Pub Date : 2025-12-18DOI: 10.1016/j.theriogenology.2025.117791
Mingyue Sui , Zhihao Wang , Jianqiang Sun , Ruohan Mei , Mingju Sun , Yichen Liu , Xiang Gao , Youwei Li , Yanjun Huan
Germinal vesicle breakdown (GVBD) is a pivotal event in oocyte nuclear maturation, yet its underlying molecular mechanisms remain incompletely elucidated. This study showed that GVBD initiation in porcine oocytes occurred at 19 h of in vitro maturation, and this process was characterized by a significant increase in lamin A/C phosphorylation, prominent spindle assembly, and marked phosphorylation coupled with nuclear translocation of p38α MAPK (P < 0.05). Inhibition of p38 MAPK with SB203580 substantially suppressed p-p38α MAPK level, resulting in a significant decrease in GVBD rate, accompanied by notably diminished p-lamin A/C and impaired spindle assembly (P < 0.05). Single-cell transcriptome sequencing identified Rap1b, a small GTPase, as a potential upstream regulator of p38 MAPK pathway during GVBD. Further study demonstrated that Rap1b overexpression significantly enhanced both p-p38α MAPK and p-lamin A/C, promoted spindle assembly, and increased GVBD rate. Conversely, Rap1b knockdown markedly suppressed p-p38α MAPK and p-lamin A/C, disrupted spindle formation and decreased GVBD rate (P < 0.05). Further investigation revealed that Rap1b acted by promoting p-MKK6 to activate p38α MAPK signaling pathway. Collectively, these findings demonstrate that Rap1b is an essential regulator of oocyte GVBD by activating p38α MAPK signaling pathway. This work provides novel insights into the molecular regulation of oocyte meiotic resumption and offers a strategic basis for enhancing oocyte maturation.
{"title":"Rap1b is a critical regulator of germinal vesicle breakdown in porcine oocytes","authors":"Mingyue Sui , Zhihao Wang , Jianqiang Sun , Ruohan Mei , Mingju Sun , Yichen Liu , Xiang Gao , Youwei Li , Yanjun Huan","doi":"10.1016/j.theriogenology.2025.117791","DOIUrl":"10.1016/j.theriogenology.2025.117791","url":null,"abstract":"<div><div>Germinal vesicle breakdown (GVBD) is a pivotal event in oocyte nuclear maturation, yet its underlying molecular mechanisms remain incompletely elucidated. This study showed that GVBD initiation in porcine oocytes occurred at 19 h of in vitro maturation, and this process was characterized by a significant increase in lamin A/C phosphorylation, prominent spindle assembly, and marked phosphorylation coupled with nuclear translocation of p38α MAPK (P < 0.05). Inhibition of p38 MAPK with SB203580 substantially suppressed p-p38α MAPK level, resulting in a significant decrease in GVBD rate, accompanied by notably diminished p-lamin A/C and impaired spindle assembly (P < 0.05). Single-cell transcriptome sequencing identified Rap1b, a small GTPase, as a potential upstream regulator of p38 MAPK pathway during GVBD. Further study demonstrated that Rap1b overexpression significantly enhanced both p-p38α MAPK and p-lamin A/C, promoted spindle assembly, and increased GVBD rate. Conversely, Rap1b knockdown markedly suppressed p-p38α MAPK and p-lamin A/C, disrupted spindle formation and decreased GVBD rate (P < 0.05). Further investigation revealed that Rap1b acted by promoting p-MKK6 to activate p38α MAPK signaling pathway. Collectively, these findings demonstrate that Rap1b is an essential regulator of oocyte GVBD by activating p38α MAPK signaling pathway. This work provides novel insights into the molecular regulation of oocyte meiotic resumption and offers a strategic basis for enhancing oocyte maturation.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117791"},"PeriodicalIF":2.5,"publicationDate":"2025-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145789547","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}
Pub Date : 2025-12-17DOI: 10.1016/j.theriogenology.2025.117789
Maria Eugenia Sella , Anabella Andrea Campagna , Guillermo Luis Taminelli , Mariana Carolina Fabra , Alejandro Relling , Noelia Nikoloff , Ana Cristina Carranza-Martin
This study evaluated the cytoprotective, antioxidant, and anti-inflammatory effects of Resolvin D1 (RvD1) during bovine oocyte in vitro maturation (IVM) and its impact on embryo development. Four experiments were performed: (1) cytoprotective effect by oocyte nuclear maturation, mitochondrial density, and cumulus cell (CC) DNA damage; (2) antioxidant activity by intracellular reactive oxygen species (ROS) and glutathione (GSH) concentration in CCs and oocytes; (3) anti-inflammatory action by mRNA expression of IL-1β, IL-6, and TNF-α in CCs; and (4) developmental competence by cleavage, blastocyst, and hatching rate. In Experiments 1–2, IVM supplementation were: control (0.1 % ethanol), 25, 50, or 75 pg RvD1/mL; Experiments 3–4 IVM supplementation were: control and 50 pg RvD1/mL. Cumulus oocyte-complexes underwent IVM for 24 h at 39 °C in 5 % CO2. In experiment 1, RvD1 did not alter nuclear maturation or CC DNA damage. Mitochondrial density was marginally greater at 25 pg RvD1/mL compared with control. In experiment 2, in CC, ROS were lesser with 50 and 75 pg RvD1/mL than control; in oocytes, ROS was reduced in all RvD1 groups. No difference was observed in GSH in CC; in oocytes, GSH level was lesser in 25 and 75 pg RvD1/mL than control. In experiment 3, RvD1 did not affect IL-1β, IL-6, or TNF-α mRNA expression. In experiment 4, RvD1 had no effect on cleavage, day-7 or total blastocyst, or hatching rate. Resolvin D1 may help modulate ROS during IVM. Further studies should assess its potential synergistic effects with other modulators or under inflammatory conditions.
