Transcriptome analysis of mRNA and long non-coding RNA during subsequent embryo development of porcine cloned zygotes after vitrification
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ABSTRACT:
PROVIDER: PRJNA747042 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR15168300.fastq.gz | Fastqsanger.gz | |||
| SRR15168301.fastq.gz | Fastqsanger.gz | |||
| SRR15168302.fastq.gz | Fastqsanger.gz | |||
| SRR15168303.fastq.gz | Fastqsanger.gz | |||
| SRR15168286.fastq.gz | Fastqsanger.gz |
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Project description:this study was designed to acquire mRNA and long non-coding RNA (lncRNA) profiles of 2-cells, 4-cells and blastocysts derived from vitrified porcine cloned zygotes using transcriptome sequencing. We identified 167 differentially expressed (DE) mRNAs and 516 DE lncRNAs in 2-cell stage, 469 DE mRNAs and 565 lncRNAs in 4-cell stage, and 389 DE mRNAs and 816 DE lncRNAs in blastocyst stage. Functional enrichment analysis revealed that DE mRNAs during embryo development were involved in many regulatory mechanisms related to cell cycle, cell proliferation, apoptosis, metabolism and others. Moreover, the target genes of DE lncRNAs in the three embryonic stages were also enriched in many key GO terms or pathways such as ‘defense response’, ‘linoleic acid metabolic process’, ‘embryonic axis specification’, ‘negative regulation of protein neddylation’, etc. In conclusion, the present study provided comprehensive transcriptomic data about mRNA and lncRNA for the vitrified porcine cloned zygotes during different developmental stages, which contributed to further understand the potential cryodamage mechanisms responsible for impaired embryo development.
2021-07-22 | GSE180219 | GEO
Project description:Cryopreservation of porcine cloned zygotes has important implications for biotechnology and biomedicine research; however, lower embryo developmental potential remains an urgent problem to be resolved. For exploring the sublethal cryodamages during embryo development, this study was designed to acquire the mRNA and long non-coding RNA (lncRNA) profiles of 2-cells, 4-cells and blastocysts derived from vitrified porcine cloned zygotes using transcriptome sequencing. We identified 167 differentially expressed (DE) mRNAs and 516 DE lncRNAs in 2-cell stage, 469 DE mRNAs and 565 lncRNAs in 4-cell stage, and 389 DE mRNAs and 816 DE lncRNAs in blastocyst stage. Functional enrichment analysis revealed that the DE mRNAs during embryo development were involved in many regulatory mechanisms related to cell cycle, cell proliferation, apoptosis, metabolism and others. Moreover, the target genes of DE lncRNAs in the three embryonic stages were also enriched in many key GO terms or pathways such as "defense response", "linoleic acid metabolic process", "embryonic axis specification", "negative regulation of protein neddylation", etc., In conclusion, the present study provided comprehensive transcriptomic data about mRNAs and lncRNAs for the vitrified porcine cloned zygotes during different developmental stages, which contributed to further understand the potential cryodamage mechanisms responsible for impaired embryo development.
| S-EPMC8718616 | biostudies-literature
Project description:The objective of this study was therefore to evaluate the impact of simultaneous vitrification of large numbers of embryos with the OC (20 embryos per device) system on gene expression patterns of in vivo-derived blastocysts when compared to the Superfine Open Pulled Straw (SOPS) system (5-7 embryos per device) through a microarray approach. In vivo-developed embryos were surgically retrieved from 16 sows in 4 replicates. Embryos at the blastocyst stage (n=180) were selected for the experiment and divided into three experimental groups: blastocysts vitrified with the OC system (n=60), blastocysts vitrified with the SOPS system (n=60), and a control group consisting of fresh, nonvitrified blastocysts (n=60). After in vitro culture for 24 h, six pools of 8 viable embryos (n=48 embryos per group) were prepared for transcriptome analysis. Embryos were placed in sterile tubes with 5 µL of PBS, immediately immersed in LN2 and stored at -80 °C until microarray analysis. After microarray analysis, a list of DEGs was generated for comparison of each vitrification system (OC and SOPS) with the control. Then, the two DEG lists were compared to determine the specific genes altered in each vitrification system and the DEGs shared by both systems. In addition, a list of DEGs was obtained by direct comparison of both vitrification systems. Pathway enrichment analysis was performed for each obtained list of DEGs.
