Theriogenology最新文献

{"title":"KHDC3 affects the lineage differentiation of early embryo by regulating the differential distribution of YAP signal","authors":"Qian Xu ,&nbsp;Yuling Lin ,&nbsp;Lina Yu ,&nbsp;Yang Zhang ,&nbsp;Na Kong ,&nbsp;Guijun Yan ,&nbsp;Haixiang Sun ,&nbsp;Guangyi Cao","doi":"10.1016/j.theriogenology.2025.117527","DOIUrl":"10.1016/j.theriogenology.2025.117527","url":null,"abstract":"<div><div>Hydatidiform mole (HM) is characterized by abnormal fetal tissue loss and placental villous trophoblastic hyperplasia, indicating aberrant early embryonic cell fate differentiation. Clinical observations linking KH Domain Containing 3 (KHDC3) mutations to complete hydatidiform mole (CHM) suggest a potentially unforeseen role for KHDC3 in early embryonic development. Single-cell transcriptome analysis of early embryos revealed notable heterogeneity in KHDC3 expression levels from the 8-cell to blastocyst stage. Initial investigations indicated that varying KHDC3 expression levels at the 8-cell stage could influence the Yes-associated protein (YAP) signaling pathway. Knocking down KHDC3 in individual blastomeres at the 2-cell stage resulted in reduced blastocyst formation rates. <em>Khdc3</em>-knockdown (<em>Khdc3</em>-KD) embryos exhibited disruptions in embryonic cell lineage differentiation, characterized by decreased expression of the inner cell mass (ICM) marker organic cation/carnitine transporter 4 (OCT4) and diminished differential expression of the trophectoderm (TE) marker caudal type homeobox 2 (CDX2). The nuclear entry of phosphorylated YAP in outer cells is pivotal for inducing lineage differentiation. Intriguingly, in <em>Khdc3</em>-KD morulae, a significant reduction in the nuclear entry of phosphorylated YAP in outer cells was observed. Moreover, KHDC3 knockdown simultaneously impaired cortical actin cap formation during the morula stage. Forces generated at the apical cortex during this process segregate cells into inner and outer positions within the embryo, thereby influencing ICM versus TE fate specification. Our research underscores the role of KHDC3 in modulating the YAP signaling pathway to establish distinct inner and outer cell distributions during the morula stage, consequently influencing early embryonic lineage differentiation. Nevertheless, the precise mechanism by which KHDC3 influences the YAP signaling pathway via the actin cytoskeleton remains to be fully elucidated. Given the severity of recurrent miscarriages associated with KHDC3 mutations, elucidating the distinctive role of KHDC3 in early embryonic development warrants further investigation.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"245 ","pages":"Article 117527"},"PeriodicalIF":2.4,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144241690","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of Kisspeptin on rabbit ovulation: a comprehensive study of ovulatory, endocrine and histological response","authors":"Silvia Gimeno-Martos ,&nbsp;Alicia Gómez-León ,&nbsp;Bosa Luigia ,&nbsp;Pedro L. Lorenzo ,&nbsp;Arias-Álvarez María ,&nbsp;García-García Rosa María ,&nbsp;Pilar G. Rebollar","doi":"10.1016/j.theriogenology.2025.117524","DOIUrl":"10.1016/j.theriogenology.2025.117524","url":null,"abstract":"<div><div>Mammalian reproductive function is regulated by hypothalamic neurons that secrete Kisspeptin (Kp), a neuropeptide that stimulates gonadotropin-releasing hormone (GnRH) secretion, triggering pituitary gonadotrophins (LH and FSH) and gonadal steroids. This study evaluated the effect of Kisspeptin-10 (Kp10) on ovulation induction in rabbits, comparing its efficacy with that of the GnRH analogue gonadorelin. Multiparous New Zealand White × California does were assigned to three groups: control group (0.5 % saline solution, i.v.), GnRH group (20 μg gonadorelin, i.m.), and Kp10 group (250 μg Kp10, i.v.). Kp10 induced ovulation in 87.5 % of does, matching the response observed in the GnRH group, with a comparable number of corpora lutea (CL) per ovulated doe (12.9 ± 1.4 vs 14.6 ± 1.4 CL/doe, respectively). On day 7, plasma progesterone (P4) was significantly higher in ovulated GnRH-treated does than in Kp10-treated ones (25.12 ± 4.17 vs 9.47 ± 4.17 ng/mL; p &lt; 0.0211), while non-ovulated controls exhibited minimal P4 concentrations (0.86 ± 0.12 ng/mL). Plasma estradiol (E2) levels showed no significant differences across days or groups, with mean values of 32.74 ± 4.33 pg/mL on day 0 and 37.27 ± 5.49 pg/mL on day 7, respectively. Histological analysis confirmed that Kp10 promoted preovulatory follicle development and CL formation, mirroring GnRH effects. Additionally, Kp10 enhanced angiogenesis, indicated by increased vascular endothelial growth factor (VEGF) expression in more developed follicles and CL. These results suggest that Kp10 could be an alternative to GnRH for ovulation induction in rabbits, although further studies are needed to explore optimal analogues, doses, and administration routes.