Theriogenology最新文献

{"title":"Chromosome remodeling and cytoplasmic distribution during embryonic development in fused pair embryos","authors":"Aochen Yu,&nbsp;Yang Xing,&nbsp;Fuyao Zhang,&nbsp;Deru Cao,&nbsp;Yuan Jiang,&nbsp;Xinyue Lu,&nbsp;Juan Li","doi":"10.1016/j.theriogenology.2025.117465","DOIUrl":"10.1016/j.theriogenology.2025.117465","url":null,"abstract":"<div><div>Cell fusion is now widely employed as an <em>in vitro</em> model for inducing and investigating cell fate changes. In early embryo research, fusion of embryos is also a way to probe the mechanisms of mammalian oogenesis and preimplantation development. To establish a novel model for porcine early embryo studies and investigate its developmental mechanisms, pair of zona pellucida (ZP)-free oocytes were electro fused to produce the fused pair (FP) embryos, which were further <em>in vitro</em> cultured to the blastocyst stage. Firstly, developmental competence was assessed, revealing a cleavage rate of 92.27 ± 5.59 % and a blastocyst rate of 26.12 ± 6.61 %, which were similar to the parthenogenetic activation (PA) embryos (<em>p</em> &gt; 0.05). Subsequently, nuclear and spindle staining was performed on FP embryos collected at 14–22 h, with 67.18 ± 3.18 % of the embryos spindle reorganization occurred and nuclei fusion, whereas a few displayed independent division of the two nuclei or tripolar spindle. Lastly, the distribution of lipid droplets (LDs) and mitochondria in FP embryos was assessed via fluorescent staining. Results showed that the even distribution of LDs from one oocyte was observed in each blastomere of 4-cell to blastocysts. A similar distribution pattern was observed for mitochondria, which was being observed at the 2-cell stage, a relatively earlier developmental stage than that of LDs. Results suggested that cytoplasm including mitochondria and LDs could redistribute once two oocytes fused into a single embryo. More studies are needed for the underlying mechanism and potential impact on the developmental ability of FP embryos.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117465"},"PeriodicalIF":2.4,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143899550","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":"Colony-stimulating factor 2 (CSF2) does not significantly affect cellular and molecular parameters, or blastocyst rates under in vitro culture conditions with reduced nutrient concentrations","authors":"Ana Caroline Chaves Vall Nicolás ,&nbsp;Ligiane de Oliveira Leme ,&nbsp;Amanda Oliveira Moura ,&nbsp;Nayara Ribeiro Kussano ,&nbsp;Emivaldo de Siqueira ,&nbsp;Maurício Machaim Franco ,&nbsp;Margot Alves Nunes Dode","doi":"10.1016/j.theriogenology.2025.117464","DOIUrl":"10.1016/j.theriogenology.2025.117464","url":null,"abstract":"<div><div>Colony-stimulating factor 2 (CSF2) is an embryokine used between days 5 and 7 of culture to improve embryonic development. We evaluated its use from Day 4 and throughout the culture period in a reduced nutrient concentrations culture medium. Presumptive zygotes were assigned to three treatments: Control – synthetic oviduct fluid (SOF), CSF2 D1 (SOF + CSF2 from Day 1–7), CSF2 D4 (SOF + CSF2 from Day 4–7). Cleavage rates on Day 4 and blastocyst rates on days 6 and 7 were evaluated. On Day 7, expanded blastocysts were used to assess mitochondrial activity, lipid quantification, total cell number, gene expression and DNA methylation patterns. Embryo development data were analyzed using the chi-square test (P ≤ 0.05). Data normally distributed were analyzed with ANOVA and Tukey's test, while non-normally distributed data were evaluated using the Kruskal-Wallis and Mann-Whitney tests. Cleavage and blastocyst rates on Day 7 were similar (P &gt; 0.05) among treatments. Of 11 genes evaluated, only Placenta-specific 8 presented a lower number of transcripts (P ≤ 0.05) in embryos cultured with CSF2 from Day 4. Assessments of DNA methylation, lipid droplet staining, mitochondrial activity and total cell number showed no significant differences among treatments (P &gt; 0.05). In conclusion, CSF2 had no effect on the assessed parameters when a culture medium with reduced nutrient concentrations was used, and can be used throughout the entire culture period without affecting embryo production and developmental kinetics compared to CSF2 added on Day 4.