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Electrophysiological Maturation of Cerebral Organoids Correlates with Dynamic Morphological and Cellular Development

ABSTRACT: Electrophysiological Maturation of Cerebral Organoids Correlates with Dynamic Morphological and Cellular Development

PROVIDER: PRJNA659793 | ENA |

REPOSITORIES: ENA

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Electrophysiological Maturation of Cerebral Organoids Correlates with Dynamic Morphological and Cellular Development

Project description:Cerebral organoids (COs) are rapidly accelerating the rate of translational neuroscience based on their potential to model complex features of the developing human brain. Several studies have examined the electrophysiological and neural network features of COs; however, no study has comprehensively investigated the developmental trajectory of electrophysiological properties in whole-brain COs and correlated these properties with developmentally-linked morphological and cellular features. Here, we profiled the neuroelectrical activities of COs over the span of five months with a multi-electrode array (MEA) platform and observed the emergence and maturation of several electrophysiologic properties, including rapid firing rates and network bursting events. To complement these analyses, we characterized the complex molecular and cellular development that give rise to these mature neuroelectrical properties with immunohistochemical and single-cell transcriptomic analyses. This integrated approach underscores the value of COs as an emerging model system of human brain development and neurological disease.

2020-08-28 | GSE157019 | GEO

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Morphological, electrophysiological and transcriptomic profiling of single neurons using Patch-seq

Project description:The mammalian neocortex is a layered sheet of neural tissue that mediates complex cognitive processes including perception and cognition. Despite its importance, we still lack detailed knowledge of the cellular components of the neocortex. Integrating morphological, electrophysiological and molecular classification schemes into a common framework for defining cell types is challenging due to the limited scope of most single-cell analyses. Here, we developed a protocol for high-throughput electrophysiological and transcriptomic analysis of single neurons that combines whole-cell recordings and single-cell RNA-sequencing, which we call Patch-seq. Using this approach, we fully characterized the electrophysiological and molecular profiles of ~50 neocortical neurons, and show that gene expression patterns can be used to infer the morphological and physiological properties of individual neurons and their corresponding cell type. Our results shed light on the molecular underpinnings of neuronal diversity and demonstrate a path forward for comprehensive cell type characterization in the nervous system.

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Morphological, electrophysiological and transcriptomic profiling of single neurons using Patch-seq

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Transcriptomic and morphological maturation of human astrocytes in cerebral organoids

Project description:Cerebral organoids (CerOrgs) derived from human induced pluripotent stem cells (iPSCs) are a valuable tool to study human astrocytes and their interaction with neurons and microglia. The timeline of astrocyte development and maturation in this model is currently unknown and this limits the value and applicability of the model. Therefore, we generated CerOrgs from three healthy individuals and assessed astrocyte maturation after 5, 11, 19, and 37 weeks in culture. At these four time points, the astrocyte lineage was isolated based on the expression of integrin subunit alpha 6 (ITGA6). Based on the transcriptome of the isolated ITGA6-positive cells, astrocyte development started between 5 and 11 weeks in culture and astrocyte maturation commenced after 11 weeks in culture. After 19 weeks in culture, the ITGA6-positive astrocytes had the highest expression of human mature astrocyte genes, and the predicted functional properties were related to brain homeostasis. After 37 weeks in culture, a subpopulation of ITGA6-negative astrocytes appeared highlighting the heterogeneity within the astrocytes. The morphology shifted from an elongated progenitor-like morphology to the typical bushy astrocyte morphology. Based on the morphological properties, predicted functional properties, and the similarities with the human mature astrocyte transcriptome, we concluded that ITGA6+ astrocytes have developed optimally in 19-week-old CerOrgs.

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Electrophysiological properties of human induced-pluripotent stem cell-derived three-dimensional cerebral organoids

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Coupled electrophysiological recording and single-cell transcriptome analyses revealed molecular mechanisms underlying neuronal maturation

Project description:The mammalian brain is heterogeneous, containing billions of neurons and trillions of synapses forming various neural circuitries, through which sense, movement, thought, and emotion are generated. The cellular heterogeneity of the brain has made it difficult to study the molecular logic of neural circuitry wiring, pruning, activation, and plasticity, until recently, transcriptome analyses with single-cell resolution makes decoding of gene regulatory networks underlying aforementioned circuitry properties possible. Here, we report success in performing both electrophysiological and whole-genome transcriptome analyses on single human neurons in culture. Using Weighted Gene Coexpression Network Analyses (WGCNA), we identified gene clusters highly correlated with neuronal maturation judged by electrophysiological characteristics. A tight link between neuronal maturation and genes involved in ubiquitination and mitochondrial function was revealed. Moreover, we identified a list of candidate genes, which could potentially serve as biomarkers for neuronal maturation. Coupled electrophysiological recording and single-cell transcriptome analysis will serve as powerful tools in the future to unveil molecular logics for neural circuitry functions.

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