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"245 ","pages":"Article 117524"},"PeriodicalIF":2.4,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144254300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Doppler ultrasound assessment of uterine artery in canine endometritis","authors":"Sol Arioni ,&nbsp;Marlene Huk ,&nbsp;Pablo R. Batista ,&nbsp;Daniel O. Arias ,&nbsp;Cristina Gobello ,&nbsp;Paula G. Blanco","doi":"10.1016/j.theriogenology.2025.117526","DOIUrl":"10.1016/j.theriogenology.2025.117526","url":null,"abstract":"<div><div>This study aimed to describe and compare uterine artery velocimetry in bitches suffering from endometritis, cystic endometrial hyperplasia (CEH) and healthy animals, and to evaluate the effect of clinical, ultrasonographic, bacteriological and histopathological parameters on the uterine artery resistance index in these bitches. According to clinical, two-dimensional ultrasonographic, gross and histopathological uterine evaluation, thirty-three bitches were classified as: Endometritis (END; n = 13), CEH (n = 7) and healthy bitches (NG; n = 13). Uterine ultrasonographic examinations were performed on each patient using B mode and pulsed-wave Doppler. Peak systolic velocity (PSV), end diastolic velocity (EDV) and resistance index (RI) were obtained. A generalized linear model was carried out to analyze the effect of group, clinical, ultrasonographic, bacteriological and histopathological variables on PSV, EDV and RI. EDV was higher in CEH than END (P &lt; 0.05) and NG (P &lt; 0.01). No difference was observed between END and NG (P &gt; 0.1). END presented a lower RI than NG and higher RI than CEH (P &lt; 0.01). Additionally, CEH showed lower RI than NG (P &lt; 0.01). This study showed an effect of higher values of endometrium thickness, PMNN/HPF and vascular congestion on RI (P &lt; 0.01). It was concluded that, in these bitches, uterine artery RI was a valuable parameter to distinguish endometritis from normal uterus and CEH. An increase in neutrophils and vascular congestion influenced RI, remarking the importance of angiogenesis and the inflammatory reaction on uterine perfusion increase. This study is the first to identify an ultrasound parameter that differentiates endometritis from normal uterus in dogs.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"245 ","pages":"Article 117526"},"PeriodicalIF":2.4,"publicationDate":"2025-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144264136","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Nuclearporin subcomplex Nup98/Rae1 is vital for maternal-to-zygotic transition during early embryonic development","authors":"Luo Anfeng ,&nbsp;Gu Hao ,&nbsp;Akhtar Ali ,&nbsp;Qi Mengfan ,&nbsp;Pan Kaixin ,&nbsp;Zhou Changfan ,&nbsp;Zeng Wei ,&nbsp;Liu Song ,&nbsp;Ren Hongyan ,&nbsp;Bi Yanzhen ,&nbsp;Chen Fan","doi":"10.1016/j.theriogenology.2025.117519","DOIUrl":"10.1016/j.theriogenology.2025.117519","url":null,"abstract":"<div><div>The Nup98/Rae1 nuclear pore subcomplex, a critical mediator of nucleocytoplasmic transport, orchestrates fundamental cellular processes including transcriptional regulation, mRNA surveillance, and mitotic fidelity. However, its functional significance during early embryogenesis remains incompletely understood. In this study, we employed an in vitro embryo culture system combined with embryo electroporation-mediated interference to investigate the role of Nup98/Rae1 in early mouse embryos. Our results revealed that Nup98/Rae1 deficiency led to delayed embryonic progression and a significant decline in blastocyst formation rates. Transcriptomic analysis using SMART-seq2 in two-cell stage embryos revealed that Nup98/Rae1 modulates global gene expression, particularly within pathways governing RNA splicing, protein catabolism, and the DNA damage response. Integration of transcriptomic data with established databases further confirmed that Nup98/Rae1 is indispensable for zygotic genome activation and maternal mRNA clearance—key events in the maternal-to-zygotic transition. Moreover, quantitative immunofluorescence analysis demonstrated that loss of Nup98/Rae1 resulted in heightened DNA damage and reduced H3K27ac levels. Additionally, the increased mRNA expression of apoptosis-related markers <em>BAX</em> and <em>CASPASE</em>3, alongside positive TUNEL staining, indicated the induction of early apoptosis. Immunostaining for Sox2 and Cdx2 revealed a defective inner cell mass development, highlighting the detrimental impact of Nup98/Rae1 on cell fate specification. Collectively, these findings indicate that Nup98/Rae1 deficiency disrupts maternal mRNA degradation, impairs zygotic genome activation, alters histone modifications, induces genomic instability, and ultimately compromises early embryonic development by triggering apoptosis.