Pub Date : 2025-02-22DOI: 10.1016/j.theriogenology.2025.02.017
Han Zhou, Jinxin Zhang, Wen Feng, Nuoer Chen, Talha Umar, Xinyu Feng, Wenjing Liu, Changwei Qiu, Ganzhen Deng
Plasma membrane transformation (PMT) during early pregnancy (EP) is a widely observed phenomenon across species, but its role in cows remains understudied. Yes1-associated transcriptional regulator (YAP1) plays a vital role in human embryo implantation and uterine development. However, its relationship with PMT in bovine endometrium remains unclear. This study aims to explore the potential influence of YAP1 on PMT and the proliferation of endometrial epithelial cells. We observed PMT-related changes in the endometrium of cows during EP, characterized by reduced morphometry scores and increased fully developed pinopodes. The epithelial marker cadherin-1 (CDH1, also known as E-cadherin) expression decreased, while the mesenchymal marker vimentin (VIM) expression increased. Additionally, YAP1 expression was up-regulated and activated in the endometrial epithelium during EP. Interferon-tau (IFNT) potentially promoted PMT-related changes and activated YAP1. The knockdown of YAP1 diminished the effect of IFNT on PMT. At the same time, overexpression of YAP1 might enhance PMT-related changes and promote the proliferation, migration, and invasion ability of bovine endometrial epithelial cells (bEECs). Our findings suggest that YAP1 is activated during EP and may regulate endometrial receptivity by promoting PMT and cell proliferation.
{"title":"YAP1 regulates endometrial receptivity by promoting the plasma membrane transformation and proliferation of bovine endometrial epithelial cells","authors":"Han Zhou, Jinxin Zhang, Wen Feng, Nuoer Chen, Talha Umar, Xinyu Feng, Wenjing Liu, Changwei Qiu, Ganzhen Deng","doi":"10.1016/j.theriogenology.2025.02.017","DOIUrl":"10.1016/j.theriogenology.2025.02.017","url":null,"abstract":"<div><div>Plasma membrane transformation (PMT) during early pregnancy (EP) is a widely observed phenomenon across species, but its role in cows remains understudied. Yes1-associated transcriptional regulator (YAP1) plays a vital role in human embryo implantation and uterine development. However, its relationship with PMT in bovine endometrium remains unclear. This study aims to explore the potential influence of YAP1 on PMT and the proliferation of endometrial epithelial cells. We observed PMT-related changes in the endometrium of cows during EP, characterized by reduced morphometry scores and increased fully developed pinopodes. The epithelial marker cadherin-1 (CDH1, also known as E-cadherin) expression decreased, while the mesenchymal marker vimentin (VIM) expression increased. Additionally, YAP1 expression was up-regulated and activated in the endometrial epithelium during EP. Interferon-tau (IFNT) potentially promoted PMT-related changes and activated YAP1. The knockdown of YAP1 diminished the effect of IFNT on PMT. At the same time, overexpression of YAP1 might enhance PMT-related changes and promote the proliferation, migration, and invasion ability of bovine endometrial epithelial cells (bEECs). Our findings suggest that YAP1 is activated during EP and may regulate endometrial receptivity by promoting PMT and cell proliferation.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"237 ","pages":"Pages 166-177"},"PeriodicalIF":2.4,"publicationDate":"2025-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143520133","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 : 2025-02-21DOI: 10.1016/j.theriogenology.2025.02.026
Lam Do Truc Phuong , Lam Chi Thien , Cao Dang Su Pham , Nguyen Uyen Minh , Nguyen Thai Huy Bao , Le Nguyen Thien Truc , Truong Thi Ngoc Huyen , Do Tu Minh , Nhat-Thinh Nguyen , Nguyen Van Thuan , Hong-Thuy Bui
Melatonin has been studied for its ability to improve oocyte quality and modulate cyclic adenosine monophosphate (cAMP) production. However, the effects of melatonin on the in vitro growth (IVG) of oocyte-cumulus-granulosa complexes (OCGCs) derived from early antral follicles (EAFs) have not been fully investigated. This study aimed to examine the effects of melatonin during IVG on the developmental competence and blastocyst quality of porcine oocytes isolated from EAFs. In addition, the combination of melatonin with dibutyl cAMP (Mela + dbcAMP) or hypoxanthine (Mela + HX) during IVG and pre-in vitro maturation (pre-IVM) was also investigated.
The result showed that the modified medium supplemented with 10 μM melatonin after 4-day IVG enhanced antrum formation, survival rate, and oocyte diameter, especially, the melatonin-treated group enhanced expression of histone acetylation (Ac-H3-K9) higher than the untreated group. In addition, the combination of 10 μM melatonin with dbcAMP during IVG and during 7h of pre-IVM had significantly improved meiotic competence and cumulus expansion after IVM compared to Mela + HX groups. Finally, the combination of Mela + dbcAMP improved parthenogenetic blastocyst formation rather than the untreated group, and expression of histone methylation (Me-H3-K4) and Ac-H3-K9 in blastocyst comparable group derived from oocytes of large antral follicles (LAFs). Furthermore, melatonin with concentrations of 10 μM and 100 μM during IVG enhanced expression of pluripotency gene-related (OCT4, NANOG, SOX2) and balance cell viability via apoptosis-related gene (BCL2/BAX). In conclusion, melatonin combined with dbcAMP during IVG and pre-IVM of oocytes derived from EAFs demonstrated superior efficacy in enhancing oocyte growth, maturation, and development of porcine pre-implantation embryos.
