Theriogenology最新文献

{"title":"The effect of L-carnitine on frozen-thawed rooster sperm quality and fertility potential","authors":"Lijun Jiang ,&nbsp;Yunlei Li ,&nbsp;Yunhe Zong ,&nbsp;Xintong Han ,&nbsp;Jiangpeng Guo ,&nbsp;Sihua Jin ,&nbsp;Yi Zhao ,&nbsp;Jingwei Yuan ,&nbsp;Hui Ma ,&nbsp;Jilan Chen ,&nbsp;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> &lt; 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> &lt; 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> &lt; 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}

{"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 ,&nbsp;Karina Pessoa Oliveira ,&nbsp;Leonardo Vitorino Costa de Aquino ,&nbsp;Alexsandra Fernandes Pereira ,&nbsp;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 &lt; 0.05), while Grade IV remained unchanged. COC recovery did not differ across storage times (P &gt; 0.05), with &gt;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 &lt; 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}

{"title":"Effects of estradiol on PGF2α synthesis and corpus luteum function during early pregnancy in beef heifers","authors":"Igor Garcia Motta ,&nbsp;Amanda Guimarães da Silva ,&nbsp;Isabella Rio Feltrin ,&nbsp;Samuel Volpe Souza ,&nbsp;Ana Clara Degan Mattos ,&nbsp;Karine Galhego Morelli ,&nbsp;Thadeu Castro ,&nbsp;Thiago Kan Nishimura ,&nbsp;Oliver Joseph Ginther ,&nbsp;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 &lt; 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 &lt; 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 &lt; 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 &lt; 0.05). The <em>ESR2</em> abundance was lower (P &lt; 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_2α 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}

{"title":"oar-miR-29a promotes the establishment of endometrial receptivity by targeting CDC42 in sheep","authors":"Xi Chen ,&nbsp;Luyi Tang ,&nbsp;Yizhe Pan ,&nbsp;Yanshe Xie ,&nbsp;Huijia Jin ,&nbsp;Xin Xiang ,&nbsp;Zhengguang Wang","doi":"10.1016/j.theriogenology.2025.02.010","DOIUrl":"10.1016/j.theriogenology.2025.02.010","url":null,"abstract":"<div><div>Endometrial receptivity is vital for the successful implantation of embryos in sheep. Insufficient receptivity is a major cause of implantation failure, highlighting the need to understand the underlying mechanisms. MicroRNAs (miRNAs) play an essential role in the epigenetic regulation of endometrial receptivity and embryo implantation by influencing post-transcriptional processes. However, the specific mechanisms by which various miRNAs contribute to endometrial receptivity and embryo implantation in sheep remain to be fully elucidated. Our research highlights the significant role of oar-miR-29a, which promotes the migration and proliferation of sheep endometrial epithelial cells (sEECs). We found that oar-miR-29a enhances mRNA expression associated with proliferation, migration, and epithelial-mesenchymal transition (EMT) in sEECs. Additionally, this miRNA enhances the proliferation and migration capabilities of sheep trophoblast cells (sTCs). Moreover, our findings suggest that the target genes of oar-miR-29a are involved in key biological processes and pathways essential for embryo implantation. Notably, our analysis identifies cell division cycle 42 (CDC42) as a target gene of oar-miR-29a. In summary, oar-miR-29a plays a crucial role in promoting endometrial receptivity by targeting CDC42 in sheep. These findings provide insights into the miRNA-based regulatory mechanisms governing uterine physiology during the early stages of pregnancy, emphasizing the importance of this research for improving reproductive outcomes in sheep.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"237 ","pages":"Pages 22-32"},"PeriodicalIF":2.4,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143421752","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Influence of the follicular wave on gene expression and in vitro embryo production in cattle","authors":"N.C. Silva ,&nbsp;D.N. Yokomizo ,&nbsp;A.F. Zangirolamo ,&nbsp;F.L.B. Cavalieri ,&nbsp;C.B. Costa ,&nbsp;E.A.A. Rossignolo ,&nbsp;T.T. Dellaqua ,&nbsp;F. Morotti ,&nbsp;M.M. Seneda","doi":"10.1016/j.theriogenology.2025.02.009","DOIUrl":"10.1016/j.theriogenology.2025.02.009","url":null,"abstract":"<div><div>This study evaluated oocyte competence, gene expression, and <em>in vitro</em> embryo production (IVEP) in cattle, based on follicular waves. Twenty <em>Bos taurus taurus</em> donors were subjected to ovulation synchronization, starting with intramuscular administration of 