Pub Date : 2024-10-11DOI: 10.1016/j.theriogenology.2024.10.008
Wenjing Yuan , Qi Zhang , Zhishan Yang , Yuting Zhang , Yang Zhou , Tingsheng Yan , Zhonghua Liu , Xinghong Ma , Xiaogang Weng
The gonadal primordial germ cells (PGCs) possess a unique state of pluripotency and X chromosome activity. However, extensive evidence indicates developmental variability in PGCs across different species. This study aims to evaluate the pluripotency status, specific gene expression patterns, and X chromosome reactivation (XCR) of pig gonadal PGCs. Single-cell RNA-seq revealed significant heterogeneity within the population of gonadal PGCs. Notably, these PGCs expressed high levels of pluripotency markers OCT4, PRDM14, and NANOG, while lacking SOX2 expression. Through the screening of marker genes and subsequent protein expression validation, we identified growth differentiation factor 3 (GDF3) as a specific surface marker for pig gonadal PGCs, facilitating their efficient purification for further study. Furthermore, analysis of gonadal PGCs demonstrated complete XCR. This was evidenced by the absence of repressive histone modifications (H3K27me3, H3K9me3, and H2AK119ub), the lack of X inactive specific transcript (XIST) RNA FISH signal, and the doubled expression of X-linked genes. Additionally, these PGCs expressed high levels of genes associated with epigenetic modification, chromatin remodeling, and XIST-associated RNA-binding. These factors likely play a crucial role in regulating pluripotency and X chromosome activity. In summary, this study reveals the heterogeneity in pig gonadal PGCs and identifies GDF3 as a specific surface marker. It also elucidates the expression patterns of pluripotency transcription factors and the events involved in XCR.
{"title":"Analysis of the pluripotent and germline marker gene expression, and the state of X chromosome reactivation of primordial germ cells in pig gonads","authors":"Wenjing Yuan , Qi Zhang , Zhishan Yang , Yuting Zhang , Yang Zhou , Tingsheng Yan , Zhonghua Liu , Xinghong Ma , Xiaogang Weng","doi":"10.1016/j.theriogenology.2024.10.008","DOIUrl":"10.1016/j.theriogenology.2024.10.008","url":null,"abstract":"<div><div>The gonadal primordial germ cells (PGCs) possess a unique state of pluripotency and X chromosome activity. However, extensive evidence indicates developmental variability in PGCs across different species. This study aims to evaluate the pluripotency status, specific gene expression patterns, and X chromosome reactivation (XCR) of pig gonadal PGCs. Single-cell RNA-seq revealed significant heterogeneity within the population of gonadal PGCs. Notably, these PGCs expressed high levels of pluripotency markers OCT4, PRDM14, and NANOG, while lacking SOX2 expression. Through the screening of marker genes and subsequent protein expression validation, we identified growth differentiation factor 3 (GDF3) as a specific surface marker for pig gonadal PGCs, facilitating their efficient purification for further study. Furthermore, analysis of gonadal PGCs demonstrated complete XCR. This was evidenced by the absence of repressive histone modifications (H3K27me3, H3K9me3, and H2AK119ub), the lack of <em>X inactive specific transcript</em> (<em>XIST</em>) RNA FISH signal, and the doubled expression of X-linked genes. Additionally, these PGCs expressed high levels of genes associated with epigenetic modification, chromatin remodeling, and <em>XIST</em>-associated RNA-binding. These factors likely play a crucial role in regulating pluripotency and X chromosome activity. In summary, this study reveals the heterogeneity in pig gonadal PGCs and identifies GDF3 as a specific surface marker. It also elucidates the expression patterns of pluripotency transcription factors and the events involved in XCR.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"231 ","pages":"Pages 52-61"},"PeriodicalIF":2.4,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142438099","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 aimed to evaluate the feasibility of performing multiple testicular biopsies in rams using Tru-cut® needles, assessing histological structure, gene expression, and potential complications such as effects on semen quality, testicular blood flow, and ultrasonographic echotexture. In Exp. 1, six mature rams underwent testicular biopsies at intervals (0, 3, 6, 12, 24, and 48 h) using a 16 G Tru-cut® needle, with alternating testes for each collection. Benzathine benzylpenicillin and flunixin meglumine were administered for infection and inflammation control. Local anesthesia and post-biopsy care included lidocaine, digital pressure, and ice application. Testicular samples were analyzed for gene expression related to inflammation, oxidative stress, and steroidogenesis. Semen quality was assessed pre-biopsy and 28 days post-biopsy. Ultrasonographic evaluations of the scrotum and testes were conducted before biopsies and on days 5, 9, 13, 17, and 21 post-biopsies. In Exp. 2, a second group of six mature rams underwent biopsies using 14 G needles, with two samples taken from each testis. Samples were histologically examined for structural preservation. Scrotal skin temperature was measured using infrared thermography, and testicular blood flow was assessed via color Doppler ultrasonography, with measurements taken before and on days 1, 2, 4, 6, 8, 10, 25, 50, 75, and 100 post-biopsies. Semen collection followed the same schedule as in Exp. 1. In Exp. 3, biopsies were performed on different testicular regions (upper, middle, lower) using 12 G, 14 G, and 16 G needles to compare structural preservation. Samples were histologically analyzed. No clinical signs of injury, inflammation, or fluid accumulation were observed. Scrotal pain, increased temperature, swelling, and bleeding were absent, and behavioral signs indicative of pain were not detected. Gene expression remained unchanged, and no significant alterations in seminal characteristics or testicular echogenicity were observed. A slight increase in resistivity and pulsatility indices was noted in Exp. 2. Biopsies with 14 G and 16 G needles resulted in structural disruptions, while 12 G needles better preserved testicular parenchyma. Multiple testicular biopsies using Tru-cut® needles did not cause significant morphological changes, alter transcriptional profiles, or affect semen or ultrasonographic characteristics, demonstrating that this method is viable for monitoring acute molecular changes in the testes.