{"title":"Effects of resolvin D1 on bovine oocytes, cumulus cells, and in vitro embryo development","authors":"Maria Eugenia Sella , Anabella Andrea Campagna , Guillermo Luis Taminelli , Mariana Carolina Fabra , Alejandro Relling , Noelia Nikoloff , Ana Cristina Carranza-Martin","doi":"10.1016/j.theriogenology.2025.117789","DOIUrl":"10.1016/j.theriogenology.2025.117789","url":null,"abstract":"<div><div>This study evaluated the cytoprotective, antioxidant, and anti-inflammatory effects of Resolvin D1 (RvD1) during bovine oocyte <em>in vitro</em> maturation (IVM) and its impact on embryo development. Four experiments were performed: (1) cytoprotective effect by oocyte nuclear maturation, mitochondrial density, and cumulus cell (CC) DNA damage; (2) antioxidant activity by intracellular reactive oxygen species (ROS) and glutathione (GSH) concentration in CCs and oocytes; (3) anti-inflammatory action by mRNA expression of IL-1β, IL-6, and TNF-α in CCs; and (4) developmental competence by cleavage, blastocyst, and hatching rate. In Experiments 1–2, IVM supplementation were: control (0.1 % ethanol), 25, 50, or 75 pg RvD1/mL; Experiments 3–4 IVM supplementation were: control and 50 pg RvD1/mL. Cumulus oocyte-complexes underwent IVM for 24 h at 39 °C in 5 % CO<sub>2</sub>. In experiment 1, RvD1 did not alter nuclear maturation or CC DNA damage. Mitochondrial density was marginally greater at 25 pg RvD1/mL compared with control. In experiment 2, in CC, ROS were lesser with 50 and 75 pg RvD1/mL than control; in oocytes, ROS was reduced in all RvD1 groups. No difference was observed in GSH in CC; in oocytes, GSH level was lesser in 25 and 75 pg RvD1/mL than control. In experiment 3, RvD1 did not affect IL-1β, IL-6, or TNF-α mRNA expression. In experiment 4, RvD1 had no effect on cleavage, day-7 or total blastocyst, or hatching rate. Resolvin D1 may help modulate ROS during IVM. Further studies should assess its potential synergistic effects with other modulators or under inflammatory conditions.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117789"},"PeriodicalIF":2.5,"publicationDate":"2025-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145789625","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}
Pub Date : 2025-12-17DOI: 10.1016/j.theriogenology.2025.117787
Xiaojing Chen , Haiyue Mei , Jie Liu , Yuyan Feng , Zichun Dai , Binbin Guo , Huanxi Zhu , Bin He
Photoperiodic regulation is a key technique in modern poultry farming. In geese, however, the molecular pathways by which the pituitary gland transduces photoperiodic signals remain incompletely understood. Therefore, this study aimed to identify key factors by analyzing the pituitary transcriptome of female Yangzhou geese subjected to a short photoperiod and a long photoperiod. RNA-seq analysis showed that the prolactin (PRL) gene was the most highly expressed among all DEGs in the pituitary, and its dynamic expression pattern was closely associated with reproductive status. Furthermore, the neuroactive ligand-receptor interaction pathway was significantly enriched. Within this pathway, we identified key receptor genes, including neurotransmitter receptors (e.g., Gamma-Aminobutyric Acid Type A Receptor Subunit Gamma1 (GABRG1), Gamma-Aminobutyric Acid Type B Receptor Subunit 2 (GABBR2), Glutamate Ionotropic Receptor NMDA Type Subunit 1 (GRIN1), Opioid Receptor Mu 1 (OPRM1)) as well as neuropeptides and their receptors (e.g., Adenylate Cyclase Activating Polypeptide 1 Receptor 1 (ADCYAP1R1), Galanin Receptor 1 (GALR1)). Our results established the pituitary transcriptomic profile of geese under photoperiod-mediated reproduction, underscoring its role as a critical neuroendocrine center. It regulates reproductive activity primarily through PRL signaling and through specific neural pathways that receive and integrate diverse neural signals.