{"title":"Melatonin and cyclic adenosine monophosphate enhance the meiotic and developmental competence of porcine oocytes from early antral follicles during in vitro growth and pre-maturation culture","authors":"Lam Do Truc Phuong , Lam Chi Thien , Cao Dang Su Pham , Nguyen Uyen Minh , Nguyen Thai Huy Bao , Le Nguyen Thien Truc , Truong Thi Ngoc Huyen , Do Tu Minh , Nhat-Thinh Nguyen , Nguyen Van Thuan , Hong-Thuy Bui","doi":"10.1016/j.theriogenology.2025.02.026","DOIUrl":"10.1016/j.theriogenology.2025.02.026","url":null,"abstract":"<div><div>Melatonin has been studied for its ability to improve oocyte quality and modulate cyclic adenosine monophosphate (cAMP) production. However, the effects of melatonin on the in vitro growth (IVG) of oocyte-cumulus-granulosa complexes (OCGCs) derived from early antral follicles (EAFs) have not been fully investigated. This study aimed to examine the effects of melatonin during IVG on the developmental competence and blastocyst quality of porcine oocytes isolated from EAFs. In addition, the combination of melatonin with dibutyl cAMP (Mela + dbcAMP) or hypoxanthine (Mela + HX) during IVG and pre-in vitro maturation (pre-IVM) was also investigated.</div><div>The result showed that the modified medium supplemented with 10 μM melatonin after 4-day IVG enhanced antrum formation, survival rate, and oocyte diameter, especially, the melatonin-treated group enhanced expression of histone acetylation (Ac-H3-K9) higher than the untreated group. In addition, the combination of 10 μM melatonin with dbcAMP during IVG and during 7h of pre-IVM had significantly improved meiotic competence and cumulus expansion after IVM compared to Mela + HX groups. Finally, the combination of Mela + dbcAMP improved parthenogenetic blastocyst formation rather than the untreated group, and expression of histone methylation (Me-H3-K4) and Ac-H3-K9 in blastocyst comparable group derived from oocytes of large antral follicles (LAFs). Furthermore, melatonin with concentrations of 10 μM and 100 μM during IVG enhanced expression of pluripotency gene-related (<em>OCT4, NANOG, SOX2</em>) and balance cell viability via apoptosis-related gene (<em>BCL2/BAX</em>). In conclusion, melatonin combined with dbcAMP during IVG and pre-IVM of oocytes derived from EAFs demonstrated superior efficacy in enhancing oocyte growth, maturation, and development of porcine pre-implantation embryos.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"237 ","pages":"Pages 129-142"},"PeriodicalIF":2.4,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143509725","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 : 2025-02-20DOI: 10.1016/j.theriogenology.2025.02.018
Tetiana Holumbiiovska , Małgorzata Ochota , Sylwia Prochowska , Zuzanna Ligocka-Kowalczyk , Maria Eberhardt , Vasyl Stefanyk , Wojciech Niżański
The use of deslorelin acetate implants for estrus induction is increasingly popular, but optimal timing for administration and removal remains uncertain. This study aimed to compare the effectiveness of estrus induction during early (90–120 days after heat cessation, G1: n = 7) and late anestrus (≥160 days after heat cessation, G2: n = 7; G3: n = 7) and at different implant removal times (Progesterone (P4) ≥ 3 ng/ml (G1 = 3.5 ± 0.3 ng/ml; G2 = 3.4 ± 0.4 ng/ml) vs day of Progesterone ≥5 ng/ml (G3 = 5.0 ± 1.0 ng/ml)). Studied bitches were divided in two groups and implanted with desloreline in early (G1: n = 7) and late anestrus (G2: n = 7) and removed at LH peak (G1, G2) or after it (G3: n = 7). A control group was created, which included bitches with a spontaneous estrous cycle (С: n = 8). Assessments included vaginal cytology, hormone measurements by enzyme-linked fluorescence assay (ELFA) method and ultrasound examination (US). In the group implanted during early anestrus (G1), heat signs appeared between days 5 and 7 after implant insertion, but no pregnancies were observed. Whereas implantation during late anestrus resulted in earlier onset of heat signs. In G2 and G3, cytological proestrus began two days earlier (on day 3 after implant insertion), and the estrus phase progressed more consistently compared to G1. Moreover, in G2, the cycle closely resembled a spontaneous sexual cycle and did not affect the subsequent fertility of the implanted females. Only in this group (G2) pregnancies were noted. The prolonged implant retention led to hormonal imbalance and no conception (G3). We conclude that estrus induction was less effective in early anestrus compared to late anestrus. For a successful and fertile estrous cycle, the implant should be administered during late anestrus (more than 160 days after the end of heat) and removed around the LH peak (P4 ≥ 3 ng/ml, measured with the mini VIDAS™ device).
