Biology of Reproduction最新文献

{"title":"Melatonin modulated GPX5 and PTGDS expression in Bactrian camel epididymis mainly via receptor MT1†.","authors":"Shuqin Zhao, Shipeng Wu, Shuai Ji, Yaxuan Han, Zhen Yang, Yuan Gao","doi":"10.1093/biolre/ioaf030","DOIUrl":"10.1093/biolre/ioaf030","url":null,"abstract":"<p><p>Melatonin (Mel), an important mediator of photoperiodic annual rhythm regulation and seasonal reproduction in animals, directly modulates the expression of specific genes in the epididymis and protects sperm from oxidative damage. Bactrian camel is a dominant species in desert and semi-desert areas, exhibiting the unique reproductive regulation patterns. However, the underlying regulation mechanism of Mel on Bactrian camel is still unclear. This study isolated the epididymal caput epithelial cells of Bactrian camels and investigated the expression of specific genes involving sperm protection after Mel treatment and overexpression/knockdown of Mel receptor MT1/MT2 using real-time quantitative PCR assay (qPCR), ELISA, and western blotting assay. The results showed that MT1, MT2, clock genes cryptochrome 1/2 (Cry1/Cry2) were all positively expressed in the epididymal lumen epithelial cells, peritubular myoid cells, and luminal spermatozoa. Intriguingly, Mel treatment activated receptor MT1 in epididymal caput epithelial cells, indicating that Mel treatment regulated genes expression mainly via MT1-dependent manner. Mel treatment or overexpression of MT1 both increased secretion of glutathione peroxidase 5 (GPX5) and prostaglandin D2 synthase (PTGDS), and MT1 silencing induced downregulation of GPX5 and PTGDS expression, indicating that the expression of GPX5 and PTGDS were regulated by Mel-MT1. Overexpression of MT1 or MT2 promoted Cry2 expression, and overexpression of Cry2 also activated the MT1/MT2 expression by feedback regulation. Finally, the double luciferase reports assay showed that the activation of MT1 by Cry2 occurred during transcription. These results help to understand the regulatory effect of Mel on the epididymis in Bactrian camels.</p>","PeriodicalId":8965,"journal":{"name":"Biology of Reproduction","volume":" ","pages":"895-905"},"PeriodicalIF":3.1,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143416943","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Age-related integrative transcriptomic profiling of human granulosa cells reveals mRNA-microRNA regulatory network associated with key ovulation dynamics†.","authors":"Heather M Rogers, Ahmed Gad, Gentry K Cork, Nico G Menjivar, William B Schoolcraft, Dawit Tesfaye, Ye Yuan","doi":"10.1093/biolre/ioaf034","DOIUrl":"10.1093/biolre/ioaf034","url":null,"abstract":"<p><p>Advanced maternal age (AMA) patients experience decreased success from assisted reproductive technologies (ART), attributed to the quantity and quality of oocytes, which is significantly influenced by the intrafollicular granulosa cells (GCs). In this study, we compared the mRNA and microRNA (miRNA) transcriptomes between young (< 32 years old) and AMA (> 38 years old) patients' GCs to identify potential ovarian aging-related molecular signatures. We identified 293 and 21 differentially expressed genes (DEGs) and miRNAs (DE miRNAs), respectively, between young and aged GCs. Highly expressed mitochondrial-encoded genes, MT-ND3, MT-ND6, and MT-CYB, were downregulated in aged GCs, indicating potential mitochondrial insufficiency. Additionally, pathway analysis indicates DEGs are involved in inflammation, cytokine signaling, extracellular matrix (ECM) remodeling, and angiogenesis. Key DEGs related to these processes include CXCL8, IL1B, NLRP3, SIGIRR, ANGPT2, ADAM8, and ADAMTS14. Additionally, target gene prediction and pathway analysis of DE miRNAs indicates their potential post-transcriptional regulation of genes associated with cell signaling, mitochondrial function, oxidative stress, apoptosis, and senescence pathways in addition to cytokine signaling, angiogenesis, and ECM remodeling. To investigate regulatory mechanisms further, we looked at the DEGs' convergence with the DE miRNAs predicted target genes and we identified miR-483-3p, miR-1268a, miR-4497, miR-7704, miR-135a-5p, miR-1261, and miR-4791 as potential crucial regulators of genes involved in pathways associated with inflammation, ECM, and angiogenesis. This data suggests that aged GCs have an impaired ability to elicit the same pro-inflammatory response combined with dysregulation of angiogenesis and ECM remodeling compared to young GCs, and miRNA may play a role in regulating key ovulatory processes. While this study identifies potential regulatory relationships between DE miRNAs and DEGs, experimental validation is necessary to confirm the relationships and biological relevance.