{"title":"Decoding the pituitary response to photoperiod in geese: Central role in regulating reproduction","authors":"Xiaojing Chen , Haiyue Mei , Jie Liu , Yuyan Feng , Zichun Dai , Binbin Guo , Huanxi Zhu , Bin He","doi":"10.1016/j.theriogenology.2025.117787","DOIUrl":"10.1016/j.theriogenology.2025.117787","url":null,"abstract":"<div><div>Photoperiodic regulation is a key technique in modern poultry farming. In geese, however, the molecular pathways by which the pituitary gland transduces photoperiodic signals remain incompletely understood. Therefore, this study aimed to identify key factors by analyzing the pituitary transcriptome of female Yangzhou geese subjected to a short photoperiod and a long photoperiod. RNA-seq analysis showed that the prolactin (<em>PRL</em>) gene was the most highly expressed among all DEGs in the pituitary, and its dynamic expression pattern was closely associated with reproductive status. Furthermore, the neuroactive ligand-receptor interaction pathway was significantly enriched. Within this pathway, we identified key receptor genes, including neurotransmitter receptors (e.g., Gamma-Aminobutyric Acid Type A Receptor Subunit Gamma1 (<em>GABRG1</em>), Gamma-Aminobutyric Acid Type B Receptor Subunit 2 (<em>GABBR2</em>), Glutamate Ionotropic Receptor NMDA Type Subunit 1 (<em>GRIN1</em>), Opioid Receptor Mu 1 (<em>OPRM1</em>)) as well as neuropeptides and their receptors (e.g., Adenylate Cyclase Activating Polypeptide 1 Receptor 1 (<em>ADCYAP1R1</em>), Galanin Receptor 1 (<em>GALR1</em>)). Our results established the pituitary transcriptomic profile of geese under photoperiod-mediated reproduction, underscoring its role as a critical neuroendocrine center. It regulates reproductive activity primarily through PRL signaling and through specific neural pathways that receive and integrate diverse neural signals.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117787"},"PeriodicalIF":2.5,"publicationDate":"2025-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145789548","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}
Pub Date : 2025-12-16DOI: 10.1016/j.theriogenology.2025.117788
Yujie Tang , Hong Wu , Yingang Guo , Xin Zhang , Yanbing Liu , Jinwang Liu , Mingxia Li , Chengtu Zhang , Jianmin Su
Apigenin (API), a naturally occurring flavonoid with potent antioxidant activity, has received limited attention regarding its protective effects on frozen–thawed sperm. Here, API was added to the freezing medium at 0–100 μM to determine the optimal concentration by assessing post-thaw survival and kinetic parameters. Dose–response screening identified 10 μM API as the most effective level, and this concentration was used to evaluate sperm quality, antioxidant capacity, fertilization potential and protein expression. Ten micromolar API significantly enhanced acrosomal and head membrane integrity and mitochondrial membrane potential, whereas tail membrane integrity and DNA integrity remained unchanged. Reactive oxygen species and malondialdehyde levels declined, superoxide dismutase activity and total antioxidant capacity increased; IVF cleavage rate improved, while blastocyst rate and blastocyst cell number remained unaffected, and AI pregnancy rate rose. iTRAQ profiling indicated that API primarily modulates proteins linked to energy metabolism, motility and mitochondrial function. API at 10 μM markedly improves the quality of frozen–thawed Oura-type Tibetan sheep sperm.
{"title":"Apigenin in freezing medium improves frozen-thawed sperm quality and fertilization competence of Oura-type Tibetan sheep","authors":"Yujie Tang , Hong Wu , Yingang Guo , Xin Zhang , Yanbing Liu , Jinwang Liu , Mingxia Li , Chengtu Zhang , Jianmin Su","doi":"10.1016/j.theriogenology.2025.117788","DOIUrl":"10.1016/j.theriogenology.2025.117788","url":null,"abstract":"<div><div>Apigenin (API), a naturally occurring flavonoid with potent antioxidant activity, has received limited attention regarding its protective effects on frozen–thawed sperm. Here, API was added to the freezing medium at 0–100 μM to determine the optimal concentration by assessing post-thaw survival and kinetic parameters. Dose–response screening identified 10 μM API as the most effective level, and this concentration was used to evaluate sperm quality, antioxidant capacity, fertilization potential and protein expression. Ten micromolar API significantly enhanced acrosomal and head membrane integrity and mitochondrial membrane potential, whereas tail membrane integrity and DNA integrity remained unchanged. Reactive oxygen species and malondialdehyde levels declined, superoxide dismutase activity and total antioxidant capacity increased; IVF cleavage rate improved, while blastocyst rate and blastocyst cell number remained unaffected, and AI pregnancy rate rose. iTRAQ profiling indicated that API primarily modulates proteins linked to energy metabolism, motility and mitochondrial function. API at 10 μM markedly improves the quality of frozen–thawed Oura-type Tibetan sheep sperm.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117788"},"PeriodicalIF":2.5,"publicationDate":"2025-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145789549","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}
Pub Date : 2025-12-13DOI: 10.1016/j.theriogenology.2025.117783
Fernanda T.D'el Rey Dantas , Igor F. Canisso , Lorena S. Feijó , Paula Maria Fernandes de Vasconcelos , Michel L. Campos , Alexander V. Ulanov , Zhong Li , Gino L.B.L. Pizzi , Carlos E.W. Nogueira , Bruna R. Curcio
Doxycycline is widely used in equine medicine, yet data on its pharmacokinetics and safety during late gestation are scarce. We investigated the pharmacokinetics, fetoplacental diffusion, and safety of compounded oral doxycycline in late-term pregnant mares. In the first experiment, six mares at 300 days of gestation received a single oral dose (10 mg/kg), and plasma concentrations were measured using LC–MS/MS. Pharmacokinetic analysis using non-compartmental and compartmental models showed rapid absorption, with a mean Cmax of about 6000 ng/mL reached within 0.8 h and a terminal half-life of 8.4 h. In the second experiment, seven mares received doxycycline (10 mg/kg, q12 h, orally) from 320 days of gestation until foaling, while six gestational-age-matched mares served as controls. Doxycycline was detected in fetal fluids, neonatal plasma, and synovial fluid, confirming its ability to cross the fetoplacental barrier, albeit at lower concentrations than in maternal plasma. Serial clinical examinations, complete blood counts, and blood chemistry analyses indicated no adverse effects in either mares or foals. Overall, compounded oral doxycycline was well absorbed during late gestation, safely diffused to the fetoplacental unit, and did not compromise maternal or neonatal health at the dose used herein. These findings provide baseline evidence supporting the use of doxycycline in late pregnant mares.
