Transcription profiling by array of mouse ovary development to compare the expression of mRNAs and noncoding RNAs
Ontology highlight
ABSTRACT: This experiment compared the expression of mRNAs and noncoding RNAs during the development of mouse ovary. The primary aim of the experiment was to identify ncRNAs that were differentially expressed during this process.
Project description:This experiment compared the expression of mRNAs and noncoding RNAs during the development of mouse testis. The primary aim of the experiment was to identify ncRNAs that were differentially expressed during this process.
Project description:Expression profiling of micro RNAs in mouse embryonic gonads before (11.5 days post coitum) and after (13.5 days post coitum) the critical period of sex determination
Project description:Specification of germ cell fate is fundamental in development. With a highly representative single-cell microarray and rigorous quantitative-PCR analysis, we defined the genome-wide transcription dynamics that create primordial germ cells (PGCs) from the epiblast, a process that exclusively segregates them from their somatic neighbors. We also analyzed the effect of the loss of Blimp1, a key transcriptional regulator, on these dynamics. Our analysis revealed that PGC specification involves complex, yet highly ordered regulation of a large number of genes, proceeding under the strong influence of mesoderm induction with active repression of specific programs such as epithelial-mesenchymal transition, Hox gene activation, cell-cycle progression and DNA methyltransferase machinery. Remarkably, Blimp1 is essential for repressing nearly all the genes normally down-regulated in PGCs relative to their somatic neighbors, whereas it is dispensable for the activation of approximately half of the genes up-regulated in PGCs. Experiment Overall Design: Embryo isolation, dissection, and single cell cDNA amplification were performed as described (Kurimot et al., 2006, Nucleic Acids Res 34: e42; Kurimoto et al., 2007, Nature protocols, 2007, 2:739-52). Randomly picked cells were screened with gene-specific primers for Blimp1 and fragilis to select Blimp1-positive embryonic cells (which marks lineage-restricted PGC precursors or PGCs). For Blimp1 KO embryos, non-functional transcript from the Blimp1null allele was detected with the same primer set.
Project description:The main goal of our study is to identify the molecular events that determine the gonadal identity in mammals. Although testis and ovary arise from a common embryonic primordium, they represent outcomes of opposing fate determination. This decision to differentiate into a testis or an ovary hinges upon the balance between two antagonizing factors, pro-testis SOX9 and pro-ovary β-catenin. This microarray analysis led to the identification of the genes involved in the fate of XX and XY gonads in absence of SOX9 and beta-catenin We developed mouse genetic models that lack either Sox9, β-catenin, or both specifically in the somatic cells. All embryos used in this study resulted from the crossing between Ctnnb1f/f; Sox9f/f females with Sf1-cre+/Tg ; Ctnnb1+/f; Sox9+/f males. XX and XY fetal gonads were collected at embryonic day E14.5
Project description:Prophase I of meiosis involves dynamic chromosome segregation processes including synapsis, meiotic recombination, and cohesion. Genetic defects in genes participating in these processes consistently cause reproduction failure in mice. To identify candidate genes responsible for infertility or recurrent pregnancy loss in humans, we performed expression profiling of male and female gonads of mice undergoing meiotic prophase I. Both male and female gonads at 15.5 days postcoitum, neonatal day 1 and 10 weeks postpartum were dissected from mice for RNA extraction and subjected to hybridization on Affymetrix microarrays.
Project description:Testicular cord formation in male gonadogenesis involves assembly of several cell types, the precise molecular mechanism is still not well known.With the high-throughput quantitative proteomics technology, a comparative proteomic profile of mouse embryonic male gonads were analyzed at three time points (11.5, 12.5 and 13.5 days post coitum), corresponding to critical stages of testicular cord formation in gonadal development.
Project description:This series represents murine dorsal neural tube bisected along the midline with one half from each embryo used for control and the other half treated with 10-6M RA dissolved in ethanol for 6, 12, 24 or 48 h. For 6 h exposures, the explants were cultured overnight on fibronectin coated 35mm dishes (Biocoat, Becton Dickinson Labware, Bedford, MA) in DMEM with 10% horse serum in order to allow for sufficient outgrowth of neural crest cells. The RA was added the following morning; RMA Express 0.2 used to initially normalize data; GeneSpring (Silicon Genetics, Inc.) used for subsequent analysis; Samples were analyzed at 6, 12, 24, and 48 hours.
Project description:The small non-coding RNAs (sncRNAs) are considered as postranscriptional key regulators of male germ cell development. In addition to microRNAs (miRNAs) and PIWI-interacting RNAs (piRNAs), other sncRNAs generated from small nucleolar RNAs (snoRNAs), tRNAs or rRNAs processing may also play important regulatory roles in spermatogenesis. By next generation sequencing (NGS), we characterized the different sncRNA populations detected at three milestone stages in male germ differentiation: primordial germ cells (PGCs) at 13.5 dpc, pubertal spermatogonia cells, and mature spermatozoa. In order to assess the potential transmission of the sncRNAs through the mature spermatozoa during fertilization, the sncRNA population detected in male germ cells was also compared with sncRNAs detected in unfertilized mouse oocytes and zygotes. Combining the data obtained by NGS and microarrays from whole PGC and spermatogonia transcriptome, we defined the potential regulatory roles of specific miRNAs and their validated targets. Similar to miRNAs, both the small RNAs derived from snoRNAs and the piRNAs, could be involved in the postranscriptional regulation of mRNA transcripts during the male germ development. Finally, our results strongly suggest that the small RNAs-derived from tRNAs and rRNAs are interacting with PIWI proteins, and specifically with MILI. These new classes of piRNAs are not generated by the ping-pong pathway and could be the source of primary piRNAs. mRNA analysis of Primordial Germ Cells (PGCs), Spermatogonia cells (SPG), adult testis (AdT) and Gonad-less (GL) embryos. Indirect comparisons were made across multiple arrays with raw data pulled from different channels for data analysis and comparison to the control data.
Project description:RNA expression microarray analysis of prospermatogonia in 15 day post-conceptus (dpc) fetuses, a stage when they are undergoing rapid de novo DNA methylation. For comparison, we also analysed 15 dpc pachytene oogonia, 15 dpc female and male gonadal somatic cells, and adult pachytene spermatocytes.
Project description:This study evaluates the temporal pattern of gene expression in human fetal gonads covering the periods of sex differentiation and the onset of steroidogenesis in the testis, with corresponding changes in the ovary.