</p>","PeriodicalId":8965,"journal":{"name":"Biology of Reproduction","volume":" ","pages":"916-931"},"PeriodicalIF":3.1,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143466397","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Celebrate the art of reproductive research with ReproBioArt!","authors":"Ewelina Bolcun-Filas","doi":"10.1093/biolre/ioaf093","DOIUrl":"10.1093/biolre/ioaf093","url":null,"abstract":"","PeriodicalId":8965,"journal":{"name":"Biology of Reproduction","volume":" ","pages":"783-785"},"PeriodicalIF":3.1,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143963513","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bta-miR-93 regulates the maternal-fetal tolerance in dairy cows via promoting Programmed Cell Death Ligand 1 mediated M2 macrophage polarization†.","authors":"Cheng Yang, Xinyu Feng, Yingfang Guo, Aftab Shaukat, Zhimin Wu, Yu Chen, Lu Meng, Ganzhen Deng","doi":"10.1093/biolre/ioaf046","DOIUrl":"10.1093/biolre/ioaf046","url":null,"abstract":"<p><p>Bovine embryo resorption is one of the key factors restricting the reproductive efficiency in dairy cattle, which mainly occurs in the early stage of maternal recognition of pregnancy, causing substantial economic losses to the dairy industry. Macrophages, the most numerous endometrial immune cells in cows during early pregnancy, are critical in developing maternal-fetal immune tolerance. However, the mechanism of their action on the maternal-fetal interface of dairy cows remains unknown. MicroRNAs have important roles in immune tolerance and macrophage polarization, but the underlying mechanisms still need further investigation. In previous laboratory studies, RNA-sequencing revealed that bta-miR-93 was dramatically reduced in bovine endometrial luminal epithelial cells (bEECs) upon IFN-tau stimulation. In the current study, it is revealed that the expression of bta-miR-93 was decreased substantially in the endometrium of pregnant cows and negatively correlated with that of Programmed Cell Death Ligand 1 (PD-L1). In vitro experiments displayed that suppression of bta-miR-93 promoted M2 polarization via promoting PD-L1/PD-1/AKT/mTOR signaling pathway in bEECs and bovine macrophage co-culture model, and vice versa. The 3'-untranslated region of PD-L1 is directly regulated by bta-miR-93. Furthermore, our in vivo studies revealed that overexpression of bta-miR-93 efficiently leads to embryo resorption and suppression of M2 polarization in mice.</p>","PeriodicalId":8965,"journal":{"name":"Biology of Reproduction","volume":" ","pages":"880-894"},"PeriodicalIF":3.1,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143623391","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Therapeutic targeting of the Tryptophan-Kynurenine Axis for HTR-8/SVneo trophoblast proliferation and migration in unexplained recurrent spontaneous abortion†.","authors":"Pingping Jin, Xinyi Lu, Lu Wang, Yan Chen, Lan Yang, Yongxiang Yin, Ye Shen, Xinxin Ni, Daozhen Chen, Yun Zhang, Yu Chen","doi":"10.1093/biolre/ioaf040","DOIUrl":"10.1093/biolre/ioaf040","url":null,"abstract":"<p><strong>Introduction: </strong>Recurrent spontaneous abortion (RSA) is associated with maternal-fetal interface dysfunction, particularly abnormal trophoblast invasion and proliferation. However, our understanding of the cause of RSA remains limited.</p><p><strong>Methods: </strong>Plasma Trp and Kyn levels were measured in two groups using enzyme-linked immunosorbent assay. Immunofluorescence and western blot analyses were employed to evaluate the expression of IDO1, VEGFA, and proteins associated with epithelial-mesenchymal transition (EMT) in villous and decidual tissues from patients with RSA. The effects of Tryptophan (Trp) and IDO1-driven Trp-Kynurenine (Kyn) metabolism on trophoblast proliferation, migration, EMT, and angiogenesis were investigated in the HTR-8/SVneo cell line using wound healing, transwell migration, quantitative real-time PCR (RT-qPCR), Western blotting, and tube formation assays. RNA sequencing (RNA-seq) identified differentially expressed genes in cells treated with 500 μM exogenous L-Trp.</p><p><strong>Results: </strong>RSA patients exhibited elevated plasma Trp levels and significantly reduced Kyn levels, indicating decreased IDO1 activity (as assessed by the Kyn/Trp ratio) compared to controls. IDO1, EMT-related proteins, and VEGFA were downregulated in RSA patient tissues. In vitro, L-Trp enhanced trophoblast migration, invasion, EMT, and microvasculature formation via IDO1 activation. The reduced functional capabilities induced by the IDO1 antagonist 1-MT (500 μM) were rescued by Kyn (300 μM). RNA-seq revealed that L-Trp upregulation modulates trophoblast gene expression and functional pathways associated with amino acid metabolism, angiogenesis, and vasculature development.</p><p><strong>Discussion: </strong>Our study reveals a novel molecular mechanism by which Trp metabolism regulates HTR-8 cell function, suggesting that modulating IDO1 activity may represent a therapeutic strategy to improve trophoblast function and pregnancy outcomes in RSA.