2 mg estradiol benzoate and a 1.9 g intravaginal progesterone device on a random estrous cycle day (ten days before synchronized ovulation; D-10). After 3 days, the device was removed, and 150 μg of D-cloprostenol sodium, 300 IU of equine chorionic gonadotrophin, and 1.0 mg of estradiol cypionate were administered. Day zero (D0) was defined as the day of ovulation, and the ovaries of the females in the crossover design were examined using Doppler ultrasonography. Ovum pick-up was scheduled on days D4, D8, D14, and D18, and the experimental groups were designated as G4 (n = 5), G8 (n = 5), G14 (n = 5), and G18 (n = 5), respectively. The corpus luteum (CL) increased in diameter, perimeter, and area throughout the estrous cycle, with significant differences between G4 and other groups (P &lt; 0.0001). CL vascularization scores on G4, G8, G14, and G18 revealed a gradual increase in peripheral blood flow(1.28, 1.79, 1.67, and 1.86, respectively). The central blood flow was higher in G8 (1.53) and G14 (1.57) than that in G4. Oocytes from each group were analyzed using reverse transcription and quantitative polymerase chain reaction after cumulus cell removal. The effect of group (OPU timing) on follicular growth waves was analyzed using ANOVA, followed by Tukey's post hoc test. All statistical analyses were conducted using Minitab statistical software version 18.1, with the significance level set at P ≤ 0.05. For evaluation of qPCR data, the 2^−ΔCq method was used. Analyses were conducted using SigmaStat 4.0 and MetaboAnalyst 5.0. Differences were considered significant at P &lt; 0.05 and/or FC &gt; 2.0 (upregulated) or FC &lt; 0.5 (downregulated).Total and viable oocyte numbers were lowest in G8 (13.9 and 9, respectively). The average numbers of embryos per donor were 2.58, 2.38, 2.29, and 1.69 G14, G18, G4, and G8, respectively. Gene expression analysis showed downregulation of genes related to apoptosis and lipid metabolism in oocytes retrieved from G14 compared to those from G4, G8, or G18. Oocytes from G18 showed upregulation of genes related to apoptosis control and lipid metabolism, whereas those from G4 and G8 were downregulated. In conclusion, ovum pick-up at the beginning of the second follicular wave can improve IVP efficiency.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"237 ","pages":"Pages 33-41"},"PeriodicalIF":2.4,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143421754","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Influence of the intrauterine application of a phytotherapeutic agent and other parameters on the uterine involution of sows in a free farrowing system","authors":"Julia Adam ,&nbsp;Philipp T. Egli ,&nbsp;Lukas Schulthess ,&nbsp;Gaby Hirsbrunner ,&nbsp;Gertraud Schüpbach-Regula ,&nbsp;Alexander Grahofer","doi":"10.1016/j.theriogenology.2025.02.011","DOIUrl":"10.1016/j.theriogenology.2025.02.011","url":null,"abstract":"<div><div>Endometritis and postpartum dysgalactiae syndrome are common postpartum disorders in sows revealing antimicrobial treatment.</div><div>This study evaluated the influence of the intrauterine application of a phytotherapeutic agent (EucaComp©) after farrowing containing essences of marjoram, marigold, melissa and eucalyptus and other sow-related parameters on the uterine involution.</div><div>In total, 150 Landrace x Large white sows (parity 1–9) were divided randomly into three groups (no intervention (Control), 300 ml sodium chloride (Placebo) and 300 ml of phytotherapeutic agent (Phytotherapeutic)). Several sow-related parameters were assessed such as, parity, Body Condition Score (1–5), birth induction (Yes/No), and feed intake. On days 2, 3, 5, 7 and 21 postpartum, uterine involution was assessed by measuring the diameter on three locations of the uterine horns using ultrasound.</div><div>The mean uterine diameter changed from 36.4 ± 6.5 mm on day 2–20.4 ± 5.9 mm on day 7 (reduction of 43.81 %) and 14.0 ± 3.2 mm on day 21 (reduction of 61.15 %). No significant differences between the groups and the regression of uterine involution were detected. However, birth induction had a positive effect on uterine involution, increasing the relative uterine involution by 8 %, whereas lack of appetite postpartum, lead to a decrease of 7 % until day 7. An suboptimal BCS (≤2; ≥4) and a parity ≥2 lead to a reduction of uterine involution regression on day 21 by 7 and 5 %. In conclusion, a single postpartal intrauterine application of a phytotherapeutic agent demonstrated no significant impact on uterine involution, while sow-specific and management factors remained pivotal.