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The P-body protein 4E-T regulates the balance between cell genesis and establishment of the postnatal neural stem cell pool by repressing mRNA translation [4ET_neurosphere]

ABSTRACT: To investigate the targets of 4E-T in postnatal V-SVZ precursors cultured as neurospheres We performed RNA immunoprecipitations (RIPseq) on neurosphere lysates with anti-4E-T or control anti-IgG antibodies, isolated and sequenced the resulting RNA

ORGANISM(S): Mus musculus

PROVIDER: GSE224966 | GEO | 2024/02/08

REPOSITORIES: GEO

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The P-body protein 4E-T regulates the balance between cell genesis and establishment of the postnatal neural stem cell pool by repressing mRNA translation [4ET_VSVZ]

Project description:To investigate the targets of 4E-T in the postnatal V-SVZ We performed RNA immunoprecipitations (RIPseq) on V-SVZ lysates anti-4E-T or control anti-IgG antibodies, isolated and sequenced the resulting RNA

2024-02-08 | GSE224897 | GEO

The P-body protein 4E-T regulates the balance between cell genesis and establishment of the postnatal neural stem cell pool by repressing mRNA translation [Cpeb4_neurosphere]

Project description:To investigate the targets of Cpeb4/Cpeb4-associated transcripts in postnatal V-SVZ precursors cultured as neurospheres We performed RNA immunoprecipitations (RIPseq) on neurospheres using anti-Cpeb4 or control anti-IgG antibodies, isolated and sequenced the resulting RNA

2024-02-08 | GSE224854 | GEO

The P-body protein 4E-T regulates the balance between cell genesis and establishment of the postnatal neural stem cell pool by repressing mRNA translation [Cpeb4_VSVZ]

Project description:To investigate the targets of Cpeb4 in the postnatal V-SVZ We performed RNA immunoprecipitations (RIPseq) on V-SVZ lysates anti-Cpeb4 or control anti-IgG antibodies, isolated and sequenced the resulting RNA

2024-02-08 | GSE224896 | GEO

The P-body protein 4E-T regulates the balance between cell genesis and establishment of the postnatal neural stem cell pool by repressing mRNA translation [Ribotag]

Project description:To investigate the translatome of postnatal precursors of the V-SVZ cultured as neurospheres, we employed RiboTag analysis We performed RiboTag analysis on precursors derived from NestinCreERT2;RPL22HA/HA animals using specific anti-Hemagluttinin antibodies versus non-specific IgG controls

2024-02-08 | GSE224853 | GEO

An eIF4E1/4E-T complex determines the genesis of neurons from precursors by translationally repressing a proneurogenic transcription program

Project description:Here, we have addressed the mechanisms that determine genesis of the correct numbers of neurons during development, focusing upon the embryonic cortex. We identify in neural precursors a repressive complex involving eIF4E1 and its binding partner 4E-T that coordinately represses translation of proteins that determine neurogenesis. This eIF4E1/4E-T complex is present in granules with the processing body proteins Lsm1 and Rck, and disruption of this complex causes premature and enhanced neurogenesis and neural precursor depletion. Analysis of the 4E-T complex shows that it is highly enriched in mRNAs encoding transcription factors and differentiation-related proteins. These include the proneurogenic bHLH mRNAs, which colocalize with 4E-T in granules, and whose protein products are aberrantly upregulated following knockdown of eIF4E, 4E-T, or processing body proteins. Thus, neural precursors are transcriptionally primed to generate neurons, but an eIF4E/4E-T complex sequesters and represses translation of proneurogenic proteins to determine appropriate neurogenesis. We obtained 3 biological replicates of IgG-bound RNA, 4E-T-bound RNA, and cooresponding total RNA input from mouse E12-13 cortices. RNA samples were analyzed on the Affymetrix Mouse Gene 2.0 ST Arrays.

2014-09-25 | E-GEOD-61729 | biostudies-arrayexpress

An eIF4E1/4E-T complex determines the genesis of neurons from precursors by translationally repressing a proneurogenic transcription program

