TheriogenologyPub Date : 2025-07-11DOI: 10.1016/j.theriogenology.2025.117575
E.A. Spanner, S.P. de Graaf, J.P. Rickard
{"title":"The validation and application of an ovine fertility model using standardised in vitro thresholds to predict the likelihood of pregnancy","authors":"E.A. Spanner, S.P. de Graaf, J.P. Rickard","doi":"10.1016/j.theriogenology.2025.117575","DOIUrl":"10.1016/j.theriogenology.2025.117575","url":null,"abstract":"<div><div>Deciphering a ram or ewe's reproductive potential is crucial to ensure high reproductive performance and maximise production outcomes. This study validates the accuracy of an ovine fertility model created to predict the likelihood of pregnancy occurring following laparoscopic artificial insemination (AI) and proposes <em>in vitro</em> semen standards to improve pregnancy outcomes. Semen from Merino sires (N = 26) was inseminated into synchronised Merino ewes (N = 1269) across 3 breeding seasons (2021–2023). Uterine tone and intra-abdominal fat of ewes were scored at AI, while the freezing concentration, abnormal sperm, acrosome viability (6h) and CASA motility and velocity traits (0h) of semen inseminated was assessed post-thaw (6h; 37 °C). Pregnancy predictions were compared with ultrasound-confirmed pregnancies ∼55 days post-AI, using discrimination and calibration tests to correctly assess its ability to classify pregnant and non-pregnant ewes. The model demonstrated high accuracy (77 %), precision (96 %) and recall (76 %) but lower specificity (33 %). It recorded an F1-score of 0.85, with an Area Under the Curve (AUC) of 0.62. There was no statistical difference between predicted and actual pregnancy results (P = 0.184) despite an error value of 26 %. A cutting point split the data for each <em>in vitro</em> semen predictor and calculated the average pregnancy rate above and below this point. The cutting point with the greatest difference between pregnancy rates was chosen as the semen threshold. When entered into the model, these thresholds returned a cumulative pregnancy probability of 64.3 %. These standards could be used to screen semen before AI, reducing the variability of laparoscopic AI programs for the industry.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"247 ","pages":"Article 117575"},"PeriodicalIF":2.4,"publicationDate":"2025-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144632094","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}
TheriogenologyPub Date : 2025-07-11DOI: 10.1016/j.theriogenology.2025.117573
Yueming Wang , Yu Wang , Yashi Fan , Fanglin Xiang, Yuechan Wang, Jinmei Cheng
{"title":"Astragaloside IV improves the quality of in vitro postovulatory aged oocytes by decreasing oxidative stress in mice","authors":"Yueming Wang , Yu Wang , Yashi Fan , Fanglin Xiang, Yuechan Wang, Jinmei Cheng","doi":"10.1016/j.theriogenology.2025.117573","DOIUrl":"10.1016/j.theriogenology.2025.117573","url":null,"abstract":"<div><div>Ovulated oocytes deteriorate rapidly if not fertilized within the optimal timeframe, known as postovulatory aging. Astragaloside IV (AS-IV), an active ingredient of <em>Astragalus membranaceus</em>, has been demonstrated to have anti-aging and antioxidant effects. However, it has not been elucidated whether AS-IV mitigates <em>in vitro</em> postovulatory oocyte aging. Here, AS-IV was administered into the <em>in vitro</em> culture buffer, and we found that its addition could improve the quality of <em>in vitro</em> postovulatory aged oocytes in mice. Specifically, AS-IV treatment enhanced morphological integrity, preserved spindle/chromosome structure, stabilized actin cytoskeleton, maintained centromere cohesion, and improved ovastacin dynamics in aging oocytes. These improvements collectively enhanced sperm binding capacity and subsequent developmental competence following parthenogenetic activation. Mechanistically, AS-IV appears to inhibit mRNA degradation of antioxidant enzymes and sustain mitochondrial function, thereby preventing reactive oxygen species (ROS) accumulation during <em>in vitro</em> oocyte aging. Meanwhile, reduced oxidative stress in AS-IV-treated oocytes may counteract apoptosis-related mRNA accumulation, delaying apoptotic initiation. Taken together, our findings demonstrate that AS-IV supplementation effectively improves the quality of <em>in vitro</em>-aged postovulatory oocytes. This provides a valuable reference and evidence base for its potential application in human-assisted reproductive technology and livestock embryo production.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"247 ","pages":"Article 117573"},"PeriodicalIF":2.4,"publicationDate":"2025-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144605044","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}