本研究旨在评估使用 Tru-cut® 注射针对公羊进行多次睾丸活检的可行性，评估组织学结构、基因表达以及潜在并发症（如对精液质量、睾丸血流和超声回声纹理的影响）。在实验 1 中，六只成熟公羊使用 16 G Tru-cut® 针每隔一段时间（0、3、6、12、24 和 48 小时）进行睾丸活检，每次采集交替进行。苄星青霉素和氟尼辛葡胺用于控制感染和炎症。局部麻醉和活检后护理包括利多卡因、数字加压和冰敷。对睾丸样本进行了与炎症、氧化应激和类固醇生成相关的基因表达分析。精液质量在活检前和活检后 28 天进行评估。在活组织检查前以及活组织检查后的第 5、9、13、17 和 21 天，对阴囊和睾丸进行了超声波评估。在实验 2 中，第二组的六只成熟公羊使用 14 G 的针头进行了活组织检查，每个睾丸取两个样本。对样本进行组织学检查，以确定其结构是否保存完好。使用红外热成像技术测量阴囊皮肤温度，并通过彩色多普勒超声波检查评估睾丸血流量，在活检前和活检后第 1、2、4、6、8、10、25、50、75 和 100 天进行测量。精液采集时间与实验 1 相同。在实验 3 中，使用 12 G、14 G 和 16 G 的活检针对不同的睾丸区域（上、中、下）进行活检，以比较结构保存情况。对样本进行了组织学分析。没有观察到损伤、炎症或积液的临床症状。阴囊无疼痛、温度升高、肿胀和出血，也未发现表明疼痛的行为体征。基因表达保持不变，精液特征和睾丸回声也没有明显改变。使用 14 G 和 16 G 的活检针进行活检会导致结构破坏，而 12 G 的活检针能更好地保留睾丸实质。使用Tru-cut®针进行多次睾丸活检不会导致明显的形态学变化，也不会改变转录谱，或影响精液或超声波特征，这表明这种方法可用于监测睾丸的急性分子变化。
{"title":"Serial frequent or multiple Tru-cut® testicular biopsies in rams enable assessment of histological characteristics or transcriptional profiles, with no acute or chronic adverse effects","authors":"A.G.R. Pupulim , P.Z. Rattes , H.D. Mogollón García , J.C. Carvalho , K.Z. Uzae , G.C. Ribeiro , G. Rizzoto , R. Denadai , G.P. Nogueira , F.M.N. Navolar , G.W. Di Santis , S.G. Nunes , A.C.S. Castilho , J.P. Kastelic , J.C.P. Ferreira","doi":"10.1016/j.theriogenology.2024.10.007","DOIUrl":"10.1016/j.theriogenology.2024.10.007","url":null,"abstract":"<div><div>This study aimed to evaluate the feasibility of performing multiple testicular biopsies in rams using Tru-cut® needles, assessing histological structure, gene expression, and potential complications such as effects on semen quality, testicular blood flow, and ultrasonographic echotexture. In Exp. 1, six mature rams underwent testicular biopsies at intervals (0, 3, 6, 12, 24, and 48 h) using a 16 G Tru-cut® needle, with alternating testes for each collection. Benzathine benzylpenicillin and flunixin meglumine were administered for infection and inflammation control. Local anesthesia and post-biopsy care included lidocaine, digital pressure, and ice application. Testicular samples were analyzed for gene expression related to inflammation, oxidative stress, and steroidogenesis. Semen quality was assessed pre-biopsy and 28 days post-biopsy. Ultrasonographic evaluations of the scrotum and testes were conducted before biopsies and on days 5, 9, 13, 17, and 21 post-biopsies. In Exp. 2, a second group of six mature rams underwent biopsies using 14 G needles, with two samples taken from each testis. Samples were histologically examined for structural preservation. Scrotal skin temperature was measured using infrared thermography, and testicular blood flow was assessed via color Doppler ultrasonography, with measurements taken before and on days 1, 2, 4, 6, 8, 10, 25, 50, 75, and 100 post-biopsies. Semen collection followed the same schedule as in Exp. 1. In Exp. 3, biopsies were performed on different testicular regions (upper, middle, lower) using 12 G, 14 G, and 16 G needles to compare structural preservation. Samples were histologically analyzed. No clinical signs of injury, inflammation, or fluid accumulation were observed. Scrotal pain, increased temperature, swelling, and bleeding were absent, and behavioral signs indicative of pain were not detected. Gene expression remained unchanged, and no significant alterations in seminal characteristics or testicular echogenicity were observed. A slight increase in resistivity and pulsatility indices was noted in Exp. 2. Biopsies with 14 G and 16 G needles resulted in structural disruptions, while 12 G needles better preserved testicular parenchyma. Multiple testicular biopsies using Tru-cut® needles did not cause significant morphological changes, alter transcriptional profiles, or affect semen or ultrasonographic characteristics, demonstrating that this method is viable for monitoring acute molecular changes in the testes.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"231 ","pages":"Pages 81-89"},"PeriodicalIF":2.4,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142475499","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 : 2024-10-09DOI: 10.1016/j.theriogenology.2024.10.006
Leonardo F.C. Brito , Matheus R. Felix , Renata L. Linardi , Elena V. Martinez de Andino , Nithiya Sri Balamurugan , Camilo Hernández-Avilés , Katrin Hinrichs
Intracytoplasmic sperm injection (ICSI) is a valuable assisted reproduction technology in clinical practice, especially when semen availability is limited. Since the number of sperm required per ICSI cycle is much less than the number of sperm available in a standard straw of frozen semen, refreezing semen at lower sperm concentrations could yield multiple straws for ICSI use. However, there is little data on the effect of sperm refreezing on ICSI outcomes, especially on the effect of extender used for refreezing. The objective of the present study was to evaluate the effect of refreezing extender on stallion sperm quality and embryo production after ICSI. Semen was frozen in a low egg yolk/glycerol/amide extender (Extender 1), then thawed, re-extended, and refrozen in each of three extenders. When sperm were refrozen in Extender 1, the cleavage rate was lower (P < 0.05) and the blastocyst development rate tended to be lower (P = 0.06) than for the once-frozen sperm. In contrast, when sperm were refrozen in high egg yolk/glycerol (Extender 2) or low egg yolk/milk/glycerol (Extender 3) extenders, cleavage and blastocyst development rates did not differ significantly from those for the once-frozen semen. Notably, sperm refrozen in Extender 1, which yielded the lowest ICSI outcomes, showed the highest sperm motility and viability, demonstrating that traditional measures of sperm quality were inadequate to assess the suitability of refrozen sperm for ICSI. In a follow-up experiment conducted to evaluate the effects of Extenders 1 and 3 when used for once-frozen semen, cleavage and blastocyst rates did not differ between extenders. In conclusion, the extender used to initially freeze stallion sperm may not significantly affect ICSI outcomes; however, the extender used for refreezing can significantly affect embryo production. Refrozen stallion semen can be effectively used for ICSI when low egg yolk/milk/glycerol extender is used for refreezing. Until further research is available, use of extenders without amides is recommended when refreezing stallion semen for ICSI.