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117464"},"PeriodicalIF":2.4,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143907835","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":"Biostimulation with rams improved embryo quality without modifying the ovulatory response in superovulated ewes","authors":"Ana Paula Pereira Schmidt ,&nbsp;Felipe Zandonadi Brandão ,&nbsp;Juliana Dantas Rodrigues Santos ,&nbsp;Camila Correa Roza Laeber ,&nbsp;Sandra Ribeiro da Silva ,&nbsp;Nathália Dutra Knust ,&nbsp;Ribrio Ivan Tavares Pereira Batista ,&nbsp;Maria Emília Franco Oliveira ,&nbsp;Joanna Maria G. Souza-Fabjan ,&nbsp;Mário Felipe Alvarez Balaro ,&nbsp;Jeferson Ferreira da Fonseca ,&nbsp;Rodolfo Ungerfeld","doi":"10.1016/j.theriogenology.2025.117463","DOIUrl":"10.1016/j.theriogenology.2025.117463","url":null,"abstract":"<div><div>The aim of this study was to evaluate whether extended ram exposure during FSH superovulatory treatment improves follicular growth, embryo production, and embryo quality in ewes. Twenty Santa Inês ewes were subjected to a superovulation protocol based on follicle-stimulating hormone (FSH) administration with or without male biostimulation in a crossover arrangement (n = 20 per treatment). Follicular development and corpus luteum functionality were monitored with ultrasound, and embryo collection was performed non-surgically. Recovered embryos were analyzed for gene expression, reactive oxygen species (ROS) levels, and blastomere count. Male biostimulation for 72 h did not modify the number or diameter of antral follicles, the recovery rate of structures, reactive oxygen species (ROS) levels, and blastomere count. However, it reduced the expression of genes linked to oxidative stress (PRDX1) and apoptosis (BCL2) in embryos. In summary, exposure to rams for 72 h during superovulatory treatment influenced molecular indicators of development competence, specifically the relative abundance of BCL2 and PRDX1 transcripts, which are associated with cellular apoptosis and stress. However, it did not improve the ovarian responses.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117463"},"PeriodicalIF":2.4,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143894334","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":"Correlation between PRDX6 levels and freezability properties of sheep semen","authors":"Erhan Hai ,&nbsp;Xue Bai ,&nbsp;Yukun Song ,&nbsp;Boyuan Li ,&nbsp;Jian Zhang ,&nbsp;Jiaxin Zhang","doi":"10.1016/j.theriogenology.2025.117461","DOIUrl":"10.1016/j.theriogenology.2025.117461","url":null,"abstract":"<div><div>Individual variations in the freezability properties of sheep semen may be related to the antioxidant levels of sperm. PRDX6 is a key antioxidant protein in sperm, but its relationship with the freezability properties of sheep semen remains unclear.</div><div>This study, using Hu sheep as the research subject, investigated the levels of PRDX6 in sperm with different freezability properties. A positive correlation was found between the levels of PRDX6 and sperm freezability. Furthermore, when different concentrations of PRDX6 recombinant protein were added to high-freezing-resistant (HF) and low-freezing-resistant (LF) sperm, it was revealed that 20 μg/ml of PRDX6 recombinant protein significantly improved various sperm quality parameters in both HF and LF sperm and reduced the differences in sperm freezability properties. Finally, adding a PRDX6 peroxidase activity inhibitor (Ethacrynic acid) or a calcium - independent phospholipase A2 (iPLA2) activity inhibitor (MJ33) to the diluent significantly reduced the freezability parameters of sperm. This indicates that PRDX6 protects sperm from freezing damage through two pathways. In summary, this study reveals the crucial role of PRDX6 in the freezability properties of sheep semen. It suggests that increasing the levels of PRDX6 in sperm may be an effective strategy to improve the quality of cryopreserved semen. Future research can further explore the molecular mechanisms regulating the levels of PRDX6, providing a scientific basis for the optimization of sheep semen cryopreservation techniques.