TheriogenologyPub Date : 2025-07-10DOI: 10.1016/j.theriogenology.2025.117571
Sebastian P. Arlt , Janna Hinderer , Lisa Riege , Johanna Leber , Elisabeth Müller , Corinna Weber , Beate Walter
{"title":"Serum anti-Müllerian hormone (AMH) concentrations in pregnant and lactating dogs","authors":"Sebastian P. Arlt , Janna Hinderer , Lisa Riege , Johanna Leber , Elisabeth Müller , Corinna Weber , Beate Walter","doi":"10.1016/j.theriogenology.2025.117571","DOIUrl":"10.1016/j.theriogenology.2025.117571","url":null,"abstract":"<div><div>Measurement of Anti-Müllerian hormone (AMH) in female dogs is becoming increasingly important in clinical practice. It has been shown earlier that concentrations vary throughout the estrous cycle. AMH concentrations are known to be high during proestrus and estrus, then decrease during the luteal phase and anestrus.</div><div>Aim of this study was to investigate variations during pregnancy and lactation. Therefore, blood samples were analyzed from 40 pregnant and 19 non-pregnant privately owned dogs of various breeds. One sample was collected from each dog during estrus, three samples during pregnancy, one at three weeks in lactation, and one after weaning. The measured concentrations were compared with those from the non-pregnant dogs, from which samples were collected at the same corresponding time points.</div><div>AMH concentration in pregnant dogs was highest in estrus (0.63 ng/ml, quartiles 0.40 and 0.87) and remained relatively high during pregnancy. At sampling during lactation the median concentration was significantly lower compared with all other time points (median 0.19 ng/ml, quartiles 0.14 and 0.26). After weaning, AMH increased again to 0.30 ng/ml (quartiles 0.24 and 0.50). During late pregnancy, lactation and after weaning, the AMH concentration was lower in the pregnant dogs compared with the non-pregnant ones. In addition, the significant decline of AMH concentration in pregnant dogs during lactation was not present in the non-pregnant dogs.</div><div>It can be hypothesized that higher prolactin concentrations may reduce AMH synthesis by inhibiting the secretion of GnRH, FSH, and LH in mammals. In our study we found moderate negative correlation between bodyweight or the number of suckling puppies and AMH concentration. However, since no assay was available, prolactin concentrations were not measured. These findings indicate that pregnancy and lactation status should be considered when interpreting AMH concentrations in clinical practice.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"247 ","pages":"Article 117571"},"PeriodicalIF":2.4,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144632116","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}
TheriogenologyPub Date : 2025-07-09DOI: 10.1016/j.theriogenology.2025.117569
Michal Andrzej Kosior , Valentina Longobardi , Chiara Del Prete , Riccardo Esposito , Federica Piscopo , Giorgio Antonio Presicce , Alessio Cotticelli , Tanja Peric , Natascia Cocchia