{"title":"Effect of refreezing extender on stallion sperm quality and embryo production after intracytoplasmic sperm injection","authors":"Leonardo F.C. Brito , Matheus R. Felix , Renata L. Linardi , Elena V. Martinez de Andino , Nithiya Sri Balamurugan , Camilo Hernández-Avilés , Katrin Hinrichs","doi":"10.1016/j.theriogenology.2024.10.006","DOIUrl":"10.1016/j.theriogenology.2024.10.006","url":null,"abstract":"<div><div>Intracytoplasmic sperm injection (ICSI) is a valuable assisted reproduction technology in clinical practice, especially when semen availability is limited. Since the number of sperm required per ICSI cycle is much less than the number of sperm available in a standard straw of frozen semen, refreezing semen at lower sperm concentrations could yield multiple straws for ICSI use. However, there is little data on the effect of sperm refreezing on ICSI outcomes, especially on the effect of extender used for refreezing. The objective of the present study was to evaluate the effect of refreezing extender on stallion sperm quality and embryo production after ICSI. Semen was frozen in a low egg yolk/glycerol/amide extender (Extender 1), then thawed, re-extended, and refrozen in each of three extenders. When sperm were refrozen in Extender 1, the cleavage rate was lower (P < 0.05) and the blastocyst development rate tended to be lower (P = 0.06) than for the once-frozen sperm. In contrast, when sperm were refrozen in high egg yolk/glycerol (Extender 2) or low egg yolk/milk/glycerol (Extender 3) extenders, cleavage and blastocyst development rates did not differ significantly from those for the once-frozen semen. Notably, sperm refrozen in Extender 1, which yielded the lowest ICSI outcomes, showed the highest sperm motility and viability, demonstrating that traditional measures of sperm quality were inadequate to assess the suitability of refrozen sperm for ICSI. In a follow-up experiment conducted to evaluate the effects of Extenders 1 and 3 when used for once-frozen semen, cleavage and blastocyst rates did not differ between extenders. In conclusion, the extender used to initially freeze stallion sperm may not significantly affect ICSI outcomes; however, the extender used for refreezing can significantly affect embryo production. Refrozen stallion semen can be effectively used for ICSI when low egg yolk/milk/glycerol extender is used for refreezing. Until further research is available, use of extenders without amides is recommended when refreezing stallion semen for ICSI.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"231 ","pages":"Pages 29-35"},"PeriodicalIF":2.4,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142432257","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 : 2024-10-09DOI: 10.1016/j.theriogenology.2024.10.005
Cristina Martínez-López , Carlos Manuel Martínez-Cáceres , María Cortina-Navarro , Mª José Izquierdo-Rico , Francisco Alberto García-Vázquez
Decellularized extracellular matrix (dECM) are widely utilized in regenerative medicine and tissue engineering due to their ability to promote cell growth, proliferation, and differentiation. In reproduction, research is focused on the utilization of these scaffolds to treat pathologies causing reproductive dysfunction or to improve assisted reproduction technologies (ARTs). We developed an efficient protocol employing the immersion-agitation technique to decellularize porcine oviductal and uterine sections, comparing the efficacy of fresh versus frozen treatments. Both methods successfully generated acellular matrices with less than 3 % residual DNA, effectively preserving structural and protein integrity. Scanning and transmission electron microscopy confirmed the ultrastructural integrity, whereas Masson's Trichrome staining highlighted better collagen preservation in frozen treatments. Proteomic analysis of decellularized scaffolds revealed collagen and key macromolecules such as laminin, filamin, dermatopontin, and fibronectin, which are essential for extracellular matrix structure and cell functions such as adhesion and migration. Innovatively, we assessed the biocompatibility and cytotoxicity of the scaffolds using spermatozoa, demonstrating that thorough washing ensures the scaffold biocompatibility without compromising sperm viability or motility. Our findings not only contribute to the standardization of decellularization protocols for female reproductive organs but also emphasize the importance of evaluating sperm biocompatibility to ensure the safety of dECM scaffolds.
