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Role of Tet1 and Tet3 genes and Chromatin Remodeling in Cerebellar Circuit Formation [gene expression]

ABSTRACT: Transcriptome analysis of mRNA samples purified from developing cerebellar granule cells and ES cell-derived granule cells using translating ribosome affinity purification (TRAP) method. Although mechanisms underlying early steps in cerebellar development are known, evidence is lacking on genetic and epigenetic changes during the establishment of the synaptic circuitry. Using metagene analysis, we report pivotal changes in multiple reactomes of epigenetic pathway genes in cerebellar granule cells (GCs) during circuit formation. During this stage, Tet genes are up-regulated and vitamin C activation of Tet enzymes increases the levels of 5-hydroxymethylcytosine (5hmC) at exon start sites of up-regulated genes, notably axon guidance genes and ion channel genes. Knockdown of Tet1 and Tet3 by RNA interference in ex vivo cerebellar slice cultures inhibits dendritic arborization of developing GCs, a critical step in circuit formation. These findings demonstrate a role for Tet genes and chromatin remodeling genes in the formation of cerebellar circuitry.

ORGANISM(S): Mus musculus

PROVIDER: GSE74400 | GEO | 2015/10/31

SECONDARY ACCESSION(S): PRJNA300291

REPOSITORIES: GEO

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Role of Tet1 and Tet3 genes and Chromatin Remodeling in Cerebellar Circuit Formation [gene expression]

Project description:Transcriptome analysis of mRNA samples purified from developing cerebellar granule cells and ES cell-derived granule cells using translating ribosome affinity purification (TRAP) method. Although mechanisms underlying early steps in cerebellar development are known, evidence is lacking on genetic and epigenetic changes during the establishment of the synaptic circuitry. Using metagene analysis, we report pivotal changes in multiple reactomes of epigenetic pathway genes in cerebellar granule cells (GCs) during circuit formation. During this stage, Tet genes are up-regulated and vitamin C activation of Tet enzymes increases the levels of 5-hydroxymethylcytosine (5hmC) at exon start sites of up-regulated genes, notably axon guidance genes and ion channel genes. Knockdown of Tet1 and Tet3 by RNA interference in ex vivo cerebellar slice cultures inhibits dendritic arborization of developing GCs, a critical step in circuit formation. These findings demonstrate a role for Tet genes and chromatin remodeling genes in the formation of cerebellar circuitry. We analyzed gene expression of cerebellar granule cells and ES cell-derived granule cells using the Affymetrix mouse gene 1.0 ST platform. Array data was processed by metagene analysis which was developed by the Broad Institute.

2015-10-31 | E-GEOD-74400 | biostudies-arrayexpress

Role of Tet1 and Tet3 genes and Chromatin Remodeling in Cerebellar Circuit Formation [hMe-Seal]

Project description:Although mechanisms underlying early steps in cerebellar development are known, evidence is lacking on genetic and epigenetic changes during the establishment of the synaptic circuitry. Using metagene analysis, we report pivotal changes in multiple reactomes of epigenetic pathway genes in cerebellar granule cells (GCs) during circuit formation. During this stage, Tet genes are up-regulated and vitamin C activation of Tet enzymes increases the levels of 5-hydroxymethylcytosine (5hmC) at exon start sites of up-regulated genes, notably axon guidance genes and ion channel genes. Knockdown of Tet1 and Tet3 by RNA interference in ex vivo cerebellar slice cultures inhibits dendritic arborization of developing GCs, a critical step in circuit formation. These findings demonstrate a role for Tet genes and chromatin remodeling genes in the formation of cerebellar circuitry. 5hmC-enriched genomic DNA fragments were purified with a selective labeling strategy known as hMe-Seal and carried out DNA deep sequencing using Illumina HiSeq 2500.

