Vitrification of pig embryos dysregulates the microRNA transcriptome profile
Ontology highlight
ABSTRACT:
ORGANISM(S): Sus scrofa
SUBMITTER: Cristina Cuello Medina
PROVIDER: E-MTAB-13814 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
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:Changes in gene expression induced by the Cryotop vitrification procedure in bovine blastocysts using Agilent EmbryoGENE microarray slides. Bovine in vitro-produced embryos at the blastocyst stage (144 to 156 hours post insemination) were vitrified using the Cryotop system and compared with non-vitrified (control) embryos. After vitrification, the embryos were warmed and cultured for an additional 4 hours. Embryos that re-expanded or developed to the expanded blastocyst stage were used for microarray analysis. Four pools of vitrified embryos were hybridized against four pools of control embryos in a dye-swap design.
2015-08-03 | E-GEOD-65953 | biostudies-arrayexpress
Project description:High hydrostatic pressure (HHP) has been used to pre-condition embryos before essential, yet potentially detrimental procedures such as cryopreservation. However, the mechanisms for HHP are poorly understood. We treated bovine blastocysts with three different HHP (40, 60 and 80 MPa) in combination with three recovery periods (0, 1h, 2h post HHP). Re-expansion rates were significantly higher at 40 and 60 but lower at 80 MPa after vitrification-warming in the treated groups than controls. Microarray analysis revealed 399 differentially expressed transcripts, representing 254 unique genes, among different groups. Gene ontology analysis indicated that HHP at 40 and 60 MPa promoted embryo competence through down-regulation of genes in cell death and apoptosis, and up-regulation of genes in RNA processing, cellular growth and proliferation. In contrast, 80 MPa up-regulated genes in apoptosis, and down-regulated protein folding and cell cycle-related genes. Moreover, gene expression was also influenced by the length of the recovery time after HHP. The significantly over-represented categories were apoptosis and cell death in the 1h group, and protein folding, response to unfolded protein and cell cycle in the 2h group compared to 0h. Taken together, HHP promotes competence of vitrified bovine blastocysts through modest transcriptional changes. 4*3*2 design experiment. The following treatments were included: (1) Control embryos were left untreated in the incubator (one atmospheric pressure or 0.1 MPa); (2) treatment groups were assigned to 40, 60 and 80 MPa HHP for 1h at either 24°C (room temperature) or 39°C (body temperature), followed by three different recovery time periods (0, 1 and 2h) post-HHP in the holding medium. Biological replicates: 3 control replicates. Technical replicates: dye-swap.
2015-12-10 | E-GEOD-75559 | biostudies-arrayexpress
Project description:Studies on embryo cryopreservation efficiency had been focused mainly on technical and embryo factors. While structural damages can be easily evaluated, physiological damages only can be estimated by analyzing their in vitro and in vivo development to later stages. In order to determine how cryopreservation process affect embryo pre-implantory development, a transcriptional microarray study has been performed comparing gene expression of 6 days old rabbit embryos, previously vitrified or frozen and transferred into recipients rabbit females, to their in vivo counterparts. For each experimental group, control, vitrified and frozen late blastocysts, total RNA was extracted from 3 pools of approximately 10 embryos and labeled with Cy3 or Cy5 dyes. Then, six competitive hybridizations were carried out including two dye-swaps to compensate dye-bias. A specifically microarray designed to study rabbit gene expression profiling, the Rabbit 44K oligonucleotide array (Agilent Technologies), was used in this study. Identification of differentially expressed transcripts from 6 day old blastocysts was achieved using the Limma algorithm, and functional annotation was performed by Blast2GO software. Compared to 6 day old in vivo derived embryos, viable vitrified embryos only present 3 differentially expressed genes, in contrast to frozen viable embryos with 24 genes upregulated and 46 genes downregulated. These results reveal that effects of cryopreservation still remain in late blastocyst pre-implantatory gene expression, and potential damage and alterations produced by vitrification and slow-freezing procedures are differentially resolved after 3 days of in vivo development. Transcriptional microarray study that compares gene expression of viable 6 day old rabbit embryos, previosly vitrified or frozen and tranferred into recipient rabbit females, to their in vivo counterparts. Experiment 1: Control embryos vs. Vitrified embryos and Experiment 2: Control embryos vs. Frozen embryos. Biological replicates used: 3 replicates for control embryos, 3 replicates for vitrified embryos and 3 replicates for frozen embryos.
2011-09-22 | E-GEOD-32263 | biostudies-arrayexpress
Project description:Nowadays there is a need to improve cryopreservation protocols in both human assisted reproduction and domestic animal field. We wanted to compare the transcriptomic changes induced by two commonly used cryopreservation procedures (slow freezing and vitrification) on rabbit late blastocyst before the onset of implantation. We recovered rabbit morulae at day 3 of development, freezing or vitrified and transferred them into recipient maternal tracts till day 6 of development. In this way, we wanted to analyse if different cryopreservation techniques produce different effects on gene expression that embryos are unable to get over after three days of in vivo development. After artificially insemination morulae were recovered at day 3 of development, frozen or vitrified, and then transferred to recipient does. To assess gene expression differences between frozen and vitrified the recipients were slaughtered three days after the transfer, at day 6.