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117461"},"PeriodicalIF":2.4,"publicationDate":"2025-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143894335","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":"Improvement of cryopreserved epididymal ram sperm quality and fertility through curcumin nanoparticles","authors":"Ramin Farhadi ,&nbsp;Abbas Farshad ,&nbsp;Abouzar Najafi ,&nbsp;Jalal Rostamzadeh","doi":"10.1016/j.theriogenology.2025.117462","DOIUrl":"10.1016/j.theriogenology.2025.117462","url":null,"abstract":"<div><div>Artificial insemination (AI) in rams relies on effective sperm preservation; however, freezing and thawing processes induce oxidative stress and sperm damage. This study evaluated the antioxidant effects of curcumin, in both free and nanoparticles (NPs) forms, on cryopreserved ram sperm. Curcumin was incorporated into the sperm extender at concentrations of 0, 25, and 50 μM. The curcumin NPs formulation, prepared using the nanoliposome technique, improved curcumin solubility and bioavailability. The 25 μM curcumin NPs group showed significantly higher sperm motility (total motility: 72.67 ± 1.15 %, progressive motility: 59.2 ± 0.96 %) compared to the control (total motility: 59.75 ± 0.68 %, progressive motility: 46.23 ± 1.50 %) (p &lt; 0.05). Membrane (75.77 ± 0.87 %) and acrosome integrity (80.67 ± 0.87 %) were significantly improved in the 25 μM NPs group compared to those in the control (61.76 ± 0.97 %, 66.77 ± 1.50 %) (p &lt; 0.05). Sperm viability was higher in the 25 μM NPs group (76.76 ± 1.22 %) vs. control (64.93 ± 0.80 %) (p &lt; 0.05), and early apoptosis was reduced (5.06 ± 0.36 % vs. 9.7 ± 0.7 %) (p &lt; 0.05). Oxidative stress markers were also improved, with lower MDA and ROS levels and enhanced antioxidant enzyme activity. The 25 μM curcumin NPs did not alter CYTb copy number. The 25 μM curcumin NPs group had higher fertility outcomes, including pregnancy (58.33 %), birth (58.33 %), and lambing (83.33 %) rates, than the control and 50 μM curcumin groups. These findings indicate that curcumin nanoparticles enhance sperm quality and fertility in cryopreserved ram sperm.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117462"},"PeriodicalIF":2.4,"publicationDate":"2025-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143899632","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":"Histological alterations in placentas of normal and anomalous pups: correlation with neonatal outcomes and fetal health","authors":"Laura M. Lovo ,&nbsp;Marina Vilela Estevam ,&nbsp;Samara Beretta ,&nbsp;Amanda Marmol ,&nbsp;Loren M. Pasqui ,&nbsp;Fernanda A. Rodrigues-Silva ,&nbsp;Renata O. Barreto ,&nbsp;Alessandre Hataka ,&nbsp;Gilson Hélio Toniollo ,&nbsp;Maria Lúcia G. Lourenço ,&nbsp;Maricy Apparício","doi":"10.1016/j.theriogenology.2025.117458","DOIUrl":"10.1016/j.theriogenology.2025.117458","url":null,"abstract":"<div><div>Given that histological evaluation is a key method for identifying placental alterations potentially linked to congenital and neonatal abnormalities, this study evaluates histological alterations in 89 placentas from pups across three groups: Gelet (elective c-sections with healthy offspring), GDist (therapeutic c-sections with healthy offspring), and GAnom (stillborn offspring or those with fetal abnormalities, regardless of the type of c-section). The placentas were evaluated for the presence of congestion, hemorrhage, inflammatory infiltrate, necrosis, and calcification, and these alterations were graded on a scale: 0 – no alterations, 1 – up to 25 % of the region affected, 2 – up to 50 % affected, and 3 – up to 75 % affected. Histological analysis revealed varying degrees of alterations across groups, with the GAnom group exhibiting the most significant changes. Only 19 % of GAnom placentas were normal, while 81 % displayed notable alterations such as grade 1, 2 and 3 hemorrhage, frequently accompanied