Transcriptomics reveals molecular differences in equine oocytes vitrified before and after in vitro maturation
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ABSTRACT:
ORGANISM(S): Equus caballus
PROVIDER: GSE225950 | GEO | 2023/05/03
REPOSITORIES: GEO
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Project description:Maturation of oocytes under in-vitro conditions (IVM) results in impaired developmental competence compared to oocytes matured in-vivo. Oocytes are closely coupled to their cumulus complex (COC) with a bidirectional exchange of metabolites. Therefore, elucidation of aberrations in cumulus metabolism in-vitro is crucial for a better mimicking of physiological maturation conditions. The aim of this study was the analysis of the equine cumulus cells proteome of single cumulus complexes of metaphase II oocytes matured either under in-vivo (n=8) or in-vitro (n=7) conditions. For in-vivo COC collection mares were slaughtered 30 hours after injection and cumulus complexes from the dominant follicle were harvested for analysis. For in-vitro maturation COCs were recovered from mares out of oestrous and matured for 30 hours in-vitro. COCs were separated in cumulus complexes and oocytes, and only cumulus of successfully matured oocytes was analyzed for this study. All cumulus samples were washed, snap frozen and stored in liquid nitrogen until preparation for analysis.
2019-11-12 | PXD011086 | Pride
Project description:In vitro maturation (IVM) of the oocytes is a routine method in bovine embryo production. The competence of bovine oocytes to develop into embryo after IVM and in vitro fertilization (IVF) is lower as compared to in vivo preovulatory oocytes. Cumulus cells (CC) that enclose an oocyte are involved in the acquisition of oocyte quality during maturation. Using transcriptomic approach we compared cumulus cells gene expression during IVM with that in vivo preovulatory period. Global transcriptional profiling was performed using cumulus cells collected from mature bovine oocytes (metaphase-II stage) after maturation performed either in vivo or in vitro. In vivo matured cumulus cells were collected from ovulatory follicles of Montbeliard adult cows by ovum pick-up in vivo (OPU, n=4). In vitro matured cumulus cells were recovered from the oocytes after 22h of in vitro culture of cumulus-oocyte complexes (50 COC per experiment) from 2-6 mm ovarian follicles of adult cows (MIV, n=4). Gene expression analysis was carried out between in vivo and in vitro matured cumulus representing a total of 8 slides (dye swap protocol)
2011-08-08 | E-GEOD-31261 | 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: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.
2015-08-03 | GSE65953 | GEO
Project description:In cattle, almost all fully grown vesicle stage oocytes (GV) have the ability to resume meisos, develop to Metaphase II stage (MII), support fertilization and progress through the early embryonic cycles in vitro. Yet without intensive selection, the majority fail to develop to the blastocyst stage. Using the Affymetrix Bovine Genome Array, global mRNA expression analysis of immature (GV) and in vitro matured (IVM) bovine oocytes was carried out to characterize the transcriptome of the bovine oocyte and to identify the key pathways associated with oocyte meiotic maturation and developmental potential. Immature and in vitro matured bovine oocytes were collected for RNA extraction and hybridization on Affymetrix GeneChip Bovine Genome array. Careful removal of cumulus and selection of oocytes was carried out under the stereo microscope in order to examine the actual cumulus-free temporal oocyte gene expression profiles. Immature oocytes at time 0 h and in vitro matured oocytes at 24 h were collected for analysis.
2011-04-25 | E-GEOD-23449 | biostudies-arrayexpress
Project description:In vitro oocyte maturation (IVM) holds great promise as a tool for enhancing clinical treatment of infertility, enhancing availability of non human primates for development of disease models, and facilitating endangered species preservation. However, IVM outcomes have remained significantly below success rates obtained using in vivo matured (VVM) oocytes from humans and non human primates. A cDNA array based analysis is presented, comparing the transcriptomes of VVM oocytes with IVM oocytes. We observe a small set of just 59 mRNAs that are differentially expressed between the two cell types. These mRNAs are related to cellular homeostasis, cell-cell interactions including growth factor and hormone stimulation and cell adhesion, and other functions such as mRNA stability and translation. Additionally, we observe in IVM oocytes overexpression of PLAGL1 and MEST, two maternally imprinted genes, indicating a possible interruption or loss of correct epigenetic programming. These results indicate that, under certain IVM conditions, oocytes that are molecularly highly similar to VVM oocytes can be obtained, however the interruption of normal oocyte-somatic cell interactions during the final hours of oocyte maturation may preclude the establishment of full developmental competence. Keywords: oocyte maturation Comparison of in vitro matured MII-stage oocytes (4 biological replicates) with in vivo matured MII-stage oocytes (4 biological replicates)
2009-05-20 | E-GEOD-11895 | biostudies-arrayexpress
Project description:In cattle, almost all fully grown vesicle stage oocytes (GV) have the ability to resume meisos, develop to Metaphase II stage (MII), support fertilization and progress through the early embryonic cycles in vitro. Yet without intensive selection, the majority fail to develop to the blastocyst stage. Using the Affymetrix Bovine Genome Array, global mRNA expression analysis of immature (GV) and in vitro matured (IVM) bovine oocytes was carried out to characterize the transcriptome of the bovine oocyte and to identify the key pathways associated with oocyte meiotic maturation and developmental potential.
