Molecular Reproduction and Development最新文献

{"title":"Jujuboside a Improves Polycystic Ovary Syndrome Through SIRT3-Mediated PINK1/Parkin-Dependent Mitochondrial Autophagy in Ovarian Granulosa Cells","authors":"Nan Zhou,&nbsp;Jiahao Xu,&nbsp;Wenqiang Lv,&nbsp;Wei Tai,&nbsp;Guangyan Xie,&nbsp;Qing He,&nbsp;Ying Zhou,&nbsp;Nan Hu,&nbsp;Xueyan Zhou","doi":"10.1002/mrd.70150","DOIUrl":"https://doi.org/10.1002/mrd.70150","url":null,"abstract":"<div>\u0000 \u0000 <p>Polycystic ovary syndrome (PCOS) is a common endocrine disorder, and its treatment remains a significant challenge. This study investigated the mechanism of Jujuboside A (JuA) in the treatment of PCOS through mitophagy. Granulosa cells (GCs) were collected from follicular fluid derived from PCOS women. The effects of JuA on PCOS were determined by measuring body weight, ovarian weight, estrous cycle, glucose level, insulin tolerance, and ovarian morphology in dehydroepiandrosterone-induced mice. SIRT3 expression levels were quantified in GCs, KGN cells, and mouse ovaries. The effects of JuA on SIRT3-related oxidative stress and mitochondrial damage were investigated in KGN cells and mice. Transmission electron microscopy was used to observe mitochondrial damage and mitophagy in GCs. The expression of mitophagy-related proteins (PINK1 and Parkin) was analyzed. JuA significantly improved body weight, estrous cycle, ovarian morphology, glucose tolerance, and insulin resistance, partially restoring ovarian function in PCOS mice. Moreover, JuA significantly activated SIRT3 expression and alleviated SIRT3-related oxidative stress and mitochondrial damage in mouse ovaries, GCs, and KGN cells. Mechanistically, JuA upregulated SIRT3 and inhibited excessive activation of the PINK1/Parkin pathway to reduce mitophagy. These findings demonstrated JuA protected GCs from mitochondrial damage by activating SIRT3 and inhibiting PINK1/Parkin-mediated mitophagy.</p></div>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 10","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149099852","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":"Global Proteomic Analysis of Sperm and Seminal Plasma From Tropically-Adapted Rams With Contrasting Parameters of Frozen-Thawed Semen","authors":"Wallisson Bruno de Morais Pacheco,&nbsp;Iran Aguila Maciel,&nbsp;Suane Santos Alves dos Santos,&nbsp;Alan Martins de Araújo,&nbsp;Francisca Nágila Matos Almeida,&nbsp;Denise Damasceno Guerreiro,&nbsp;Sheheryar Sheheryar,&nbsp;Lais Lacerda Brasil de Oliveira,&nbsp;Felipe Domingos de Sousa,&nbsp;Mohibullah Shah,&nbsp;Maria Elisabete Amaral Moraes,&nbsp;Carlos Roberto Koscky Paier,&nbsp;Ana Cristina de Oliveira Monteiro,&nbsp;Carminda Sandra Brito Salmito-Vanderley,&nbsp;Arlindo A. Moura","doi":"10.1002/mrd.70148","DOIUrl":"https://doi.org/10.1002/mrd.70148","url":null,"abstract":"<p>The aim of this study was to characterize the proteome of sperm and seminal plasma (SP) from rams with high semen freezability (HF) and low semen freezability (LF). Eleven ejaculates were collected from 12 rams, at 3-day intervals, and cryopreserved (132 semen samples). For proteomic analysis, sperm and SP samples were pooled separately for each ram. Pools 1 and 2 included ejaculates obtained on odd-numbered and even-numbered collection days, respectively. This generated 24 sperm and 24 SP pools. Sperm and SP proteins were analyzed by mass spectrometry, and computational biology platforms, used for calculation of differentially abundant proteins (DAPs; <i>p</i> &lt; 0.05) in HF (<i>n</i> = 5) and LF (<i>n</i> = 7) rams. Total sperm motility decreased from 91.87% ± 0.63% in fresh to 65.65% ± 2.58% in frozen-thawed semen of HF rams; and decreased from 89.47% ± 1.15% to 50.05% ± 2.33% in LF rams. Reduction in total sperm motility after cryopreservation was lower in HF (26.22% ± 2.39%) compared to LF rams (37.70% ± 2.23%; <i>p</i> = 0.006). Sperm progressive motility decreased less in HF rams (from 87.20% ± 0.50% to 58.12% ± 2.82%) than in LF rams (from 85.54% ± 1.41% to 44.00 ± 2.13; <i>p</i> = 0.006). Rams with the HF phenotype had better plasma membrane (39.00% ± 2.79% vs. 15.14% ± 0.92%) and acrosome integrity (37.54% ± 2.72% vs. 15.40% ± 1.92%; <i>p</i> &lt; 0.001) than LF rams. Sperm motility was similar in fresh semen of HF and LF rams. There were 1599 and 1677 sperm proteins identified in HF and LF rams, with 73 and 76 DAPs, respectively. SP had 1127 proteins, with 25 and 45 DAPs in HF and LF rams. This is the greatest coverage ever published about the ram SP proteome. Sperm proteins with greater abundance in HF rams were extracellular and immune-related proteins, antiproteases, chaperones and structural regulators, contributing to cytoskeletal integrity, apoptotic resistance and maintenance of proteostasis. Sperm proteins with greater abundance in LF rams were mainly mitochondrial subunits, axonemal components, and bioenergetic enzymes, indicating greater reliance on oxidative processes. Major components of HF SP participate in sperm protection, stabilization of membrane lipids, calcium fluxes, epididymal maturation and ATP availability. The major LF SP proteome was associated with cellular stress, inflammatory responses, oxidative damage and ATP production. Thus, the nature of sperm and SP proteome atlas potentially defines ram sperm cryoresilience.