by necrosis and inflammatory infiltrates. Statistical analysis showed no significant difference in pup and placenta weights among groups (p &gt; 0.1), but Apgar scores were considerably lower in the GAnom group (p &lt; 0.001). A significant correlation was found between the type of placental alteration and pup health, with anomalous pups showing increased incidences of grade 3 hemorrhage (p &lt; 0.001) and grade 2 necrosis (29 % vs. 6 % in the normal group; p = 0.0002). The presence of necrosis negatively correlated with Apgar scores in normal pups (p = 0.02), indicating its potential impact on neonatal outcomes. These findings highlight the critical association between severe placental alterations and adverse fetal outcomes, emphasizing the need for further investigation into the mechanisms underlying these changes.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117458"},"PeriodicalIF":2.4,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143899631","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":"Reproductive outcomes of prepubertal Bos indicus beef heifers raised in a feedlot system submitted or not to ovulation induction strategies prior to a timed-artificial insemination treatment regimen","authors":"Rodrigo L.O.R. Alves ,&nbsp;Lucas O. e Silva ,&nbsp;Carlos E.C. Consentini ,&nbsp;Oseias P. Santos ,&nbsp;Andrés A.C. Narváez ,&nbsp;Leandro D. da Cunha ,&nbsp;Rafael A. Braz ,&nbsp;Maria E.R. e Silva ,&nbsp;Rodrigo Brüner ,&nbsp;Leonardo F. Melo ,&nbsp;Roberto Sartori","doi":"10.1016/j.theriogenology.2025.117457","DOIUrl":"10.1016/j.theriogenology.2025.117457","url":null,"abstract":"&lt;div&gt;&lt;div&gt;Reproductive outcomes were evaluated in prepubertal Nelore (&lt;em&gt;Bos indicus&lt;/em&gt;) heifers submitted or not to two ovulation induction treatment regimens based on progesterone (P4) and estradiol treatments prior to the timed-artificial insemination (TAI). A total of 583 heifers (11.3 ± 1.3 mo old and 272.2 ± 1.6 kg of body weight [BW]) were randomly assigned to one of two treatments: &lt;strong&gt;0IND&lt;/strong&gt; (n = 284): no treatment; or &lt;strong&gt;2IND&lt;/strong&gt; (n = 299): two ovulation induction treatment regimens. On Day −47, heifers from 2IND received an intravaginal P4 device (0.5 g), kept until Day −40, when there was treatment with 0.5 mg of estradiol cypionate (EC). On Day −19, in heifers from 2IND, the same treatment regimen was imposed. On Day 0, all heifers were submitted to the same TAI treatment regimen, starting with a P4 device (0.5 g), 0.53 mg of cloprostenol sodium (PGF&lt;sub&gt;2α&lt;/sub&gt;), and 1.5 mg of estradiol benzoate. On Day 7, the P4 device was removed, and 0.53 mg of PGF&lt;sub&gt;2α&lt;/sub&gt;, 0.5 mg of EC, and 200 IU of equine chorionic gonadotropin (eCG) were administered. The TAI was conducted 2 d later (Day 9, 46–50 h after P4 device withdrawal), concurrently with the administration of 8.4 μg of buserelin acetate. Ultrasound evaluations were performed on Days −47, −19 and 0 to evaluate uterine (UT) and largest follicle (LF) diameters and the presence of CL; on Day 40 to detect the presence of CL in nonpregnant heifers; and on Days 40, 70, and 150 (final pregnancy per AI [P/AI]) to assess P/AI and pregnancy losses (PL). Statistical analyses were conducted using SAS 9.4 (&lt;sup&gt;a-c&lt;/sup&gt;P ≤ 0.05; &lt;sup&gt;A,B&lt;/sup&gt;0.05 &lt; P ≤ 0.10). A greater proportion of 2IND-treated heifers had CL on Days −19 (73.9 and 18.3%) and 0 (80.3 and 32.4%) compared to heifers of the 0IND group. On Day 0, the LF diameter was larger in heifers of the 0IND group than 2IND-treated heifers (9.9 ± 0.1 and 8.7 ± 0.1 mm). However, the UT diameter was larger in 2IND-treated heifers on both Days −19 (14.2 ± 0.1 and 13.4 ± 0.1 mm) and 0 (14.0 ± 0.1 and 13.7 ± 0.1 mm). The 2IND-treated heifers had greater expression of behavioral estrus near TAI compared to heifers of the 0IND group (88.0 and 77.8%), although there was no difference in P/AI on Days 40 (39.5 and 41.6%), 70 (33.1 and 37.7%) and final P/AI (32.1 and 35.9%), nor on total PL (18.6 and 13.6%), considering 2IND-treated and heifers of the 0IND