Similar Datasets
Project description:We report the application of single cell transcriptome sequencing technology for high-throughput profiling of the brilliant cresyl blue test-positive porcine oocytes had higher rates of meiotic maturation, lower death rates, and better cleavage and blastocyst rates as well. Single oocyte transcriptome sequencing on porcine germinal vesicle (GV) stage oocytes that differentially stained by BCB identified 155 genes with significant abundance differences, including CDC5L, LDHA, SPATA22, RGS2, PAIP1, WEE1B and HSP27, which enriched in functionally important signaling pathways, such as spliceosome, cell cycle, oocyte meiosis, and nucleotide excision repair.
2019-01-17 | GSE101181 | GEO
Project description:Cytoplasmic and nuclear maturation of oocytes as well as interaction with the surrounding cumulus cells are important features relevant to the acquisition of developmental competence. We used brilliant cresyl blue (BCB) to distinguish oocytes with low activity of the enzyme Glucose-6-Phosphate Dehydrogenase, and thus separated fully grown (BCB+) oocytes from those in the growing phase (BCB-). The BCB+ oocytes are twice as likely to produce a blastocyst in vitro compared to BCB- oocytes (P<0.01). We analyzed mitochondrial DNA (mtDNA) copy number in single oocytes and determined that BCB- oocytes have 1.3-fold more copies than BCB+ oocytes (P=0.004). We also interrogated the transcriptome of oocytes and surrounding cumulus cells of BCB+ versus BCB- oocytes. There was no differential transcript abundance of genes expressed in oocytes, but we identified 172 genes in cumulus cells with differential transcript abundance (FDR<0.05) based on the BCB staining of their oocyte. Differential gene co-expression analysis between BCB+ and BCB- oocytes and their surrounding cumulus cells revealed dynamic coordination of transcript abundance between both compartments of the cumulus-oocyte complex. We identified a subset of genes whose co-expression in fully grown oocytes (n=75) and their surrounding cumulus cells (n=108) compose a unique profile of the cumulus-oocyte complex. In conclusion, as oocytes transition from growing to fully grown, there is an increase in the likelihood of producing a blastocyst, a reduction of mtDNA copies and no systematic variation of transcript abundance. Cumulus cells present changes in transcript abundance, which reflects in a dynamic co-expression between oocyte and cumulus cells.
2022-03-28 | GSE199210 | GEO
Project description:The molecular characterisation of mitochondrial DNA deficient oocytes using a pig model
| PRJNA429045 | ENA
Project description:Comparison of mitochondrial DNA (mtDNA) methylation patterns in oocytes, blastocysts and ovarian granulosa cells indicates hitherto unsuspected dynamics. Oocytes and blastocysts recovered from animals subjected to ovarian stimulation and from abattoir ovaries were analyzed using bisulphite transformation of DNA followed by whole genome sequencing. The number of mtDNA copies is quite high in oocytes (200–400,000) and early embryos, resulting in very high coverage (> 3000 X) and very low p values for each of 716 cytosine-based nucleosides. Methylation was lowest in oocytes, following by blastocysts then granulosa cells and was not restricted to CpG sites but found also at CHG and CHH sites not distributed randomly. Analysis of the corresponding genomic DNA also revealed numerous methylated CHG and CHH sites, particularly in oocytes. In association with ovarian stimulation, mtDNA methylation was lower in oocytes than in blastocysts, suggesting a developmental function. RNA analysis of mitochondria encoded genes showed a significant inverse correlation between methylation and expression level for all sequences. Methylation was more extensive in mature animals than in pre-pubertal animals and the profile in granulosa cells differed strikingly from those in oocytes and blastocysts. Although preliminary, the results suggest that mtDNA methylation might play a role during gametogenesis and might be subject to regulation according to animal environment and/or age.
2019-08-26 | GSE122599 | GEO
Project description:Flag-PKM2 was expressed in H1299 cells. Then the H1299 cells were treated with DMSO or 100nM MLN8237. Then, cell lysate was incubated with anti-Flag M2-agarose. Elutes were separated with 10% SDS-PAGE. The gel was stained with Coomassie Brilliant Blue. Visualized bands were excised and subjected to LC-MS/MS analysis. Then the phosphorylation sites of PKM2 were identified.