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 10","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mrd.70148","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149099782","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":"JMJD3 Deficiency Alters H3K27me3 Landscapes and Adhesion-Related Transcriptional Programs in Aged Spermatogonia","authors":"Yusuke Sakurai,&nbsp;Hozumi Nomura,&nbsp;Kazumasa Aoyama,&nbsp;Sakurako Shima,&nbsp;Aya Yoshida,&nbsp;Tokuko Iwamori,&nbsp;Atsushi Iwama,&nbsp;Naoki Iwamori","doi":"10.1002/mrd.70147","DOIUrl":"10.1002/mrd.70147","url":null,"abstract":"<div>\u0000 \u0000 <p>Spermatogonial stem cells (SSCs) sustain spermatogenesis throughout life by generating differentiating spermatogonia, spermatocytes, and spermatozoa. During this process, male germ cells are interconnected by intercellular bridges (ICBs). Although ICB fragmentation has been implicated in the replenishment of the SSC pool, the molecular mechanisms underlying this process remain poorly understood. Our previous study demonstrated that JMJD3 deficiency promotes spermatogonial fragmentation and preserves spermatogenesis during ageing. To investigate the molecular basis underlying these phenotypes, we performed H3K27me3 ChIP-seq using a spermatogonia-enriched CD9-positive cell fraction together with RNA-seq of SSC-enriched spermatogonial populations from JMJD3-deficient mice. Cross-dataset comparison revealed extensive alterations in H3K27me3 landscapes accompanied by age- and JMJD3-dependent transcriptional changes. RNA-seq demonstrated downregulation of multiple signaling pathways, including NF-κB and TNF signaling, together with dysregulation of extracellular matrix (ECM)- and cell adhesion-related genes in JMJD3-deficient spermatogonia. Consistent with these transcriptional changes, JMJD3-deficient spermatogonia exhibited excessive ECM accumulation and altered expression of adhesion-associated genes in vivo. Although several mesenchymal-associated genes were upregulated, canonical cadherin switching was not observed, indicating the presence of partial epithelial-mesenchymal transition (EMT)-like transcriptional features rather than a complete EMT program. Cross-dataset comparison further identified Ptk2b as a candidate gene associated with JMJD3-dependent H3K27me3 changes. These findings suggest that JMJD3 deficiency is associated with extensive epigenomic remodeling and adhesion-related transcriptional alterations in aged spermatogonia and provide candidate molecular pathways that may contribute to the maintenance of spermatogenesis during ageing.</p>\u0000 </div>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 9","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148930276","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":"Follicular Fluid Exosomes From Diminished Ovarian Reserve Patients Impair Granulosa Cell Function via miR-135a-5p-Mediated Suppression of BIRC5","authors":"Ziyun Ruan,&nbsp;Zhengda Li,&nbsp;Jinhui Shu,&nbsp;Huadan Lin,&nbsp;Tian Li,&nbsp;Yun Feng,&nbsp;Xu Wang,&nbsp;Caizhu Wang,&nbsp;Zhulian Wu,&nbsp;Yulin Huang,&nbsp;Meng Xia,&nbsp;Xin Zhao","doi":"10.1002/mrd.70143","DOIUrl":"https://doi.org/10.1002/mrd.70143","url":null,"abstract":"<div>\u0000 \u0000 <p>Diminished ovarian reserve (DOR) is associated with poor reproductive outcomes; however, the underlying molecular mechanisms—particularly those involving the regulation of granulosa cell (GC) function by exosomal microRNAs (miRNAs) derived from follicular fluid—remain unclear. This study integrated clinical analysis with molecular profiling to investigate the regulatory effects of DOR patient-derived follicular fluid exosomes on GC function. Clinical data confirmed significantly poorer embryological and clinical outcomes in DOR patients. Follicular fluid was collected from patients with normal ovarian reserve (NOR) and those with DOR. Exosomes and GCs were subsequently isolated from the respective follicular fluid samples. Molecular profiling of follicular