TheriogenologyPub Date : 2024-10-26DOI: 10.1016/j.theriogenology.2024.10.025
{"title":"Dexamethasone and azithromycin enhance goat sperm preservation quality by regulating lipid metabolism","authors":"","doi":"10.1016/j.theriogenology.2024.10.025","DOIUrl":"10.1016/j.theriogenology.2024.10.025","url":null,"abstract":"<div><div>Phospholipase A (PLA) in goat semen aggregates with egg yolk in semen diluent, leading to sperm death. The aim of this study is to address the issue of sperm death caused by the interaction between PLA and egg yolk, and to explore the protective effect and metabolic regulation mechanism of the combination of dexamethasone (DXMS) and azithromycin (AZM) on goat sperm under low temperature conditions. At a low temperature of 4 °C, different concentrations of DXMS were added to semen diluents containing 30 μg/mL AZM to detect the quality of goat sperm. The optimal concentration of DXMS was determined to be 20 μg/mL. On the 5th day of storage, antioxidant capacity, total cholesterol (TC) levels, energy metabolism, and metabolomics analysis were performed on the sperm of the 20 μg/mL DXMS group. The results showed that there was no aggregation caused by the interaction between PLA and egg yolk in the group containing 30 μg/mL AZM at 4 °C. 20 μg/mL DXMS significantly improved sperm motility, plasma membrane integrity, acrosome integrity, glutathione peroxidase (GPX) (P < 0.05), catalase (CAT) (P < 0.01), and superoxide dismutase (SOD) activity (P < 0.01). The content of reactive oxygen species (ROS) and Fe<sup>2+</sup> significantly decreased (P < 0.01), while the content of ATP (P < 0.01) and TC (P < 0.05) significantly increased. Through metabolomics analysis, a total of 56 differential metabolites (P < 0.05) were screened, including 5a, 6-Anhydrotetracycline, Betamethasone, and 11-Dehydrocorticosterone, mainly enriched in 8 metabolic pathways (P < 0.05), including steroid hormone biosynthesis, glycerophospholipid metabolism, and choline metabolism in cancer. Among them, 5 metabolic pathways are related to lipid metabolism. The results indicate that AZM effectively inhibits the aggregation of PLA and yolk, and the combination of AZM and DXMS enhances the preservation quality of goat sperm during low-temperature preservation by regulating lipid metabolism.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142547635","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":"NAGK regulates the onset of puberty in female mice.","authors":"Wei Zhang, Ping Qin, Mengxian Li, Zhihao Pan, Zhuoya Wu, Yanyun Zhu, Ya Liu, Yunsheng Li, Fugui Fang","doi":"10.1016/j.theriogenology.2024.10.023","DOIUrl":"https://doi.org/10.1016/j.theriogenology.2024.10.023","url":null,"abstract":"<p><p>This study examines the role of N-acetylglucosamine kinase (NAGK) in initiating puberty in female mice. We employed real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and immunofluorescence to measure NAGK expression in the hypothalamic-pituitary-ovarian axis across various developmental stages: infant, prepuberty, puberty, and adult. We further investigated the impact of Nagk gene knockdown on puberty in female mice. This included assessing the expression of puberty-related genes both in vivo and in vitro, GT1-7 cells proliferation and apoptosis, concentrations of GnRH and Kisspeptin, puberty onset timing, serum levels of progesterone (P<sub>4</sub>) and estradiol (E<sub>2</sub>), and ovarian morphology. Results revealed that Nagk mRNA is present in the hypothalamus, pituitary, and ovaries throughout different developmental stages in female mice. In the hypothalamus, Nagk mRNA levels were comparable during infant and prepuberty, lowest during puberty, and highest in adult. In the pituitary, Nagk mRNA peaked in adult, with no significant variation between infant, prepuberty, and puberty. In the ovaries, Nagk mRNA levels increased during puberty and peaked in adult. NAGK is predominantly located in the arcuate nucleus (ARC), periventricular nucleus (PeN), dorsomedial hypothalamic nucleus (DMH), paraventricular nucleus (PVN), adenohypophysis, and in the ovarian oocytes, interstitium, and granulosa cells across all developmental stages in female mice. Nagk knockdown in GT1-7 cells decreased the transcriptional level of Gnrh, Kiss1, Gpr54, Igf1 and Mapk14 mRNA and cell proliferation but increased the level of β-catenin mRNA and cell apoptosis, while reducing GnRH secretion. Following ICV injection, Nagk gene knockdown mice exhibited delayed the timing of vaginal opening (VO) and reduced hypothalamic levels of Gnrh, Kiss1, Gpr54, Igf1, Mapk14, and β-catenin mRNA. Additionally, serum concentrations of E<sub>2</sub> in Nagk gene knockdown mice were significantly lower compared to the control group. These findings indicate that Nagk regulates the expression of Gnrh and Kiss1 mRNA in GT1-7 cells, affects hypothalamus Gnrh mRNA levels and serum E<sub>2</sub> concentration, and that its knockdown can delay puberty onset in female mice.</p>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142565188","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-10-24DOI: 10.1016/j.theriogenology.2024.10.024