{"title":"Characterization of decellularized porcine oviduct- and uterine-derived scaffolds evaluated by spermatozoa-based biocompatibility and biotoxicity","authors":"Cristina Martínez-López , Carlos Manuel Martínez-Cáceres , María Cortina-Navarro , Mª José Izquierdo-Rico , Francisco Alberto García-Vázquez","doi":"10.1016/j.theriogenology.2024.10.005","DOIUrl":"10.1016/j.theriogenology.2024.10.005","url":null,"abstract":"<div><div>Decellularized extracellular matrix (dECM) are widely utilized in regenerative medicine and tissue engineering due to their ability to promote cell growth, proliferation, and differentiation. In reproduction, research is focused on the utilization of these scaffolds to treat pathologies causing reproductive dysfunction or to improve assisted reproduction technologies (ARTs). We developed an efficient protocol employing the immersion-agitation technique to decellularize porcine oviductal and uterine sections, comparing the efficacy of fresh versus frozen treatments. Both methods successfully generated acellular matrices with less than 3 % residual DNA, effectively preserving structural and protein integrity. Scanning and transmission electron microscopy confirmed the ultrastructural integrity, whereas Masson's Trichrome staining highlighted better collagen preservation in frozen treatments. Proteomic analysis of decellularized scaffolds revealed collagen and key macromolecules such as laminin, filamin, dermatopontin, and fibronectin, which are essential for extracellular matrix structure and cell functions such as adhesion and migration. Innovatively, we assessed the biocompatibility and cytotoxicity of the scaffolds using spermatozoa, demonstrating that thorough washing ensures the scaffold biocompatibility without compromising sperm viability or motility. Our findings not only contribute to the standardization of decellularization protocols for female reproductive organs but also emphasize the importance of evaluating sperm biocompatibility to ensure the safety of dECM scaffolds.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"231 ","pages":"Pages 36-51"},"PeriodicalIF":2.4,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142432255","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-08DOI: 10.1016/j.theriogenology.2024.10.004
P.S. Baruselli , L.A. Abreu , A. Menchaca , G.A. Bó
South American beef production varies due to diverse climates, environmental conditions, animal breeds (Bos indicus, Bos taurus and crossbreeds), management strategies, and nutritional sources. Applying technology in the South American beef production system can significantly enhance efficiency, sustainability, and profitability. Reproductive efficiency is a significant challenge, especially in cow-calf operation systems conducted under adverse conditions. Consequently, implementing effective assisted reproduction technologies (ART) can make a significant contribution. In the last two decades, the development of fixed-time artificial insemination (FTAI) protocols permitted the widespread application of artificial insemination for breeding management and genetic improvement in beef herds in South America. Nowadays, FTAI is being applied in South America in large-scale programs, with around 20 % of heifers and cows receiving this technology every year. This results in a greater calving rate and significant genetic gain occurring in this territory. Also, in vitro embryo production, mainly using sex-selected sperm has been widely applied in this region, leading to significant improvements in herd genetics and productivity. Recently, 94 % of all embryo transfers in South America consist of in vitro-produced embryos (41,429 being in vivo-derived and 650,782 being in vitro-produced embryos), mainly using fixed-time embryo transfer technology (FTET). Genomic selection combined with in vitro embryo production with oocytes from heifer calves provides a powerful technology platform to reduce generation interval and significantly increase the rate of genetic gain in beef cattle. Emerging biotechnologies, such as genome editing via the CRISPR/Cas system, are being developed to enhance productivity, confer resilience to adverse environmental conditions, increase disease resistance, and control pest species that affect livestock. Finally, while all these technologies offer significant potential, further progresses are needed to transform livestock production. The vast geographical scale and diverse climates of South America make regional knowledge crucial for aligning beef production with sustainability goals and supporting global food security.
{"title":"The future of beef production in South America","authors":"P.S. Baruselli , L.A. Abreu , A. Menchaca , G.A. Bó","doi":"10.1016/j.theriogenology.2024.10.004","DOIUrl":"10.1016/j.theriogenology.2024.10.004","url":null,"abstract":"<div><div>South American beef production varies due to diverse climates, environmental conditions, animal breeds (<em>Bos indicus</em>, <em>Bos taurus</em> and crossbreeds), management strategies, and nutritional sources. Applying technology in the South American beef production system can significantly enhance efficiency, sustainability, and profitability. Reproductive efficiency is a significant challenge, especially in cow-calf operation systems conducted under adverse conditions. Consequently, implementing effective assisted reproduction technologies (ART) can make a significant contribution. In the last two decades, the development of fixed-time artificial insemination (FTAI) protocols permitted the widespread application of artificial insemination for breeding management and genetic improvement in beef herds in South America. Nowadays, FTAI is being applied in South America in large-scale programs, with around 20 % of heifers and cows receiving this technology every year. This results in a greater calving rate and significant genetic gain occurring in this territory. Also, <em>in vitro</em> embryo production, mainly using sex-selected sperm has been widely applied in this region, leading to significant improvements in herd genetics and productivity. Recently, 94 % of all embryo transfers in South America consist of <em>in vitro</em>-produced embryos (41,429 being <em>in vivo</em>-derived and 650,782 being <em>in vitro</em>-produced embryos), mainly using fixed-time embryo transfer technology (FTET). Genomic selection combined with <em>in vitro</em> embryo production with oocytes from heifer calves provides a powerful technology platform to reduce generation interval and significantly increase the rate of genetic gain in beef cattle. Emerging biotechnologies, such as genome editing via the CRISPR/Cas system, are being developed to enhance productivity, confer resilience to adverse environmental conditions, increase disease resistance, and control pest species that affect livestock. Finally, while all these technologies offer significant potential, further progresses are needed to transform livestock production. The vast geographical scale and diverse climates of South America make regional knowledge crucial for aligning beef production with sustainability goals and supporting global food security.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"231 ","pages":"Pages 21-28"},"PeriodicalIF":2.4,"publicationDate":"2024-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142406911","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 : 2024-10-05DOI: 10.1016/j.theriogenology.2024.09.031