fluid exosomes and GCs from both groups identified differentially expressed miRNAs and mRNAs via next-generation sequencing. Integrated bioinformatics analysis and dual‑luciferase reporter assays confirmed that exosomal hsa‑miR‑135a‑5p directly targets and suppresses BIRC5 expression in GCs. Functional experiments demonstrated that incubating NOR GCs with DOR exosomes not only significantly downregulated BIRC5 but also induced marked cellular autophagy and mitochondrial dysfunction. These findings reveal a novel exosome‑mediated regulatory pathway in which DOR follicular fluid exosomes deliver miR‑135a‑5p to recipient GCs, repress the anti‑apoptotic gene BIRC5, and disrupt cellular homeostasis, thereby providing a potential mechanistic explanation for the compromised follicular microenvironment and adverse clinical outcomes in DOR.</p></div>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 9","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148878227","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":"STOX1 Promotes Pyroptosis of Hypoxic Trophoblast Cells Via the Nrf2/Caspase-1 Axis","authors":"Huiqiu Xiang,&nbsp;Xianping Huang,&nbsp;Zilian Jiang,&nbsp;Xiaoting Yu","doi":"10.1002/mrd.70145","DOIUrl":"https://doi.org/10.1002/mrd.70145","url":null,"abstract":"<div>\u0000 \u0000 <p>This study aimed to investigate the role of STOX1 in trophoblast pyroptosis during preeclampsia (PE) and elucidate its underlying molecular mechanisms. A trophoblast hypoxia cell model was established by exposing HTR-8/SVneo cells to hypoxic conditions. The results showed that STOX1 messenger RNA expression was significantly elevated in placental tissues from PE patients as well as in hypoxia-treated HTR-8/SVneo cells. Hypoxia markedly induced pyroptosis in HTR-8/SVneo cells, whereas STOX1 silencing enhanced cell proliferation and migration while suppressing programmed cell death. Moreover, STOX1 knockdown reduced the levels of reactive oxygen species and malondialdehyde, increased superoxide dismutase (SOD) activity, and decreased the expression of pyroptosis-associated inflammatory cytokines, including interleukin-1β (IL-1β) and interleukin-18 (IL-18). In addition, silencing STOX1 downregulated the expression of inflammasome-related proteins, including NLRP3, ASC, cleaved caspase-1, and GSDMD-N. Mechanistically, STOX1 knockdown activated the Nrf2/HO-1 signaling pathway, as evidenced by increased expression of Nrf2 and HO-1 and decreased expression of Keap1. Rescue experiments further demonstrated that co-silencing Nrf2 partially reversed the inhibitory effects of STOX1 knockdown on pyroptosis in hypoxia-treated HTR-8/SVneo cells. Collectively, these findings indicate that STOX1 is upregulated in PE and that STOX1 silencing alleviates oxidative stress and inhibits caspase-1-mediated trophoblast pyroptosis through activation of the Nrf2 signaling pathway. Consequently, STOX1 may serve as a promising therapeutic target for the prevention and treatment of preeclampsia.</p></div>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 9","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148878345","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":"Acute Dietary Zinc Deficiency Impairs Sperm Motility but Not Gonadal mRNA Methylation in Mice","authors":"Ngozi O. Ibadin,&nbsp;Francisco J. Diaz","doi":"10.1002/mrd.70144","DOIUrl":"10.1002/mrd.70144","url":null,"abstract":"<p>Zinc deficiency is associated with impaired male fertility and with reduced DNA and histone methylation in oocytes, but the temporal relationship between acute dietary zinc deficiency and sperm function is unclear. To fill in these gaps, the effects of acute dietary zinc deficiency on sperm function were tested. Five-week-old F1 (DBA × C57BL/6) male and female mice were fed a control diet (29 mg Zn/kg) or zinc-deficient diet (&lt; 1 mg Zn/kg) for 6 days (<i>n</i> = 11/group), a duration selected to model acute zinc depletion based on previous studies demonstrating rapid reproductive defects within a similar timeframe (Tian and Diaz 2012). Sperm number and motility were evaluated by computer-assisted sperm analysis in all animals (<i>n</i> = 11), and testicular histology and DDX4 and YBX2 (MSY2) immunostaining and global m6A levels in testes and liver were measured in a subset of males (<i>n</i> = 6) and in ovaries and liver in females (<i>n</i> = 6) by an ELISA-based assay. Zinc deficiency did not alter body weight but significantly reduced total sperm number recovered from the cauda epididymis and impaired total and progressive motility, straight-line velocity, and hyperactivation. Testes from zinc-deficient males showed reduced thickness of germinal epithelium and increased basement membrane thickness, disorganized seminiferous tubules, and fewer DDX4-positive germ cells, while YBX2-positive cells were largely preserved. Despite these structural and functional defects, zinc deficiency did not significantly change global m6A levels in testes or ovaries, although hepatic m6A methylation was increased in females. These findings demonstrate that a short-term zinc-deficient diet rapidly compromises sperm production, motility, and testicular morphology without measurably altering global gonadal mRNA methylation.