{"title":"Analyzing estrous cycle dynamics with different protocols of 4.7 mg deslorelin implant aplication in bitches","authors":"Tetiana Holumbiiovska , Małgorzata Ochota , Sylwia Prochowska , Zuzanna Ligocka-Kowalczyk , Maria Eberhardt , Vasyl Stefanyk , Wojciech Niżański","doi":"10.1016/j.theriogenology.2025.02.018","DOIUrl":"10.1016/j.theriogenology.2025.02.018","url":null,"abstract":"<div><div>The use of deslorelin acetate implants for estrus induction is increasingly popular, but optimal timing for administration and removal remains uncertain. This study aimed to compare the effectiveness of estrus induction during early (90–120 days after heat cessation, G1: n = 7) and late anestrus (≥160 days after heat cessation, G2: n = 7; G3: n = 7) and at different implant removal times (Progesterone (P4) ≥ 3 ng/ml (G1 = 3.5 ± 0.3 ng/ml; G2 = 3.4 ± 0.4 ng/ml) vs day of Progesterone ≥5 ng/ml (G3 = 5.0 ± 1.0 ng/ml)). Studied bitches were divided in two groups and implanted with desloreline in early (G1: n = 7) and late anestrus (G2: n = 7) and removed at LH peak (G1, G2) or after it (G3: n = 7). A control group was created, which included bitches with a spontaneous estrous cycle (С: n = 8). Assessments included vaginal cytology, hormone measurements by enzyme-linked fluorescence assay (ELFA) method and ultrasound examination (US). In the group implanted during early anestrus (G1), heat signs appeared between days 5 and 7 after implant insertion, but no pregnancies were observed. Whereas implantation during late anestrus resulted in earlier onset of heat signs. In G2 and G3, cytological proestrus began two days earlier (on day 3 after implant insertion), and the estrus phase progressed more consistently compared to G1. Moreover, in G2, the cycle closely resembled a spontaneous sexual cycle and did not affect the subsequent fertility of the implanted females. Only in this group (G2) pregnancies were noted. The prolonged implant retention led to hormonal imbalance and no conception (G3). We conclude that estrus induction was less effective in early anestrus compared to late anestrus. For a successful and fertile estrous cycle, the implant should be administered during late anestrus (more than 160 days after the end of heat) and removed around the LH peak (P4 ≥ 3 ng/ml, measured with the mini VIDAS™ device).</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"237 ","pages":"Pages 143-152"},"PeriodicalIF":2.4,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143509726","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}
Neddylation is a biological process that covalently links neuronal precursor cell-expressed developmentally downregulated protein 8 (NEDD8) to its substrate. Its main function is to activate the cullin ubiquitin junction complex, thereby promoting substrate protein degradation. The research of neddylation mainly focus on cell senescence, Alzheimer's disease progression, and tumor treatment. However, the neddylation mechanism underlying follicular granulosa cells (GCs) proliferation and steroidogenesis remains unclear. Therefore, this study aimed to reveal the functions of neddylation in sheep follicular GCs. Here, immunohistochemistry results revealed that NEDD8 was expressed in sheep follicular GCs. The neddylation-specific inhibitor MLN4924 and NEDD8 small interfering (si) RNA (si-NEDD8) were used to inhibit neddylation, significantly attenuating sheep follicular GC proliferation. The RNA sequencing (RNA-Seq) showed that cytochrome P450 11A1 (CYP11A1) and steroidogenic acute regulator (StAR) were significantly downregulated via the inhibition of neddylation. Additionally, qRT-PCR results showed that CYP11A1, StAR and 3β-hydroxysteroid dehydrogenase (3β-HSD) mRNA abundance decreased significantly following the addition of various MLN4924 doses and si-NEDD8, however, CYP19A1 mRNA levels did not significantly differ. Western blotting results showed that inhibition of neddylation also significantly decreased the expression of CYP11A1, 3β-HSD, cytochrome P450 19A1 (CYP19A1), StAR, and androgen receptor (AR). The current study revealed that the inhibition of neddylation affects sheep follicular GC function by inhibiting proliferation and the enzymes involved in synthesis in sheep.
