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A subpopulation of deep cerebellar nuclei glutamatergic neurons contributes to cerebellar control of satiation

ABSTRACT: A subpopulation of deep cerebellar nuclei glutamatergic neurons contributes to cerebellar control of satiation

PROVIDER: PRJNA764260 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15948289_1.fastq.gz	Fastqsanger.gz
	SRR15948289_2.fastq.gz	Fastqsanger.gz
	SRR15948290_1.fastq.gz	Fastqsanger.gz
	SRR15948290_2.fastq.gz	Fastqsanger.gz
	SRR15948291_1.fastq.gz	Fastqsanger.gz

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A subpopulation of deep cerebellar nuclei glutamatergic neurons contributes to cerebellar control of satiation

Project description:We profiled micro-dissected deep cerebellar nuclei (DCN) from 2-3 month old mice for single-nucleus RNAseq to study the cellular populations present in the region and in particular, to define the molecular subtype identity of glutamatergic neurons. From our data, we isolated vGluT2-expressing transcriptomes and identified a graded and anti-correlated expression of two sets of genes associated with the first principal component (PC1) in our dataset, defining separate cell populations. Multiplexed in situ hybridization analysis confirmed the nonoverlapping expression patterns and revealed their topography within the DCN. We then probed the function of these neurons in further experiments and found that one of the two populations was associated with food intake. Our broader study defines a conserved satiation centre and neural mechanism that may represent a novel therapeutic target for the management of excessive eating.

2021-09-26 | GSE184385 | GEO

Transcriptional responses to deep cerebellar nuclei (DCN) and entorhinal cortex (EC) protein homogenates in cultured microglia

Project description:We hypothesised that enriched cytokines in the microenvironment of the deep cerebellar nuclei (DCN) contribute to the inflammatory gene expression profiles of DCN microglia and thus might induce a similar response in cultured microglia. To test this hypothesis, we treated BV2 microglia cell lines with DCN or entorhinal cortex (EC) protein homogenates and analysed RNA expression 4h post-stimulation, the peak timepoint of LPS-induced inflammatory gene expression (Das et al, 2016). To enrich for extracellular secreted proteins and to prevent effects of extraction buffer components on cell cultures, proteins were gently homogenised in DPBS on ice in the absence of detergent and protease inhibitors. As a positive control for inflammatory response gene induction, we concurrently treated BV2 cultures with LPS.

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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:In the cerebellum, the excitatory neurons of the three cerebellar nuclei (eCN) form the primary output for the circuit. The medial eCN (eCNm) were recently divided into molecularly defined subdomains in the adult, however how they are established during development is not known. We define molecular subdomains of the eCNm using scRNA-seq and spatial expression analysis during embryogenesis. Our analysis shows that the eCNm are transcriptionally divergent from the rest of the eCN by E14.5. We previously showed that conditional loss of the homeobox genesEn1andEn2leads to death of a subset of embryonic eCNm. We demonstrate that loss ofEn1/2in embryonic eCNm results in cell death of a specific posterior eCNm molecular subdomain and loss of TBR2 expression in an anterior subdomain, as well as reduced synaptic gene expression. We further reveal a similar function for EN1/2 in mediating TBR2 expression, neuron differentiation and survival in the two other cerebellar excitatory neuron types. Thus, our work defines embryonic eCNm molecular diversity and reveals conserved roles for EN1/2 in the cerebellar excitatory neuron lineage.

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Project description:Cerebellar organoids differentiated from human induced pluripotent stem cells (iPSCs) contain key cerebellar cell types and are increasingly used to study cerebellar diseases. In this study, we demonstrate the potential of cerebellar organoids for studying features of early human cerebellar development. Forkhead box protein P2 (FOXP2) is a transcription factor associated with speech and language development that is highly expressed in the developing brain. However, little attention has been directed to the study of FOXP2 in the early developing cerebellum. Here we used CRISPR gene editing in human iPSCs to generate a fluorescent FOXP2-reporter line. By combining transcriptomic analysis of iPSC-derived cerebellar organoids with published cerebellar datasets, we describe the expression and identify potential downstream targets of FOXP2 in the early developing human cerebellum. Our results highlight expression of FOXP2 in early human Purkinje cells and cerebellar nuclei neurons, and the vulnerability of these cell populations to neurodevelopmental disorders. Our study demonstrates the power of cerebellar organoids to model early human developmental processes and disorders.

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Project description:During cerebellar development, the main portion of the cerebellar plate neuroepithelium (NE) gives birth to Purkinje cells and interneurons, while the germinal zone at its dorsal edge, called the rhombic lip (RL), generates granule cells and cerebellar nuclei neurons. However, it remains elusive how these components work together to generate the intricate structure of the cerebellar anlage. In this study, we found that a polarized cerebellar anlage structure self-organizes in three-dimensional (3D) human ES cell (hESC) culture. This NE is capable of differentiating into electrophysiologically functional Purkinje cells. The addition of FGF19 promotes spontaneous generation of dorsoventrally polarized NE structures containing cerebellar and basal plates. Furthermore, further addition of SDF1 promoted the generation of stratified cerebellar plate NE with RL-like germinal zones self-forming at the edge. Thus, hESC-derived cerebellar progenitors exhibit substantial self-organizing potential for generating a polarized structure reminiscent of the early human cerebellar anlage at the first trimester. Examination of mRNA profile in two different treated human ES cells .

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Project description:Movements synchronized with external rhythms are ubiquitous in our daily lives. Despite the involvement of the cerebellum, the underlying mechanism remains unclear. In monkeys performing synchronized saccades to periodically alternating visual stimuli, we found that neuronal activity in the cerebellar dentate nucleus correlated with the timing of the next saccade and the current temporal error. One-third of the neurons were active regardless of saccade direction and showed greater activity for synchronized than for reactive saccades. During the transition from reactive to predictive saccades in each trial, the activity of these neurons coincided with target onset, representing an internal model of rhythmic structure rather than a specific motor command. The behavioural changes induced by electrical stimulation were explained by activating different groups of neurons at various strengths, suggesting that the lateral cerebellum contains multiple functional modules for the acquisition of internal rhythms, predictive motor control, and error detection during synchronized movements.

| S-EPMC9076601 | biostudies-literature

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