⁹ M), and cleavage, blastocyst rates, blastomere number, apoptosis, mitochondrial activity, lipid accumulation, and expression of genes related to metabolism, oxidative stress and embryo quality were evaluated. Under high O₂ tension, melatonin increased blastocyst rate (39.8% vs. 34.0%), raised blastomere number, reduced apoptosis, and decreased lipid accumulation. It also upregulated SOD2, KRT8, IFN-τ, and PLAC8. Under low O₂ tension, melatonin increased cleavage rate and mitochondrial activity but did not affect blastocyst rate; only SOD2 was upregulated. Embryos cultured without melatonin under low O₂ showed higher IFN-τ and PLAC8 expression. In conclusion, melatonin improves embryo quality and viability mainly under high oxygen tension, acting as an antioxidant, gene modulator and apoptosis inhibitor. Its effects are more limited under low O₂, likely because this environment is closer to physiological conditions and requires fewer responses to oxidative stress.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 4","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-04-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13051412/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147623222","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":"Early Fertilization Dynamics in Lytechinus variegatus: Ionic Specificity, Voltage-Dependent Entry, and Cortical Granule Activation","authors":"Gabriela C. Powers,&nbsp;Pedro I. Ivonnet","doi":"10.1002/mrd.70101","DOIUrl":"10.1002/mrd.70101","url":null,"abstract":"<p>Sea urchin fertilization is a rapid and coordinated cascade of electrical, ionic, and biochemical events that transform the egg into a developmentally competent zygote. While many of these transitions have been described, several remain obscure, misinterpreted, or lost to time. This review reexamines early fertilization events with emphasis on electrophysiological and calcium-dependent signatures, integrating voltage-clamp and unclamped recordings with cytoskeletal and metabolic remodeling. We delineate a biphasic response—excitation and activation—initiated by sperm-derived conductance and sustained through egg-intrinsic sodium channels. By mapping these transitions onto a unified timeline, we highlight transient but critical events such as the cortical flash, calcium-wave propagation, and actin-mediated cytoskeletal remodeling, restoring mechanistic clarity to the ionic and structural transitions that define successful fertilization. We also revisit voltage-dependent sperm entry and propose a mechanism by which excessive calcium influx may destabilize the fusion interface, potentially resulting in unfusion. Ion substitution experiments further reveal that although Sr and Ba can support sperm entry at negative potentials, they elicit only a diminished early activation current, underscoring calcium's unique specificity in triggering downstream signaling. This framework invites renewed attention to the electrical and ionic foundations of monospermy, egg activation, and zygotic competence, and highlights unresolved questions in how early ionic events drive subsequent structural transitions.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 4","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mrd.70101","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147593338","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":"Ovarian Fibrosis Through Comparative Analysis of Collagen Architecture.","authors":"Caroline J Aitken, Lanna Kadhim, Alyssa Murray, David A Landry","doi":"10.1002/mrd.70107","DOIUrl":"https://doi.org/10.1002/mrd.70107","url":null,"abstract":"<p><p>Ovarian fibrosis is a known pathology of reproductive aging, becoming a growing concern for infertility and complex ovarian diseases. In research, mouse and human ovary samples are utilized, though distinct differences between species warrant validation of architectural phenotypes to accurately define its pathology. Using picrosirius red staining under polarized light microscopy, this study characterize fibrillar collagen distribution and organization across defined ovarian regions, specifically the cortex and medulla, in aging human ovary and the equivalent region in the aging mouse ovary. Our results reveal cortex-specific collagen accumulation with anisotropic organization in aging human ovaries, in contrast to the diffuse, isotropic collagen deposition observed in the whole ovary of aging mice. Our findings support age-associated ovarian fibrosis as the accumulation and/or cortical anisotropic organization of fibrillar collagen within the ovary.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 4","pages":"e70107"},"PeriodicalIF":3.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13113214/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147776755","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":"PIP₂ Accumulation in the Spongiotrophoblast Drives Trophoblast Apoptosis in Intrahepatic Cholestasis of Pregnancy.","authors":"Zhou-Tian Xu, Lu-Xing Ge, Li Luo, Hong-Ying Ma, Hong Xiao, Yu-Bin Ding","doi":"10.1002/mrd.70105","DOIUrl":"https://doi.org/10.1002/mrd.70105","url":null,"abstract":"<p><p>Intrahepatic cholestasis of pregnancy (ICP) is a pregnancy-specific liver disorder characterized by elevated maternal bile acids and an increased risk of adverse fetal outcomes. Although placental dysfunction is a key contributor to ICP pathogenesis, the underlying mechanisms remain incompletely understood. Here, we combined a Sprague-Dawley rat model of ICP with spatial metabolomics to delineate region-specific metabolic alterations within the placenta. Multivariate analysis revealed distinct metabolic signatures between ICP and control