</p>","PeriodicalId":8965,"journal":{"name":"Biology of Reproduction","volume":" ","pages":"969-980"},"PeriodicalIF":3.1,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143717647","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Equine in vitro fertilization with frozen-thawed semen is associated with shortened pre-incubation time and modified capacitation-related changes.","authors":"Matheus R Felix, Tamara Dobbie, Elizabeth Woodward, Renata Linardi, Carolina Okada, Rebeca Santos, Katrin Hinrichs","doi":"10.1093/biolre/ioaf043","DOIUrl":"10.1093/biolre/ioaf043","url":null,"abstract":"<p><p>We recently reported successful equine in vitro fertilization using fresh semen pre-incubated for a prolonged period (22 h) before co-culture with oocytes. In this study, we evaluated the feasibility of equine in vitro fertilization with frozen-thawed sperm and evaluated capacitation-related changes in these sperm over the pre-incubation period. Sperm selected via a commercial sperm separation device yielded significantly higher fertilization than did sperm selected by swim-up or by colloid centrifugation. Using the sperm separation device method, fertilization rates with sperm pre-incubated for 15 min, 3, 6, and 9 h were 7.1, 22.2, 38.5, and 73.3% respectively (9 h vs. 15 min or 3 h, P < 0.05). Fertilization rates differed significantly (45.9% vs. 85.5%) between freezing extenders. Blastocysts were produced using frozen-thawed semen from each of three stallions and transfer of nine vitrified-warmed blastocysts to mares yielded seven embryonic vesicles. Anti-protein tyrosine phosphorylation staining of the entire sperm tail increased over pre-incubation, and sperm both with and without staining in the tail bound to the oocyte cumulus after co-incubation. Using the stain DiSC3(5) and flow cytometric analysis, a population of apparently hyperpolarized sperm was identified at 22 h in fresh sperm that was not seen at any time in frozen-thawed sperm. We conclude that frozen-thawed equine sperm can successfully fertilize oocytes after a shortened pre-incubation time of 9 h, suggesting that the freeze-thawing process induces capacitation-related changes. Our findings on evaluation of pre-incubated sperm indicate that the mechanisms by which frozen-thawed sperm become capable of fertilization may differ from those found in fresh sperm.</p>","PeriodicalId":8965,"journal":{"name":"Biology of Reproduction","volume":" ","pages":"867-879"},"PeriodicalIF":3.1,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12078078/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143584499","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"SATINN v2: automated image analysis for mouse testis histology with multi-laboratory data integration†.","authors":"Ran Yang, Fritzie T Celino-Brady, Jessica E M Dunleavy, Katinka A Vigh-Conrad, Georgia R Atkins, Rachel L Hvasta, Christopher R X Pombar, Alexander N Yatsenko, Kyle E Orwig, Moira K O'Bryan, Ana C Lima, Donald F Conrad","doi":"10.1093/biolre/ioaf033","DOIUrl":"10.1093/biolre/ioaf033","url":null,"abstract":"<p><p>Analysis of testis histology is fundamental to the study of male fertility, but it is a slow task with a high skill threshold. Here, we describe new neural network models for the automated classification of cell types and tubule stages from whole-slide brightfield images of mouse testis. The cell type classifier recognizes 14 cell types, including multiple steps of meiosis I prophase, with an external validation accuracy of 96%. The tubule stage classifier distinguishes all 12 canonical tubule stages with external validation accuracy of 63%, which increases to 96% when allowing for ±1 stage tolerance. We addressed generalizability of SATINN, through extensive training diversification and testing on external (non-training population) wildtype and mutant datasets. This allowed us to use SATINN to successfully process data generated in multiple laboratories. We used SATINN to analyze testis images from eight different mutant lines, generated from three different labs with a range of tissue processing protocols. Finally, we show that it is possible to use SATINN output to cluster histology images in latent space, which, when applied to the eight mutant lines, reveals known relationships in their pathology. This work represents significant progress towards a tool for robust, automated testis histopathology that can be used by multiple labs.