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"245 ","pages":"Article 117519"},"PeriodicalIF":2.4,"publicationDate":"2025-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144231542","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Brain-derived neurotrophic factor (BDNF), an autocrine/paracrine regulator of basic ovarian functions and antagonist of kisspeptin","authors":"Alexander V. Sirotkin ,&nbsp;Zuzana Fabová ,&nbsp;Barbora Loncová ,&nbsp;Abdel Halim Harrath ,&nbsp;Sofía Elisa Alzuri ,&nbsp;María Belén Montiel ,&nbsp;Beata Fuchsová","doi":"10.1016/j.theriogenology.2025.117520","DOIUrl":"10.1016/j.theriogenology.2025.117520","url":null,"abstract":"<div><div>The aim of the present study was to examine whether brain-derived neurotrophic factor (BDNF) and BDNF-related molecules, such as the neuronal membrane glycoprotein M6a (GPM6A) and myocyte enhancer factor 2C (MEF2C), can be produced by ovarian cells and regulate ovarian cell functions as well as elucidate its functional interrelationships with the known regulator of ovarian cell functions kisspeptin (KP). For this purpose, we analyzed the expression of BDNF and GPM6A in rat ovaries and of BDNF, GPM6A and MEF2C in porcine ovarian granulosa cells; the influence of KP addition on the expression of BDNF, GPM6A and MEF2C; and the effects of BDNF, KP and the combination of BDNF + KP treatment on the proliferation, apoptosis and release of progesterone and estradiol by cultured porcine granulosa cells. The expression of BDNF, GPM6A and MEF2C was quantified by qPCR, and proliferation, apoptosis and hormone release were analyzed by BrdU incorporation, TUNEL and ELISA.</div><div>Our results demonstrate that both rat and porcine ovarian cells express BDNF and related molecules. Furthermore, the addition of BDNF stimulates the proliferation and release of progesterone and reduces the degree of apoptosis and estradiol output by porcine cells. KP has the opposite effect on these parameters. Furthermore, KP reduces the expression of BDNF and GPM6A but not MEF2C and suppresses the effects of BDNF on porcine granulosa cell apoptosis as well as progesterone and estradiol release. These observations are the first to demonstrate (1) the production of BDNF, GPM6A and MEF2C in the ovary, (2) the ability of BDNF to regulate ovarian cell proliferation, apoptosis and steroid hormone release by porcine ovarian cells, and (3) the antagonism between BDNF and KP production and their control of ovarian functions.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"246 ","pages":"Article 117520"},"PeriodicalIF":2.4,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144272112","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel method for parturition induction in cattle using vaginal wall stimulation","authors":"Shin Nakayama ,&nbsp;Daisuke Nakamitsu ,&nbsp;Daiki Aomori ,&nbsp;Shintaro Ohigashi ,&nbsp;Osamu Nishino ,&nbsp;Shinichi Tose","doi":"10.1016/j.theriogenology.2025.117522","DOIUrl":"10.1016/j.theriogenology.2025.117522","url":null,"abstract":"<div><div>Induction of parturition in cattle is used to reduce labor in calving management, prevent dystocia due to oversized calves, and treat prolonged gestation. Conventional pharmacological induction methods rely on prostaglandin F2α (PGF2α), dexamethasone, estrogens, and oxytocin (OXT), either individually or in combination. However, variability in the induction-to-calving time (ICT), including a prolonged ICT, remains a major challenge. In this study, we developed a novel method for inducing parturition by combining conventional pharmacological induction with vaginal wall stimulation (VWS), which artificially activates the Ferguson reflex and enhances endogenous OXT secretion. We compared its effects on the ICT, the incidence of dystocia, and neonatal vitality with those of pharmacological induction alone to evaluate the efficacy and safety of this new approach. Japanese Black breeding cows (n = 15 per group) in the control group received PGF2α, dexamethasone, and estriol 1 day before expected calving, while the VWS group received the same agents followed by VWS approximately 24 h later. Compared with the control group, the VWS group showed a statistically significant reduction in the ICT and decreased ICT variability. Given the statistically significant promotion of labor observed in the VWS group, this improvement in ICT may be attributable to the labor-inducing effect of VWS. Additionally, no significant differences were observed in obstetrical intervention rates or neonatal calf vitality. This novel approach may facilitate more rapid and consistent parturition while maintaining safety for both cows and newborn calves, and may contribute to improving calving management efficiency and reducing the workload of livestock producers.