{"title":"Compounded oral doxycycline in late-term pregnant mares: pharmacokinetics, fetoplacental diffusion, and neonatal safety","authors":"Fernanda T.D'el Rey Dantas , Igor F. Canisso , Lorena S. Feijó , Paula Maria Fernandes de Vasconcelos , Michel L. Campos , Alexander V. Ulanov , Zhong Li , Gino L.B.L. Pizzi , Carlos E.W. Nogueira , Bruna R. Curcio","doi":"10.1016/j.theriogenology.2025.117783","DOIUrl":"10.1016/j.theriogenology.2025.117783","url":null,"abstract":"<div><div>Doxycycline is widely used in equine medicine, yet data on its pharmacokinetics and safety during late gestation are scarce. We investigated the pharmacokinetics, fetoplacental diffusion, and safety of compounded oral doxycycline in late-term pregnant mares. In the first experiment, six mares at 300 days of gestation received a single oral dose (10 mg/kg), and plasma concentrations were measured using LC<strong>–</strong>MS/MS. Pharmacokinetic analysis using non-compartmental and compartmental models showed rapid absorption, with a mean Cmax of about 6000 ng/mL reached within 0.8 h and a terminal half-life of 8.4 h. In the second experiment, seven mares received doxycycline (10 mg/kg, q12 h, orally) from 320 days of gestation until foaling, while six gestational-age-matched mares served as controls. Doxycycline was detected in fetal fluids, neonatal plasma, and synovial fluid, confirming its ability to cross the fetoplacental barrier, albeit at lower concentrations than in maternal plasma. Serial clinical examinations, complete blood counts, and blood chemistry analyses indicated no adverse effects in either mares or foals. Overall, compounded oral doxycycline was well absorbed during late gestation, safely diffused to the fetoplacental unit, and did not compromise maternal or neonatal health at the dose used herein. These findings provide baseline evidence supporting the use of doxycycline in late pregnant mares.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117783"},"PeriodicalIF":2.5,"publicationDate":"2025-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145757742","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}
Pub Date : 2025-12-13DOI: 10.1016/j.theriogenology.2025.117785
Hong Xu , Tian Tian , Yu Wang , Ying Liu , Yuqing Zhang , Jinghong Liang , Wenjie Yu , Qiang Sun , Guangying Zhu , Chang Liu , Mingkai Yu , Shuang Liang , Chengzhen Chen , Yuanshun Feng , Yan Li , Guangqiang Ji , Jiabao Zhang , Bao Yuan , Hao Jiang
Copper is an important trace element required by mammals and is widely involved in regulating various life activities. Overload or imbalance of copper ions can activate specific signaling pathways, leading to impaired oocyte function, decreased ovulation efficiency, and impaired embryo implantation. However, whether these changes are related to cuproptosis, a copper-driven, mitochondrial form of regulated cell death triggered by proteotoxic stress, a newly discovered phenomenon in recent years, remains unclear. In this study, combination treatment with a specific concentration of CuSO4 and elesclomol, a copper ionophore, delivered Cu+/Cu2+ to the mitochondrial matrix and potentiated cuproptosis, which triggered cuproptosis, leading to a decreased ability of cumulus cells to synthesize progesterone and estrogen. Moreover, during the maturation of oocytes, the expansion of cumulus cells and the levels of electron transport chain-related proteins decreased, whereas the calcium ion level, CaMKIIγ protein level, and phosphorylation level of CX43Ser255 significantly increased. In addition, although cuproptosis in cumulus cells did not directly induce cuproptosis in oocytes, it decreased the quality and further maturation of oocytes. During this process, abnormalities in mitochondrial distribution and function, cortical granule distribution, spindle morphology, and intracellular cAMP levels were detected in oocytes. The protein level of CDK1 decreased, whereas that of Emi1 increased. These results suggest that in cumulus cells, cuproptosis impairs mitochondrial function and triggers abnormal intracellular calcium homeostasis, which subsequently leads to overactivation of CaMKIIγ, blockage of Cx43 channels between COCs, and abnormal accumulation of cAMP in oocytes. Consequently, aberrant expression of the cell cycle regulatory factors CDK1 and Emi1 ultimately inhibits the in vitro maturation of oocytes. These findings clarify the effect of cuproptosis on bovine oocyte maturation, which contributes to a deeper understanding of the regulatory mechanisms governing oocyte maturation and may provide new directions for optimizing in vitro oocyte culture systems.