{"title":"Neddylation inhibition affects cell proliferation and steroidogenesis in sheep follicular granulosa cells","authors":"Lihua Lyu , Xiaowei Qin , Haoyu Xiu , Yuhan Qu , Yipin Wang , Xiaofeng Yang , Wenqing Dang , Ermias Kebreab","doi":"10.1016/j.theriogenology.2025.02.020","DOIUrl":"10.1016/j.theriogenology.2025.02.020","url":null,"abstract":"<div><div>Neddylation is a biological process that covalently links neuronal precursor cell-expressed developmentally downregulated protein 8 (NEDD8) to its substrate. Its main function is to activate the cullin ubiquitin junction complex, thereby promoting substrate protein degradation. The research of neddylation mainly focus on cell senescence, Alzheimer's disease progression, and tumor treatment. However, the neddylation mechanism underlying follicular granulosa cells (GCs) proliferation and steroidogenesis remains unclear. Therefore, this study aimed to reveal the functions of neddylation in sheep follicular GCs. Here, immunohistochemistry results revealed that NEDD8 was expressed in sheep follicular GCs. The neddylation-specific inhibitor MLN4924 and NEDD8 small interfering (si) RNA (si-NEDD8) were used to inhibit neddylation, significantly attenuating sheep follicular GC proliferation. The RNA sequencing (RNA-Seq) showed that <em>cytochrome P450 11A1</em> (<em>CYP11A1</em>) and <em>steroidogenic acute regulator</em> (<em>StAR</em>) were significantly downregulated via the inhibition of neddylation. Additionally, qRT-PCR results showed that <em>CYP11A1</em>, <em>StAR</em> and <em>3β-hydroxysteroid dehydrogenase</em> (<em>3β-HSD</em>) mRNA abundance decreased significantly following the addition of various MLN4924 doses and si-NEDD8, however, <em>CYP19A1</em> mRNA levels did not significantly differ. Western blotting results showed that inhibition of neddylation also significantly decreased the expression of CYP11A1, 3β-HSD, cytochrome P450 19A1 (CYP19A1), StAR, and androgen receptor (AR). The current study revealed that the inhibition of neddylation affects sheep follicular GC function by inhibiting proliferation and the enzymes involved in synthesis in sheep.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"237 ","pages":"Pages 99-109"},"PeriodicalIF":2.4,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143453709","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 : 2025-02-18DOI: 10.1016/j.theriogenology.2025.02.022
Marwa El-Sheikh , Ahmed Atef Mesalam , Ahmed F. El-Sayed , Ayman Mesalam , Heba M. Metwally , Seo-Hyun Lee , Il-Keun Kong
Nicotinamide (NAM) is a widely utilized compound in cell culture systems, yet its role during the morula-to-blastocyst transition remains underexplored. This study investigates the effects of NAM supplementation during both in vitro maturation (IVM) of oocytes and late-stage embryo culture (IVC3; the morula stage) on blastocyst development, metabolic flux, mitochondrial bioenergetics, and pluripotency of cells. Bovine oocytes were subjected to dual NAM treatment during IVM and IVC3 and its impact was assessed through cleavage and blastocyst development rates, mitochondrial membrane potential (ΔΨm), and the expression of key metabolic and pluripotency markers using RT-qPCR and immunofluorescence. Additionally, molecular docking was performed to evaluate NAM's interaction with Zonula Occludens-1 (ZO-1) protein. Dual NAM administration significantly increased both blastocyst formation and hatching rates. Computational modeling revealed a strong binding affinity (−6.44 kcal/mol) between NAM and the ZO-1 protein, associated with the morula-to-blastocyst transition. Quantitative RT-PCR analysis showed upregulation of genes related to NAD + biosynthesis (NAMPT, MDH1), glycolysis (PFK1), glycogenesis (GSK-3A), and mitochondrial bioenergetics (SDHA, ND2, ATPase8, TFAM) in NAM-treated group. Additionally, mitochondrial profiling demonstrated enhanced polarization, and OCT4 expression was elevated in NAM-treated embryos. These findings underscore NAM's potential role in enhancing morula-to-blastocyst transition, improving embryonic development through metabolic and mitochondrial regulation, as well as pluripotency factor enhancement.
{"title":"Nicotinamide dual treatment enhances morula-to-blastocyst transition through binding to Zonula Occludens-1 protein","authors":"Marwa El-Sheikh , Ahmed Atef Mesalam , Ahmed F. El-Sayed , Ayman Mesalam , Heba M. Metwally , Seo-Hyun Lee , Il-Keun Kong","doi":"10.1016/j.theriogenology.2025.02.022","DOIUrl":"10.1016/j.theriogenology.2025.02.022","url":null,"abstract":"<div><div>Nicotinamide (NAM) is a widely utilized compound in cell culture systems, yet its role during the morula-to-blastocyst transition remains underexplored. This study investigates the effects of NAM supplementation during both <em>in vitro</em> maturation (IVM) of oocytes and late-stage embryo culture (IVC3; the morula stage) on blastocyst development, metabolic flux, mitochondrial bioenergetics, and pluripotency of cells. Bovine oocytes were subjected to dual NAM treatment during IVM and IVC3 and its impact was assessed through cleavage and blastocyst development rates, mitochondrial membrane potential (ΔΨm), and the expression of key metabolic and pluripotency markers using RT-qPCR and immunofluorescence. Additionally, molecular docking was performed to evaluate NAM's interaction with Zonula Occludens-1 (ZO-1) protein. Dual NAM administration significantly increased both blastocyst formation and hatching rates. Computational modeling revealed a strong binding affinity (−6.44 kcal/mol) between NAM and the ZO-1 protein, associated with the morula-to-blastocyst transition. Quantitative RT-PCR analysis showed upregulation of genes related to NAD + biosynthesis (<em>NAMPT</em>, <em>MDH1</em>), glycolysis (<em>PFK1</em>), glycogenesis (<em>GSK-3A</em>), and mitochondrial bioenergetics (<em>SDHA</em>, <em>ND2</em>, <em>ATPase8</em>, <em>TFAM</em>) in NAM-treated group. Additionally, mitochondrial profiling demonstrated enhanced polarization, and OCT4 expression was elevated in NAM-treated embryos. These findings underscore NAM's potential role in enhancing morula-to-blastocyst transition, improving embryonic development through metabolic and mitochondrial regulation, as well as pluripotency factor enhancement.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"237 ","pages":"Pages 110-119"},"PeriodicalIF":2.4,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143464047","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 : 2025-02-17DOI: 10.1016/j.theriogenology.2025.02.019