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"237 ","pages":"Pages 13-21"},"PeriodicalIF":2.4,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143421751","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mogroside III improves bovine oocyte in vitro maturation by regulating autophagy in cumulus cells","authors":"Xinru Tian ,&nbsp;Peng Xiao ,&nbsp;Mengqi Li ,&nbsp;Nannan Li ,&nbsp;Yilin Huang ,&nbsp;Chunyan Yang ,&nbsp;Haiying Zheng ,&nbsp;Xiaogan Yang ,&nbsp;Jianghua Shang ,&nbsp;Xingwei Liang","doi":"10.1016/j.theriogenology.2025.02.006","DOIUrl":"10.1016/j.theriogenology.2025.02.006","url":null,"abstract":"<div><div><em>In vitro</em> maturation (IVM) of oocytes is pivotal for successful embryo production. Cumulus cells (CCs) contribute to oocyte maturation through the secretion of hormones and nutrients, with proper autophagic activity being crucial for this process. However, the role of autophagy in CCs remains underexplored. <em>Siraitia grosvenorii</em> extract Mogroside III (MIII), known for its antioxidant properties, has yet to be extensively studied for its impact on bovine oocyte IVM and its potential regulatory effects on autophagy. This study assessed the influence of MIII on autophagic activity in CCs and its subsequent effects on oocyte developmental potential. The results demonstrated that MIII enhanced bovine oocyte IVM, promoted CC expansion, and supported embryonic development. Transcriptomic analysis indicated that MIII upregulated the expression of autophagy-related genes. <em>In vitro</em> experiments on CCs revealed that MIII increased LC3B protein levels, reduced SQSTM1 accumulation, and upregulated the gene expression of <em>LC3</em>, <em>Beclin1</em>, and <em>ATG5</em>. In co-culture systems, autophagy inhibition in CCs impaired oocyte IVM and embryonic development, but MIII alleviated these effects, restoring oocyte developmental capacity compromised by 3-MA-induced autophagy inhibition. Mechanistically, MIII facilitated the degradation of WT1 by upregulating LC3B, influencing CC differentiation, enhancing FSHR synthesis, and increasing estrogen and progesterone secretion. In conclusion, MIII enhances oocyte developmental potential by modulating autophagy in CCs.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"237 ","pages":"Pages 1-12"},"PeriodicalIF":2.4,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143421743","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Revealing the effects of lactate on bovine SCNT embryo development through transcriptome sequencing analyses","authors":"Nannan Li ,&nbsp;Huan Chen ,&nbsp;Haiying Zheng ,&nbsp;Chunyan Yang ,&nbsp;Mengqi Li ,&nbsp;Peng Xiao ,&nbsp;Yilin Huang ,&nbsp;Xinru Tian ,&nbsp;Xingwei Liang ,&nbsp;Jianghua Shang ,&nbsp;Xiaogan Yang","doi":"10.1016/j.theriogenology.2025.02.007","DOIUrl":"10.1016/j.theriogenology.2025.02.007","url":null,"abstract":"<div><div>As a major energy source in the metabolism of early embryos, lactate also participates in regulating zygotic gene expression and subsequently contributes to preimplantation embryonic development. Nevertheless, the influence of lactate on embryonic development in bovine cloned embryos remains elusive. For this reason, this research explored the differences in metabolic pathways between in vitro fertilization (IVF) embryos and somatic cell nuclear transfer (SCNT) embryos during zygotic genome activation (ZGA) using Smart-seq and observed changes in SCNT embryo development in response to lactate supplementation. Weighted gene coexpression network analysis (WGCNA) revealed that <em>LDHA</em> functions as a hub gene that significantly influences the gene expression profile of 8-cell SCNT embryos. Compared with those of IVF embryos, SCNT embryos showed lower LDHA levels and lactate contents. Lactate supplementation was found to increase the developmental potential and blastocyst quality of SCNT embryos. Furthermore, the addition of lactate significantly increased the immunofluorescence intensity of both Pan Kla and H3K18la as well as the expression levels of the zygotic genes <em>ZSCAN5B</em> and <em>SUPT4H1</em> in SCNT embryos. These results indicate that lactate deficiency leading to the downregulation of histone lactylation may be an important factor affecting the in vitro development of SCNT embryos.