Theriogenology最新文献

{"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}

{"title":"Is platelet-rich plasma effective in treating uterine inflammation in jennies inseminated with cryopreserved donkey semen?","authors":"","doi":"10.1016/j.theriogenology.2024.10.009","DOIUrl":"10.1016/j.theriogenology.2024.10.009","url":null,"abstract":"<div><div>Despite frozen donkey semen demonstrating high quality after thawing and achieving suitable pregnancy rates in mares, it yields unsatisfactory results in jennies, likely due to a stronger uterine inflammatory response. This study assessed the effects of platelet-rich plasma (PRP) on uterine inflammation and pregnancy rates in jennies inseminated with frozen donkey semen. Estrous cycles from 11 jennies were assigned to three groups: Control (CTR, n = 22) with no treatment; Single PRP infusion (S-PRP, n = 22) administered 30 h after ovulation induction, prior to artificial insemination (AI); and Double PRP infusion (D-PRP, n = 21) with the first infusion at 30 h after ovulation induction and the second 4 h after AI. Insemination was performed with frozen donkey semen (1 billion sperm) deposited deeply in the uterine horn immediately after ovulation. Endometrial edema, intrauterine fluid (IUF), uterine vascularization, and endometrial cytology were evaluated pre-AI (TCt) and post-AI (6, 24, and 48 h). Uterine biopsies were taken at T48 for histopathological and collagen evaluation. Peripheral blood samples were collected on D5 for serum progesterone measurement, and pregnancy was evaluated via ultrasonography on D14. Data were analyzed using GLMMs, ANOVA, Friedman, and Kruskal-Wallis tests in SAS and GraphPad Prism, with significance set at p &lt; 0.05. The S-PRP group showed less IUF accumulation than the CTR group at T6. Other parameters showed no significant differences among the groups. Cytology revealed a high percentage of inflammatory cells at T6 in all groups, which decreased in subsequent evaluations. In the CTR group, neutrophil percentages were similar to TCt at T24, while treated groups reached this similarity only by T48. Eosinophil percentages increased over time only in the treated groups. Pregnancy rates showed no differences among the groups (CTR: 0 %, S-PRP: 0 %, D-PRP: 10 %). Results indicate that PRP treatments were ineffective in modulating uterine inflammation and did not enhance pregnancy rates in jennies inseminated with frozen donkey semen.</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":"142508616","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":"Identification of an osteopontin structural element for the restoration of a normal endometrial epidermal growth factor (EGF) profile determined by the EGF concentration on day 3 of estrous cycle and pregnancy outcome in repeat breeder dairy cows","authors":"","doi":"10.1016/j.theriogenology.2024.10.013","DOIUrl":"10.1016/j.theriogenology.2024.10.013","url":null,"abstract":"<div><div>The loss of a cyclic change with two peaks of increased endometrial epidermal growth factor (EGF) concentration on days 2–4 and 13−14 during the estrous cycle has been linked to low fertility in repeat breeder (RB) cows. We have shown that an intravaginal infusion of osteopontin (OPN) restored the EGF profile in RB cows. The present study aimed to determine a structural element of OPN to restore the normal EGF profile and fertility. Holstein RB cows were diagnosed the EGF profile by a single examination of the endometrial EGF concentration on day 3 of the estrous cycle. Those with an altered EGF profile were intravaginally infused with OPN and its fragments on the day of insemination (day 0); the concentration of endometrial EGF was measured on day 3, and pregnancy was diagnosed on days 30–35. In Study 1, recombinant OPN (rOPN) (16 nmol), thrombin-cleaved N- and C-terminal fragments of rOPN (N-rOPN and C-rOPN, respectively), and a combination of these fragments (Th-rOPN) were infused (n = 13–20). The restoration rate of the normal EGF profile of the N-rOPN group (25.0 %) was a level in between the C-rOPN group (7.7 %) and both the rOPN (55.6 %) and Th-rOPN (64.3 %) groups. In Study 2, PBS (n = 47), rOPN (9.5 nmol, n = 83), and peptides of integrin binding motifs, GRGDSVAYGLK (peptide 1; 32, 320, and 1600 nmol), GRGDS (peptide 2; 320 and 1600 nmol), and SVAYGLK (peptide 3; 320 and 1600 nmol), were infused (n = 20–25). Restoration rates of the normal EGF profile of peptide 1 (320 and 1600 nmol) and peptide 3 (1600 nmol) groups (44.0−56.3 %) were comparable with those of the rOPN group (63.9 %) and higher than those of the PBS group (15.6 %). Restoration rates of the other groups were similar to those of the PBS group. Additional cows received infusions to determine the effect on fertility. Conception rates of the peptide 1 (320 and 1600 nmol; n = 50 each), peptide 3 (1600 nmol; n = 55), and rOPN (n = 111) groups (41.8−50.0 %) were comparable and higher than that of the PBS group (21.6 %, n = 75). In Study 3, PBS (n = 24), peptide 1 (320 nmol; n = 78), and GRGESVAYGLK peptide (peptide 4; 320 and 1600 nmol; n = 50 and 26, respectively) were infused. Restoration rates of the normal EGF profile of peptide 4 and PBS groups (16.0−19.2 %) were comparable and lower than those of the peptide 1 group (44.9 %). Thus, the SVAYGLK motif may be an OPN structural element to restore the normal EGF profile and fertility in RB cows, and the RGD motif may enhance its effect.