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"245 ","pages":"Article 117522"},"PeriodicalIF":2.4,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144241580","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cryopreservation at −80 °C impacts sperm integrity and fertility in a mouse strain-dependent manner","authors":"Manon Peltier ,&nbsp;Marcello Raspa ,&nbsp;Sabrina Putti ,&nbsp;Renata Paoletti ,&nbsp;Ferdinando Scavizzi ,&nbsp;Esther Mahabir","doi":"10.1016/j.theriogenology.2025.117523","DOIUrl":"10.1016/j.theriogenology.2025.117523","url":null,"abstract":"<div><div>We previously developed a simple method to cryopreserve and maintain B6N spermatozoa at −80 °C, instead of using liquid nitrogen (LN₂), for up to one year without detrimental effects on fertility. The goal of the present study was to test the efficacy of this method in six different mouse strains (129/SvImJCnrm; 129, C57BL/6JCnrm; B6J, C57BL/6NTacCnrm; B6N, BALB/cByJCnrm; BALB/c, Crl:CD1(ICR); CD-1, and FVBN/JCnrm; FVB). At different time points up to one year, we evaluated the integrity and the fertility of the spermatozoa cryopreserved and stored at −80 °C in comparison to the classical LN₂ method. After 1 year, no differences in the <em>in vitro</em> fertilisation (IVF) rate and in the number of dead spermatozoa were observed in 129, B6N, CD-1 and FVB strains. In contrast, spermatozoa from B6J and BALB/c cryopreserved at −80 °C showed a significant reduction in the IVF rate, an increased number of dead spermatozoa and increased morphological abnormalities after 3 months (BALB/c) and 6 months (B6J). In all strains, the total sperm motility decreased after 1 month at −80 °C and increased ultrastructural damage was found in the −80 °C group. The present results indicate that spermatozoa from 129, B6N, CD-1 and FVB can be cryopreserved and stored at −80 °C for at least one year without impacting their fertility. However, spermatozoa from B6J and BALB/c are the most sensitive to temperature. Therefore, cryopreservation and storage of 129, B6N, CD-1 and FVB strains at −80 °C for up to one year is feasible, thereby circumventing the use of LN₂.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"245 ","pages":"Article 117523"},"PeriodicalIF":2.4,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144262186","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Towards the best enzymatic treatment prior to ovarian tissue cryopreservation. A statistical and model-based analysis","authors":"Alodia Lacueva-Aparicio ,&nbsp;Jacobo Ayensa-Jiménez ,&nbsp;María Francisca Perulero ,&nbsp;Dácil Diaz ,&nbsp;Sigrid Zalaya ,&nbsp;Emilio Ignacio Abecia ,&nbsp;Clara Malo","doi":"10.1016/j.theriogenology.2025.117521","DOIUrl":"10.1016/j.theriogenology.2025.117521","url":null,"abstract":"<div><div>The objective was to study an innovative approach based on a mathematical model to analyze the potential benefits of using enzymatic pre-treatment of ovarian tissue to increase its permeability. Our approach aims to elucidate whether enzymatic pre-treatment would enhance the penetration of cryoprotectants in the ovarian tissue, leading to better preservation and giving best results for obtaining primordial and primary follicles without damage. The effect of five enzymatic treatments (TrypLE, collagenase, dispase, accutase, hyaluronidase) over time on sheep ovarian tissue was evaluated, focusing on penetration depth and tissue damage. First results showed that up to 45 min of incubation, all enzymes penetrated similarly, but longer treatments revealed differences. Dispase showed the greatest penetration at 180 min but was highly aggressive, causing rapid tissue lysis. TrypLE and hyaluronidase were found to be the most effective in minimizing follicular damage while ensuring sufficient tissue penetration, particularly when applied for 10 min. TrypLE caused significant edema after 10 min, but this was reduced after 15 min. Hyaluronidase caused minimal edema and had excellent penetration, making it ideal for enhancing ovarian tissue permeability. The present study showed that pre-treatments with hyaluronidase and TrypLe for 10 min are potential candidates for enhancing ovarian tissue permeabilization, offering the best balance between penetration and minimal tissue damage. These findings could improve fertility preservation techniques and future cryopreservation strategies.