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"236 ","pages":"Pages 137-146"},"PeriodicalIF":2.4,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143403543","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhancing spermatogonial stem cell differentiation: The role of alpha-ketoglutarate in in-Vitro cultures","authors":"Mahdi Jahanbakhsh ,&nbsp;Tooba Mirzapour ,&nbsp;Fatemeh Asgari ,&nbsp;Hediyeh Fadakar ,&nbsp;Fatemeh Ghasemian ,&nbsp;Morteza Koruji","doi":"10.1016/j.theriogenology.2025.02.005","DOIUrl":"10.1016/j.theriogenology.2025.02.005","url":null,"abstract":"<div><div>Spermatogonial stem cells (SSCs) have the unique ability to self-renew and differentiate into mature spermatozoa. In vitro culture of SSCs, however, presents several challenges, particularly in promoting efficient differentiation. This study investigates the role of metabolic intermediates, such as alpha-ketoglutarate (AKG), on the differentiation of SSCs isolated from the testes of 3–6 day-old mice. SSCs and sertoli cells were extracted using collagenase ІV and trypsin and co-cultured in DMEM/F12 supplemented with 20 % fetal bovine serum (FBS) and glial cell-derived neurotrophic factor (GDNF) for one week. The survival rate of cells was evaluated under influence of different dosages of AKG (0.04, 0.1, 0.4, 4, 10 mM) after 1 and 7 days of culture using the MTT test. The cell viability was significantly increased at the 0.1 mM dose of AKG rather than other groups. This dosage was selected for adding to culture system.</div><div>In the control group, the cells cultured for three weeks in DMEM/F12 with 10 % FBS, 10⁻6 M retinoic acid, and 40 ng/mL bone morphogenetic protein-4 (BMP-4). The treatment group received the same medium with the addition of 0.1 mM AKG. The presence of Sertoli cell in the culture system was confirmed by SOX9-positive immunocytochemistry. The Colonies that formed on the Sertoli cells exhibited positive alkaline phosphatase activity and reacted positively for Oct4 and GFRa1 immunocytochemistry. The expression of testicular-specific genes (Acrosin and Sycp3) and anti-apoptosis-related genes (Nrf2 and Bcl2) was evaluated after 7 days (as the zero group) and again after 21 days of culture, in treatment (0.1 mM AKG) and control groups. A high expression of Acrosin and Sycp3 expression was observed in AKG-treated group compared to control and zero groups (p ≤ 0.05). The expression of Nrf2 and Bcl2 genes was also significantly increased in the treatment group (p ≤ 0.05). These findings suggest that AKG activates mechanisms of cellular antioxidant response and subsequently increase the expression of differentiation genes in SSCs.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"237 ","pages":"Pages 61-69"},"PeriodicalIF":2.4,"publicationDate":"2025-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143437446","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Editing the growth differentiation factor 9 gene affects porcine oocytes in vitro maturation by inactivating the maturation promoting factor","authors":"Yafei Jiao ,&nbsp;Alian Liao ,&nbsp;Xintong Jiang,&nbsp;Jinming Guo,&nbsp;Bingqian Mi,&nbsp;Chang Bei,&nbsp;Xinran Li,&nbsp;Tiantuan Jiang,&nbsp;Xiaohong Liu,&nbsp;Yaosheng Chen,&nbsp;Peiqing Cong,&nbsp;Zuyong He","doi":"10.1016/j.theriogenology.2025.02.004","DOIUrl":"10.1016/j.theriogenology.2025.02.004","url":null,"abstract":"<div><div>Growth differentiation factor 9 (GDF9), an oocyte-secreted factor, plays a vital role in porcine oocyte development. However, its function during oocyte <em>in vitro</em> maturation (IVM) remains unclear. In this study, we achieved GDF9 editing in approximately 59 % of cultured oocytes by cytoplasmic injection of a pre-assembled crRNA-tracrRNA-Cas9 ribonucleoprotein complex into porcine oocytes at the germinal vesicle (GV) stage. GDF9 editing caused significant damage to porcine oocytes during IVM. Additionally, GDF9 editing impaired mitochondrial function, increased reactive oxygen species (ROS) accumulation, and decreased glutathione (GSH) levels. The impaired IVM of GDF9-edited porcine oocytes was primarily driven by active cAMP-PKA signaling, which inhibited MOS expression, leading to the activation of the WEE1B/MYT1 kinase and inactivation of CDC25B phosphatase. This cascade resulted in the inactivation of CDK1, thereby preventing the activation of maturation-promoting factor (MPF) and inhibiting first polar body (PB1) extrusion. Our findings enhance the understanding of GDF9's regulatory role in porcine oocyte IVM and provide a theoretical foundation for improving porcine reproductive performance.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"236 ","pages":"Pages 120-136"},"PeriodicalIF":2.4,"publicationDate":"2025-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143379376","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}