2023-03-11 | PXD036575 | Pride
Project description:We aim to find out the Kindlin-2 interaction proteins. We transfected plasmids Flag or Flag-Kindlin-2 to HKC cell lines and then used Flag-M2 beads to do the coimmunoprecipitation experiments and followed by NuPAGE Tris-Acetate Mini Gels detection and stained by Coomassie brilliant blue. Compared the two groups,and then we will find the proteins interact with Kindlin-2.
2019-12-16 | PXD016008 | Pride
Project description:The maternal tract plays a critical role in the success of early embryonic development providing an optimal environment for establishment and maintenance of pregnancy. Preparation of this environment requires an intimate dialogue between the embryo and her mother. To advance our understanding of the process by which a foreign blastocyst is accepted by the maternal endometrium and better address the clinical challenges of infertility and pregnancy failure, it is imperative to decipher this complex molecular dialogue. The objective of the present work is to define the local response(s) of the maternal tract towards the embryo during the earliest stages of pregnancy. In order to identify the uterine gene expression modified in the presence of the embryo when compared to the oocyte we used a novel model that eliminated genetic variability. Using laparoscopic insemination one oviduct was inseminated with spermatozoa and the contralateral oviduct was injected with diluent. This model allowed us to obtain samples from the oviduct and the uterine horn containing either embryos or oocytes from the same sow. Uterine horn pig samples containing embryos at blastocyst stage and and their contralateral uterine horn containing oocytes from individual sows were collected for RNA isolation and hybridization on Affymetrix microarrays. Three biological replicates were performed (n= 3 sows) and a total of 6 arrays were used for microarrays study (3 arrays for uterine horn samples containing embryos (inseminated-side) and 3 arrays for samples containing oocytes (non-inseminated side).
2012-04-18 | E-GEOD-33262 | biostudies-arrayexpress
Project description:Conservation and species specificity are both unveiled in the reprogramming process of epigenetic modifications in early embryos. Pig is important agricultural livestock that closely related to human life and understanding the mechanisms of epigenetic reprogramming in porcine embryos could be assistive to the pig breeding industry and also informative to human embryo research. Nevertheless, current knowledge of epigenetic reprogramming in porcine embryos is elusive. We profile H3K4me3, H3K27me3, and H3K27ac modifications in porcine oocytes and preimplant embryos. Our data reveal the existence of unusual broad H3K4me3 domains not only in oocytes but also expanded in pre-ZGA embryos and demonstrates the extensive transitions of unusual broad H3K4me3 domains and their roles during embryogenesis. Notably, broad H3K27me3 and H3K27ac enriched domains are found to be preferably colocalized in these broad H3K4me3 domains when comparing to it in mice and human. By knocking down KDM5B in pig embryos, cleavage and developments were disrupted as demonstrated in mouse.
2022-07-08 | GSE163709 | GEO
Project description:Oocyte defects lie at the heart of some forms of infertility and could potentially be addressed therapeutically by alternative routes for oocyte formation. Here, we describe the generation of functional human oocytes following nuclear transfer of first polar body (PB1) genomes from metaphase II (MII) oocytes into enucleated donor MII cytoplasm (PBNT). The reconstructed oocytes supported the formation of de novo meiotic spindles and, after fertilization with sperm, meiosis completion and formation of normal diploid zygotes. While PBNT zygotes developed to blastocysts less frequently (42%) than controls (75%), genome-wide genetic, epigenetic, and transcriptional analyses of PBNT and control ESCs indicated comparable numbers of structural variations and markedly similar DNA methylation and transcriptome profiles. We conclude that rescue of PB1 genetic material via introduction into donor cytoplasm may offer a source of oocytes for infertility treatment or mitochondrial replacement therapy for mtDNA disease.
2016-12-07 | GSE79309 | GEO