2013-09-19 | E-GEOD-50979 | biostudies-arrayexpress
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:Use of combined estrus synchronization/superovulation treatments (SS) alters ovarian and en-dometrial gene expression patterns, impairing follicle and oocyte growth, fertilization, and em-bryo development. Since the impact of SS-treatments on the transcriptome of the surviving em-bryos remains undisclosed, we hereby examined gene expression changes in day 6 blastocysts that survived a brief regimen of synchronization treatment combined with superovulation. Embryos were surgically collection at day 6 after AI and transcriptome analysis performed on blas-tocyst-stage embryos with good morphology to disclose differentially expressed genes (DEGs) between groups at P-value < 0.05 and </> 1.5-fold change. Compared with the blastocysts from control (untreated) sows, the blastocysts from SS treated sows had moderate gene expression changes, with 7 pathways disrupted with a total of 10 transcripts affected, including metabolic RDH10 and SPTLC2 gene upregulation, and downregulation of oxidation related GSTK1 and GSTO1 genes. These gene expression alterations may suggest suboptimal embryo quality and could depress the embryos' response to oxidative stress, thereby impairing subsequent embryo development. These and previous findings call for avoidance of SS-treatments in embryo transfer programs.
2023-03-22 | E-MTAB-12820 | biostudies-arrayexpress
Project description:A long-standing question in developmental and reproductive biology is when the mammalian embryo becomes sufficiently distinct from its oocyte precursor. Myriads of studies examined the messenger RNAs that change during the oocyte-to-embryo transition, whereas proteins have been much less studied, in spite of their greater vicinity to phenotype. In the present study we modified the widely used embryo culture medium KSOM (PMID 12470333, PMID 10859270) to make it apt for our application. We replaced the serum albumin with polyvinylpyrrolidone and also replaced the natural Arginine and Lysine with their “heavy” isotopic variants Arginine 13C 15N and Lysine 13C 15N. Fertilized oocytes were retrieved from oviducts of gonadotropin-primed B6C3F1 females mated to CD1 males, and cultured at 37 degrees Celsius under 5% CO2 in KSOM containing 0.3 mM Arginine 13C 15N and 0.2 mM Lysine 13C 15N, which are the regular concentrations of these two amino acids in KSOM medium (PMID 12470333; PMID 10859270). After 4 days of culture, the embryos of the isotopic group had undergone blastocyst formation just like the control embryos cultured in normal medium. Samples of approx. 500 “heavy”-labeled blastocysts were collected zona-free and subjected to mass spectrometric analysis. The median labeling rate was 83%, ranging from 0% in proteins that did not incorporate any Arginine 13C 15N and Lysine 13C 15N, to 100% in proteins that were completely labeled. Our study demonstrates that a commonly used, chemically defined medium can be adapted for Stable Isotope Labeling by/with Amino acids in Cell culture (SILAC) and combined with high-resolution mass spectrometry, in a preimplantation embryo setting. This allows to tackle long-standing questions in developmental and reproductive biology, such as the identification of putative maternal (0% labeled), putative embryonic (100% labeled) or shared proteins in live mammalian embryos.
2023-10-19 | PXD035570 | Pride
Project description:Vitrification is commonly used in the cryopreservation of mammalian blastocysts to overcome the temporal and spatial limitations of embryo transfer. Previous studies have shown that the implantation ability of vitrified blastocysts is impaired and that microRNAs (miRNAs) regulate the critical gene for embryo implantation. However, little information is available about the effect of vitrification on the miRNA transcriptome in blastocysts, which partially explains the impaired implantation ability of vitrified blastocysts. In the present study, the miRNA transcriptomes in fresh and vitrified mouse blastocysts were analyzed by microRNA Taqman assay based method, and the results were validated using quantitative real-time PCR (qRT-PCR). Then, the differentially expressed miRNAs were assessed using the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Overall, 760 known mouse miRNAs were detected in the vitrified and fresh mouse blastocysts. Of these, the expression levels of five miRNAs differed significantly: in the vitrified blastocysts, four miRNAs (mmu-miR-199a-5p, mmu-miR-329-3p, mmu-miR-136-5p and mmu-miR-16-1-3p) were upregulated, and one (mmu-miR-212-3p) was downregulated. The expression levels of all miRNAs measured by the microRNA Taqman assay based method and qRT-PCR were consistent. The four upregulated miRNAs were predicted to regulate 877 candidate target genes, and the downregulated miRNA was predicted to regulate 231 genes. The biological analysis further showed that the differentially expressed miRNAs mainly regulated the implantation of embryos. In conclusion, the results of our study showed that vitrification significantly altered the miRNA transcriptome in mouse blastocysts, which may decrease the implantation potential of vitrified blastocysts.
2016-12-31 | GSE62581 | GEO
Project description:Inner cell mass (ICM) cells of two independent E3.5 blastocysts were profiled by transcriptome sequencing to assess the expression of extracellular matrix components.
2014-05-15 | E-MTAB-2555 | biostudies-arrayexpress