Zeb2 is Essential for Bergmann Glial Development and Cerebellar Organization
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE84098 | GEO | 2018/06/25
REPOSITORIES: GEO
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Project description:Bergmann glial cells of the vertebrate cerebellum play essential roles in the development and maintenance of cerebellar structure and function. During development, Bergmann glia provide structural support to the expanding cerebellar anlage and also serve as guides for migrating neurons (granule cells). As the cerebellum matures, Bergmann glia become important in dendritic arborization, synapse maintenance and synaptic function. The molecular mechanisms underlying these diverse and important functions of Bergmann glia remain largely unknown. We used microarray analysis to examine global gene expression in individual Bergmann glial cells derived at P6 (a time of extensive neuronal migration and cerebellar growth) and at P30 (when cerebellar development is complete and Bergmann glia play important roles at synapses). After gentle dissociation of cerebellar tissue derived from mice expressing GFP under the GFAP gene promoter (GFAP-GFP mice) (Zhuo, L., et al., 1997, Developmental biology), single GFP-positive Bergmann glial cells were aspirated into microcapillary tubes. Amplified cDNAs were prepared from single cells using RT-PCR and hybridized to Affymetrix GeneChip Mouse Genome 430 2.0 expression arrays (one array per cell). Five P6 cells and five P30 cells were used to generate the data presented in this study.
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Project description:Bergmann glial cells of the vertebrate cerebellum play essential roles in the development and maintenance of cerebellar structure and function. During development, Bergmann glia provide structural support to the expanding cerebellar anlage and also serve as guides for migrating neurons (granule cells). As the cerebellum matures, Bergmann glia become important in dendritic arborization, synapse maintenance and synaptic function. The molecular mechanisms underlying these diverse and important functions of Bergmann glia remain largely unknown. We used microarray analysis to examine global gene expression in individual Bergmann glial cells derived at P6 (a time of extensive neuronal migration and cerebellar growth) and at P30 (when cerebellar development is complete and Bergmann glia play important roles at synapses).
2010-02-12 | GSE18617 | GEO
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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Project description:Zeb2 is Essential for Bergmann Glial Development and Cerebellar Organization
| PRJNA328019 | ENA
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 | GSE155872 | GEO
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.
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Project description:Neurogenic astroglia-like cells of the prospective white matter (PWM) generate interneurons and astroglia during cerebellar development. However, the characterization of PWM subpopulations is still insufficient. Here we show that ACSA-2 (Astrocyte Cell Surface Antigen-2) is an intrinsic marker of astrocyte-committed precursors. In the developing cerebellum, the ontogeny of ACSA-2+ astrocytes revealed this epitope to be exclusively expressed by PWM cells. By contrast, in the adult brain, ACSA-2 is observed on parenchymal astrocytes, fibrous astrocytes of the white matter and at low level on Bergmann glia. In combination with GLAST, a marker for astroglia-like progenitors, we addressed the characteristics of neurogenic (ACSA-2-/GLAST+) and astrocyte-committed (ACSA-2+/GLAST+) precursors using transcriptomics and cell transplantation assays. Gene expression analyses identified major differences in genes related to the maturation status of astrocytes, their potential to generate interneurons and genes required for Bergmann glia specification. Transplantation assays further confirmed a divergent differentiation potential: ACSA-2+/GLAST+ cells, on the one side, differentiate into astrocytes and oligodendrocytes but not into Bergmann glia or neurons when transplanted into the neonatal cerebellum. On the other side, ACSA-2-/GLAST+ progenitors were able to generate interneurons and all the glial phenotypes. Moreover, ACSA-2-/GLAST+ progenitors even maintained the neurogenic potential when transplanted into the adult cerebellum. In conclusion, this work reports about ACSA-2, a marker for glial-restricted precursors of the PWM that in combination with GLAST enables for the discrimination and isolation of neurogenic and astrocyte-committed progenitors of the neonatal cerebellum.