placentas, with pronounced reprogramming in trophoblast-enriched regions. Differential metabolite and pathway analyses identified significant perturbations in glycerophospholipid metabolism, pyruvate metabolism, phospholipase D signaling, and the tricarboxylic acid cycle. Notably, phosphatidic acid (PA) was broadly elevated, whereas its downstream product phosphatidylinositol 4,5-bisphosphate (PIP₂) exhibited spatially restricted accumulation in trophoblasts. Mechanistically, taurocholic acid exposure in HTR-8/SVneo cells induced upregulation of PIP5K1, leading to PIP₂ accumulation and increased trophoblast apoptosis via disruption of the Bcl-2/Bax balance. Silencing of PIP5K1 restored PIP₂ homeostasis and attenuated apoptosis. Importantly, these molecular alterations were recapitulated in placental tissues from ICP patients, confirming activation of the PIP5K1-PIP2 axis in vivo. Collectively, our findings identify a spatially resolved PA-PIP5K1-PIP₂ lipid signaling cascade that links bile acid-induced metabolic stress to trophoblast apoptosis and placental dysfunction in ICP.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 4","pages":"e70105"},"PeriodicalIF":3.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147776774","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":"Thermal Adaptation Inhibits the Wnt Signaling Pathway Regulating Autophagy and Improving the Development of Embryos","authors":"Runze Song,&nbsp;Zhongshun Zhang,&nbsp;Shiyong Zhu,&nbsp;Kaiqiang Fu,&nbsp;Zhongling Jiang,&nbsp;Rongfeng Cao,&nbsp;Huatao Li","doi":"10.1002/mrd.70100","DOIUrl":"10.1002/mrd.70100","url":null,"abstract":"<div>\u0000 \u0000 <p>Heat stress impairs the reproductive capacity of organisms. In contrast, thermal adaptation can have beneficial effects. In this study, we investigated the mechanism through which thermal adaptation (39°C for 1 h followed by recovery at 37°C for 3 h) improves the quality of in vitro cultured mouse morula-stage embryos. Thermal adaptation resulted in several key changes: increased expression of genes associated with embryo quality (<i>Cdh1, Gja1, Dnmt1</i>); decreased expression of Wnt and mTOR genes and proteins; and upregulated expression of <i>GSK3β</i> and autophagy-related markers (<i>LC3B, Beclin-1</i>). Consequently, thermal adaptation increased the blastocyst rate and improved embryo quality. These findings suggest that thermal adaptation during early embryo development enhances developmental competence by inhibiting the Wnt signaling pathway and activating autophagy.</p>\u0000 </div>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 3","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147521421","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":"Mitochondrial and Ribosomal Stress Underlying Pronuclear Envelope Breakdown Failure: Insights From Single-Cell Transcriptomics in Human Zygotes","authors":"Chaoying Wang,&nbsp;Junnan Fang,&nbsp;Guang Yang,&nbsp;Ran Jiang,&nbsp;Haixia Jin,&nbsp;Wenyan Song,&nbsp;Senlin Shi,&nbsp;Jun Zhai,&nbsp;Huihui Wang,&nbsp;Tongwei Zhang,&nbsp;Guidong Yao","doi":"10.1002/mrd.70096","DOIUrl":"10.1002/mrd.70096","url":null,"abstract":"<div>\u0000 \u0000 <p>Pronuclear Envelope Breakdown (PNEB) failure is a critical factor contributing to the early developmental arrest of intracytoplasmic sperm injection (ICSI) embryos; however, its molecular mechanism remains inadequately understood. This study aimed to elucidate the core regulatory network underlying PNEB failure occurrence and its impact on embryonic development. Single-cell sequencing was used to identify differentially expressed genes (DEGs) between PNEB‑type two pronuclei (2PN) zygotes and 3PN control zygotes, and weighted gene coexpression network analysis (WGCNA) was performed to identify module genes associated with PNEB failure. The least absolute shrinkage and selection operator was applied to model disease types and screen core genes, thereby establishing a multiomics integration strategy. Reactive oxygen species (ROS) and 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolocarbocyanine iodide (JC-1) fluorescence staining were conducted to analyze oxidative stress and mitochondrial function. A total of 1294 DEGs were identified, including genes involved in the oxidative phosphorylation pathway. Mitochondrially encoded reduced nicotinamide adenine dinucleotide dehydrogenase 1 (MT-ND1) and mitochondrially encoded cytochrome <i>c</i> oxidase III (MT-CO3) were significantly upregulated, whereas genes associated with the DNA repair pathway were downregulated. WGCNA revealed a light-green module strongly associated with PNEB failure (<i>r</i> = 0.89, <i>P</i> = 1.2e − 5). Hub genes, ribosomal protein L10a (RPL10A) and ribosomal protein L38 (RPL38), within this module were implicated in ribosome biogenesis. The PPI network confirmed functional interactions between MT-ND1 and RPL10A, suggesting that dysregulated mitochondrial function and ribosomal assembly are central to PNEB failure. ROS and JC-1 fluorescence staining showed a significant decrease in the JC-1 red/green fluorescence intensity ratio in the PNEB failure group (<i>p</i> &lt; 0.05), while ROS levels were significantly elevated (<i>p</i> &lt; 0.01). This study reveals that mitochondrial metabolic dysfunction and ribosomal assembly abnormalities contribute to PNEB failure, thereby disrupting nuclear envelope stability. Furthermore, it identifies the MT-ND1-RPL10A/RPL38 axis as a potential novel molecular marker for assessing ICSI embryo quality.</p></div>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 3","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147366105","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}