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 8","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13501185/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148808816","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":"Embryonic Mediators of Embryo–Uterus Communication, Implantation and Pregnancy","authors":"Keith E. Latham","doi":"10.1002/mrd.70137","DOIUrl":"10.1002/mrd.70137","url":null,"abstract":"<p>Successful reproduction in eutherian mammals requires intimate connections between the embryo and uterus, involving adhesion, attachment and placentation, with or without invasion of the endometrium. The complex dialogue between embryo and uterus involves numerous molecules and pathways. The study of the mechanisms driving early embryo–uterus interactions has historically been challenging, particularly discovering the embryo-expressed molecules. Some components of the interactions have been known for decades, but many have been revealed in recent years through innovative transcriptome studies, studies of the contents and functions of extracellular particles, use of in vitro three-dimensional organoids, genetic studies, approaches for dissecting the regulation of maternal immune system cells, and many other approaches. This review focuses on embryo-expressed molecules that support implantation and embryo attachment, implantation and pregnancy establishment. Both classical and recent discoveries are examined, encompassing results from human, murine, livestock and comparative studies. Opportunities for further mechanistic insights are considered. Additionally, the review considers the potential for agricultural and clinical applications of the more recently identified molecular players and mechanisms for enhancing reproductive success or for providing new approaches to contraception.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 8","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13429183/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148664304","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":"MK-4 Ameliorates Post-Ovulatory Aging of Mouse Oocytes by Alleviating Iron Metabolism Disorder-Induced Oxidative Stress","authors":"Yangzi Zheng,&nbsp;Xiangning Xu,&nbsp;Yanbing Zhang,&nbsp;Yuying Yang,&nbsp;Bicheng Wang,&nbsp;Shuo Lou,&nbsp;Jingyi Kang,&nbsp;Jingyu Li,&nbsp;Kai Xue,&nbsp;Yuzhu Jin,&nbsp;Jieping Liu,&nbsp;Yuanjing Liang,&nbsp;Wei Ma,&nbsp;Jing Weng","doi":"10.1002/mrd.70141","DOIUrl":"10.1002/mrd.70141","url":null,"abstract":"<div>\u0000 \u0000 <p>Post-ovulatory aging (POA) reduces oocyte quality, yet effective interventions remain limited. Here we demonstrate that early intervention with vitamin K2 (menaquinone-4 isoform, MK-4) alleviates POA by inhibiting iron-disorder-induced oxidative stress. Western blot revealed that the expressions of ferroptosis suppressor protein 1 (FSP1) and vitamin K epoxide reductase-like protein (VKORC1L1) were significantly decreased in mouse POA oocytes; however, MK-4 attenuated these changes. Further, MK-4 inhibited ferritinophagy by downregulating NCOA4 and LC3II, markedly restored ferritin level, reduced intracellular Fe²⁺ amount, thus attenuating oxidative stress. These results were characterized by reducing lipid peroxidation marker malondialdehyde (MDA) and acyl-CoA synthetase long chain family member 4 (ACSL4) expression, decreasing γ-H2AX and 8-Hydroxy-2′-deoxyguanosine (8-OHdG), and enhancing expression of the mitochondrial antioxidant enzyme superoxide dismutase 1 (SOD1). Additionally, MK-4 suppressed mitophagy by downregulating PINK1 and Parkin, and upregulating Rab7 improving mitochondrial membrane potential. Consequently, abnormalities in chromosomes, spindles, and the cytoskeleton were significantly ameliorated in POA oocytes receiving early MK-4 intervention. Notably, in vitro-aging oocytes expressed higher FSP1 and VKORC1L1 but lower ACSL4 and MDA versus the in vivo counterparts, suggesting a milder lipid peroxidation in vitro. Collectively, our findings uncover a novel anti-aging mechanism triggered by MK-4, highlighting its potential to establish a preventive strategy for female reproductive aging.