{"title":"Cuproptosis-mediated mitochondrial dysfunction in cumulus cells inhibits bovine oocyte maturation","authors":"Hong Xu , Tian Tian , Yu Wang , Ying Liu , Yuqing Zhang , Jinghong Liang , Wenjie Yu , Qiang Sun , Guangying Zhu , Chang Liu , Mingkai Yu , Shuang Liang , Chengzhen Chen , Yuanshun Feng , Yan Li , Guangqiang Ji , Jiabao Zhang , Bao Yuan , Hao Jiang","doi":"10.1016/j.theriogenology.2025.117785","DOIUrl":"10.1016/j.theriogenology.2025.117785","url":null,"abstract":"<div><div>Copper is an important trace element required by mammals and is widely involved in regulating various life activities. Overload or imbalance of copper ions can activate specific signaling pathways, leading to impaired oocyte function, decreased ovulation efficiency, and impaired embryo implantation. However, whether these changes are related to cuproptosis, a copper-driven, mitochondrial form of regulated cell death triggered by proteotoxic stress, a newly discovered phenomenon in recent years, remains unclear. In this study, combination treatment with a specific concentration of CuSO<sub>4</sub> and elesclomol, a copper ionophore, delivered Cu<sup>+</sup>/Cu<sup>2+</sup> to the mitochondrial matrix and potentiated cuproptosis, which triggered cuproptosis, leading to a decreased ability of cumulus cells to synthesize progesterone and estrogen. Moreover, during the maturation of oocytes, the expansion of cumulus cells and the levels of electron transport chain-related proteins decreased, whereas the calcium ion level, CaMKIIγ protein level, and phosphorylation level of CX43<sup>Ser255</sup> significantly increased. In addition, although cuproptosis in cumulus cells did not directly induce cuproptosis in oocytes, it decreased the quality and further maturation of oocytes. During this process, abnormalities in mitochondrial distribution and function, cortical granule distribution, spindle morphology, and intracellular cAMP levels were detected in oocytes. The protein level of CDK1 decreased, whereas that of Emi1 increased. These results suggest that in cumulus cells, cuproptosis impairs mitochondrial function and triggers abnormal intracellular calcium homeostasis, which subsequently leads to overactivation of CaMKIIγ, blockage of Cx43 channels between COCs, and abnormal accumulation of cAMP in oocytes. Consequently, aberrant expression of the cell cycle regulatory factors CDK1 and Emi1 ultimately inhibits the in vitro maturation of oocytes. These findings clarify the effect of cuproptosis on bovine oocyte maturation, which contributes to a deeper understanding of the regulatory mechanisms governing oocyte maturation and may provide new directions for optimizing in vitro oocyte culture systems.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117785"},"PeriodicalIF":2.5,"publicationDate":"2025-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145783039","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}
Pub Date : 2025-12-12DOI: 10.1016/j.theriogenology.2025.117784
Laura Rohde Brondani , Ana Paula Martini , Camila Rohde Brondani , Daniela dos Santos Brum , Gilson Antônio Pessoa , Fábio Gallas Leivas
The aim of this study was to evaluate the use of buserelin acetate, a GnRH analog, in combination with estradiol benzoate (EB) at the beginning of an early resynchronization protocol (R22) in Bos taurus beef heifers. A total of 656 heifers underwent a first fixed-time artificial insemination (FTAI; designated as D0) and received a reused intravaginal progesterone (P4; 1 g) device on D22. At that time, heifers were randomized into two treatment groups: EB (1 mg; n = 329) or GnRH + EB (1 mg of EB plus 10 μg of buserelin acetate; n = 327). On D30, the P4 device was removed, and a pregnancy diagnosis was performed. Non-pregnant heifers (EB = 152/329; GnRH + EB = 134/327) received 200 IU of equine chorionic gonadotropin, 1 mg of estradiol cypionate and 0.5 mg of cloprostenol sodium, followed by a second FTAI on D32. A subset of heifers underwent ovarian ultrasonography on D22 (n = 463), D30 (n = 193), and D32 (n = 165); a second pregnancy diagnosis was performed on D62. Heifers treated with EB alone had larger dominant follicles on both D30 and D32 (P = 0.0001) and a higher incidence of early ovulation before FTAI compared to GnRH + EB group (18.9 % vs. 8.8 %; P = 0.03). The GnRH + EB group achieved a superior pregnancy rate at R22 (EB = 40.8 % vs. GnRH + EB = 56.0 %; P = 0.008) and a higher cumulative pregnancy rate (EB = 68.4 % vs. GnRH + EB = 79.2 %; P = 0.01). In conclusion, the administration of 10 μg of buserelin acetate combined with 1 mg of EB at the beginning of the R22 protocol improved conception rates in Bos taurus beef heifers.