Xianhong Mo , Rong Liang , Cheng Guo , Zengyuan Zhao , Jing Zhao , Yi Fang , Zhenjun Xu
Intracellular calcium (Ca2+) homeostasis is primarily maintained by the endoplasmic reticulum (ER). Fluctuations in intracellular Ca2+ concentrations can induce ER stress, ultimately causing cellular apoptosis. Vitrification can increase Ca2+ concentrations and reduce oocyte vitality. However, 2-Aminoethyl Diphenylborinate (2-APB), a Ca2+ release regulator, can maintain ER calcium homeostasis. In this study, after in vitro maturation, MII oocytes were randomly allocated into 4 group in a 4 groups in a 2 x 2 factorial study (fresh or vitrified and with or without exposure to 2-APB before vitrification). There were fewer ER clusters in the cortical region of oocytes after vitrification in the Control group, but they were protected by pre-treated with 2-APB. In addition, 2-APB significantly increased survival and cleavage of vitrified-warmed parthenogenetically activated blastocysts. Expression of the ER stress proteins GRP78 and CHOP, and the apoptotic proteins caspase-9, caspase-3, and cytochrome c (cytC) were all significantly increased in vitrified oocytes. However, with 2-APB pretreatment, expression of those proteins were not significant different from the Control group. In conclusion, 2-APB effectively alleviated ER stress by maintaining Ca2+ homeostasis to mitigate damage in vitrified-warmed bovine oocytes.
{"title":"2-Aminoethyl diphenylborinate mitigates damage from the endoplasmic reticulum of vitrified bovine oocyte","authors":"Xianhong Mo , Rong Liang , Cheng Guo , Zengyuan Zhao , Jing Zhao , Yi Fang , Zhenjun Xu","doi":"10.1016/j.theriogenology.2025.02.019","DOIUrl":"10.1016/j.theriogenology.2025.02.019","url":null,"abstract":"<div><div>Intracellular calcium (Ca<sup>2+</sup>) homeostasis is primarily maintained by the endoplasmic reticulum (ER). Fluctuations in intracellular Ca<sup>2+</sup> concentrations can induce ER stress, ultimately causing cellular apoptosis. Vitrification can increase Ca<sup>2+</sup> concentrations and reduce oocyte vitality. However, 2-Aminoethyl Diphenylborinate (2-APB), a Ca<sup>2+</sup> release regulator, can maintain ER calcium homeostasis. In this study, after <em>in vitro</em> maturation, MII oocytes were randomly allocated into 4 group in a 4 groups in a 2 x 2 factorial study (fresh or vitrified and with or without exposure to 2-APB before vitrification). There were fewer ER clusters in the cortical region of oocytes after vitrification in the Control group, but they were protected by pre-treated with 2-APB. In addition, 2-APB significantly increased survival and cleavage of vitrified-warmed parthenogenetically activated blastocysts. Expression of the ER stress proteins GRP78 and CHOP, and the apoptotic proteins caspase-9, caspase-3, and cytochrome <em>c</em> (cytC) were all significantly increased in vitrified oocytes. However, with 2-APB pretreatment, expression of those proteins were not significant different from the Control group. In conclusion, 2-APB effectively alleviated ER stress by maintaining Ca<sup>2+</sup> homeostasis to mitigate damage in vitrified-warmed bovine oocytes.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"237 ","pages":"Pages 93-98"},"PeriodicalIF":2.4,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143453708","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 : 2025-02-17DOI: 10.1016/j.theriogenology.2025.02.016
Lijun Jiang , Yunlei Li , Yunhe Zong , Xintong Han , Jiangpeng Guo , Sihua Jin , Yi Zhao , Jingwei Yuan , Hui Ma , Jilan Chen , Yanyan Sun
Semen cryopreservation technology plays a crucial role in enhancing animal reproductive efficiency and genetic resources preservation. However, the remarkable decline in fertility caused by sperm damage during the freezing process restricts its application in the poultry industry. The addition of antioxidants in semen is an effective approach to mitigate oxidative damage and enhance fertility. L-carnitine (LC) is an antioxidant and previous study indicated that it was related with sperm motility regulation. To explore its proper application in rooster semen cryopreservation, this study explored the effect of different levels of LC (0, 0.5 1.0, 2.5, 5.0, 7.5 mM) on post-tawed sperm motility, morphology, mitochondrial function, antioxidant activity, and fertility potential. The results demonstrated that sperm motility parameters including motility and motion parameters did not differ between groups. The sperm abnormality of 5.0 mM LC was lower than that in the control group (0 mM LC in the basic extender) (P < 0.05). The sperm plasma membrane integrity with 0.5, 2.5 and 5.0 mM LC was significantly higher than that in the control group (P < 0.05). All the treatment groups showed lower sperm reactive oxygen species than the control group. The 2.5 and 5.0 mM LC groups showed higher fertility (P < 0.05). Overall, this study provides empirical evidence supporting the effectiveness of LC for improving the efficiency of roosters’ semen cryopreservation in a dose dependent manner, based on the results that supplementation of 2.5 and 5.0 mM LC in the basic extender improved the post-thawed sperm quality and fertility of roosters. In general, our study indicated that the addition of LC exhibited a notable impact on the freezing of rooster semen.