2023-03-06 | E-MTAB-12772 | biostudies-arrayexpress
Project description:Vitrification cryopreservation of oocytes is an enabling technology for assisted reproductive technology. However, many factors in the vitrification process, such as the toxicity of cryoprotectant, osmotic stress, ice crystal formation during rewarming, will cause fatal damage to oocytes, thus affecting the embryo developmental potential and subsequent clinical outcomes. Therefore, oocyte vitrification still faces significant challenges. Recent studies have shown that LEA protein has the potential to improve oocyte cryopreservation, while the molecular mechanism by which it exerts protective effects is still unclear. Therefore, we have systematically investigated the effects of LEA proteins on the vitrification cryopreservation and their molecular mechanisms. Results revealed that the proteins improved the developmental potential of human oocytes following cryopreservation, mostly by downregulating FOS genes and reducing oxidative stress. Moreover, the synergistic cryoprotection of LEA proteins by inhibiting the formation of ice crystals was given full play. This study provides a new strategy for high-quality cryopreservation of human oocytes.
2024-04-01 | GSE239302 | GEO
Project description:Cryopreservation of embryos has been confirmed to cause oxidative stress as a factor responsible for impaired developmental competence. Currently, astaxanthin (Ax) raises considerable interest as a strong exogenous antioxidant and for its potential in reproductive biology. The present study aimed to investigate the beneficial effects of Ax supplementation during in vitro culture of vitrified porcine zygotes and the possible underlying mechanisms. First, the parthenogenetic zygotes were submitted to vitrification and then cultured in the medium added with various concentrations of Ax (0, 0.5, 1.5, and 2.5 μM). Supplementation of 1.5 μM Ax achieved the highest blastocyst yield and was considered as the optimal concentration. This concentration also improved the blastocyst formation rate of vitrified cloned zygotes. Moreover, the vitrified parthenogenetic zygotes cultured with Ax exhibited significantly increased mRNA expression of CDX2, SOD2, and GPX4 in their blastocysts. We further analyzed oxidative stress, mitochondrial and lysosomal function in the 4-cell embryos and blastocysts derived from parthenogenetic zygotes. For the 4-cell embryos, vitrification disturbed the levels of reactive oxygen species (ROS) and glutathione (GSH), and the activities of mitochondria, lysosome and cathepsin B, and Ax supplementation could fully or partially rescue these values. The blastocysts obtained from vitrified zygotes showed significantly reduced ATP content and elevated cathepsin B activity, which also was recovered by Ax supplementation. There were no significant differences in other parameters mentioned above for the resultant blastocysts. Furthermore, the addition of Ax significantly enhanced mitochondrial activity and reduced lysosomal activity in resultant blastocysts. In conclusion, these findings revealed that Ax supplementation during the culture period improved subsequent embryonic development and quality of porcine zygotes after vitrification and might be used to ameliorate the recovery culture condition for vitrified embryos.
| S-EPMC9011099 | biostudies-literature
Project description:Faulty epigenetic reprogramming of somatic nuclei is thought to be the main reason for low cloning efficiency by somatic cell nuclear transfer (SCNT). Histone deacetylase inhibitors (HDACi), such as Scriptaid, improve developmental competence of SCNT embryos in several species. Another HDACi, Oxamflatin, is about 100 times more potent than Scriptaid in the ability to inhibit nuclear-specific HDACs. The present study determined the effects of Oxamflatin treatment on embryo development, DNA methylation, and gene expression. Oxamflatin treatment enhanced blastocyst formation of SCNT embryos in vitro. Embryo transfer produced more pigs born and fewer mummies from the Oxamflatin-treated group compared to the Scriptaid-treated positive control. Oxamflatin also decreased DNA methylation of POU5F1 regulatory elements and centromeric repeat elements in day-7 blastocysts. When compared to in vitro-fertilized (IVF) embryos, the methylation status of POU5F1, NANOG, and centromeric repeat was similar in the cloned embryos, indicating these genes were successfully reprogrammed. However, compared to the lack of methylation of XIST in day-7 IVF embryos, a higher methylation level in day-7 cloned embryos was observed, implying that X chromosomes were activated in day-7 IVF blastocysts, but were not fully activated in cloned embryos, i.e., reprogramming of XIST was delayed. A time-course analysis of XIST DNA methylation on day-13, -15, -17, and -19 in vivo embryos revealed that XIST methylation initiated at about day 13 and was not completed by day 19. The methylation of the XIST gene in day-19 control cloned embryos was delayed again when compared to in vivo embryos. However, methylation of XIST in Oxamflatin-treated embryos was comparable with in vivo embryos, which further demonstrated that Oxamflatin could accelerate the delayed reprogramming of XIST gene and thus might improve cloning efficiency.