Mayuko Anazawa , Shiori Ashibe , Yoshikazu Nagao
The generation of mammalian embryos by in vitro culture is hampered by the failure of many of the embryos to develop to the blastocyst stage. This problem occurs even when cumulus–oocyte complexes (COCs) with good morphology are visually selected and used for culture. Because cumulus cells are important for oocyte maturation and subsequent embryo development, here we compared gene expression patterns in cumulus cells of COCs that developed in vitro to the blastocyst stage with those of COCs that failed to develop. Cumulus cells were aspirated from bovine COCs selected for in vitro culture. Oocyte developmental competence was evaluated by screening for cleavage and development to the blastocyst stage. The collected cumulus cells were used to quantify mRNA levels of FSH receptor (FSHR), insulin-like growth factor-1 receptor (IGF-1R), anti-Müllerian hormone (AMH), AMH receptor II (AMHRII), epidermal growth factor receptor (EGFR), estrogen receptor β (ERβ), B cell lymphoma/leukemia-2 associated X (Bax), and cysteine-aspartic acid protease-3 (Caspase-3). We found that the expression levels of FSHR, IGF-1R, AMH, and EGFR were higher in cumulus cells from COCs that developed to blastocysts as compared with those that failed to develop, whereas expression levels of Bax and Caspase-3 were lower in cumulus cells of COCs that matured to the blastocyst stage. Positive correlations were found between FSHR and IGF-1R expression (r = 0.59) and between ERβ and EGFR expression (r = 0.43) in cumulus cells from COCs that developed to the blastocyst stage. Our findings indicate that gene expression levels in cumulus cells are correlated with the developmental competence of bovine oocytes. Measurement of gene expression in cumulus cells therefore offers a non-invasive means of predicting oocyte developmental competence.
{"title":"Gene expression levels in cumulus cells are correlated with developmental competence of bovine oocytes","authors":"Mayuko Anazawa , Shiori Ashibe , Yoshikazu Nagao","doi":"10.1016/j.theriogenology.2024.09.031","DOIUrl":"10.1016/j.theriogenology.2024.09.031","url":null,"abstract":"<div><div>The generation of mammalian embryos by in vitro culture is hampered by the failure of many of the embryos to develop to the blastocyst stage. This problem occurs even when cumulus–oocyte complexes (COCs) with good morphology are visually selected and used for culture. Because cumulus cells are important for oocyte maturation and subsequent embryo development, here we compared gene expression patterns in cumulus cells of COCs that developed in vitro to the blastocyst stage with those of COCs that failed to develop. Cumulus cells were aspirated from bovine COCs selected for in vitro culture. Oocyte developmental competence was evaluated by screening for cleavage and development to the blastocyst stage. The collected cumulus cells were used to quantify mRNA levels of FSH receptor (FSHR), insulin-like growth factor-1 receptor (IGF-1R), anti-Müllerian hormone (AMH), AMH receptor II (AMHRII), epidermal growth factor receptor (EGFR), estrogen receptor β (ERβ), B cell lymphoma/leukemia-2 associated X (Bax), and cysteine-aspartic acid protease-3 (Caspase-3). We found that the expression levels of <em>FSHR</em>, <em>IGF-1R</em>, <em>AMH</em>, and <em>EGFR</em> were higher in cumulus cells from COCs that developed to blastocysts as compared with those that failed to develop, whereas expression levels of <em>Bax</em> and <em>Caspase-3</em> were lower in cumulus cells of COCs that matured to the blastocyst stage. Positive correlations were found between <em>FSHR</em> and <em>IGF-1R</em> expression (r = 0.59) and between <em>ERβ</em> and <em>EGFR</em> expression (r = 0.43) in cumulus cells from COCs that developed to the blastocyst stage. Our findings indicate that gene expression levels in cumulus cells are correlated with the developmental competence of bovine oocytes. Measurement of gene expression in cumulus cells therefore offers a non-invasive means of predicting oocyte developmental competence.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"231 ","pages":"Pages 11-20"},"PeriodicalIF":2.4,"publicationDate":"2024-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142401392","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-03DOI: 10.1016/j.theriogenology.2024.09.033
Amir Abbas Shams , Soheil Vesal , Danial Hashemi Karoii , Samira Vesali , AliReza Alizadeh , Maryam Shahhoseini
Numerous studies have shown that an improper diet in parents has a negative impact on offspring's health. Furthermore, the negative effects of trans fatty acids (TFA) in maternal diets on fertility and health and their impact on future generations have been documented. However, there is limited research on the negative effects of TFA in paternal diets on male children. The current work used qRT-PCR to investigate the effects of trans fatty acids and vitamin E in the paternal diet on the expression pattern of androgen signaling pathway genes such as STAR, CYP11a1, HSD3B, SRD5a2, and SCARB1 in offspring testes. In this experiment, parental rats were randomly separated into four groups, each with ten father rats, and were fed for eight weeks (60 days) as follows. 1: Standard diet group plus liquid sunflower oil (control). 2: Standard diet group containing trans fatty acids (CTH). 3: The regular diet group received 2.5 times the recommended quantity of vitamin E supplement. 4: Standard diet group with vitamin E and trans fatty acid supplementation (ETH). The testis tissue samples from 35 offspring were then used. Following RNA extraction from tissues and cDNA synthesis, quantitative real-time PCR was used to evaluate the expression levels of androgen signaling pathway genes such as STAR, CYP11A1, HSD3B, SCARB1, and SRD5A2. Our findings showed that the expression of CYP11A1 was considerably reduced in the progeny of paternal rats given ETH compared to the CTH group. The expression levels of the STAR gene were significantly lower in the progeny of paternal rats administered TFA, ETH, and vitamin E compared to the controls. Although the CTH group had lower SCARB1 expression than the other groups, the difference was not statistically significant. Paternal vitamin E consumption substantially affected SRD5A2 expression when compared to offspring of paternal rats fed vitamin E + trans fatty acid or those fed a conventional diet containing trans fatty acid. Furthermore, the vitamin E group showed a statistically significant increase in HSD3B expression compared to the other groups. Bioinformatics analyses, such as protein-protein interaction networks and gene ontology term enrichment, revealed that these genes play roles in lipid biosynthesis, hormone metabolism, male sex differentiation, reproductive development, and steroid biosynthesis. Our data indicate that paternal trans fatty acid consumption influences the expression of particular androgen signaling pathway genes in offspring testis, with vitamin E potentially mitigating some of these effects.