</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":"142508615","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":"Reduced NET1 adversely affects early embryonic development in mice","authors":"","doi":"10.1016/j.theriogenology.2024.10.012","DOIUrl":"10.1016/j.theriogenology.2024.10.012","url":null,"abstract":"<div><div>Neuroepithelial transforming gene 1 (NET1) is a RhoA subfamily-specific guanine/nucleotide-exchange factor that exhibits critical roles in diverse biological processes. However, the functions in mouse preimplantation embryonic development have not yet determined. In the present study we demonstrated that NET1 is a key factor in the outcome of early mouse embryonic development. Immunofluorescence detection showed that NET1 is principally localized to the nucleus during mouse pre-implantation embryonic development. Silencing <em>Net1</em> at the zygote stage using a specific siRNA impaired the developmental competence of early mouse embryos, and <em>Net1</em>-knockdown (<em>Net1</em>-KD) induced mitotic spindle-assembly defects and chromosomal alignment abnormalities at the first embryonic cleavage. In addition, reduced NET1 exacerbated reactive oxygen species production and DNA lesions in two-cell stage embryos, further augmenting cellular apoptosis in the preimplantation blastocyst. In summary, our data display key roles for NET1 in mitotic spindle assembly, oxidative stress, and DNA damage during early mouse embryonic development.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142475498","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":"Expression of TLR7/8 in canine sperm and evaluation of the effect of ligand R848 on the sorting of canine X/Y sperm","authors":"","doi":"10.1016/j.theriogenology.2024.10.015","DOIUrl":"10.1016/j.theriogenology.2024.10.015","url":null,"abstract":"<div><div>The aim of this study was to analyze the expression pattern of Toll receptor 7/8 (TLR7/8) in canine sperm, and explore the feasibility of using TLR7/8 ligand resiquimod(R848)to separate canine X and Y sperm. In this study, cellular immunofluorescence was used to analyze the expression of TLR7/8 in canine sperm, real-time fluorescence quantitative PCR was used to calculate the proportion of X sperm in the lower layer of the incubation solution with R848 to evaluate the sorting effect of R848 on canine X/Y sperm, and sperm quality detection system was used to analyze the effect of R848 on the motility of canine sperm. The mechanism of effect of R848 on canine sperm motility was analyzed by Western blot. The results showed that TLR8 was not expressed in all canine sperm, while TLR7 was expressed in all canine sperm and was localized in the head and tail of sperm. When 0.4 μM R848 was incubated with canine sperm for 1 h, the total motility, average path velocity (VAP), average straight-line velocity (VSL), and average curved-line velocity (VCL) of canine sperm were significantly decreased(<em>P</em> &lt; 0.05). There was no significant difference between the lower and upper layers of the R848 treatment group and the control group(<em>P</em> &gt; 0.05), and the proportion of X sperm was nearly half. The levels of NF-κB and GSK3α/β phosphorylation of sperm in R848 treatment group were significantly increased compared with control group(<em>P</em> &lt; 0.05). The above results showed that TLR7/8 was not differentially expressed in canine X and Y sperm. R848 could decrease the motility of canine spermatozoa and inhibit sperm motility by the GSK3α/β-hexokinase pathway through the phosphorylation of NFκB and GSK3α/β, while could not separate X and Y spermatozoa. The method of sorting X/Y sperm based on TLR7/8 is not feasible for dogs.