</p></div>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 8","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148663281","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":"Mussel Spermatozoa Express a Specific Isoform of the Catalytic Subunit of cAMP-Dependent Protein Kinase","authors":"Izaskun Ibarguren,&nbsp;Saskia-Camille Flament-Simon,&nbsp;Ana P. Losada,&nbsp;Susana. B. Bravo,&nbsp;José Antonio Villamarín","doi":"10.1002/mrd.70139","DOIUrl":"https://doi.org/10.1002/mrd.70139","url":null,"abstract":"<p>cAMP-dependent protein kinase (PKA) plays a central role in regulating sperm motility in both invertebrates and mammals. The sea mussel <i>Mytilus galloprovincialis</i> provides a valuable model for investigating the molecular mechanisms underlying gamete activation in externally fertilizing organisms. In previous work, several catalytic (C) subunit isoforms of PKA were isolated from this species. Here, we report that one of these isoforms appears to be specifically expressed in spermatozoa, indicating the presence of a sperm-specific C-subunit isoform in a non-mammalian species. The protein was purified from male gonadal tissue and identified by LC-MS/MS as the isoform corresponding to UniProt entry A0A8B6GIP6. Sequence analysis revealed that this mussel C-isoform shares strong homology with mammalian sperm-specific Cα2-isoform within the catalytic core but possesses a unique N-terminal region. Whereas mammalian Cα2 contains a short six-residue N-terminal extension, the mussel sperm isoform features a longer acidic extension of 17 amino acids. Moreover, phosphoproteomic analysis reveals the presence of two phosphorylation sites (Ser5 and Thr30) in the N-terminal tail of the mussel C-subunit that are absent in Cα2. These structural differences suggest specific regulatory mechanisms associated with sperm activation in externally fertilizing bivalves. Immunolocalization studies show that the mussel sperm C-subunit is distributed in the acrosomal region, mitochondria, and along the flagellum, where it is predominantly associated with R_myt2, a type II regulatory subunit previously characterized in the mussel.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 8","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mrd.70139","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148617377","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":"Epididymosome-Supplemented Extender Induces Changes in Morpho-Functional Traits and microRNA Levels of Post-Thaw Sperm and Improves Embryo Developmental Potential","authors":"Laura Gabrielli Haupenthal,&nbsp;Maria Alice de Almeida,&nbsp;Cibele Maria Prado,&nbsp;Amanda Nespolo Silva,&nbsp;Gabriela Melendes Schneider,&nbsp;Paola Maria da Silva Rosa,&nbsp;Flávio Vieira Meirelles,&nbsp;Juliano Coelho da Silveira,&nbsp;Felipe Perecin,&nbsp;Maíra Bianchi Rodrigues Alves","doi":"10.1002/mrd.70138","DOIUrl":"https://doi.org/10.1002/mrd.70138","url":null,"abstract":"<p>While molecular signature contributes to sperm quality and fertility potential, sperm may lost key molecules during cryopreservation. Since sperm cells are transcriptionally inert and rely on interactions with epididymosomes (epEVs) to acquire molecules, herein, we investigated the effects of the addition of epEVs to the cryopreservation extender on post-thaw sperm quality and fertility potential. Epididymal sperm were obtained from six bulls (<i>n </i>= 6) and a pool of epEVs was formed from the epididymal fluid of 15 bulls. After, a total of 30 × 10⁶ sperm was cryopreserved in the presence (epEVs) or not (Control) of PKH67 green-labeled epEVs. Post-thaw, sperm were evaluated for interaction with epEVs by flow cytometry (FC) and fluorescence microscopy (FM); motility, vigor, and morphology by microscopy; membrane integrity by FC and FM; microRNA levels by qPCR; and fertility potential by in vitro embryo production (IVEP). EpEVs-sperm showed interaction with green-epEVs, and higher motility and plasma membrane integrity compared to Control-sperm. Levels of 28 microRNAs were higher or uniquely detected in epEVs-sperm. IVEP with epEVs-sperm resulted in a higher rate of hatched blastocyst. Thus, epEVs-supplemented to the sperm cryopreservation extender improved post-thaw sperm quality and embryo developmental potential. These findings provide new insights into the molecular modulation of sperm fertility.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"93 8","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mrd.70138","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148616810","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}