{"title":"The effect of L-carnitine on frozen-thawed rooster sperm quality and fertility potential","authors":"Lijun Jiang , Yunlei Li , Yunhe Zong , Xintong Han , Jiangpeng Guo , Sihua Jin , Yi Zhao , Jingwei Yuan , Hui Ma , Jilan Chen , Yanyan Sun","doi":"10.1016/j.theriogenology.2025.02.016","DOIUrl":"10.1016/j.theriogenology.2025.02.016","url":null,"abstract":"<div><div>Semen cryopreservation technology plays a crucial role in enhancing animal reproductive efficiency and genetic resources preservation. However, the remarkable decline in fertility caused by sperm damage during the freezing process restricts its application in the poultry industry. The addition of antioxidants in semen is an effective approach to mitigate oxidative damage and enhance fertility. L-carnitine (LC) is an antioxidant and previous study indicated that it was related with sperm motility regulation. To explore its proper application in rooster semen cryopreservation, this study explored the effect of different levels of LC (0, 0.5 1.0, 2.5, 5.0, 7.5 mM) on post-tawed sperm motility, morphology, mitochondrial function, antioxidant activity, and fertility potential. The results demonstrated that sperm motility parameters including motility and motion parameters did not differ between groups. The sperm abnormality of 5.0 mM LC was lower than that in the control group (0 mM LC in the basic extender) (<em>P</em> < 0.05). The sperm plasma membrane integrity with 0.5, 2.5 and 5.0 mM LC was significantly higher than that in the control group (<em>P</em> < 0.05). All the treatment groups showed lower sperm reactive oxygen species than the control group. The 2.5 and 5.0 mM LC groups showed higher fertility (<em>P</em> < 0.05). Overall, this study provides empirical evidence supporting the effectiveness of LC for improving the efficiency of roosters’ semen cryopreservation in a dose dependent manner, based on the results that supplementation of 2.5 and 5.0 mM LC in the basic extender improved the post-thawed sperm quality and fertility of roosters. In general, our study indicated that the addition of LC exhibited a notable impact on the freezing of rooster semen.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"237 ","pages":"Pages 70-75"},"PeriodicalIF":2.4,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143437450","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}
Alterations during the early stages of embryo development have been associated with long-term effects on the fetus, neonate, and adult, but this has not been investigated in horses. In recent years, intracytoplasmic sperm injection (ICSI) has gained in commercial popularity in the equine population. Research suggests an association between ICSI-produced embryos and placental malformations, but there exists little understanding of the physiology involved. Therefore, we aim to produce a complete transcriptomic analysis of chorioallantois and provide potential pathways that may be impacted following pregnancies associated with in vitro-produced equine embryos. To do so, seventeen warmblood mares were bred either naturally to produce in vivo-produced pregnancies that were carried by self (in vivo; n = 8) or with in vitro-produced pregnancies created via intracytoplasmic sperm injection (ICSI) that were transferred to a recipient (in vitro; n = 9). Mares were monitored throughout gestation to ensure the health of the pregnancy, and impending parturition was monitored for progress. Chorioallantois was collected immediately postpartum and placed in RNALater for future extraction. RNA was isolated using Trizol, and RNASeq was performed by Novogene, with 93.3 % total mapping and 40 million read depth. The false discovery rate (FDR) was set to <0.05. When comparing groups (in vivo vs. in vitro-produced embryos), 1589 genes were differentially expressed. This included an upregulation of 626 genes, alongside a downregulation of 963 genes. Impacted gene ontology included aspects of the central dogma of molecular biology, including ribosome biogenesis, RNA polymerase activity, and spliceosome function. Additional biological processes that were impacted included aspects of the immune system relating to auto-immunity and disordered antigen response, such as the IL-17 signaling pathway, rheumatoid arthritis, and lupus. Additionally, pathways relating to hypoxia and ribosome biogenesis were associated with in vitro-produced pregnancies. Overall, it appears that the in vitro production of pregnancies is associated with placental dysregulation during pregnancy, which may be related to poor fetal and neonatal outcomes that have been associated with ART in other species.