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142508614","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":"Perspectives in milk production in India","authors":"","doi":"10.1016/j.theriogenology.2024.10.001","DOIUrl":"10.1016/j.theriogenology.2024.10.001","url":null,"abstract":"<div><div>India is the largest milk producer contributing 25 % to the global production. From a mere 17 million tons (MTs) in 1951, milk production increased to an inconceivable 230 MTs in 2023. Nondescript cows and buffaloes were the main milk animals. Since 1965, nationwide programs for cross breeding native cattle with exotic milk breeds such as Jersey, Holstein Friesian (HF) and Brown Swiss (BS) and upgradation of buffalo breeds along with disease eradication, feed and fodder development and provision of veterinary and extension services were implemented. Now 75 % of the milk in India is produced by the native, exotic and crossbred cows (43 %) and native breeds of buffaloes (32 %). National Dairy Development Board (NDDB) created dairy cooperatives, an Indian innovation to connect rural milk producers to urban and semi-urban consumers. These cooperatives transformed subsistence dairy farming into a profitable rural enterprise and created millions of jobs. Due to the success of the above activities, since 1998 India continues to produce the highest volumes of milk in the world.</div><div>It is envisioned to increase (i) milk production to 450 million tons (MTs) (one-third of the global production) (ii) gain a share of 15 % in global milk and milk products exports and (iii) build sustainable green practices to match global standards during the next 25 years.</div><div>India is increasingly deploying artificial insemination (AI) with sex sorted-frozen semen, embryo transfer and <em>in vitro</em> production of embryos using sex sorted semen, progeny testing and genomic selection to improve reproduction and productivity among dairy animals. Mitigation of disease burden is undertaken through nationwide vaccination against diseases such as Foot and Mouth, TB, Brucellosis etc. Nutritious fodder crops and feed additives that support higher productivity and reduce enteric methane emissions are promoted. Management and interventional practices such as better nutrition, amelioration of summer stress, oestrus synchronisation etc., are increasingly implemented. Use of internet enabled technology services (IETS) for extension activities, mobile veterinary and animal husbandry services are rapidly becoming commonplace. All these practices portend greater achievements in milk production in the fore-seeable future. Achievements, aspirations, accosts, approaches and appraisal are presented.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142508617","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":"Mitoquinone improves porcine embryo development through modulating oxidative stress and mitochondrial function","authors":"","doi":"10.1016/j.theriogenology.2024.10.011","DOIUrl":"10.1016/j.theriogenology.2024.10.011","url":null,"abstract":"<div><div>Oxidative stress caused by excess reactive oxygen species (ROS) is one of the main causes of low efficiency in <em>in vitro</em> production of embryos. These ROS can cause mitochondrial dysfunction and apoptosis, resulting in poor embryo development. Therefore, to prevent mitochondrial damage and apoptosis caused by ROS, we investigated the effects of mitoquinone (MitoQ), a mitochondrial-targeted antioxidant, on the <em>in vitro</em> culture (IVC) of porcine embryos. Various concentrations of MitoQ (0, 0.01, 0.1, or 1 nM) were supplemented during the entire period of IVC. The results showed that supplementation with 0.1 nM MitoQ significantly increased the blastocyst formation rate, with a higher total cell number including trophectoderm cell number and higher transcript expression of lineage-specific transcription factors in blastocysts. In addition, the 0.1 nM MitoQ-treated group showed a significantly lower percentage and number of apoptotic cells in blastocysts with positively regulated transcript expression of apoptosis-related genes. Therefore, 0.1 nM MitoQ was suggested as optimal concentration for porcine IVC and used for further investigations. MitoQ treatment significantly reduced intracellular ROS levels and increased glutathione levels in Day 2 embryos, with upregulated the transcript expression of antioxidant enzymes-related genes. Furthermore, the MitoQ group exhibited a significantly higher mitochondrial quantity, mitochondrial membrane potential, and ATP content in Day 2 embryos, with increased transcript expression of mitochondrial biogenesis-related genes. Taken together, these findings reveal that MitoQ supplementation can enhance the developmental competence of porcine embryos by decreasing oxidative stress and improving mitochondrial function.