大量研究表明，父母饮食不当会对后代的健康产生负面影响。此外，母体饮食中的反式脂肪酸（TFA）对生育和健康的负面影响及其对后代的影响也有文献记载。然而，关于父亲饮食中的反式脂肪酸对男性子女的负面影响的研究却很有限。本研究采用 qRT-PCR 技术研究父代膳食中反式脂肪酸和维生素 E 对子代睾丸中 STAR、CYP11a1、HSD3B、SRD5a2 和 SCARB1 等雄激素信号通路基因表达模式的影响。本实验将亲代大鼠随机分为四组，每组十只，按以下方法喂养八周（60 天）。1：标准饮食加液体葵花籽油组（对照组）。2：标准饮食组，含反式脂肪酸（CTH）。3：普通饮食组添加 2.5 倍于推荐量的维生素 E 补充剂。4：补充维生素 E 和反式脂肪酸的标准饮食组（ETH）。然后使用 35 个后代的睾丸组织样本。提取组织中的 RNA 并合成 cDNA 后，使用实时定量 PCR 评估 STAR、CYP11A1、HSD3B、SCARB1 和 SRD5A2 等雄激素信号通路基因的表达水平。我们的研究结果表明，与CTH组相比，给予ETH的父代大鼠后代中CYP11A1的表达明显降低。与对照组相比，给予反式脂肪酸、ETH和维生素E的父代大鼠后代中STAR基因的表达水平明显较低。虽然CTH组的SCARB1表达量低于其他组，但差异无统计学意义。与喂食维生素 E + 反式脂肪酸或喂食含有反式脂肪酸的传统饮食的父代大鼠的后代相比，父代食用维生素 E 会大大影响 SRD5A2 的表达。此外，与其他组相比，维生素 E 组的 HSD3B 表达有显著的统计学增长。生物信息学分析（如蛋白质-蛋白质相互作用网络和基因本体术语富集）显示，这些基因在脂质生物合成、激素代谢、雄性性别分化、生殖发育和类固醇生物合成中发挥作用。我们的数据表明，父亲摄入反式脂肪酸会影响后代睾丸中特定雄激素信号通路基因的表达，而维生素 E 有可能减轻其中的一些影响。
{"title":"Paternal trans fatty acid and vitamin E diet affect the expression pattern of androgen signaling pathway genes in the testis of rat offspring","authors":"Amir Abbas Shams , Soheil Vesal , Danial Hashemi Karoii , Samira Vesali , AliReza Alizadeh , Maryam Shahhoseini","doi":"10.1016/j.theriogenology.2024.09.033","DOIUrl":"10.1016/j.theriogenology.2024.09.033","url":null,"abstract":"<div><div>Numerous studies have shown that an improper diet in parents has a negative impact on offspring's health. Furthermore, the negative effects of trans fatty acids (TFA) in maternal diets on fertility and health and their impact on future generations have been documented. However, there is limited research on the negative effects of TFA in paternal diets on male children. The current work used qRT-PCR to investigate the effects of trans fatty acids and vitamin E in the paternal diet on the expression pattern of androgen signaling pathway genes such as STAR, CYP11a1, HSD3B, SRD5a2, and SCARB1 in offspring testes. In this experiment, parental rats were randomly separated into four groups, each with ten father rats, and were fed for eight weeks (60 days) as follows. 1: Standard diet group plus liquid sunflower oil (control). 2: Standard diet group containing trans fatty acids (CTH). 3: The regular diet group received 2.5 times the recommended quantity of vitamin E supplement. 4: Standard diet group with vitamin E and trans fatty acid supplementation (ETH). The testis tissue samples from 35 offspring were then used. Following RNA extraction from tissues and cDNA synthesis, quantitative real-time PCR was used to evaluate the expression levels of androgen signaling pathway genes such as STAR, CYP11A1, HSD3B, SCARB1, and SRD5A2. Our findings showed that the expression of CYP11A1 was considerably reduced in the progeny of paternal rats given ETH compared to the CTH group. The expression levels of the STAR gene were significantly lower in the progeny of paternal rats administered TFA, ETH, and vitamin E compared to the controls. Although the CTH group had lower SCARB1 expression than the other groups, the difference was not statistically significant. Paternal vitamin E consumption substantially affected SRD5A2 expression when compared to offspring of paternal rats fed vitamin E + trans fatty acid or those fed a conventional diet containing trans fatty acid. Furthermore, the vitamin E group showed a statistically significant increase in HSD3B expression compared to the other groups. Bioinformatics analyses, such as protein-protein interaction networks and gene ontology term enrichment, revealed that these genes play roles in lipid biosynthesis, hormone metabolism, male sex differentiation, reproductive development, and steroid biosynthesis. Our data indicate that paternal trans fatty acid consumption influences the expression of particular androgen signaling pathway genes in offspring testis, with vitamin E potentially mitigating some of these effects.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"231 ","pages":"Pages 1-10"},"PeriodicalIF":2.4,"publicationDate":"2024-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142393615","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 : 2024-10-03DOI: 10.1016/j.theriogenology.2024.10.003
Esther Collantes-Fernández , Pilar Horcajo , Julio Benavides , Roberto Sánchez-Sánchez , Javier Blanco-Murcia , Sandra Montaner-Da Torre , Yanina P. Hecker , Luis Miguel Ortega-Mora , Iván Pastor-Fernández
Bovine abortions, often caused by infectious agents like Neospora caninum, inflict substantial economic losses. Studying host-pathogen interactions in pregnant cows is challenging, and existing cell cultures lack the intricate complexity of real tissues. To bridge the gap between in vitro and in vivo models, we explored the use of cryopreserved bovine placental explants. Building upon our successful development of protocols for obtaining, culturing, and cryopreserving sheep placental explants, we applied these methods to bovine tissues. Here, we compared fresh and cryopreserved bovine explants, evaluating their integrity and functionality over culture time. Additionally, we investigated their susceptibility to N. caninum infection. Our findings revealed that bovine explants deteriorate faster in culture compared to sheep explants, exhibiting diminished viability and function. Cryopreservation further exacerbated this deterioration. While fresh explants were successfully infected with N. caninum, parasite replication was limited. Notably, cryopreservation reduced infection efficiency. This pioneering work paves the way for developing ex vivo models to study reproductive pathogens in cattle. However, further optimization of the model is essential. These improved models will have the potential to significantly reduce the reliance on animals in research.