{"title":"Unveiling the equine placental transcriptome: A novel study on ICSI-derived pregnancies","authors":"C.E. Fedorka , K.E. Scoggin , S.J. Coleman , J.N. Hatzel , M.D. Burleson , M.H.T. Troedsson","doi":"10.1016/j.theriogenology.2025.02.013","DOIUrl":"10.1016/j.theriogenology.2025.02.013","url":null,"abstract":"<div><div>Alterations during the early stages of embryo development have been associated with long-term effects on the fetus, neonate, and adult, but this has not been investigated in horses. In recent years, intracytoplasmic sperm injection (ICSI) has gained in commercial popularity in the equine population. Research suggests an association between ICSI-produced embryos and placental malformations, but there exists little understanding of the physiology involved. Therefore, we aim to produce a complete transcriptomic analysis of chorioallantois and provide potential pathways that may be impacted following pregnancies associated with <em>in vitro</em>-produced equine embryos. To do so, seventeen warmblood mares were bred either naturally to produce <em>in vivo</em>-produced pregnancies that were carried by self (<em>in vivo</em>; n = 8) or with <em>in vitro</em>-produced pregnancies created via intracytoplasmic sperm injection (ICSI) that were transferred to a recipient (<em>in vitro</em>; n = 9). Mares were monitored throughout gestation to ensure the health of the pregnancy, and impending parturition was monitored for progress. Chorioallantois was collected immediately postpartum and placed in RNALater for future extraction. RNA was isolated using Trizol, and RNASeq was performed by Novogene, with 93.3 % total mapping and 40 million read depth. The false discovery rate (FDR) was set to <0.05. When comparing groups (<em>in vivo</em> vs. <em>in vitro-</em>produced embryos), 1589 genes were differentially expressed. This included an upregulation of 626 genes, alongside a downregulation of 963 genes. Impacted gene ontology included aspects of the central dogma of molecular biology, including ribosome biogenesis, RNA polymerase activity, and spliceosome function. Additional biological processes that were impacted included aspects of the immune system relating to auto-immunity and disordered antigen response, such as the IL-17 signaling pathway, rheumatoid arthritis, and lupus. Additionally, pathways relating to hypoxia and ribosome biogenesis were associated with <em>in vitro</em>-produced pregnancies. Overall, it appears that the <em>in vitro</em> production of pregnancies is associated with placental dysregulation during pregnancy, which may be related to poor fetal and neonatal outcomes that have been associated with ART in other species.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"237 ","pages":"Pages 120-128"},"PeriodicalIF":2.4,"publicationDate":"2025-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143487571","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}
This study evaluated the effects of storing queen ovaries in saline at 22 °C for up to 12 h. Two experiments were conducted. In the first experiment, ovaries from five queens were sectioned into four fragments; stored for 4, 8, or 12 h; and histologically analyzed to assess follicular morphology (Grades I–IV). In the second experiment, ovaries from 15 additional queens were stored under the same conditions, after which cumulus–oocyte complexes (COCs) were retrieved and graded (I–IV); only Grades I and II underwent in vitro maturation to assess cumulus expansion and meiotic status. The effects of storage on follicle morphology, COC quality, and maturation were analyzed using the chi-square test, while Spearman's correlation assessed the relationship between storage time and follicle morphology. The results showed that early-stage follicles were more sensitive to short-term storage, with 56 %, 22 %, 2 %, and 0 % of primordial, primary, secondary, and antral follicles, respectively, classified as Grade IV after 4 h. Grade I follicles declined to 0 % across all developmental stages after 12 h. Significant correlations were found between storage duration and follicle morphology (Grade I: r = −0.92, Grade II: r = −0.65, Grade III: r = 0.68; P < 0.05), while Grade IV remained unchanged. COC recovery did not differ across storage times (P > 0.05), with >60 % classified as Grade III or IV at all time points. After 12 h, there was a significant reduction in Grade I COCs, cumulus expansion, and the percentage of oocytes reaching metaphase II (P < 0.05). In conclusion, while storing queen ovaries in saline at 22 °C negatively affects follicular morphology – particularly preantral follicles – oocytes can still be recovered up to 8 h post-storage, without compromising COC quality or maturation potential.
{"title":"Effects of storing queen ovaries in saline solution at 22 C on ovarian follicle integrity and oocyte quality and maturation","authors":"Franciely Santos Feijó , Karina Pessoa Oliveira , Leonardo Vitorino Costa de Aquino , Alexsandra Fernandes Pereira , Diogo Ribeiro Câmara","doi":"10.1016/j.theriogenology.2025.02.015","DOIUrl":"10.1016/j.theriogenology.2025.02.015","url":null,"abstract":"<div><div>This study evaluated the effects of storing queen ovaries in saline at 22 °C for up to 12 h. Two experiments were conducted. In the first experiment, ovaries from five queens were sectioned into four fragments; stored for 4, 8, or 12 h; and histologically analyzed to assess follicular morphology (Grades I–IV). In the second experiment, ovaries from 15 additional queens were stored under the same conditions, after which cumulus–oocyte complexes (COCs) were retrieved and graded (I–IV); only Grades I and II underwent <em>in vitro</em> maturation to assess cumulus expansion and meiotic status. The effects of storage on follicle morphology, COC quality, and maturation were analyzed using the chi-square test, while Spearman's correlation