{"title":"Effect of a single administration of FSH delivered in hyaluronic acid on oocyte competence and hormonal concentrations in Italian Mediterranean buffaloes undergoing ovarian stimulation prior to ovum pick-up","authors":"Michal Andrzej Kosior , Valentina Longobardi , Chiara Del Prete , Riccardo Esposito , Federica Piscopo , Giorgio Antonio Presicce , Alessio Cotticelli , Tanja Peric , Natascia Cocchia","doi":"10.1016/j.theriogenology.2025.117569","DOIUrl":"10.1016/j.theriogenology.2025.117569","url":null,"abstract":"<div><div>This study evaluated the effectiveness of a single intramuscular administration of follicle-stimulating hormone (FSH) dissolved in 1 % hyaluronic acid (HA) for ovarian stimulation in Italian Mediterranean buffaloes (IMB) undergoing ovum pick-up (OPU). The aim was to simplify hormonal protocols, reduce handling stress, and improve oocyte competence. In Experiment 1, multiparous IMB (n = 24) were enrolled in a crossover design comparing the standard six-dose FSH protocol (FSH-6) with a single FSH-HA injection. Transrectal ultrasonography was used to record the number and the size of follicles, in vitro embryo production to assess oocyte developmental competence, and an in-house radioimmunoassay (RIA) method to measure plasma cortisol levels. In Experiment 2, plasma FSH profile during follicular growth was evaluated in IMB (n = 12) subjected to FSH-HA, FSH-6, or no stimulation (control) protocols. The FSH-HA treatment resulted in a higher (P < 0.05) proportion of medium-sized follicles compared to FSH-6 (3.4 ± 2.7 and 2.0 ± 2.3, respectively) as well as a significantly greater number of cleaved oocytes (3.3 ± 0.4 and 2.4 ± 0.4 respectively), although the total embryo yield remained similar (30.1 ± 7.2 and 29. 2 ± 4.5, respectively). Cortisol concentrations increased after OPU in all groups, but the FSH-HA group showed a trend (P < 0.10) toward lower stress levels compared to FSH-6. In Experiment 2, FSH-HA maintained higher plasma FSH concentrations for a longer period, with peak values observed between 30 and 72 h post-administration, suggesting the importance of optimizing the timing of OPU in HA-based protocols.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"247 ","pages":"Article 117569"},"PeriodicalIF":2.4,"publicationDate":"2025-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144605043","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":"Colostrum and milk production in sows housed in free-farrowing versus crated systems under tropical conditions: Associations with sow metabolic state, oxidative stress, and piglet survival","authors":"Natchanon Dumniem , Junpen Suwimonteerabutr , Padet Tummaruk","doi":"10.1016/j.theriogenology.2025.117572","DOIUrl":"10.1016/j.theriogenology.2025.117572","url":null,"abstract":"<div><div>Modern prolific sows experience substantial metabolic demands while nursing large litter, often leading to fat mobilization and oxidative stress. This study investigated the impacts of farrowing systems (free-farrowing pens vs. crates) and lactational backfat loss on colostrum yield, milk production, metabolic indicators (serum insulin, IGF-1, NEFA), and plasma MDA levels, in 76 crossbred sows raised under tropical conditions. From farrowing to weaning, sows were monitored for backfat loss, farrowing duration, litter traits, piglet preweaning mortality, and metabolic profiles. All sows received routine treatment with anti-inflammatory and antibiotic medications to manage <em>postpartum</em> uterine infection and inflammation, and to reduce the severity of health issues under tropical conditions. Sows were measured for backfat thickness, loin muscle depth, and their lactational loss, along with metabolic state, and oxidative stress profiles from farrowing to weaning. On average, sows lost 28.6 % of backfat and 15.9 % of loin muscle during lactation, with no differences between housing systems. Sows in free-farrowing pens produced more colostrum (5.6 vs. 5.0 kg; <em>P</em> = 0.035) and exhibited higher IGF-1 levels on day 1 <em>postpartum</em> (<em>P</em> = 0.018). NEFA concentrations tended to be higher at weaning in free-farrowing sows (<em>P</em> = 0.070), while insulin, MDA, and milk yield did not differ between groups. Preweaning mortality tended to be higher among piglets from free-farrowing sows than those from crated sows (24.7 % vs. 16.6 %, <em>P</em> = 0.074). Sows with high backfat loss produced more milk during both days 3–10 and 10–17 of lactation (<em>P</em> < 0.05), and colostrum yield was positively associated with subsequent milk production. Total preweaning mortality from days 0–21 was higher among piglets born to sows with moderate backfat loss (26.2 %) compared to those with low backfat loss (15.6 %, <em>P</em> = 0.037), but not different from those with high backfat loss (20.5 %, <em>P</em> = 0.263). In summary, the housing system influenced early lactation metabolism, with free-farrowing enhancing colostrum production and IGF-1 secretion, potentially due to increased physical activity and reduced stress, which may improve mammary blood flow and support hormonal regulation. High backfat loss was linked to greater milk output and reflected increased energy demand. These findings highlight the need for tailored management strategies, based on individual body condition and farrowing environment, to support sow metabolic status, oxidative resilience, and piglet survival in tropical production systems.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"247 ","pages":"Article 117572"},"PeriodicalIF":2.4,"publicationDate":"2025-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144605639","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}
TheriogenologyPub Date : 2025-07-09DOI: 10.1016/j.theriogenology.2025.117570
Li-Ying Liu , Xian-Feng Yu , Zhi-Chao Chi , Shu-Ming Shi , Tian Lan , Jia-Hui Leng , Jing-hang Li , Yu-Yan He , Lin-Yi Qu , Guan-Lin Jia , Ilkeun Kong , Ming-Jun Zhang , Yong-Xun Jin
{"title":"Salidroside improves bovine embryo quality by mitigating oxidative stress and regulating mitochondrial lipid metabolism","authors":"Li-Ying Liu , Xian-Feng Yu , Zhi-Chao Chi , Shu-Ming Shi , Tian Lan , Jia-Hui Leng , Jing-hang Li , Yu-Yan He , Lin-Yi Qu , Guan-Lin Jia , Ilkeun Kong , Ming-Jun Zhang , Yong-Xun Jin","doi":"10.1016/j.theriogenology.2025.117570","DOIUrl":"10.1016/j.theriogenology.2025.117570","url":null,"abstract":"<div><div>In vitro production techniques for bovine embryos can improve reproductive efficiency and expand quality breeding stock, but lipid metabolism disturbances during in vitro embryo culture can decrease embryo quality. Salidroside (SAL) is a glycoside extracted from the rhizome of the medicinal plant <em>Rhodiola rosea</em> that has antioxidant, antiaging, anti-inflammatory, and lipid metabolism-regulating effects. This study demonstrated that the addition of SAL to the culture medium of bovine embryos during in vitro culture increased the blastocyst rate and number of blastocyst cells and improved bovine blastocyst totipotency and proliferation. SAL reduced the lipid droplet content in bovine blastocysts and increased the levels of lipolysis-related genes (<em>PNPLA2, LIPE,</em> and <em>MGLL</em>). Fatty acids serve as ligands to activate PPARα and promote the transcription and expression of downstream fatty acid β-oxidation-related genes (<em>CPT1A, CPT2, ACOX1,</em> and <em>ACOX2</em>). SAL reduced the ROS level, increased the GSH level, increased the expression of antioxidant-related proteins (Nrf2 and downstream HO-1), and increased the levels of antioxidant enzyme-related genes (<em>GPx1, SOD1, SOD2,</em> and <em>CAT</em>) in bovine embryos. SAL increased the mitochondrial membrane potential and mitochondrial function, number, and distribution; facilitated mitochondria‒lipid droplet interactions; increased fatty acid availability to mitochondria; and further enhanced fatty acid β-oxidation. In conclusion, SAL not only acts as an antioxidant to reduce oxidative stress generated during in vitro bovine embryo culture but also promotes lipolysis to produce free fatty acids (FFA) to activate PPARα, enhances fatty acid β-oxidation, regulates lipid metabolism, and reduces the lipid content in blastocysts, thereby improving embryo developmental competence.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"247 ","pages":"Article 117570"},"PeriodicalIF":2.4,"publicationDate":"2025-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144605042","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}
TheriogenologyPub Date : 2025-07-08DOI: 10.1016/j.theriogenology.2025.117568
Yiwen Sha , Xue Qin , Weijun Pang
