Mitochondrial supplementation at fertilization enhances early developmental outcome
Ontology highlight
ABSTRACT:
ORGANISM(S): Sus scrofa
PROVIDER: GSE65310 | GEO | 2016/09/01
SECONDARY ACCESSION(S): PRJNA273654
REPOSITORIES: GEO
Similar Datasets
Project description:Competent oocytes can be discriminated by BCB staining. Positive stained oocytes are considered more competent than BCB negative oocyte, and injection of BCB+ oocyte extracted mitochondria into BCB negative oocytse can increase fertilisation and blastocyst rate. Here we have analysed the impact of mitochondrial supplementation on subsequent blastocyst transcriptome using agilent one color microarray that is specificly design to study the porcine embryo preimplantation period. Blastocysts were produced by intra cytoplasmic sperm injection (ICSI) from BCB positive and BCB negative oocytes as well as BCB negative oocytes supplemented with mitochondrial extract during ICSI (mICSI), and 3-4 single blatocyst transcriptomes were analysed for each group. 3-4 single blastocysts were analysed at the RNA level after whole transcriptome amplification, and level of gene expression was compared between groups, i.e ICSI BCB+ blastocysts (4), ICSI BCB- blastocysts (3) and mICSI BCB- blastocysts (4).
2016-09-01 | E-GEOD-65310 | biostudies-arrayexpress
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: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:Oocyte competence is progressively acquired during follicle growth, with large follicles containing more often competent oocytes than small follicles. A competent oocyte is able to mature in vitro, be fertilized, and develop to the blastocyst stage. Extracellular vesicles (EVs) and their cargo micro RNAs (miRNAs) are critical in mediating intercellular communication inside the follicle and facilitating oocyte maturation. Whether follicular EVs are also involved in the acquisition of oocyte competence in the growing follicle remains unclear. Here, we have been using an individual culture system to distinguish follicular fluid and their corresponding bovine oocytes as competent or noncompetent. We used a qEV single-column method to isolate EVs from follicular fluid. RNA sequencing revealed 16 differentially expressed (DE) miRNAs in EVs from follicular fluid derived from either competent or noncompetent oocytes . Among the up-regulated miRNAs, we selected bta-miR-34c to further validate its effect on oocyte competence during in vitro maturation using mimics and inhibitors. By supplementing miR-34c mimics to the oocyte maturation medium, we could significantly improve blastocyst quality, as evidenced by higher cell numbers, while supplementation of miR-34c inhibitors produced the opposite effect. Taken together, the up-regulation of miR-34c in EVs derived from follicular fluid from competent oocytes is indicative of its regulatory role, and, when added during in vitro maturation of unselected oocytes, is able to increase total cell number and inner cell mass of resulting embryos, thus contributing to the development of healthy offspring
2024-09-03 | GSE250281 | GEO
Project description:Somatic cells surrounding the oocyte were sampled at the following stages: developmentally incompetent or poorly competent prophase I oocytes (NC1 oocytes), developmentally competent prophase I oocytes (C1 oocytes), and developmentally competent metaphase II oocytes (C2 oocytes). NC1 samples were collected from late vitellogenic females (LV), C1 samples from post-vitellogenic females (PV), and C2 samples from females undergoing meiotic maturation (Germinal Vesicle Breakdown) Global transcriptional profiling was performed using somatic cells collected from rainbow trout ovarian follicles during in vivo oocyte developmental competence acquisition. Somatic cells were collected at 3 stages of oogenesis: NC1 stage follicles (LV, late vitellogenic, prophase I arrested oocytes, meiotically incompetent and developmentally incompetent, n=6), C1 stage follicles (PV, post-vitellogenic, prophase I arrested oocytes, meiotically competent and developmentally competent, n=16). Ovulatory follicles were also collected during oocyte maturation after in vivo induction (metaphase II arrested oocytes, developmentally fully competent, n=6).
2012-10-14 | E-GEOD-36602 | biostudies-arrayexpress
Project description:Following intracytoplasmic sperm injection (ICSI), in vitro culture (IVC) in ART media causes imbalance in the blastocyst's inner cell mass (ICM)with reduction of the primitive endoderm compartment compared to non-IVC blastocystsFollowing embryo transfer (ET), these imbalances do not influence birth rates, and the postnatal effects are inconspicuous. ICSI alters gene expression in mouse embryos. Choice of embryo culture media has been correlated with birth and body growth rates in human studies. No analysis has yet been made combining ICSI and IVC in the mouse. We produced mouse concepti by ICSI followed by IVC and ET so as to mimic,at least in part, the way human embryos are produced and handled in the clinical practice. Fertilized oocytes were cultured in two sets of sequential ART media, compared with an optimized mouse medium and with the mouse oviduct. Embryos were analyzed or implanted to produce newborns. In total, 3706 fertilized mouse oocytes, 1672 blastocysts and 78 newborns were examined during the course of the study. Mice of the B6C3F1, C57Bl/6 and CD1 strains were used as oocyte donors, sperm donors and embryo recipients, respectively. B6C3F1 oocytes were fertilized by intracytoplasmic C57Bl/6 sperm injection and were allowed to cleave in HTF/MultiBlast and in ISM1/ISM2 sequential culture protocols as well as in optimized mouse medium (KSOM(aa)) and in vivo in the CD1 mouse oviduct. Cleavage and blastocyst rates were analyzed by Wilcoxon / Kruskal-Wallis test and the blastocysts were split in two subgroups:one subgroup was analyzed for cell lineage allocation and for transcriptome, the other subgroup was transferred to genital tract, allowed to develop to term and characterized for birth rate, litter size, neonatal bodyweight and behavior using ANOVA F-test and t-test. The effects of ART