A G Curcio, T I S Ribeiro, H F Gomes, C S Paes de Carvalho, M C C Bussiere, A J B Dias
{"title":"Increased in vitro production of bovine embryos resulting from oocyte maturation in the presence of triciribine, a specific inhibitor of AKT.","authors":"A G Curcio, T I S Ribeiro, H F Gomes, C S Paes de Carvalho, M C C Bussiere, A J B Dias","doi":"10.1016/j.theriogenology.2024.10.024","DOIUrl":"https://doi.org/10.1016/j.theriogenology.2024.10.024","url":null,"abstract":"<p><p>The aim of this study was to evaluate the effect of different concentrations of triciribine, a selective Akt inhibitor, on various aspects of oocyte maturation and on the IVF of bovine embryos. Cumulus-oocyte complexes (COCs) were matured in vitro in medium supplemented with: 0 (control), 1, 5, 10, and 20 μM of triciribine. The nuclear maturation was assessed by staining with acetic orcein, while the cytoplasmic maturation was evaluated by mitochondrial (MitoTracker® Red CMXRos) and lipid droplets distribution (LipidTOX). COCs were fertilized in vitro and cultured for nine days. Cleavage rates, blastocyst production, and hatching rates were determined on days three, seven, and nine of in vitro culture, respectively. Oocytes from COCs treated with 1 μM of triciribine were stained at 3, 6, and 9 h of IVM to determine the inhibitor's involvement in germinal vesicle breakdown. Analysis of variance (ANOVA) of the data was performed and the means were compared using the SNK test at a 5 % significance level. Exposure of COCs to 1, 5, and 10 μM of triciribine did not alter the number of matured oocytes (P < 0.05), a concentration of 20 μM reduced the number of oocytes in MII with a consequent increase in oocytes in MI (P < 0.05). This concentration markedly reduced the number of oocytes with peripheral cortical granules and the rates of cleavage and blastocysts (P < 0.05). On the other hand, when COCs were matured in the presence of 1 μM, there was an increase in the blastocyst rate (P < 0.05), but without altering the timing of meiosis resumption (P < 0.05). It is concluded that the Akt pathway participates in the nuclear and cytoplasmic events of in vitro maturation of bovine oocytes, but through mechanisms that do not interfere with germinal vesicle breakdown. Modulation of Akt activity in bovine COCs during IVM with 1 μM of triciribine increases the in vitro production of bovine embryos.</p>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142565169","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-10-23DOI: 10.1016/j.theriogenology.2024.10.019
{"title":"Taste receptor T1R3 regulates testosterone synthesis via the cAMP-PKA-SP1 pathway in testicular Leydig cells","authors":"","doi":"10.1016/j.theriogenology.2024.10.019","DOIUrl":"10.1016/j.theriogenology.2024.10.019","url":null,"abstract":"<div><div>Taste receptor type 1 subunit 3 (T1R3) is a G protein-coupled receptor encoded by the <em>TAS1R3</em> gene that can be specifically activated by certain sweeteners or umami agents for sweet/umami recognition. T1R3 is a potential target for regulating male reproduction. However, studies on the impact of non-nutritive sweeteners on reproduction are limited. In the present study, we evaluated the impact of the non-nutritive sweeteners (saccharin sodium, sucralose and acesulfame-K) on testosterone synthesis in testicular Leydig cells of Xiang pigs by comparing the relative abundance of mRNA transcripts and protein expression of T1R3, steroidogenic related factors, and intracellular cyclic adenosine monophosphate (cAMP), protein kinase A (PKA), as well as testosterone levels using Western blotting, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA). To clarify the specific mechanism, a dual luciferase assay was used to uncover the relationship between the transcription factors and steroidogenic enzyme. The acute intratesticular injection of a typical non-nutritive sweeteners was conducted to verify this impact in mouse. The results showed that saccharin sodium not only enhanced T1R3 expression in Leydig cells of Xiang pigs, but also caused significant increases in testosterone, cAMP, PKA, phosphorylation of specificity protein 1 (p-SP1), total protein of specificity protein 1 (SP1), steroidogenic acute regulatory protein (StAR), and 3β-hydroxysteroid