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Project description:The multi-domain transcription factor ZEB2 controls embryonic and adult cell fate decisions and maturation in many stem/progenitor cell types. Defects in these processes in specific cell types underlie Mowat-Wilson syndrome (MOWS), which is caused by ZEB2 haplo-insufficiency. Human ZEB2, like mouse Zeb2, is located on chromosome 2 downstream of an approximately 3.5 Mb-long gene-desert, lacking any protein-coding gene sequence. Using temporal high-resolution Targeted Chromatin Capture (T2C), we show major chromatin structural changes based on mapping in-cis proximities between the ZEB2 gene promoter and this gene desert during neural differentiation of induced pluripotent cells (iPSCs), including at early neuroprogenitor cell (NPC)/rosette state, where ZEB2 mRNA levels increase significantly. Combining T2C with histone-3 acetylation mapping, we identified three novel candidate enhancers about 500 kb upstream of the ZEB2 transcription start site (TSS). Functional luciferase-based assays in heterologous cells and NPCs reveal co-operation between these three enhancers. This study is the first to document in-cis regulatory elements (REs) located in ZEB2’s gene desert. The results further show the usability of T2C for future studies of ZEB2 REs in differentiation and maturation of relevant cell types, and the molecular characterization of identified MOWS patients that lack mutations in ZEB2 protein-coding exons.
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Project description:Familial amyotrophic lateral sclerosis (ALS) represents about 10% of ALS cases. In about 20% of familial ALS patients, a mutation in superoxide dismutase-1 (SOD1) can be found. The ubiquitous SOD1 protein converts superoxide radical anions to oxygen and hydrogen peroxide. Patients with familial ALS caused by mutations in SOD1 can show comorbidity with frontotemporal dementia and develop cognitive impairment, including apathy, inattention, verbal deficits, and hypersexuality. At the cellular level, pathological signs of ALS may include tau immunoreactive astrocytic and neuronal inclusions, suggesting that cognitive dysfunction in ALS may also reflect abnormal protein metabolism of the microtubule associated protein (MAP), tau. To identify cell-specific expression changes, we performed laser capture microdissection (LCM) to isolate anterior horn motor neurons and surrounding cells. To determine common pathways for the development of ALS due to dysfunction of SOD1 or TAU, we performed whole transcriptome analysis in SOD1 and TAU mouse models. Because ALS is a neurodegenerative disease that specifically affects motor neurons, these cells were the main investigative target. Global transcriptomes of glial cells surrounding the motor neurons were also assessed, because these cells have been implicated as triggers of neurodegeneration. Mouse experiments were performed at the presymptomatic stage, prior to the onset of cell loss, in order to reduce false positive signals due to tissue reactive changes. Lumbar anterior horn anterior horn motor neurons and surrounding cells (glia) were isolated from presymptomatic TAU-P301L and SOD1-G93A transgenic mice using laser capture microdissection (LCM; PixCell® IIe LCM System, Arcturus, Molecular Devices, CA). On average, 500 motor neuron bodies or surrounding glial cells from 20 tissue sections per animal were collected. RNA isolation from LCM collected cells was performed using the Qiagen RNeasy Micro Kit. Only RNA with RNA Integrity Numbers (RIN) above 7.5 were taken for further analysis (Agilent Bioanalyzer 2100). A two-round T7-based amplification and labeling protocol (Agilent Low RNA Input Linear Amplification Kit PLUS) was used to generate high quality labeled cRNA. Agilent Whole Mouse Genome Microarray comparisons of motor neurons and surrounding glia were performed between transgenic (SOD1G93A and TAUP301L) and corresponding nontransgenic (control) littermate animals, producing four independent comparison groups: SOD1G93A motor neurons versus control motor neurons (SOD1mn), SOD1G93A motor neuron surrounding glia versus control glia (SOD1gl), TAUP301L motor neurons versus control motor neurons (TAUmn) and TAUP301L glia versus control glia (TAUgl). Each comparison used four pairs of transgenic (SOD1G93A or TAUP301L) vs. corresponding control littermate animals, producing four biological replicates. Raw microarray data were acquired using the Agilent DNA Microarray scanner and processed with the accompanying Agilent