{"title":"Association of buserelin acetate with estradiol benzoate in early resynchronization of Bos taurus beef heifers improved conception rates","authors":"Laura Rohde Brondani , Ana Paula Martini , Camila Rohde Brondani , Daniela dos Santos Brum , Gilson Antônio Pessoa , Fábio Gallas Leivas","doi":"10.1016/j.theriogenology.2025.117784","DOIUrl":"10.1016/j.theriogenology.2025.117784","url":null,"abstract":"<div><div>The aim of this study was to evaluate the use of buserelin acetate, a GnRH analog, in combination with estradiol benzoate (EB) at the beginning of an early resynchronization protocol (R22) in <em>Bos taurus</em> beef heifers. A total of 656 heifers underwent a first fixed-time artificial insemination (FTAI; designated as D0) and received a reused intravaginal progesterone (P4; 1 g) device on D22. At that time, heifers were randomized into two treatment groups: EB (1 mg; n = 329) or GnRH + EB (1 mg of EB plus 10 μg of buserelin acetate; n = 327). On D30, the P4 device was removed, and a pregnancy diagnosis was performed. Non-pregnant heifers (EB = 152/329; GnRH + EB = 134/327) received 200 IU of equine chorionic gonadotropin, 1 mg of estradiol cypionate and 0.5 mg of cloprostenol sodium, followed by a second FTAI on D32. A subset of heifers underwent ovarian ultrasonography on D22 (n = 463), D30 (n = 193), and D32 (n = 165); a second pregnancy diagnosis was performed on D62. Heifers treated with EB alone had larger dominant follicles on both D30 and D32 (<em>P</em> = 0.0001) and a higher incidence of early ovulation before FTAI compared to GnRH + EB group (18.9 % vs. 8.8 %; <em>P</em> = 0.03). The GnRH + EB group achieved a superior pregnancy rate at R22 (EB = 40.8 % vs. GnRH + EB = 56.0 %; <em>P</em> = 0.008) and a higher cumulative pregnancy rate (EB = 68.4 % vs. GnRH + EB = 79.2 %; <em>P</em> = 0.01). In conclusion, the administration of 10 μg of buserelin acetate combined with 1 mg of EB at the beginning of the R22 protocol improved conception rates in <em>Bos taurus</em> beef heifers.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117784"},"PeriodicalIF":2.5,"publicationDate":"2025-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145763895","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}
Pub Date : 2025-12-10DOI: 10.1016/j.theriogenology.2025.117782
Yuqi Li , Yunxian Zhou , Yujiao Jin , Yongjing Li , Xi Shi , Qing Su , Xuejun Li , Limin Wu , Linyan Zhou
The Qi River crucian carp (Carassius auratus), a gynogenetic all-female fish, reproduces both unisexually and sexually, but the underlying molecular mechanisms remain unclear. The Wnt/β-catenin pathway influences mammalian ovarian development via cytochrome P450 family 19 subfamily A member 1 (CYP19A1) activation, however, its role in teleosts is poorly understood. Here, we investigated β-catenin's involvement in gonadal differentiation through histological and molecular analyses. Histological examination with Hematoxylin and Eosin (HE) staining and immunohistochemistry showed that oogonia emerged in female gonads at 15 days after hatching (dah), undergoing mitotic proliferation between 20 and 25 dah, and transitioning to meiosis from 25 to 30 dah. Significantly, β-catenin exhibited synchronized upregulation with cyp19a1a during the gonadal differentiation window, indicating functional synergy. Cellular co-localization studies further revealed β-catenin and cyp19a1a co-expressed in both germline (oogonia/oocytes) and somatic compartments, while pharmacological inhibition of β-catenin with quercetin and RNA interference via injection of β-catenin double-stranded RNA both significantly suppressed cyp19a1a expression. Mechanistically, promoter analysis demonstrated that β-catenin-Lef1 complexes directly activate cyp19a1a transcription through conserved T-cell factor (TCF)/lymphoid enhancer factor (LEF) response elements. The study identifies β-catenin as a key regulator of ovarian differentiation in Qi River crucian carp by coordinating cyp19a1a expression, offering initial experimental evidence for the role of Wnt/β-catenin signaling in sustaining unisexual reproduction. This work advances our understanding of evolutionary conserved versus species-specific mechanisms in teleost sex differentiation systems.