media on blastocyst rates exceed those of ICSI. Protein and mRNA analysis reveal that IVC has a marked effect on the primitive endoderm lineage of ICSI blastocysts. In contrast to preimplantation, birth rates after ET are indistinguishable not only among ICSI but also in comparison to non-injected controls. At birth, ICSI-IVC-ET concepti are outwardly normal and weigh the same as non-ICSI controls. Confounding effects of subfertility, lifestyle and genetic heterogeneity are reduced to a minimum in the hybrid inbred mouse model compared to human patients. Confounding effects of polyspermy and parthenogenetic activation are prevented by ICSI. There is more to ART than ICSI, IVC and ET, and only a small number of ART media were examined. Preimplantation effects of ART culture media can effectively be modeled in the mouse because the in vitro environment is suboptimal for both mouse and human. However, in the optimal uterine environment of their own species, mouse and human ART blastocysts may be supported and selected for differently, depending on species-specific features such as the kinetics of implantation and the endometrium. 14 samples were analyzed. HTF/MultiBlast: Mouse multiblastocysts in HTF medium, 3 biological rep ISM1/ISM2: Mouse blastocysts in ISM1 and ISM2 medium, 3 biological rep KSOM(aa): Mouse blastocysts in KSOM(aa) medium, 2 biological rep oviduct: Mouse oviduct, 3 biological rep micromanipulation control: Mouse micromanipulation control, 3 biological rep
2016-08-25 | E-GEOD-50193 | biostudies-arrayexpress
Project description:Somatic cells surrounding the oocyte were sampled at the following stages: developmentally incompetent or poorly competent prophase I oocytes (NC1 oocytes), developmentally competent prophase I oocytes (C1 oocytes), and developmentally competent metaphase II oocytes (C2 oocytes). NC1 cumulus cells (CC) were sampled from immature calf oocytes, C1 samples from immature cow oocytes, and C2 samples from in vivo matured cow oocytes. Global transcriptional profiling was performed using cumulus cells collected from bovine ovarian follicles during in vivo oocyte developmental competence acquisition. Cumulus cells were collected at 3 stages: early stage follicles (prophase I arrested oocytes, meiotically competent but developmentally incompetent, n=6), late stage follicles (prophase I arrested oocytes, meiotically competent and developmentally competent, n=6) and ovulatory follicles collected by ovum pick-up (OPU) in vivo (metaphase II arrested oocytes, developmentally fully competent, n=5).
2012-10-14 | E-GEOD-36605 | biostudies-arrayexpress
Project description:Cumulus cells surrounding the oocyte were sampled at the following stages: developmentally incompetent or poorly competent prophase I oocytes (NC1 oocytes), developmentally competent prophase I oocytes (C1 oocytes), and developmentally competent metaphase II oocytes (C2 oocytes). NC1 samples were collected from immature, unexpanded cumulus-oocytes complexes (COC) from prepubertal (3-week-old) mice, C1 samples from immature, unexpanded cumulus-oocytes complexes (COC) from adult (8-week-old) and C2 samples from mature, expanded COCs obtained from the oviduct from 8-week-old mice after standard superovulation protocol. Global transcriptional profiling was performed using cumulus cells collected from murine ovarian follicles during in vivo oocyte developmental competence acquisition. Cumulus cells were collected at 3 stages: early stage follicles (prophase I arrested oocytes, meiotically competent but developmentally incompetent, n=5), late stage follicles (prophase I arrested oocytes, meiotically competent and developmentally competent, n=5) and ovulatory follicles collected in vivo (metaphase II arrested oocytes, developmentally fully competent, n=5).
2012-10-14 | E-GEOD-36604 | biostudies-arrayexpress
Project description:Somatic cells surrounding the oocyte were sampled at the following stages: developmentally incompetent or poorly competent prophase I oocytes (NC1 oocytes), developmentally competent prophase I oocytes (C1 oocytes), and developmentally competent metaphase II oocytes (C2 oocytes). NC1 samples were collected from immature stage IV follicles, C1 samples from immature stage VI follicles, and C2 samples from in vitro matured stage VI follicles. Global transcriptional profiling was performed using somatic cells collected from xenopus ovarian follicles during in vivo oocyte developmental competence acquisition. Somatic cells were collected at 3 stages of oogenesis: early stage follicles (stage IV, vitellogenic, prophase I arrested oocytes, meiotically competent but developmentally incompetent, n=5), late stage follicles (stage VI, post-vitellogenic, prophase I arrested oocytes, meiotically competent and developmentally competent, n=5) and ovulatory follicles collected after in vitro maturation induction with hCG of post-vitellogenic follicles (metaphase II arrested oocytes, developmentally fully competent, n=5).
2012-10-14 | E-GEOD-36603 | biostudies-arrayexpress
Project description:Ten volunteers underwent 13 research IVM cycles. Immature oocytes were retrieved after mild ovarian stimulation with gonadotropins but no hCG ovulation trigger, in vitro matured and fertilized using ICSI. Oocyte maturation rate in vitro was 47% (94/199), the mean fertilization rate was 71% (67/94) and 37% (25/67) of mature oocytes developed into a morphologically good quality embryo on day 3 after ICSI, after which the blastomeres were analysed by single-cell aCGH. Sixty-seven of the 111 blastomeres analysed showed at least one chromosomal abnormality. Two out of seventeen embryos had only normal cells while one embryo carried a meiotic abnormality, 11 were mosaic and three were chaotic. Although the number of embryos tested as well as the data available in the literature are too small to allow any statistical analysis, these data are comparable to the few data available on the total chromosome complement of day 3 ICSI embryos. This would indicate that other mechanisms than chromosomal abnormalities explain the low efficiency and high miscarriage rate in IVM cycles.
2014-11-12 | GSE56799 | GEO