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142475496","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 PPARG on the proliferation, apoptosis, and estrogen secretion in goat granulosa cells","authors":"","doi":"10.1016/j.theriogenology.2024.10.010","DOIUrl":"10.1016/j.theriogenology.2024.10.010","url":null,"abstract":"<div><div>As a member of peroxisome proliferator-activated receptor (PPAR) family, PPARG has been reported to be involved in glucolipid metabolism in various species. However, the function of PPARG in estradiol (E2) synthesis, apoptosis, and proliferation in goat ovarian granulosa cells (GCs) is unclear. In this study, we found that goat PPARG was expressed in GCs of all grades of follicles, and localized in the cytoplasm and nucleus of GCs. Transfection of small interfering RNA-PPARG2 (si-PPARG2) decreased E2 synthesis and the abundances of HSD3B and CYP19A1 mRNA and protein. It also promoted cell apoptosis with significant increases in the ratio of BAX/BCL-2 and Caspase3 mRNA and protein. Meanwhile, cell cycle was inhibited by si-PPARG2 transfection, accompanied by decreased mRNA levels of <em>CDK4</em>, <em>CKD6</em>, <em>MYC</em>, <em>CCND1</em>, <em>CCND2</em>, <em>PCNA</em>, and <em>CCNB</em>, increased mRNA level of <em>P53</em>, and decreased protein levels of CDK4, MYC, and CCND1. Furthermore, PPARG interference affected the mRNA and protein abundances of CREB as well as the phosphorylation of CREB but not PKA. In conclusion, our data suggest that PPARG plays an important role in regulating E2 synthesis, cell apoptosis, and proliferation of goat GCs, including the CREB expression and phosphorylation. These results provide evidences for the in-depth study of PPARG in the regulation of goat GCs function.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142442546","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":"Sperm collection in the domestic cat: A comparison of two techniques","authors":"","doi":"10.1016/j.theriogenology.2024.09.024","DOIUrl":"10.1016/j.theriogenology.2024.09.024","url":null,"abstract":"<div><div>Semen collection in cats in the clinic setting can be difficult. However, semen analysis is vital when evaluating breeding soundness of a male. Electroejaculation (EEJ) is currently the most reliable semen collection method but requires specialized equipment and training of the operator. Chemical ejaculation followed by urethral catheterization (UC) is a technique that allows semen collection without special equipment: a catheter is placed into the urethra of a sedated tom and semen is collected passively into the catheter. Earlier studies used the sedative medetomidine at high doses for this procedure. However, medetomidine has been replaced with dexmedetomidine in some countries. This study sought to compare the results of EEJ and UC for semen collection in the domestic cat using dexmedetomidine, a potent α2-adrenoceptor agonist (α2A), as a substitute for medetomidine at the equivalent dose to that used in earlier studies. Twelve domestic cats were collected thrice at weekly intervals. All cats received intramuscular ketamine (5 mg/kg) and intramuscular dexmedetomidine (30 μg/kg) for initial cleanout via EEJ, then randomly underwent either EEJ or UC one week apart. The EEJ was performed under the same anesthetic protocol as the initial cleanout. The UC was performed using intramuscular dexmedetomidine at a dose of 60 μg/kg. Success of collection, total sperm number, sperm morphology, and motility characteristics were analyzed. Sperm was collected successfully from all 12 cats via EEJ and from 11/12 via UC. There were no significant differences in the percentage of total motile, progressively motile, or morphologically normal sperm between ejaculate types when averaged across all cats or individual cats. Although UC yielded a lower volume and higher concentration ejaculate, it consistently produced a lower total sperm number than ejaculates retrieved via EEJ (17.91 x 10⁶ total sperm for UC versus 46.51 x 10⁶ total sperm for EEJ). These results indicated that dexmedetomidine is a very effective sedative and performed satisfactorily in both procedures at the doses used in this study. It was also safe with no adverse effects on healthy toms. EEJ remained the most reliable in terms of assessing semen quality and retrieving semen with adequate number of sperm for breeding purposes. However, UC with dexmedetomidine at this dose demonstrated a 92 % success rate, presenting itself as a remarkably consistent alternative.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142508620","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":"Analysis of the pluripotent and germline marker gene expression, and the state of X chromosome reactivation of primordial germ cells in pig gonads","authors":"","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":null,"pages":null},"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}