{"title":"Evaluating the suitability of placental bovine explants for ex vivo modelling of host-pathogen interactions in Neospora caninum infections","authors":"Esther Collantes-Fernández , Pilar Horcajo , Julio Benavides , Roberto Sánchez-Sánchez , Javier Blanco-Murcia , Sandra Montaner-Da Torre , Yanina P. Hecker , Luis Miguel Ortega-Mora , Iván Pastor-Fernández","doi":"10.1016/j.theriogenology.2024.10.003","DOIUrl":"10.1016/j.theriogenology.2024.10.003","url":null,"abstract":"<div><div>Bovine abortions, often caused by infectious agents like <em>Neospora caninum</em>, inflict substantial economic losses. Studying host-pathogen interactions in pregnant cows is challenging, and existing cell cultures lack the intricate complexity of real tissues. To bridge the gap between in vitro and in vivo models, we explored the use of cryopreserved bovine placental explants. Building upon our successful development of protocols for obtaining, culturing, and cryopreserving sheep placental explants, we applied these methods to bovine tissues. Here, we compared fresh and cryopreserved bovine explants, evaluating their integrity and functionality over culture time. Additionally, we investigated their susceptibility to <em>N. caninum</em> infection. Our findings revealed that bovine explants deteriorate faster in culture compared to sheep explants, exhibiting diminished viability and function. Cryopreservation further exacerbated this deterioration. While fresh explants were successfully infected with <em>N. caninum</em>, parasite replication was limited. Notably, cryopreservation reduced infection efficiency. This pioneering work paves the way for developing ex vivo models to study reproductive pathogens in cattle. However, further optimization of the model is essential. These improved models will have the potential to significantly reduce the reliance on animals in research.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"230 ","pages":"Pages 305-313"},"PeriodicalIF":2.4,"publicationDate":"2024-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142378335","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-02DOI: 10.1016/j.theriogenology.2024.10.002
S. Cánovas, S. Heras, J. Romero-Aguirregomezcorta, A.A. Quintero-Moreno, J. Gadea, P. Coy, R. Romar
The aim of the present work was to gain insight into the metabolism of pigs derived from assisted reproductive technologies during their adulthood. Approximately 4h after feeding, a blood sample was taken from 3.5 year old sows born by artificial insemination (AI group, n = 7) and transfer of in vitro produced embryos (IVP group, n = 11) to determine the physiological concentrations of the main biomarkers of carbohydrates (glucose and lactate), proteins (albumin, creatinine and urea) and lipids (cholesterol and triglycerides). Four weeks later, an oral glucose tolerance test (OGTT; 1.75g glucose/kg body weight) was performed after an overnight fast and 1h of water withdrawal. Blood samples were obtained prior (T = 0 min; fasting conditions) and 15, 30, 45, 60, 90, 120, 150, 180, 210 and 240 min after glucose intake. At each time point, glycemia was measured immediately using glucometer test strips, and serum was collected to determine the above metabolites along with insulin and glucagon. After OGTT, the area under the curve (AUC) between sampling times and homeostasis model assessment of insulin resistance (HOMA) indices were calculated. Under physiological conditions, the concentration of metabolites studied was similar between AI and IVP sows. In both groups, fasting decreased cholesterol and increased triglycerides and urea (P < 0.001). However, creatinine and lactate were similar in both groups under physiological and fasting conditions. The expected increase in albuminemia and decrease in glycaemia after fasting was only observed in IVP sows. OGTT revealed a different glucose curve pattern (monophasic in AI and biphasic in IVP group), a lower mean concentration of cholesterol, glucose, lactate, triglycerides in IVP compared to AI pigs (P < 0.01), and a higher mean concentration of albumin, creatinine and insulin in IVP compared to AI group (P < 0.05). On the contrary, no differences were found between groups for mean serum glucagon and urea levels, nor for glucose homeostasis indices HOMA-IR and HOMA-%B. The AUC differed between groups at several time points with larger AUC for creatinine, and smaller AUC for glucose, glucagon, and triglycerides, in IVP pigs than in AI pigs at 180–210 min (P < 0.05). In conclusion, under physiological conditions the metabolic profile of fully-grown AI and IVP sows is similar and within normal ranges. Glucose challenge revealed differences in metabolic and insulin responses between groups but with normal glucose tolerance in both cases.
