Project description:Oocyte-derived paracrine factors and estrogens cooperate to regulate the function and development of mouse cumulus cells. Cumulus oocyte complexes (COCs) were isolated from ovaries of mice. Oocytes were removed from some complexes (OOX). Groups were cultured with/without oocytes with/without estrogen and then cumulus cell transcriptome analyzed by microarrays.
Project description:PURPOSE: To provide a detailed gene expression profile of the normal postnatal mouse cornea. METHODS: Serial analysis of gene expression (SAGE) was performed on postnatal day (PN)9 and adult mouse (6 week) total corneas. The expression of selected genes was analyzed by in situ hybridization. RESULTS: A total of 64,272 PN9 and 62,206 adult tags were sequenced. Mouse corneal transcriptomes are composed of at least 19,544 and 18,509 unique mRNAs, respectively. One third of the unique tags were expressed at both stages, whereas a third was identified exclusively in PN9 or adult corneas. Three hundred thirty-four PN9 and 339 adult tags were enriched more than fivefold over other published nonocular libraries. Abundant transcripts were associated with metabolic functions, redox activities, and barrier integrity. Three members of the Ly-6/uPAR family whose functions are unknown in the cornea constitute more than 1% of the total mRNA. Aquaporin 5, epithelial membrane protein and glutathione-S-transferase (GST) omega-1, and GST alpha-4 mRNAs were preferentially expressed in distinct corneal epithelial layers, providing new markers for stratification. More than 200 tags were differentially expressed, of which 25 mediate transcription. CONCLUSIONS: In addition to providing a detailed profile of expressed genes in the PN9 and mature mouse cornea, the present SAGE data demonstrate dynamic changes in gene expression after eye opening and provide new probes for exploring corneal epithelial cell stratification, development, and function and for exploring the intricate relationship between programmed and environmentally induced gene expression in the cornea. Keywords: other
Project description:The oocyte forms a complex with their somatic cumulus cells within the follicle throughout the preovulatory maturation steps. Cumulus cells support their oocyte not only through mechanical protection but also with a close bidirectional exchange of metabolites. Analysis of the oocytes cumulus gives the opportunity to explore non-invasively oocytal well-being and quality. In vitro maturation (IVM) is the first rate-limiting step in in vitro embryo production. Analysis of protein expression in cumulus cells around this critical step helps to explore the impact of maturation conditions and to examine an influence on maturational competence of the oocyte. The goal of this study was the comparison of the cumulus proteome of oocytes with and without maturational competence matured under in vivo and in vitro conditions. Therefore twenty cumulus samples corresponding to single oocytes were analysed. Half of the samples were matured in vivo and the other half in vitro. For each maturation group, cumulus from oocytes matured successfully (SM; n=5) and failed to mature (FM; n=5) were analysed.
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.
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.
Project description:Cumulus cells, surrounding the oocyte, play a key role in the acquisition of oocyte competence to be fertilized and to sustain early embryo development. Cumulus cells contribute to oocyte development by metabolizing energy substrates such as glutathione that may protect the oocyte from oxidative stress damages. The aim of our study was to compare transcriptomics profiles of cumulus enclosed (CEO) and cumulus denuded (CDO) oocytes after in vitro maturation. Global transcriptional profiling was performed using cumulus enclosed and cumulus denuded oocytes after in vitro maturation. Matured oocytes were obtained after 22h of maturation with (CEO) or without (CDO) cumulus cells and four replicates of 25 oocytes were collected for RNA extraction. Gene expression analysis was performed by comparing CDO versus CEO oocytes that represents a total of 8 slides using a dye swap hybridisation protocol.
Project description:The maturation of an oocyte, the female gamete, during folliculogenesis is highly dependent on the molecular interaction with the somatic cells. Here we generated transcriptome data of single oocytes and the surrounding cumulus cells from antral follicles, and developed an analytical framework to unveil their inter-cellular communication. We identified hundreds of ligand-receptors pairs that can transduce paracrine signaling between oocyte and cumulus. Several of the ligand-coding genes expressed in oocytes and cumulus cells are also functionally associated with regulation of transcription. Ligand-coding genes expressed in oocytes or cumulus showed distinct enrichment for biological functions that are likely associated with a coordinated formation of the transzonal projections from the cumulus that reach the oocyte’s membrane. Finally, there were thousands of gene pairs with non-trivial linear co-expression pattern between oocytes and cumulus cells. Our analyses revealed a complex and functional regulatory circuit between the oocyte and surrounding cumulus cells.