2014-09-25 | GSE61729 | GEO

Culture protocol and transcriptomic analysis of murine SVZ NPCs and OPCs

Project description:The mammalian adult brain contains two neural stem and precursor (NPC) niches: subventricular zone [SVZ] lining the lateral ventricles and subgranular zone [SGZ] in the hippocampus. From these SVZ NPCs represent the largest NPC pool. Notably, while SGZ NPCs typically only produce neurons and astrocytes, SVZ NPCs produce neurons, astrocytes and oligodendrocytes throughout life. Of particular importance is the generation and replacement of oligodendrocytes, the only myelinating cells of the central nervous system (CNS). SVZ NPCs contribute to myelination by regenerating oligodendrocyte precursor cell (OPC) pool and by differentiating into oligodendrocytes in the developing and demyelinated brain. The neurosphere assay has been widely adopted by the scientific community to facilitate the study of NPCs in vitro. Here, we present a streamlined protocol for culturing postnatal and adult SVZ NPCs and OPCs from primary neurosphere cells. We characterize the purity and differentiation potential as well as provide RNA-sequencing profiles of postnatal SVZ NPCs, postnatal SVZ OPCs and adult SVZ NPCs. We show that primary neurospheres cells generated from postnatal and adult SVZ differentiate into neurons, astrocytes and oligodendrocytes concurrently and at comparable levels. SVZ OPCs are generated by subjecting primary neurosphere cells to OPC growth factors fibroblast growth factor (FGF) and platelet-derived growth factor-AA (PDGF-AA). We further show SVZ OPCs can differentiate into oligodendrocytes in the absence and presence of thyroid hormone T3. Transcriptomic analysis confirmed the identities of each cell population and revealed novel immune and signalling pathways expressed in an age and cell type specific manner.

2022-12-19 | GSE217507 | GEO

Project description:Eukaryotic translation initiation factor 4E (eIF4E)–binding protein 1 (4E-BP1) inhibits cap-dependent translation in eukaryotes by competing with eIF4G for an interaction with eIF4E. Phos-phorylation at Ser-83 of 4E-BP1 occurs during mitosis through the activity of cyclin-dependent kinase 1 (CDK1)/cyclin B rather than through canonical mTOR kinase activity. Here, we investi-gated the interaction of eIF4E with 4E-BP1 or eIF4G during interphase and mitosis. We observed that 4E-BP1 and eIF4G bind eIF4E at similar levels during interphase and mitosis. The most highly phosphorylated mitotic 4E-BP1 isoform (δ) did not interact with eIF4E, whereas a distinct 4E-BP1 phospho-isoform, EB-γ—phosphorylated at Thr-70, Ser-83, and Ser-101—bound to eIF4E during mitosis. Two-dimensional gel electropho-retic analysis corroborated the identity of the phosphorylation marks on the eIF4E-bound 4E-BP1 isoforms and uncovered a population of phosphorylated 4E-BP1 molecules lacking Thr-37/Thr-46–priming phosphorylation. Moreover, proximity ligation assays for phospho–4E-BP1 and eIF4E revealed different in situ interactions during interphase and mitosis. The eIF4E:eIF4G interaction was not inhibited, but rather increased in mitotic cells, consistent with active translation initiation during mitosis. Phospho-defective substitution of 4E-BP1 at Ser-83 did not change global translation or individual mRNA translation profiles as measured by single-cell nascent protein synthesis and eIF4G RNA-immunoprecipitation sequencing. Mitotic 5’-terminal oligopyrimidine RNA translation was active and, unlike interphase translation, resistant to mTOR inhibition. Our findings reveal the phosphorylation profiles of 4E-BP1 isoforms and their interactions with eIF4E throughout the cell cycle and indicate that 4E-BP1 does not specifically inhibit translation initiation during mitosis.

2019-06-14 | GSE131668 | GEO

Gene Set Enrichment Analysis reveals lineage relationships between mouse and human gliomas and identifies stem cells and PNET as distinct entities

Project description:Murine and human genome-wide microarray expression profiles (including GSE19404) were compared using Partek and Gene Set Enrichment Analysis (GSEA) in order to identify similarities. A murine PNET-like tumour by morphology was found to resemble human ATRT, whereas experimental glioma resembled a particular human glioblastoma subclass. Tumour material was resected or neurosphere cultures pelleted. Part of the material was processed for histological analysis, another was snap-frozen for isolation of RNA. RNA was then labelled and hybridised to Affymetrix MoEx-v1 arrays and analysed using Partek. This array dataset comprises Affymetrix MoEx-v2 microarrays hybridised with RNA from brain-derived tumour material (CA), subventricular zone (SVZ)-derived growth transformed neurospheres (CATR) and control neurospheres (CS, GFP).

2013-07-29 | E-GEOD-42515 | biostudies-arrayexpress

The translational protein 4E-T regulates the balance between differentiation and establishment of postnatal neural stem cell pools

Project description:Microarrays were performed (N=3) on postnatal precursors grown as neurospheres to determine level of gene expression To determine the transcript targets of the translational regulator 4E-T and whether these targets are repressed, we performed RIPs (GSE224966) for these targets as well as Ribotag (GSE224853) in addition to the following microarray as control to determine gene expression in posntatal precursors and ensure the differences observed in Ribotag were not simply due to differences in gene expression levels

2024-02-12 | GSE225351 | GEO

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