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"245 ","pages":"Article 117521"},"PeriodicalIF":2.4,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144222498","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhanced maturation of mouse immature oocytes using a novel three-dimensional culture system","authors":"Xinyang Zhao (First author) ,&nbsp;Xudong Zhang ,&nbsp;Shanshan Wu ,&nbsp;Siwen Zhang ,&nbsp;Hsun-Ming Chang ,&nbsp;Peter C.K. Leung ,&nbsp;Jichun Tan","doi":"10.1016/j.theriogenology.2025.117518","DOIUrl":"10.1016/j.theriogenology.2025.117518","url":null,"abstract":"<div><div>The effective utilization of immature oocytes holds promise for improving conception rates in women with low ovarian reserve. The oocyte microenvironment, comprising the extracellular matrix and intercellular communication with somatic cells, is crucial for supporting oocyte maturation. This study hypothesizes that a three-dimensional (3D) culture system, constructed with hydrogels modified by laminin-mimetic peptides—a major component of the extracellular matrix—and granulosa cells (GCs) from mature mouse oocytes, can replicate the biomechanical microenvironment required for oocyte maturation, thereby promoting the development of immature mouse oocytes. To validate this hypothesis, we initially assessed the physical properties of hydrogels and determined the optimal concentration for in vitro oocyte culture. A total of 304 germinal vesicle (GV) phase mouse oocytes were retrieved and randomly allocated to two-dimensional (2D), 3D, and 3D-GCs culture groups. The 3D-GCs group exhibited the highest rate of first polar body extrusion (83.1 ± 5.0 %). Oxidative stress levels and mitochondrial membrane potential were evaluated using DCFH-DA and JC-1 staining, confirming that 3D culture significantly reduced oxidative stress and enhanced mitochondrial activity. Cytoplasmic maturation, assessed by cortical granule distribution and endoplasmic reticulum organization, further highlighted the superiority of the 3D-GCs system. The 3D-GCs group demonstrated 87.0 % of oocytes with grade III cortical granule distribution and 84.6 % with mature endoplasmic reticulum, significantly surpassing the other groups (p &lt; 0.05). In conclusion, the 3D-GCs culture system effectively supports synchronized nuclear and cytoplasmic maturation in immature oocytes. This approach provides a promising platform for improving the developmental competence of oocytes, potentially benefiting women with diminished ovarian reserve.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"245 ","pages":"Article 117518"},"PeriodicalIF":2.4,"publicationDate":"2025-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144254299","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Nitric oxide regulates spindle dynamics to modulate the maturation of goat oocytes","authors":"Rui Xu ,&nbsp;Zhi Zheng ,&nbsp;Weizhao Bai ,&nbsp;Minghui Liu ,&nbsp;Sihai Lu ,&nbsp;Sha Peng ,&nbsp;Menghao Pan ,&nbsp;Baohua Ma","doi":"10.1016/j.theriogenology.2025.117517","DOIUrl":"10.1016/j.theriogenology.2025.117517","url":null,"abstract":"<div><div>Oocyte maturation is a complex and tightly regulated process. Nitric oxide (NO) has been implicated in the regulation of oocyte maturation, but its precise mechanism of action remains unclear. In this study, the inhibition of NO synthase activity using L-NMMA was found to impair meiosis and inhibit the maturation of goat oocytes. Further analysis revealed that the meiotic arrest induced by L-NMMA was primarily due to disruptions in metaphase I (MI) spindle dynamics. Specifically, L-NMMA treatment led to disorganized MI spindle assembly, abnormal chromosome alignment, elevated levels of microtubule acetylation, and sustained activation of the spindle assembly checkpoint. These defects were partially rescued by supplementation with the NO donor sodium nitroprusside (SNP). Proteomic analysis of oocytes from the CON and L-NMMA-treated groups identified a significant downregulation of KIF15 in the L-NMMA group. KIF15 has previously been established as a key regulator of spindle dynamics during oocyte maturation. Moreover, we demonstrated that the RhoA–ROCK signaling pathway modulated KIF15 protein expression in goat oocytes. Our results further showed that reduced NO levels increased RhoA phosphorylation, which in turn suppressed KIF15 expression and disrupted MI spindle dynamics. In conclusion, this study provides new insights into the role of NO in goat oocyte maturation, revealing that it regulates spindle dynamics through the RhoA–ROCK-KIF15 signaling axis.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"245 ","pages":"Article 117517"},"PeriodicalIF":2.4,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144189605","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}