{"title":"The role of β-catenin in ovarian differentiation by upregulation cyp19a1a in a natural triploid teleost, Qi River crucian carp (Carassius auratus)","authors":"Yuqi Li , Yunxian Zhou , Yujiao Jin , Yongjing Li , Xi Shi , Qing Su , Xuejun Li , Limin Wu , Linyan Zhou","doi":"10.1016/j.theriogenology.2025.117782","DOIUrl":"10.1016/j.theriogenology.2025.117782","url":null,"abstract":"<div><div>The Qi River crucian carp (<em>Carassius auratus</em>), a gynogenetic all-female fish, reproduces both unisexually and sexually, but the underlying molecular mechanisms remain unclear. The Wnt/β-catenin pathway influences mammalian ovarian development via cytochrome P450 family 19 subfamily A member 1 (CYP19A1) activation, however, its role in teleosts is poorly understood. Here, we investigated β-catenin's involvement in gonadal differentiation through histological and molecular analyses. Histological examination with Hematoxylin and Eosin (HE) staining and immunohistochemistry showed that oogonia emerged in female gonads at 15 days after hatching (dah), undergoing mitotic proliferation between 20 and 25 dah, and transitioning to meiosis from 25 to 30 dah. Significantly, β<em>-catenin</em> exhibited synchronized upregulation with <em>cyp19a1a</em> during the gonadal differentiation window, indicating functional synergy. Cellular co-localization studies further revealed β<em>-catenin</em> and <em>cyp19a1a</em> co-expressed in both germline (oogonia/oocytes) and somatic compartments, while pharmacological inhibition of β<em>-catenin</em> with quercetin and RNA interference via injection of β<em>-catenin</em> double-stranded RNA both significantly suppressed <em>cyp19a1a</em> expression. Mechanistically, promoter analysis demonstrated that β-catenin-Lef1 complexes directly activate <em>cyp19a1a</em> transcription through conserved T-cell factor (TCF)/lymphoid enhancer factor (LEF) response elements. The study identifies β-catenin as a key regulator of ovarian differentiation in Qi River crucian carp by coordinating <em>cyp19a1a</em> expression, offering initial experimental evidence for the role of Wnt/β-catenin signaling in sustaining unisexual reproduction. This work advances our understanding of evolutionary conserved versus species-specific mechanisms in teleost sex differentiation systems.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117782"},"PeriodicalIF":2.5,"publicationDate":"2025-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145782949","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}
Pub Date : 2025-12-09DOI: 10.1016/j.theriogenology.2025.117781
Henry D. Mogollón-García , Antônio G.R. Pupulim , Gabriella C. Ribeiro , Paula Z. Rattes , Guilherme Rizzoto , John P. Kastelic , Ramanathan Kasimanickam , Fabiana F. Souza , Joao C.P. Ferreira
The objective was to assess effects of acute epididymal heat stress (HS) on bovine sperm and seminal plasma proteomes. Fourteen Nelore bulls, ∼18 mo old, with ≥70 % motile sperm and ≥70 % morphologically normal sperm were allocated into three groups: 12H (n = 5; 12 h of scrotal insulation [SI]); 24H (n = 5; 24 h of SI); and 48H (n = 4; 48 h of SI). Semen was collected before (control samples) and immediately after SI and differentially abundant proteins (DAPs) were identified in seminal plasma and sperm. Intratesticular temperature (at the end of HS; °C) was highest in 48H (mean ± SEM; 38.6 ± 0.5a, 37.3 ± 0.3ab, 35.6 ± 0.7b, and 35.2 ± 0.2b for 48H, 24H, 12H, and Control, respectively; P < 0.05). Sperm motility and morphology were not significantly different among groups. Scrotal insulation resulted in 172 and 518 DAPs in seminal plasma and sperm, respectively. Epididymal HS caused significant proteomic changes, in both seminal plasma and sperm. In seminal plasma, downregulated proteins were primarily associated with ATP production, oxidative stress protection, immune function, and sperm cytoskeleton integrity. In sperm, proteomic alterations were strongly linked to suppression of mitochondrial electron transport (oxidative phosphorylation) and estrogen signaling, along with increased proteins involved in glycolysis/gluconeogenesis pathways. Despite these changes, sperm motility remained unaffected.
目的是评估急性附睾热应激（HS）对牛精子和精浆蛋白质组的影响。14头Nelore公牛，年龄为18月龄，精子活动率≥70%，形态正常精子≥70%，分为3组：12H (n = 5；阴囊隔离12 h [SI])；24H (n = 5; SI 24H)；48H （n = 4； 48H的SI）。采集精液之前（对照样本）和之后立即收集精液，并在精浆和精子中发现差异丰富蛋白（DAPs）。48H、24H、12H和对照时，睾丸内温度（HS末；℃）在48H时最高(平均±SEM； 38.6±0.5a, 37.3±0.3ab, 35.6±0.7b, 35.2±0.2b
{"title":"Acute scrotal insulation in Nelore bulls impairs oxidative phosphorylation and estrogen signaling in epididymal sperm","authors":"Henry D. Mogollón-García , Antônio G.R. Pupulim , Gabriella C. Ribeiro , Paula Z. Rattes , Guilherme Rizzoto , John P. Kastelic , Ramanathan Kasimanickam , Fabiana F. Souza , Joao C.P. Ferreira","doi":"10.1016/j.theriogenology.2025.117781","DOIUrl":"10.1016/j.theriogenology.2025.117781","url":null,"abstract":"<div><div>The objective was to assess effects of acute epididymal heat stress (HS) on bovine sperm and seminal plasma proteomes. Fourteen Nelore bulls, ∼18 mo old, with ≥70 % motile sperm and ≥70 % morphologically normal sperm were allocated into three groups: 12H (n = 5; 12 h of scrotal insulation [SI]); 24H (n = 5; 24 h of SI); and 48H (n = 4; 48 h of SI). Semen was collected before (control samples) and immediately after SI and differentially abundant proteins (DAPs) were identified in seminal plasma and sperm. Intratesticular temperature (at the end of HS; °C) was highest in 48H (mean ± SEM; 38.6 ± 0.5<sup>a</sup>, 37.3 ± 0.3<sup>ab</sup>, 35.6 ± 0.7<sup>b</sup>, and 35.2 ± 0.2<sup>b</sup> for 48H, 24H, 12H, and Control, respectively; P < 0.05). Sperm motility and morphology were not significantly different among groups. Scrotal insulation resulted in 172 and 518 DAPs in seminal plasma and sperm, respectively. Epididymal HS caused significant proteomic changes, in both seminal plasma and sperm. In seminal plasma, downregulated proteins were primarily associated with ATP production, oxidative stress protection, immune function, and sperm cytoskeleton integrity. In sperm, proteomic alterations were strongly linked to suppression of mitochondrial electron transport (oxidative phosphorylation) and estrogen signaling, along with increased proteins involved in glycolysis/gluconeogenesis pathways. Despite these changes, sperm motility remained unaffected.