2015-10-31 | E-GEOD-74401 | biostudies-arrayexpress

Role of Tet1 and Tet3 genes and Chromatin Remodeling in Cerebellar Circuit Formation [hMe-Seal]

2015-10-31 | GSE74401 | GEO

Histone bivalency regulates the timing of cerebellar granule cell development in vivo [RNA-seq]

Project description:Developing neurons undergo a progression of morphological and gene expression changes as they transition from neuronal progenitors to mature, multipolar neurons. Here we use RNA-seq and ChIP-seq to analyze how chromatin modifications control gene expression in a specific type of CNS neuron, the mouse cerebellar granule cell (GC). We find that in proliferating GC progenitors, H3K4me3/H3K27me3 bivalency is common at neuronal genes and undergoes dynamic changes that correlate with gene expression during migration and circuit formation. Inhibiting H3K27 methyltransferases EZH1 and EZH2 in vitro and in organotypic cerebellar slices dramatically altered the expression of bivalent genes and induced the downregulation of migration-related genes and upregulation of synaptic genes, inhibited glial-guided migration and accelerated dendrite formation. These data show that histone bivalency is required to regulate the timing of the progression from progenitor cells to mature neurons.

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Histone bivalency regulates the timing of cerebellar granule cell development in vivo [ChIP-seq]

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Zeb2 is Essential for Bergmann Glial Development and Cerebellar Organization

Project description:Cerebellar circuitry is critical for balance and motor control among a wide array of functions and largely consists of granule and Purkinje neurons. Bergmann glia in the cerebellum form distinct morphological structures that facilitate granule neuron migration during development and that maintain the cerebellar organization and functional integrity. At present, molecular control of the formation and morphogenesis of Bergmann glia remains obscure. In this study, we found that Zeb2 (a.k.a. Sip1 or Zfhx1b), a Mowat-Wilson syndrome-associated transcriptional regulator, is highly restricted to Bergmann glia and is essential for their development and maturation. The mice with Zeb2 ablation in the cerebellar neural progenitor exhibit dysgenesis of cerebellar cortical lamination and locomotion defects. Deletion of Zeb2 markedly reduced Bergmann glial proliferation, differentiation and the establishment of the normal radial scaffold, disrupting migration of granule cell progenitors from external to internal granular layers. Transcriptome profiling indicated that Zeb2 regulates multiple pathways including FGF and Notch signaling as well as axonal guidance cues including Netrin G2 and Gdf10 to control Bergmann glial development. Our data reveal that Zeb2 acts as a transcriptional integrator of diverse signaling pathways to regulate the formation and morphogenesis of Bergmann glia ensuring maintenance of cerebellar integrity, suggesting that Zeb2 dysfunction in Bergmann glia might contribute to motor deficits in Mowat-Wilson syndrome.

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Notch signaling between cerebellar granule cell progenitors

Project description:During cerebellar development, maternal granule cell progenitors (GCPs) divide to produce not only postmitotic granule cells (GCs) but also sister GCPs. However, molecular machinery to proportionally produce distinct sister cell types from seemingly uniform GCPs is still elusive. Here we report Notch signaling makes a difference among GCPs, leading to their proportional differentiation.

2020-06-27 | GSE153313 | GEO

Temporarily epigenetic repression in Bergmann glia regulates the migration of Granule cells (RNA-seq)

Project description:Forming tight interaction with both Purkinje and granule cells (GCs), Bergmann glia (BG) are essential for cerebellar morphogenesis and neuronal homeostasis. However, how BG act in this process is unclear without comprehensively transcriptomic landscape of BG. Here, high temporal-resolution investigation of transcriptomes with FACS-sorted BG revealed the dynamic of genes within given functions and pathways enabled BG to assist neural migration and construct neuron-glia network. We found the peak time of GCs migration (P7-10) was strikingly coincided with the downregulation of extracellular matrix (ECM) related genes by prevailing transcriptional repression, and the disrupting of which by Setdb1 ablation at P7-10 in BG led to the significant migration defect of GCs by abnormal ECM expression emphasizing the criticality of Nfix-Setdb1 mediated H3K9me3 repressive complex for the precise regulation of GCs migration in vivo. Thus, BG’s transcriptomic landscapes offer an insight into the mechanism by which BG are in-depth integrated in cerebellar neural network.

2021-04-14 | GSE155871 | GEO

Temporarily epigenetic repression in Bergmann glia regulates the migration of Granule cells (ChIP-seq)

2021-04-14 | GSE155872 | GEO

Human iPSC-derived organoids recapitulate development of the cerebellum.

Project description:Single-cell profiling of stem cell-derived cerebellar organoids revealed transcriptionally-discrete populations encompassing the major cerebellar neuronal cell types including granule cells, roof plate, choroid plexus, Bergmann glia, Purkinje cells and glutamatergic deep cerebellar nuclei. Cellular identity and maturity were confirmed through comparison to an atlas of developing murine cerebellar cell types.

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