group, respectively. Moreover, among heifers without CL on Day 0, 2IND-treated heifers were less fertile than heifers of the 0IND group (final P/AI: 20.3 and 33.9%). Regardless of treatment, presence of CL on Day 0 and expression of behavioral estrus were positively associated with P/AI. Regardless of treatment, BW class (measured on Day 0) positively affected LF and UT diameters, and presence of CL on Days −19 and 0. Moreover, age class positively affected LF and UT diameters, and presence of CL on Days −47, −19, 0 and 40. Age class had no effect on fertility, but BW class","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117457"},"PeriodicalIF":2.4,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143895267","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":"Phoenixin-14 and its receptor GPR173 in the bovine endometrium: Expression and regulation during the estrous cycle and impact on secretory activity","authors":"Aleksandra Mówińska,&nbsp;Robert Rękawiecki,&nbsp;Paulina Zając,&nbsp;Magdalena K. Kowalik","doi":"10.1016/j.theriogenology.2025.117456","DOIUrl":"10.1016/j.theriogenology.2025.117456","url":null,"abstract":"<div><div>The endometrium remodels during the estrous cycle due to many endogenous factors. Phoenixin (PNX)-14 is a relatively recently discovered neuropeptide with a pivotal role in reproduction, specifically in follicle development and the functioning of the corpus luteum and uterus. Our study aimed to determine (1) the expression of PNX-14 and G protein-coupled receptor 173 (GPR173) in the bovine endometrium during the estrous cycle, (2) the cellular localization of PNX-14 and GPR173, (3) the effect of luteotropic and luteolytic factors on PNX-14 and GPR173 expression in the endometrium, and (4) the impact of PNX-14 on the secretory activity of the endometrium. The ex vivo experiments used bovine uterine tissues collected from the four stages of the estrous cycle (days 2–5, 6–10, 11–16, and 17–20). The in vitro experiments used uteri collected from days 10–12 of the estrous cycle. The mRNA expression of small integral membrane protein 20 (<em>SMIM20</em>) and protein expression of PNX-14 was upregulated on days 6–10 of the estrous cycle. GRP173 protein expression was significantly higher on days 11–16. The concentration of PNX-14 in endometrial tissue homogenate was significantly lower on days 17–20 compared to all other days of the estrous cycle. In immunofluorescence analysis, luminal and glandular epithelium and endothelial cells of blood vessels were positive for PNX-14 and GPR173. Progesterone upregulated the mRNA expression of <em>SMIM20</em> in the bovine endometrium, whereas progesterone and oxytocin upregulated <em>GPR1</em>73 mRNA and protein expression. Treating endometrial slices with PNX-14 enhanced prostaglandin E₂ secretion. In conclusion, these findings indicate that PNX-14 targets the bovine endometrium and should be considered a locally acting regulator of endometrium function in non-pregnant cows.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117456"},"PeriodicalIF":2.4,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143886926","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":"Specificity protein 1 (SP1) plays an essential role in early bovine embryo development","authors":"Anup K. Talukder ,&nbsp;Abdullah A. Naib ,&nbsp;Solomon Mamo ,&nbsp;Laura Thompson ,&nbsp;Juliana G. Ferst ,&nbsp;Maria B. Rabaglino ,&nbsp;John A. Browne ,&nbsp;Trudee Fair ,&nbsp;Pat Lonergan","doi":"10.1016/j.theriogenology.2025.117455","DOIUrl":"10.1016/j.theriogenology.2025.117455","url":null,"abstract":"<div><div>A previous RNA-Seq study revealed that the transcript abundance of specificity protein 1 (SP1) was significantly higher in Day 7 bovine blastocysts compared to conceptuses on Days 10, 13, 16, and 19, suggesting a stage-specific role in early bovine embryo development. The present study aimed to characterize the mRNA expression of SP1 and associated candidate genes (<em>ACSS1</em>, <em>C1QBP</em>, <em>ATF3</em>, <em>MAT2A</em>, and <em>POLD1</em>) during early bovine embryo development from the 2-cell to blastocyst stage. Further, the