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117781"},"PeriodicalIF":2.5,"publicationDate":"2025-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145775863","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}
Pub Date : 2025-12-08DOI: 10.1016/j.theriogenology.2025.117779
M. Molina , M.E. Odriozola , S. Cainelli , U.S. Notaro , E.M. Belotti , L. Cattaneo , A.F. Stassi , N.R. Salvetti , H.H. Ortega , A.N. Amweg , P.U. Díaz
Gonadotropins exert angiogenic and steroidogenic effects in the ovary. Likewise, recombinant equine chorionic gonadotropin (reCG) and GnRH through FSH and LH actions, may affect follicular perfusion, steroidogenesis, and the uterine milieu. The aim of the study was to determine dominant follicle irrigation at the onset of proestrus and to evaluate the associated effects on follicular perfusion, steroidogenic activity, and endometrial receptors abundance, following reCG and GnRH administration in beef cows subjected to estradiol (E2)/progesterone (P4)-based fixed-time artificial insemination (FTAI) treatment regimen. At P4 device removal, dominant follicles were classified as “Irrigated” or “Non-Irrigated” and randomly assigned to a 2 × 4 factorial design to be administered reCG, GnRH, reCG + GnRH, or no additional treatment. Follicular irrigation, follicular diameter, and plasma E2 and P4 concentrations during proestrus were assessed by Doppler ultrasonography and electrochemiluminescence. Follicular fluid E2 and P4, and endometrial abundance of estrogen (ESR1, ESR2) and progesterone (PR) receptors at final proestrus were evaluated using electrochemiluminescence and immunohistochemistry. Repeated-measures variables were analyzed using GLMM, and endpoint variables using one-way ANOVA. In females treated with GnRH, there was increase in follicular diameter compared with those of the irrigated control group on day 9 (P < 0.05), whereas in females treated with reCG and reCG + GnRH there were no differences (P > 0.05). Neither blood perfusion status nor treatments modified follicular irrigation, plasma P4 and E2 concentrations, follicular fluid E2 concentrations, or endometrial receptor abundance in females of any group (P > 0.05). In conclusion, dominant follicle blood flow at proestrus onset does not predict subsequent ovarian or uterine responses.
{"title":"Dominant follicle blood flow at the onset of proestrus does not affect steroid hormones, endometrial receptors, or follicular perfusion in beef cows under FTAI treatment regimens","authors":"M. Molina , M.E. Odriozola , S. Cainelli , U.S. Notaro , E.M. Belotti , L. Cattaneo , A.F. Stassi , N.R. Salvetti , H.H. Ortega , A.N. Amweg , P.U. Díaz","doi":"10.1016/j.theriogenology.2025.117779","DOIUrl":"10.1016/j.theriogenology.2025.117779","url":null,"abstract":"<div><div>Gonadotropins exert angiogenic and steroidogenic effects in the ovary. Likewise, recombinant equine chorionic gonadotropin (reCG) and GnRH through FSH and LH actions, may affect follicular perfusion, steroidogenesis, and the uterine milieu. The aim of the study was to determine dominant follicle irrigation at the onset of proestrus and to evaluate the associated effects on follicular perfusion, steroidogenic activity, and endometrial receptors abundance, following reCG and GnRH administration in beef cows subjected to estradiol (E<sub>2</sub>)/progesterone (P<sub>4</sub>)-based fixed-time artificial insemination (FTAI) treatment regimen. At P<sub>4</sub> device removal, dominant follicles were classified as “Irrigated” or “Non-Irrigated” and randomly assigned to a 2 × 4 factorial design to be administered reCG, GnRH, reCG + GnRH, or no additional treatment. Follicular irrigation, follicular diameter, and plasma E<sub>2</sub> and P<sub>4</sub> concentrations during proestrus were assessed by Doppler ultrasonography and electrochemiluminescence. Follicular fluid E<sub>2</sub> and P<sub>4,</sub> and endometrial abundance of estrogen (ESR1, ESR2) and progesterone (PR) receptors at final proestrus were evaluated using electrochemiluminescence and immunohistochemistry. Repeated-measures variables were analyzed using GLMM, and endpoint variables using one-way ANOVA. In females treated with GnRH, there was increase in follicular diameter compared with those of the irrigated control group on day 9 (P < 0.05), whereas in females treated with reCG and reCG + GnRH there were no differences (P > 0.05). Neither blood perfusion status nor treatments modified follicular irrigation, plasma P<sub>4</sub> and E<sub>2</sub> concentrations, follicular fluid E<sub>2</sub> concentrations, or endometrial receptor abundance in females of any group (P > 0.05). In conclusion, dominant follicle blood flow at proestrus onset does not predict subsequent ovarian or uterine responses.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117779"},"PeriodicalIF":2.5,"publicationDate":"2025-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145737471","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}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号