dehydrogenase type 1 (3β-HSD1) (<em>P</em> < 0.05). Similarly, treatment of Leydig cells with sucralose and acesulfame-K also increased testosterone level, protein expression of T1R3, 17-α-hydroxylase/17, 20-lyase (CYP17A1), and 3β-HSD1 (<em>P</em> < 0.05). Treatment with SQ22536 (an adenylate cyclas inhibitor) or H89 (a PKA inhibitor) significantly reduced saccharin sodium-induced protein levels of p-SP1, StAR, CYP17A1, and 3β-HSD1 (<em>P</em> < 0.05). In addition, a dual luciferase assay further demonstrated that <em>SP1</em> significantly increased the promoter activity of <em>CYP17A1</em> (<em>P</em> < 0.05). When mouse testes were injected with saccharin sodium, T1R3, p-SP1, CYP17A1, and 3β-HSD1 were upregulated, leading to a significant testicular increase in testosterone and cAMP levels (<em>P</em> < 0.05). These results suggest a mechanism by which the taste receptor T1R3 regulates testosterone production, and this mechanism may be linked to the cAMP-PKA pathway. Understanding the interrelationship between T1R3 and the cAMP-PKA-SP1 pathway contributes to clarify the regulatory mechanisms of male reproduction.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142547636","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-10-22DOI: 10.1016/j.theriogenology.2024.10.020
{"title":"Detecting Leptospira spp. infection in cows by PCR: What is the best sample to test?","authors":"","doi":"10.1016/j.theriogenology.2024.10.020","DOIUrl":"10.1016/j.theriogenology.2024.10.020","url":null,"abstract":"<div><div>Bovine leptospirosis is a major reproductive disease. As cows can be leptospiral carriers both on the renal and genital tract, diagnosis can be challenging, with an underlying risk of misdiagnosis. Traditionally, the infection has been diagnosed by culturing or PCR from urine samples. Nevertheless, recent studies have suggested testing genital samples rather than urine, particularly for the diagnosis of genital colonization and reproductive disorders. The present study aimed to compare urine versus genital samples to detect leptospiral carriers in naturally infected cows with poor reproductive performance under field conditions. Five herds presenting >20 % of seroreactive animals against the Sejroe serogroup were selected. Of these, 106 cows with poor reproductive performance were studied, and urine, uterine fragment (UF), and cervicovaginal mucus (CVM) were obtained and tested by <em>lip</em>L32-PCR. A total of 73 (68.9 %) cows were infected; 64 of which (87.7 %) were diagnosed via positive genital samples (UF and/or CVM), while only 14 (19.2 %) by urine (<em>p</em> ≤ 0.001). Therefore, if the study had been limited to urine samples, as largely recommended, less than 20 % of the infected cows would have been detected, representing a huge misdiagnosis of the disease that could undermine the efficacy of control programs. In this context, the present study reinforces prior findings that testing genital samples, particularly CVM, is crucial to effectively diagnosing infected subfertile cows.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142508601","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-10-22DOI: 10.1016/j.theriogenology.2024.10.021
{"title":"Duration of efficacy and effect of implant location in adult queens treated with a 9.4 mg deslorelin subcutaneous implant","authors":"","doi":"10.1016/j.theriogenology.2024.10.021","DOIUrl":"10.1016/j.theriogenology.2024.10.021","url":null,"abstract":"<div><div>Deslorelin is a GnRH-agonist used off-label for contraception in female cats. Little is known about duration and safety of the 9.4 mg subcutaneous implant in the queen as well as its efficacy when placed periumbilically. Fourteen female cats were administered the 9.4 mg deslorelin implant (during interestrus or anestrus) either in interscapular (N = 8) or periumbilical (N = 6) sites, following general and reproductive examination, vaginal cytology, hematology, biochemistry and progesterone assay to ensure health status. All above procedures (except for progesterone assay) were repeated weekly during the first month, then every 2 months until 6 months and then every 6 months until treated cats regained full reproductive function. No side effects were observed in any treated queen. Post implantation estrus occurred in 40 % of the subjects. A significant increase in body weight was observed during treatment (12/14 queens gained weight), particularly at the end of the study. At the end of the study some queens mated, conceived and kittened, proving reversibility of the treatment. The average duration of action of the 9.4 mg deslorelin implant was 790 ± 155 days (range 525–1140 days) with no significant difference in duration or efficacy depending on implantation sites. The 9.4 mg deslorelin implant causes pharmacological sterilization for about 2 years in female cats, is fully reversible and caused no clinically relevant side effects when administered at both interscapular and periumbilical sites.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142523205","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}