effects of SP1 inhibition on embryo development were evaluated by culturing embryos with the SP1 inhibitor, mithramycin A (MT) at varying concentrations (0, 25, 50, 100, and 1000 nM). As further validation, we examined expression of <em>SP1</em> and associated genes by interrogating transcriptomic data from Day 4 (16-cell stage) embryos cultured <em>in vitro</em> or <em>in vivo</em> in the oviducts of lactating or nonlactating dairy cows. The relative abundance of <em>SP1</em> peaked at the time of embryonic genome activation, being higher (P &lt; 0.05) in 8- and 16-cell embryos compared to the 2-cell stage, and decreasing thereafter (at the morula and blastocyst stages). Similarly, transcript abundance for most of the selected candidate genes involved in the SP1 network were upregulated (P &lt; 0.05) at the 8- and 16-cell stage, but not at other stages investigated. Inhibition of SP1 with MT did not affect embryo development up to the 8-cell stage but reduced (P &lt; 0.05) the proportion of embryos reaching the 16-cell and blastocyst stages in a dose-dependent manner. Moreover, blastocysts produced in the presence of MT contained fewer (P &lt; 0.05) cells than blastocysts developed without MT. Expression of <em>SP1</em> and associated genes in 16-cell stage (Day 4) embryos produced either <em>in vitro</em> or <em>in vivo</em> was higher (P &lt; 0.05) compared to <em>in vitro</em>-produced 2- to 4- cell stage (Day 2) embryos. These findings suggest an essential role of SP1 during early embryo development, particularly around the time of embryonic genomic activation.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"242 ","pages":"Article 117455"},"PeriodicalIF":2.4,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143868758","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":"Transcriptomic profiling of miRNA-mRNA interactions in canine oocytes and cumulus cells during in vitro maturation: a sequencing analysis","authors":"Phillip Dettleff ,&nbsp;Jaime Palomino ,&nbsp;Fabiola González-Coppia ,&nbsp;Monica De los Reyes","doi":"10.1016/j.theriogenology.2025.117454","DOIUrl":"10.1016/j.theriogenology.2025.117454","url":null,"abstract":"<div><div>Oocyte maturation is a critical process for successful fertilization and early embryonic development. In this study, we investigated the molecular mechanisms underlying oocyte maturation in dogs by analyzing the transcriptomic profiles of miRNAs and mRNAs in canine cumulus cells (CCs) and oocytes during <em>in vitro</em> maturation (IVM). RNA sequencing identified 285 miRNAs expressed in oocytes and 310 in CCs, with 282 miRNAs shared between the two cell types, highlighting the role of intercellular communication in maintaining miRNA expression equilibrium. Differential expression analysis revealed 222 mRNAs with significant differences between CCs and oocytes, including genes involved in transcriptional regulation and nuclear structure. Enrichment analyses identified pathways such as actin cytoskeleton regulation, mTOR signaling, cAMP signaling, and calcium signaling, all critical to oocyte maturation. Network analysis revealed 643 significant miRNA-mRNA coexpression relationships, suggesting miRNAs play pivotal roles in regulating mRNA expression during oocyte maturation. Notably, key miRNAs such as miR-30b, miR-375, and miR-503 were implicated in regulating genes involved in oocyte maturation pathways, while others like miR-378 and miR-21 aligned with known roles in suppressing cumulus expansion and influencing maturation. The absence of differential miRNA expression between CCs and oocytes suggests the miRNA transfer through gap junctions. These findings provide new insights into the transcriptional and post-transcriptional regulation of oocyte maturation in dogs, offering valuable knowledge to improve reproductive biotechnologies such as <em>in vitro</em> fertilization and embryo development in this species.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"242 ","pages":"Article 117454"},"PeriodicalIF":2.4,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143874859","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}