| S-EPMC4312790 | biostudies-literature
Project description:Vitrification is replacing slow freezing as the most popular method for human embryo cryopreservation in clinics world-wide. Several studies demonstrated that cryopreservation alters gene expression of mammalian embryos, but none of them analysed what happen with those embryos that get implanted and follow with the gestation. The aim of this study was to evaluate the effect of vitrification technique on rabbit embryonic and fetal development by performing a transcriptomic analysis of 6 day old embryos and 14 days old fetal placentas. Effect of vitrification on late blastocyst and fetal placenta transcriptome. Four indepent replicates were performed for each condition (control and vitrified) and for both tissues (embryo and fetal placenta).
2013-09-26 | E-GEOD-51208 | biostudies-arrayexpress
Project description:Vitrification is a method for long-term biological sample cryopreservation without causing intra- and extra-cellular ice formation. We recently established a novel closed vitrification system to cryopreserve mouse ovarian follicles. Using the 3D alginate hydrogel encapsulated in vitro follicle growth (eIVFG) method, we have demonstrated that compared to freshly-harvested follicles, vitrified follicles have normal follicle and oocyte reproductive outcomes. Our recent study further demonstrated the faithful preservation of molecular signatures of follicle-stimulating hormone (FSH)-stimulated follicle maturation in vitrified follicles. However, it is unknown whether ovulation, another crucial gonadotropin-dependent follicular event, and involved ovulatory gene regulatory pathways are well conserved in vitrified follicles. Fresh and vitrified follicles grown for 8 days by eIVFG were collected at 0, 1, 4 and 8-hour post human chorionic gonadotropin (hCG) treatment for the single-follicle RNA sequencing. Principal component analysis (PCA) and Pearson’s correlation analysis revealed that vitrified follicles have similar transcriptomic profiles to fresh follicles. Furthermore, the expression of several genes essential for ovulation were comparable between vitrified and fresh follicles. In summary, these results demonstrate that our closed vitrification system preserves follicular transcriptomic dynamics during ex vivo ovulation. Together with our previous findings that vitrification preserves FSH-stimulated follicle maturation, the integration of vitrification of immature follicles and eIVFG has a great potential to serve as an additional fertilization preservation approach for young cancer patients and endangerers species. Moreover, follicle vitrification enables a high-content ovarian follicle biobank, which can greatly improve the throughput of eIVFG for studying ovulation biology, anovulatory disease, toxicology, and novel contraceptive drug development targeting ovulation.
2023-09-20 | GSE227091 | GEO
Project description:To study the efficiency of a semi-automated technique of oocyte vitrification as compared with a manual method and transcriptomic landscape associated with the oocyte vitrification.
2022-12-01 | GSE205417 | GEO
Project description:Embryo vitrification involves exposure to high concentrations of cryoprotectants and osmotic stress during cooling and warming. A multitude of these factors have the potential of affecting gene expression. In this study, in vitro produced bovine embryos at the blastocyst stage were submitted to vitrification. Four recipients each were used for transferring non-vitrified (80) and vitrified (80) embryos. Twelve non-vitrified and nine vitrified viable day-14 (D14) embryos were recovered by uterine flushing. RNA-seq analysis of whole embryo or isolated trophectoderm (TE) from vitrified and fresh recovered D14 embryos revealed a total of 927 genes changed their expression as a result of vitrification, among them 782 and 145 genes were up- and down-regulated, respectively. In TE isolates, vitrification resulted in 4,096 and 280 up- and down-regulated genes, respectively. In addition, we found 671 and 61 genes commonly up- and down-regulated in both vitrified whole embryo and TE. Commonly up-regulated pathways by vitrification included epithelial adherens junctions, sirtuin signaling, germ cell-sertoli cell junction, ATM signaling, NER, and protein ubiquitination pathways. The commonly down-regulated pathways included EIF2 signaling, oxidative phosphorylation, mitochondrial dysfunction, regulation of eIF4 and p70S6K signaling, mTOR signaling, sirtuin singling, and NER pathways. Our analysis identified specific pathways and implicated specific gene expression patterns important to cryopreservation affecting embryo developmental competence.
2021-03-01 | GSE159891 | GEO