2011-04-25 | GSE23449 | GEO
Project description:Oocyte quality largely determines the embryo development after fertilization. High-quality oocyte requires the competence of both cytoplasm and nucleus. However, clinically, the treatment of developmentally competent oocytes was limited. Here, in order to improve the embryo development efficiency of developmentally incompetent oocytes, we performed spindle-chromosome complex (SCC) transfer between in vitro matured (IVM) and in vivo matured (IVO) oocytes of the non-human primate. We observed that the blastocyst rate of embryos derived by transferring the SCC of IVM (IVM-SCC) oocytes into enucleated IVO oocytes was comparable with that of embryos derived by IVO oocytes. After transferring the reconstructed embryos into the uterus of surrogate mothers, two live rhesus monkeys were obtained, indicating that the nuclei of IVM oocytes support both the pre-and post-implantation embryo development of non-human primates.
2023-07-15 | GSE233232 | GEO
Project description:Oocyte quality largely determines the embryo development after fertilization. High-quality oocyte requires the competence of both cytoplasm and nucleus. However, clinically, the treatment of developmentally competent oocytes was limited. Here, in order to improve the embryo development efficiency of developmentally incompetent oocytes, we performed spindle-chromosome complex (SCC) transfer between in vitro matured (IVM) and in vivo matured (IVO) oocytes of the non-human primate. We observed that the blastocyst rate of embryos derived by transferring the SCC of IVM (IVM-SCC) oocytes into enucleated IVO oocytes was comparable with that of embryos derived by IVO oocytes. After transferring the reconstructed embryos into the uterus of surrogate mothers, two live rhesus monkeys were obtained, indicating that the nuclei of IVM oocytes support both the pre-and post-implantation embryo development of non-human primates.
2022-12-31 | GSE221785 | GEO
Project description:The present study mainly aimed to investigate the effect of vitrification temperature (VT) and cryoprotective agent concentrations (CPAs) on the mRNA transcriptome of bovine immature oocytes (BIOs). Cumulus oocyte complexes (COCs) were randomly divided into the following five groups: fresh oocytes (control), oocytes vitrified in liquid helium (LHe; -269℃) with 5.6 M CPAs (LHe 5.6 M), oocytes vitrified in LHe with 6.6 M CPAs (LHe 6.6 M), oocytes vitrified in liquid nitrogen (LN; -196℃) with 5.6 M CPAs (LN 5.6 M), and oocytes vitrified in LN with 6.6 M CPAs (LN 6.6 M). mRNA transcriptomes of each sample were analyzed by Smart-seq4, and the differentially expressed genes (DGEs) were detected by edgeR (p≤0.05; fold-change≥2). Compared with the control group, 505 DGEs were detected with 342 up-regulated and 163 down-regulated genes in LHe 5.6 M; 609 DGEs were detected with 493 up-regulated and 116 down-regulated gene in LHe 6.6 M; 218 DGEs were determined with 101 up-regulated and 117 down-regulated genes in LN 5.6 M; and 221 DGEs were detected with 104 up-regulated and 117 down-regulated genes in LN 6.6 M. LHe vitrification affected the mRNA transcriptome of vitrified BIOs mainly by up-regulating gene expression. Reduced CPAs (5.6 M) decreased the effect of vitrification on mRNA transcriptome when LHe vitrification was used. Among all the DGEs closely related to bovine immature oocytes, the genes possibly related to VT were ND2, MPV17L2, PIF1, LPIN1, IMP3, BRD1, DCTN3, DERA, ATP7B, NEK5, HVCN1 and MARK2. The gene that may be associated with CPAs is CC2D2A. Genes that may be affected by both VT and CPAs were PGK1, SLC7A3, FITM2, NPM3, ISCU, CWC15 and PSAP. DERA was up-regulated when reduced VT (-269 °C) was used, which may have contributed to the recovery and stress prevention of vitrified oocyte.
2022-04-02 | GSE133550 | GEO