{"title":"Metabolic profile and glycemic response in fully-grown sows born using assisted reproductive technologies","authors":"S. Cánovas, S. Heras, J. Romero-Aguirregomezcorta, A.A. Quintero-Moreno, J. Gadea, P. Coy, R. Romar","doi":"10.1016/j.theriogenology.2024.10.002","DOIUrl":"10.1016/j.theriogenology.2024.10.002","url":null,"abstract":"<div><div>The aim of the present work was to gain insight into the metabolism of pigs derived from assisted reproductive technologies during their adulthood. Approximately 4h after feeding, a blood sample was taken from 3.5 year old sows born by artificial insemination (AI group, n = 7) and transfer of in vitro produced embryos (IVP group, n = 11) to determine the physiological concentrations of the main biomarkers of carbohydrates (glucose and lactate), proteins (albumin, creatinine and urea) and lipids (cholesterol and triglycerides). Four weeks later, an oral glucose tolerance test (OGTT; 1.75g glucose/kg body weight) was performed after an overnight fast and 1h of water withdrawal. Blood samples were obtained prior (T = 0 min; fasting conditions) and 15, 30, 45, 60, 90, 120, 150, 180, 210 and 240 min after glucose intake. At each time point, glycemia was measured immediately using glucometer test strips, and serum was collected to determine the above metabolites along with insulin and glucagon. After OGTT, the area under the curve (AUC) between sampling times and homeostasis model assessment of insulin resistance (HOMA) indices were calculated. Under physiological conditions, the concentration of metabolites studied was similar between AI and IVP sows. In both groups, fasting decreased cholesterol and increased triglycerides and urea (<em>P</em> < 0.001). However, creatinine and lactate were similar in both groups under physiological and fasting conditions. The expected increase in albuminemia and decrease in glycaemia after fasting was only observed in IVP sows. OGTT revealed a different glucose curve pattern (monophasic in AI and biphasic in IVP group), a lower mean concentration of cholesterol, glucose, lactate, triglycerides in IVP compared to AI pigs (P < 0.01), and a higher mean concentration of albumin, creatinine and insulin in IVP compared to AI group (P < 0.05). On the contrary, no differences were found between groups for mean serum glucagon and urea levels, nor for glucose homeostasis indices HOMA-IR and HOMA-%B. The AUC differed between groups at several time points with larger AUC for creatinine, and smaller AUC for glucose, glucagon, and triglycerides, in IVP pigs than in AI pigs at 180–210 min (P < 0.05). In conclusion, under physiological conditions the metabolic profile of fully-grown AI and IVP sows is similar and within normal ranges. Glucose challenge revealed differences in metabolic and insulin responses between groups but with normal glucose tolerance in both cases.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"230 ","pages":"Pages 314-321"},"PeriodicalIF":2.4,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142378336","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-29DOI: 10.1016/j.theriogenology.2024.09.032
Yimeng Wei , Xiyu Zhao , Yao Zhang , Can Cui , Shunshun Han , Chaowu Yang , Huadong Yin
Granulosa cell (GC) death, which leads to follicular atresia, primarily occurs through apoptosis and autophagy. miRNAs are known to be key regulators of autophagy and apoptosis. Although miR-7 acting as a key regulator of follicular atresia, its precise role in granulosa cell autophagy and apoptosis remains to be fully elucidated. In this study, we found that miR-7 was highly expressed in the follicle based on qPCR analysis. Subsequently, transfection of miR-7 inhibitors and mimics downregulated or upregulated the expression of miR-7 and promoted autophagic and apoptotic processes in chicken follicle granulosa cells. Mechanistically, through dual-luciferase reporter gene assays, we validated that KLF4 is a target gene of miR-7. Contrarily, KLF4 was found to negatively regulate autophagy and apoptosis in follicular granulosa cells as evidenced by genetic intervention of KLF4 silencing and overexpression. Furthermore, JAK/STAT3 signaling pathway was confirmed to mediate the regulation of miR-7-KLF4 axis on GC autophagy and apoptosis. These findings offer evidences of the crucial involvement of the miR-7-KLF4 signaling axis in determining autophagy and apoptosis of GCs. This study could offer an important theoretical basis for the use of molecular-assisted breeding in chickens.
{"title":"miR-7 promotes apoptosis and autophagy of granulosa cells by targeting KLF4 via JAK/STAT3 signaling pathway in chickens","authors":"Yimeng Wei , Xiyu Zhao , Yao Zhang , Can Cui , Shunshun Han , Chaowu Yang , Huadong Yin","doi":"10.1016/j.theriogenology.2024.09.032","DOIUrl":"10.1016/j.theriogenology.2024.09.032","url":null,"abstract":"<div><div>Granulosa cell (GC) death, which leads to follicular atresia, primarily occurs through apoptosis and autophagy. miRNAs are known to be key regulators of autophagy and apoptosis. Although miR-7 acting as a key regulator of follicular atresia, its precise role in granulosa cell autophagy and apoptosis remains to be fully elucidated. In this study, we found that miR-7 was highly expressed in the follicle based on qPCR analysis. Subsequently, transfection of miR-7 inhibitors and mimics downregulated or upregulated the expression of miR-7 and promoted autophagic and apoptotic processes in chicken follicle granulosa cells. Mechanistically, through dual-luciferase reporter gene assays, we validated that KLF4 is a target gene of miR-7. Contrarily, KLF4 was found to negatively regulate autophagy and apoptosis in follicular granulosa cells as evidenced by genetic intervention of KLF4 silencing and overexpression. Furthermore, JAK/STAT3 signaling pathway was confirmed to mediate the regulation of miR-7-KLF4 axis on GC autophagy and apoptosis. These findings offer evidences of the crucial involvement of the miR-7-KLF4 signaling axis in determining autophagy and apoptosis of GCs. This study could offer an important theoretical basis for the use of molecular-assisted breeding in chickens.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"230 ","pages":"Pages 322-329"},"PeriodicalIF":2.4,"publicationDate":"2024-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142381700","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}
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