Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Expression data from wildtype and Dicer-deficient mouse embryonic stem (ES) cells at different stages of the cell cycle.

ABSTRACT: The protein Dicer is required for microRNA (miRNA) biogenesis. Dicer-deficient cells therefore lack almost all mature, functional miRNAs. We investigated the role of miRNAs in regulation of gene expression in mouse ES cells by analysing gene expression in either wildtype or Dicer-deficient cells grown either under asynchronous growth conditions, or cells in the early G1 phase of the cell cycle. RNA was extracted from three biological replicates of both wildtype and Dicer-deficient ES cells grown under asynchronous culture conditions. To obtain cells in early G1, populations (in triplicate for both cell types) were cultured in 20ng/ml demecolcine for 4 hours, then mitotic cells were isolated using the mitotic shake-off technique (Terasima & Tolmach, Experimental Cell Research (30) 1963). Mitotic cells were replated in demecolcine-free media for 2 hours, allowing them to progress in a highly synchronised manner into early G1.

ORGANISM(S): Mus musculus

SUBMITTER: Gopuraja Dharmalingam

PROVIDER: E-GEOD-60096 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

MicroRNAs of the miR-290-295 Family Maintain Bivalency in Mouse Embryonic Stem Cells.

Graham Bryony B Marcais Antoine A Dharmalingam Gopuraja G Carroll Thomas T Kanellopoulou Chryssa C Graumann Johannes J Nesterova Tatyana B TB Bermange Anna A Brazauskas Pijus P Xella Barbara B Kriaucionis Skirmantas S Higgs Douglas R DR Brockdorff Neil N Mann Matthias M Fisher Amanda G AG Merkenschlager Matthias M

Stem cell reports 20160414 5

Numerous developmentally regulated genes in mouse embryonic stem cells (ESCs) are marked by both active (H3K4me3)- and polycomb group (PcG)-mediated repressive (H3K27me3) histone modifications. This bivalent state is thought to be important for transcriptional poising, but the mechanisms that regulate bivalent genes and the bivalent state remain incompletely understood. Examining the contribution of microRNAs (miRNAs) to the regulation of bivalent genes, we found that the miRNA biogenesis enzyme ...[more]

PMID: 27150236

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Project description:The endoribonuclease, Dicer, is indispensible for generating the majority of mature microRNAs (miRNAs), which are posttranscriptional regulators of gene expression involved in a wide range of developmental and pathological processes in mammalian central nervous system. While functions of Dicer-dependent miRNA pathways in neurons and oligodendrocytes have been extensively investigated, little is known about the role of Dicer in astrocytes. Here we report the effect of Cre-loxP mediated conditional deletion of Dicer selectively from postnatal astroglia on brain development. Dicer-deficient mice exhibited normal motor development and neurological morphology prior to postnatal week 5. Thereafter mutant mice invariably developed a rapidly fulminant neurological decline characterized by ataxia, severe progressive cerebellar degeneration, seizures, uncontrollable movements and premature death by postnatal week 9-10. Integrated transcription profiling, histological and functional analyses of cerebella showed that deletion of Dicer in cerebellar astrocytes altered the transcriptome of astrocytes to be more similar to an immature or reactive-like state prior to the onset of neurological symptoms or morphological changes. As a result, critical and mature astrocytic functions including glutamate uptake and antioxidant pathways were substantially impaired, leading to massive apoptosis of cerebellar granule cells and degeneration of Purkinje cells. Collectively, our study demonstrates the critical involvement of Dicer in normal astrocyte maturation and maintenance. Our findings also reveal non-cell autonomous roles of astrocytic Dicer-dependent pathways in regulating proper neuronal functions and implicate that loss of or dysregulation of astrocytic Dicer-dependent pathways may be involved in neurodegeneration and other neurological disorders. Four replicate experiments using samples derived from biologically independent pairs of control (mGfap-Cre; Dicer +/flox) and Dicer mutant (mGfap-Cre; Dicer flox/flox) littermate mice (postnatal day 30) were performed with a dye-swap experimental design.

Dataset Information

Expression data from wildtype and Dicer-deficient mouse embryonic stem (ES) cells at different stages of the cell cycle.

Publications

MicroRNAs of the miR-290-295 Family Maintain Bivalency in Mouse Embryonic Stem Cells.

Similar Datasets

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