</p>","PeriodicalId":8965,"journal":{"name":"Biology of Reproduction","volume":" ","pages":"996-1014"},"PeriodicalIF":3.1,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143439663","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Whole-genome transcriptome and DNA methylome analyses reveal molecular abnormalities during the oocyte-to-embryo transition in preimplantation embryos derived from prepubertal lamb oocytes†.","authors":"Qi Qi, Jiangyue Bian, Junjin Li, Kexiong Liu, Fengxiang Yan, Jian Hou","doi":"10.1093/biolre/ioaf045","DOIUrl":"10.1093/biolre/ioaf045","url":null,"abstract":"<p><p>The juvenile in vitro embryo transfer technology holds the potential to accelerate livestock breeding. However, its application is limited due to the weak in vitro development of oocytes and embryos from prepubertal lambs. To dissect the regulatory networks of gene expression of sheep embryos and identify the defects in gene expression in prepubertal lamb embryos during the oocyte-to-embryo transition, full-length RNA sequencing and whole-genome bisulfite sequencing based on trace cells were conducted on in vitro-derived embryos generated from adult sheep and prepubertal lamb oocytes. We found that the maternal transcript degradation occurred selectively in adult sheep embryos in multiple waves and was most completed until the morula stage. Major embryonic genome activation was found to occur at the morula stage. By comparing with the patterns of adult embryos, we observed incomplete maternal transcript degradation and abnormal embryonic genome activation in lamb embryos and analyzed their potential molecular mechanisms. Furthermore, we explored dynamic DNA methylation concerning the paternal and maternal genomes during the preimplantation development of sheep embryos, revealing the negative regulatory role of promoter DNA methylation on embryonic genome activation process. Lamb embryos generally displayed higher DNA methylation levels than adults, potentially repressing the embryonic genome activation gene expression, especially the genes associated with ribosomal and mitochondrial organization. We also found abnormalities in the methylation status of imprinted genes in lamb embryos. Our findings advance the understanding of sheep in vitro embryo development and offer insights for improving the juvenile in vitro embryo transfer technology in livestock.</p>","PeriodicalId":8965,"journal":{"name":"Biology of Reproduction","volume":" ","pages":"824-839"},"PeriodicalIF":3.1,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143584502","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Duplex sequencing identifies unique characteristics of ENU-induced mutations in male mouse germ cells†.","authors":"Danielle P M LeBlanc, Gu Zhou, Andrew Williams, Matthew J Meier, Charles C Valentine, Jesse J Salk, Carole L Yauk, Francesco Marchetti","doi":"10.1093/biolre/ioaf029","DOIUrl":"10.1093/biolre/ioaf029","url":null,"abstract":"<p><p>Germ cell mutagenicity testing is increasingly required for chemical risk assessment. Duplex sequencing is rapidly gaining acceptance as a method to assess in vivo mutagenesis, and as a valid alternative to transgenic rodent mutation models such as the MutaMouse. We used a duplex sequencing panel of 20 genomic targets and the transgenic rodent assay to measure mutations in the germ cells of MutaMouse males exposed to 0, 1, 2, or 5 mg/kg N-ethyl-N-nitrosourea for 28 days. Germ cells from the seminiferous tubules were collected 28 days post-exposure. The transgenic rodent assay showed a significant increase in mutant frequencies at the high (P < 0.001) and medium (P = 0.01) N-ethyl-N-nitrosourea doses relative to controls, while duplex sequencing revealed a significant increase (P < 0.001) in N-ethyl-N-nitrosourea-induced mutations only at the high dose. Duplex sequencing mutation frequencies were lower in genic than in intergenic targets, suggesting a protective role for transcription-coupled repair. Interestingly, we observed several unique germ cell characteristics with respect to duplex sequencing data from rodent somatic tissues: 1) larger inter-animal variability in clonally expanded mutations that affects the ability to detect significant increases in mutation frequency; 2) a target on chromosome 2 showing much higher susceptibility to spontaneous and chemical-induced mutagenesis than other targets; and 3) a mutation spectrum consistent with that observed in the offspring of N-ethyl-N-nitrosourea-treated males but not with the spectrum in bone marrow of directly-exposed males. These results suggest that duplex sequencing is a promising approach for characterizing germ cell mutagenesis and that mutagenic mechanisms operating in germ cells differ from those in somatic tissues.</p>","PeriodicalId":8965,"journal":{"name":"Biology of Reproduction","volume":" ","pages":"1015-1027"},"PeriodicalIF":3.1,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12078080/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143398064","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Loss of progesterone receptor in smooth muscle cells has no impact on oviductal contractions and preimplantation embryo development†.","authors":"Emily A McGlade, Jiude Mao, Kalli K Stephens, Cayce N Rose, John P Lydon, Wipawee Winuthayanon","doi":"10.1093/biolre/ioaf042","DOIUrl":"10.1093/biolre/ioaf042","url":null,"abstract":"","PeriodicalId":8965,"journal":{"name":"Biology of Reproduction","volume":" ","pages":"786-788"},"PeriodicalIF":3.1,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12078075/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143584184","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}