assessed the relationship between storage time and follicle morphology. The results showed that early-stage follicles were more sensitive to short-term storage, with 56 %, 22 %, 2 %, and 0 % of primordial, primary, secondary, and antral follicles, respectively, classified as Grade IV after 4 h. Grade I follicles declined to 0 % across all developmental stages after 12 h. Significant correlations were found between storage duration and follicle morphology (Grade I: r = −0.92, Grade II: r = −0.65, Grade III: r = 0.68; P < 0.05), while Grade IV remained unchanged. COC recovery did not differ across storage times (P > 0.05), with >60 % classified as Grade III or IV at all time points. After 12 h, there was a significant reduction in Grade I COCs, cumulus expansion, and the percentage of oocytes reaching metaphase II (P < 0.05). In conclusion, while storing queen ovaries in saline at 22 °C negatively affects follicular morphology – particularly preantral follicles – oocytes can still be recovered up to 8 h post-storage, without compromising COC quality or maturation potential.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"237 ","pages":"Pages 42-48"},"PeriodicalIF":2.4,"publicationDate":"2025-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143427957","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 : 2025-02-14DOI: 10.1016/j.theriogenology.2025.02.014
Igor Garcia Motta , Amanda Guimarães da Silva , Isabella Rio Feltrin , Samuel Volpe Souza , Ana Clara Degan Mattos , Karine Galhego Morelli , Thadeu Castro , Thiago Kan Nishimura , Oliver Joseph Ginther , Guilherme Pugliesi
This study investigated the effects of estradiol benzoate (EB) administered 13 days post-ovulation on PGF2α release and corpus luteum function in pregnant and non-pregnant heifers. In Exp. 1, Nelore (Bos indicus) heifers, either inseminated or non-inseminated, were randomly assigned on Day 13 (D13) to receive 0, 1, or 2 mg of EB. Blood samples were collected at baseline (H0) and hourly from H3 to H12 to assess plasma P4 and PGFM concentrations. In a subgroup of pregnant heifers, blood samples were also collected to determine plasma E2 concentrations. Doppler ultrasonography was performed daily from D13 to D19 for monitoring the luteal function, and pregnancy was determined on D28. Luteolysis was earlier (P < 0.05) in non-inseminated heifers treated with 1 or 2 mg EB than in the controls (16.3 ± 0.2 vs. 17.3 ± 0.6 days). Pregnancy rate was lower (P < 0.05) in the EB-1 (50 %; 8/16) and EB-2 (29.2 %; 7/24) groups than in the EB-0 group (90 %; 9/10). The average PGFM concentrations were greater (P < 0.05) in the EB-1 and EB-2 groups than in the EB-0 group, regardless of gestational status. In Exp. 2, inseminated (n = 39) and non-inseminated (n = 21) Nelore heifers received either 0 or 1 mg of EB on D13. Three hours later, endometrial cytology was performed and samples were evaluated by qPCR. Expression of OXTR and PGR was greater and IL1β was lower in EB-treated heifers (P < 0.05). The ESR2 abundance was lower (P < 0.05) in pregnant heifers, regardless of EB treatment. In conclusion, an elevation of circulating E2 at late diestrus upregulates the OXTR and PGR expression in the endometrium, inducing PGF2α release and luteolysis, which negatively impact on pregnancy establishment, especially in heifers treated with 2 mg EB.
{"title":"Effects of estradiol on PGF2α synthesis and corpus luteum function during early pregnancy in beef heifers","authors":"Igor Garcia Motta , Amanda Guimarães da Silva , Isabella Rio Feltrin , Samuel Volpe Souza , Ana Clara Degan Mattos , Karine Galhego Morelli , Thadeu Castro , Thiago Kan Nishimura , Oliver Joseph Ginther , Guilherme Pugliesi","doi":"10.1016/j.theriogenology.2025.02.014","DOIUrl":"10.1016/j.theriogenology.2025.02.014","url":null,"abstract":"<div><div>This study investigated the effects of estradiol benzoate (EB) administered 13 days post-ovulation on PGF2α release and corpus luteum function in pregnant and non-pregnant heifers. In Exp. 1, Nelore (<em>Bos indicus</em>) heifers, either inseminated or non-inseminated, were randomly assigned on Day 13 (D13) to receive 0, 1, or 2 mg of EB. Blood samples were collected at baseline (H0) and hourly from H3 to H12 to assess plasma P4 and PGFM concentrations. In a subgroup of pregnant heifers, blood samples were also collected to determine plasma E2 concentrations. Doppler ultrasonography was performed daily from D13 to D19 for monitoring the luteal function, and pregnancy was determined on D28. Luteolysis was earlier (P < 0.05) in non-inseminated heifers treated with 1 or 2 mg EB than in the controls (16.3 ± 0.2 <em>vs.</em> 17.3 ± 0.6 days). Pregnancy rate was lower (P < 0.05) in the EB-1 (50 %; 8/16) and EB-2 (29.2 %; 7/24) groups than in the EB-0 group (90 %; 9/10). The average PGFM concentrations were greater (P < 0.05) in the EB-1 and EB-2 groups than in the EB-0 group, regardless of gestational status. In Exp. 2, inseminated (n = 39) and non-inseminated (n = 21) Nelore heifers received either 0 or 1 mg of EB on D13. Three hours later, endometrial cytology was performed and samples were evaluated by qPCR. Expression of <em>OXTR</em> and <em>PGR</em> was greater and <em>IL1β</em> was lower in EB-treated heifers (P < 0.05). The <em>ESR2</em> abundance was lower (P < 0.05) in pregnant heifers, regardless of EB treatment. In conclusion, an elevation of circulating E2 at late diestrus upregulates the <em>OXTR</em> and <em>PGR</em> expression in the endometrium, inducing PGF<sub>2α</sub> release and luteolysis, which negatively impact on pregnancy establishment, especially in heifers treated with 2 mg EB.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"237 ","pages":"Pages 49-60"},"PeriodicalIF":2.4,"publicationDate":"2025-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143427958","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