{"title":"Histone modifications: key mechanisms in embryo implantation of domestic animal","authors":"Yiwen Sha , Xue Qin , Weijun Pang","doi":"10.1016/j.theriogenology.2025.117568","DOIUrl":"10.1016/j.theriogenology.2025.117568","url":null,"abstract":"<div><div>Embryo implantation is a critical event in the reproductive development of domestic animals, where research on this process holds significant practical importance for enhancing animal production efficiency and reducing embryo loss. As an essential epigenetic regulatory mechanism, histone modifications can profoundly influence cell function and developmental processes by altering chromatin structure and gene expression patterns. In recent years, with the continuous development of epigenetic technologies, more and more studies have begun to reveal the important roles of histone modifications in embryonic development. This review specifically highlights the regulation of embryo activation, endometrial receptivity and decidualization by histone modifications in farm animals, aiming to provide a comprehensive understanding of their impact on embryo implantation. From this perspective, in-depth research on histone modifications in livestock species can not only offer new insights into the molecular regulatory mechanisms of embryo implantation but also enhance our understanding of reproductive development in farm animals.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"247 ","pages":"Article 117568"},"PeriodicalIF":2.4,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144605045","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}
TheriogenologyPub Date : 2025-07-07DOI: 10.1016/j.theriogenology.2025.117567
Guangzheng Liu , Xilin Zhu , Zeyu Li , Liqian Zhu , Guohong Chen , Yingjie Niu , Hongyan Sun , Qisheng Zuo , Jiuzhou Song , Wei Han , Wanhong Wei , Bichun Li , Kai Jin
{"title":"Generation of chicken induced pluripotent stem cells (iPSCs) from somatic cells using multi-lentiviral transduction and OAC2","authors":"Guangzheng Liu , Xilin Zhu , Zeyu Li , Liqian Zhu , Guohong Chen , Yingjie Niu , Hongyan Sun , Qisheng Zuo , Jiuzhou Song , Wei Han , Wanhong Wei , Bichun Li , Kai Jin","doi":"10.1016/j.theriogenology.2025.117567","DOIUrl":"10.1016/j.theriogenology.2025.117567","url":null,"abstract":"<div><div>The chicken iPSCs have been successfully established and exhibit significant potential for the recovery and conservation of endangered avian species. However, the efficiency of induction remains quite limited. To enhance the induction system, a six-factor reprogramming approach incorporating OCT4, SOX2, NANOG, LIN28, KLF4, and C-MYC (OSNLKM) was employed via lentiviral infection, in combination with the small-molecule compound OAC2 and mouse embryonic fibroblast (MEF) feeder layers. The results demonstrated that the six-factor system significantly accelerated the reprogramming process and improved induction efficiency compared to the conventional four-factor system (OCT4, SOX2, NANOG, and LIN28; OSNL). Additionally, the MEF feeder layers were found to be essential for the successful reprogramming of chicken somatic cells. The inclusion of Oct4-activating compound 2 (OAC2) further optimized the reprogramming conditions, enabling the successful induction of iPSCs from chicken embryonic fibroblasts (CEFs) and Sertoli cells. Collectively, this study establishes an optimized reprogramming system for chicken somatic cells, enhancing iPSCs induction efficiency and expanding the potential applications of iPSCs technology in avian conservation and genetic modification.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"247 ","pages":"Article 117567"},"PeriodicalIF":2.4,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144588362","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":"DUXA regulates the early development of porcine cloned embryos","authors":"Zhenhu Shi, Yelian Yan, Ruiqing Zhu, Xinyue Zhu, Kunlong Hu, Yingying Yue, Wenhuan Xu, Mengqing Xuan, Xinqi Gan, Zhiyuan Yang, Yunhai Zhang , Zubing Cao","doi":"10.1016/j.theriogenology.2025.117563","DOIUrl":"10.1016/j.theriogenology.2025.117563","url":null,"abstract":"<div><div>Abnormal zygotic genome activation (ZGA) during the early development of somatic cell nuclear transfer (SCNT) embryos is one of the main reasons for the low cloning efficiency. The double homeobox (DUX) family, which includes important transcription factors in mammals, has been shown to play an important role in the ZGA process in mice. However, the role of DUXA, a member of the DUX family, in the early development of porcine somatic cloned embryos is unknown. Here, CRISPR/Cas9 gene editing and lentiviral infection technologies were used to construct stable DUXA knockout and overexpression cell lines for the production of SCNT embryos. Compared with that of wild-type (WT) SCNT embryos, the blastocyst rate of <em>DUXA</em> knockout embryos was significantly lower (P < 0.05), whereas the blastocyst rate of <em>DUXA</em>-overexpressing embryos was significantly greater (P < 0.05). Moreover, RT‒qPCR results revealed that <em>DUXA</em> knockout significantly reduced the expression levels of ZGA-related genes (<em>TDG</em>, <em>SNAI1</em>, <em>RSRP1</em>, <em>TFAP2C, ZSCAN4, LEUTX, and KLF17)</em> (P < 0.05). Additionally, in <em>DUXA-</em>overexpressing embryos, the mRNA levels of <em>TDG</em>, <em>SNAI1</em>, <em>RSRP1</em>, and <em>TFAP2C</em> significantly decreased (P < 0.05), whereas the <em>ZSCAN4</em>, <em>LEUTX</em>, and <em>KLF1</em>7 mRNA levels increased (P < 0.05). These findings suggest that <em>DUXA</em> regulates the early development of porcine SCNT embryos by modulating the expression of ZGA-related genes. This research provides significant insights into the potential mechanisms of early embryo loss in porcine SCNT.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"247 ","pages":"Article 117563"},"PeriodicalIF":2.4,"publicationDate":"2025-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144605638","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":"Antioxidant and antiapoptotic effect of melatonin on cat vitrified oocytes","authors":"Martina Colombo, Alessandra Mascaro, Alessandro Pecile, Jasmine Fusi, Gaia Cecilia Luvoni","doi":"10.1016/j.theriogenology.2025.117566","DOIUrl":"10.1016/j.theriogenology.2025.117566","url":null,"abstract":"<div><div>The development of cryopreservation protocols for feline gametes is a crucial part of conservation efforts. Preservation of oocytes is particularly challenging, since they often degenerate after warming or develop into embryos poorly. Two of the main causes are thought to be oxidative stress and apoptotic cell death. Thus, melatonin was employed in this study for its well-known antioxidant and antiapoptotic properties. After determining suitable concentrations of melatonin (Experiment I), the oxidative status (Experiment II), apoptotic status (Experiment III), and developmental competence (Experiment IV) of cat immature oocytes following vitrification and/or in vitro maturation (IVM) with the supplementation of melatonin were assessed. In Experiment I, 10<sup>−9</sup> M was the lowest melatonin concentration with favorable maturation of fresh oocytes (58.06 %, p = 0.6 vs. control, 51.48 %). In Experiment II, oocytes vitrified with melatonin showed decreased oxidative stress after IVM, close to negative control (p = 0.849). In Experiment III, the addition of melatonin to vitrification-warming and IVM medium hindered apoptotic signaling by reducing caspase activity (p = 0.9 vs. fresh control oocytes) and enhancing maturation rates (48.39 % vs. 12.12 % in control vitrified oocytes; p = 0.002). In Experiment IV, melatonin brought maturation rates of treated vitrified oocytes close to those of fresh oocytes (46.38 ± 10.68 % and 68.42 ± 10.88 %, respectively, p = 0.145), but there were no differences in embryo development or morphological quality (cleavage rates: melatonin-treated oocytes: 33.29 ± 17.86 %; control vitrified oocytes: 31.59 ± 21.63 %; p = 0.992). In summary, the use of the antioxidant and antiapoptotic melatonin during vitrification-warming and IVM mitigated oxidative and apoptotic stress in vitrified oocytes, though its impact on developmental competence remained limited.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"247 ","pages":"Article 117566"},"PeriodicalIF":2.4,"publicationDate":"2025-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144588364","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}