TheriogenologyPub Date : 2024-10-22DOI: 10.1016/j.theriogenology.2024.10.018
{"title":"Periovulatory anticoagulant therapy enhances embryo recovery rates in superovulated mares","authors":"","doi":"10.1016/j.theriogenology.2024.10.018","DOIUrl":"10.1016/j.theriogenology.2024.10.018","url":null,"abstract":"<div><div>Although protocols for superovulation have been described in horses, this technique has been discouraged due to the low embryo recovery rates in superovulated mares. The reason for these poor results is poorly understood, but the formation of a blood clot in the ovulation fossa following ovulations has been hypothesized. Therefore, this study aimed to assess the safety and effect of periovulatory anticoagulant therapy on embryo recovery of superovulated mares. In experiment 1, five mares were assigned to receive five anticoagulant treatments in a crossover design: intravenous injections of 150 (H1), 300 (H2), 400 (H3), 450 (H4), 600 (H5) IU/kg of unfractionated heparin (UFH, heparin sodium); and had blood samples sequentially collected for up to 48 h post-treatment to test Prothrombin (PT) and activated partial thromboplastin time (aPTT). In experiment 2, four mares were treated in a crossover design with intravenous injection of 450 IU/kg of UFH and 1 mg/kg of low molecular weight heparin (LMWH, enoxaparin) and had blood collected as previously for analysis of plasma anti-Xa activity. In experiment 3, eleven mares had four cycles randomly assigned to four groups. In the control group, mares did not receive any treatment. In contrast, in groups G1, G2, and G3, mares were superovulated with equine pituitary extract and treated 34 h after the induction of ovulation with a placebo (NaCl 0.9 %, G1), 450 IU/kg of UFH (G2), or 1 mg/kg of LMWH. Mares in all groups had ovulation induced with hCG plus histrelin acetate and were bred with fresh semen from one stallion. Embryo flushing was performed nine days post-ovulation. In experiment 1, only mares in groups H4 and H5 had increased aPTT and PT for up to 12 h, and in all groups, aPTT and PT values returned to baselines at 24 h post-treatment. In experiment 2, plasma anti-Xa activity was increased by both therapies for up to 12 h after treatment and was at baseline levels 24 h post-treatment. In experiment 3, periovulatory therapy with anticoagulants increased embryo recovery rates per cycle (G2, 250 %; G3, 260 %) compared to control-assigned cycles (60 %; P < 0.05), whereas G1-assigned cycles (160 %) had intermediate embryo recovery. In conclusion, periovulatory anticoagulant therapies may be an alternative to improve embryo recovery in superovulated mares.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142530197","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-10-21DOI: 10.1016/j.theriogenology.2024.10.017
{"title":"Putrescine supplementation improves the developmental competence of in vitro produced bovine embryos","authors":"","doi":"10.1016/j.theriogenology.2024.10.017","DOIUrl":"10.1016/j.theriogenology.2024.10.017","url":null,"abstract":"<div><div>The aim of this study was to investigate the effect of putrescine, anti-apoptotic, antioxidant, and a cell proliferation stimulant, on embryo development and quality by supplementing it to in vitro culture medium. In this study, oocytes were obtained from the ovaries of Holstein cattle. Following maturation and fertilization, the presumptive zygotes were randomly assigned to two groups. The first group (Putrescine, n = 435) was supplemented with putrescine at a concentration of 0.5 mM to in vitro culture. The second group (n = 407) was maintained under standard culture conditions without any supplementations to the medium. Following the determination of the developmental stages of the embryos, only those in the blastocyst stage were subjected to differential staining and the cell numbers of the embryos were determined. Moreover, the TUNEL assay was employed to ascertain the extent of cell death and the apoptotic index in the embryos. Additionally, the levels of ROS were determined in the embryos. Furthermore, gene expression analyses were conducted on blastocyst-stage embryos to ascertain the potential of putrescine supplementation in embryo development along specific pathways. Following in vitro culture, the blastocyst formation rate was 44.37 % in the putrescine group and 32.97 % in the control group (P < 0.05). The counts of ICM (60.60 ± 15.79 vs 50.73 ± 16.74), TE (117.70 ± 23.67 vs 94.0 ± 22.46), and TCC (178.30 ± 26.15 vs 144.73 ± 26.86) were found to be statistically higher in blastocysts developing after putrescine supplementation compared to the control group. Furthermore, the number of apoptotic cells (7.69 ± 2.17 vs 9.96 ± 3.99) and the apoptotic index (5.07 % vs 8.01 %) were found to be lower in the putrescine group in comparison to the control group. Nevertheless, it was established that the ROS level in the control group was approximately two-fold higher than in the putrescine group (P < 0.05). The findings also revealed that putrescine up-regulated the gene expression of SOD, GPX4, CAT, BCL2, NANOG and GATA3 while simultaneously down-regulating the BAX expression level. In conclusion, the supplementation of putrescine to the culture medium during in vitro bovine embryo production was found to contribute to the improvement of embryo quality and early embryonic development.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142508618","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-10-21DOI: 10.1016/j.theriogenology.2024.10.016
{"title":"Rat copper transport protein 2 (CTR2) is involved in fertilization through interaction with IZUMO1 and JUNO","authors":"","doi":"10.1016/j.theriogenology.2024.10.016","DOIUrl":"10.1016/j.theriogenology.2024.10.016","url":null,"abstract":"<div><div>In mammalian reproduction, testis-specific protein IZUMO1 and its receptor JUNO on the oocyte surface are essential for sperm-oocyte recognition, binding, and membrane fusion. However, these factors alone are insufficient to accomplish cytoplasmic membrane fusion. It is believed that other gametic proteins interact with them to facilitate sperm-oocyte interaction on the head and mid-tail of rat spermatozoa as well as on the surface of oocytes. In this study, Copper Transport Protein 2 (CTR2) has been identified on the head and mid-tail of rat spermatozoa as well as on the surface of oocytes. CTR2 directly interacts with both IZUMO1 and JUNO, colocalizing with IZUMO1 on the sperm head and with JUNO on the oocyte membrane. Treatment of the capacitated sperm and zona pellucida-free oocytes with <em>anti</em>-CTR2 antibody resulted in a significant decrease in fertilization rates in IVF experiments. These findings suggest that CTR2 plays an important role in mammalian fertilization by interacting with IZUMO1 and JUNO, providing new insights into the molecular mechanisms of mammalian sperm-oocyte adhesion and fusion.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142508619","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-10-18DOI: 10.1016/j.theriogenology.2024.10.014
{"title":"Molecular characterization of MSX2 gene and its role in regulating steroidogenesis in yak (Bos grunniens) cumulus granulosa cells","authors":"","doi":"10.1016/j.theriogenology.2024.10.014","DOIUrl":"10.1016/j.theriogenology.2024.10.014","url":null,"abstract":"<div><div>Cumulus granulosa cells (CGCs) are somatic cells surrounding the oocyte that play an important role in oocyte growth, meiotic maturation, ovulation, and fertilization in mammals. Therefore, revealing the molecular mechanisms related to the development and function of CGCs is essential for further understanding the regulatory network in female reproduction. <em>MSX2</em> belongs to the highly conserved <em>msh</em> homeobox gene family and plays diverse roles in different biological processes. This study cloned the coding sequence (CDS) of the yak <em>MSX2</em> gene and detected the abundance and localization of <em>MSX2</em> in the major female reproductive organs. The results indicated that the CDS of this gene included 747 base pairs and encoded 248 amino acids. The abundance of <em>MSX2</em> mRNA was highly expressed in the luteal phase of the yak ovary during the estrous cycle, and MSX2 protein was widely expressed in different female reproductive organs, including the ovary, corpus luteum, uterus, and oviduct. Repressing <em>MSX2</em> abundance in yak CGCs declined the cell viability and defective steroidogenesis. Several genes abundances related to cell proliferation, apoptosis, and sterogenesis also changed after <em>MSX2</em> knockdown. <em>MSX2</em> overexpression had the opposite effect on cell viability in yak CGCs. These results reveal the specific mechanism by which <em>MSX2</em> regulates the development and function of yak CGCs and give novel and valuable insights into the mechanisms involved in yak reproduction.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142475497","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}