Feature Extraction 10.5 Image Analysis software using default settings. Normalized signal intensities were used to identify gene expression changes in SOD1G93A and TAUP301L motor neurons and surrounding glial cells, generating four partially overlapping sets of data. For the identification of differential expression, the genes were required to pass two conservative criteria: a ratio beyond the 99.5% confidence interval observed in homotypic comparisons, which corresponded to an approximately 1.5-fold expression change, and a paired t-test (P<0.01) computed using 100 permutations of the data for each gene. Correction for multiple comparisons was performed using the adjusted Bonferroni test. The analysis was performed in the TM4: Microarray Software Suite. These techniques identified 251 transcripts representing 186 known genes for which expression was altered in at least one of the four comparisons. Four-condition experiment, SOD1G93A motor neurons versus control motor neurons (SODmn), SOD1G93A motor neuron surrounding glia versus control glia (SODgl), TAUP301L motor neurons versus control motor neurons (TAUmn) and TAUP301L glia versus control glia (TAUgl). Each comparison used four pairs of transgenic (SOD1G93A or TAUP301L) vs. corresponding control littermate animals, producing four biological replicates. Biological replicates: 4 SOD1G93A motor neurons (SODmn), 4 non SOD littermate motor neurons (nonSODmn), 4 SOD1G93A motor neuron surrounding glia (SODgl), 4 non SOD littermate motor neuron surrounding glia (nonSODgl), 4 TAUP301L motor neurons (TAUmn), 4 non TAU littermate motor neurons (nonTAUmn), 4 TAUP301L motor neuron surrounding glia (TAUgl), 4 non TAU littermate motor neuron surrounding glia (nonTAUgl). Four independent comparison groups were generated: SOD1G93A motor neurons versus control nonSOD motor neurons, SOD1G93A motor neuron surrounding glia versus control nonSOD glia, TAUP301L motor neurons versus control nonTAU motor neurons and TAUP301L glia versus control nonTAU glia. Each comparison used four pairs of transgenic (SOD1G93A or TAUP301L) vs. corresponding control littermate animals, producing four biological replicates. Each replicate was repeated twice with dye flip to correct for unequal dye incorporation rates. Therefore, eight microarray hybridizations were performed for each biological comparison, for a total of 32 microarray slides and generating four groups of differentially expressed genes in SOD1G93A motor neurons (SODmn), SOD1G93A motor neuron surrounding glial cells (SODgl), TAUP301L motor neurons (TAUmn), and TAUP301L surrounding glial cells (TAUgl).
2010-07-30 | E-GEOD-22482 | biostudies-arrayexpress
Project description:Brain development is regulated by conserved transcriptional programs across species, but little is known about divergent mechanisms that create species-specific characteristics. Among brain regions, the cerebellum is now recognized to contribute to human cognitive evolution having a broad range of non-motor cognitive functions in addition to motor control. Emerging studies highlight the complexity of human cerebellar histogenesis, compared with non-human primates and rodents, making it important to develop methods to generate human cerebellar neurons that closely resemble those in the developing human cerebellum. Here we report a rapid and simple protocol for the directed derivation of the human ATOH1 lineage, the precursor of all excitatory cerebellar neurons, from human pluripotent stem cells (hPSC), and strategies to decrease culture variability; a common limitation in hPSC studies. Upon transplantation into juvenile mice, early postmitotic hPSC-derived cerebellar granule cells migrated along glial fibers and integrated into the cerebellar cortex. By Translational Ribosome Affinity Purification (TRAP)-seq, the ATOH1 lineage most closely resembled human cerebellar tissue in the second trimester. Unexpectedly, TRAP-seq identified a heterochronic shift in the expression of RBFOX3 (NeuN) and NEUROD1, which are classically associated with differentiated neurons, within granule cell progenitors (GCPs) in the human external granule layer. This molecular divergence may provide the mechanism by which the GCP pool persists into year two post birth in humans, but only lasts for two weeks in mice. Our approach provides a scalable in vitro model of the human ATOH1 lineage that yields cerebellar granule cells within 48 days as well as a strategy for identifying uniquely human cellular and molecular characteristics.
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