Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1101871 | ENA |
REPOSITORIES: ENA
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Project description:Profiling of transcriptional changes in rat astrocytes when co-cultured with neurons: comparison of astrocytes cultured alone with astrocytes co-cultured with mouse hippocampal neurons. Co-cultured astrocytes are isolated using cold jet, a novel tool for these neuron-glia cultures. Over the last decade, the importance of astrocyte-neuron communication in neuronal development and synaptic plasticity has become increasingly clear. Since neuron-astrocyte interactions represent highly dynamic and reciprocal processes, we hypothesized that at least part of the involved astrocyte genes may be regulated as a consequence of their interactions with maturing neurons. In order to identify such neuron-induced astrocyte genes in vitro, we tested the effectiveness of the ‘cold jet’, a new method for separation of neurons from co-cultured astrocytes. The cold jet method is performed under ice-cold conditions and avoids protease-mediated isolation of astrocytes or time-consuming centrifugation, yielding intact astrocyte mRNA with approximately 90% of neuronal RNA removed. Using this method, we executed genome-wide profiling in which RNA derived from astrocyte-only cultures was compared with astrocyte RNA derived from differentiating neuron-astrocyte co-cultures. Data analysis revealed changes in expression of a large number of mRNAs and biological processes, including novel findings. Thus, cold jet is an efficient method to separate astrocytes from neurons in co-culture, and in this study reveals that neurons induce robust gene-expression changes in co-cultured astrocytes.
2016-11-09 | GSE52481 | GEO
Project description:Communication between astrocytes and neurons plays a pivotal role in the development, maintenance and dysfunction of cellular networks within the central nervous system. Further understanding the contribution of astrocytes to the initiation and progression of neuropathology has, however, been hindered by the complexity of cell networks in traditional in vivo and ex vivo systems. We have designed and constructed a novel three-compartment microfluidic cell culture device that enables us to overcome these limitations and uncouple the roles of astrocytes and neurons in the transfer of pathology through complex cell networks. Our microfluidic device integrates a novel maze-like structure that prevents synaptic connectivity between two fluidically isolated neuron populations, while allowing astrocyte infiltration and growth throughout. We use primary neuron/astrocyte co-cultures, proteomic analysis and immunocytochemistry to validate the application of this device. Using this device, we identify and describe a novel calcium-dependent role for astrocytes in the transfer of excitotoxic pathology between the segregated neuron populations.
2026-06-08 | PXD055523 | Pride
Project description:Advancements in human induced pluripotent stem cell (hiPSC) technology have enabled co-culture models for disease modeling in physiologically relevant systems. However, co-culturing protocols face challenges in usability and consistency. Here, we introduce a robust, reproducible hiPSC-derived co-culture system integrating astrocytes, neurons, and microglia. This model leverages cryopreserved cells, enabling co-cultures within 20 days post-thaw. Comparing monocultures and tri-cultures, we show how cell-cell interactions shape transcriptional and functional states across all three cell types. Neurons in tri-culture exhibit increased spine density and activity, while astrocytes and microglia show altered responses to proinflammatory stimulation. Surprisingly, astrocyte co-culture induces upregulation of disease-associated microglia (DAM) genes, including TREM2, SPP1, APOE, and GPNMB. Additionally, while familial Alzheimer’s disease neurons induce a prototypical inflammatory response in microglia, the DAM signature is significantly downregulated. Collectively, this study establishes a versatile human tri-culture model as a valuable resource for dissecting neuron-glia interactions and their role in neurodegenerative disease.
2025-06-02 | GSE295330 | GEO
Project description:Little is known about factors that induce stem cells differentiated into astrocyte. Using double cross strategy, we effective narrow down the candidate molecules crucial for astroglial differentiation. By cross comparison the data from transcriptomic and proteomic experiments, we selected 24 candidate genes which might be involved in the neuron differentiation. The 24 genes were further cross compared with Alzheimer disease (AD) database, which regarded as a neuron degenerative database. There were 18 out of 24 candidates showed opposite expression level in our data and AD database. Among them, the combination of Heat Shock Protein 27 (HSP27) and S100 calcium-binding protein A16 (S100A16) showed predominant crucial in neuronal differentiation than other gene combination. We previously showed that the expression level of HSP27 directly related to the glutamatergic neuron differentiation under IBMX induction. In this paper, we found co-silence of HSP27 and S100A16 directly induced PDMCs differentiate into functional glutamatergic neurons and astrocytes while IBMX is no longer required in this process. The resulting astrocyte not only exhibit the classic morphology but also with astrocyte function.
2019-11-01 | GSE139656 | GEO
Project description:Protein from astrocytes (glial fibrillary acidic protein-positive cells), neurons (beta-III tubulin-positive cells), and unsorted (cell suspension without any enrichment based on astrocyte or neuron markers) from human, post-mortem Alzheimer's disease (AD) and aged-matched non-symptomatic (NS) prefrontal cortex brain samples.
2019-05-06 | MSV000083747 | MassIVE
Project description:Astrocyte-to-neuron conversion has developed into a promising avenue for neuronal replacement therapy. Neurons depend critically on mitochondria function and often die by ferroptosis during the conversion process. Here we examined the extent of adequate mitochondrial reprogramming by morphology and proteome analysis. While mitochondria profoundly changed their morphology during Neurogenin2 (Neurog2) – or Achaete-scute homolog 1 (Ascl1)-mediated astrocyte-to-neuron reprogramming, we found neuron-specific mitochondrial proteins, here identified in a comprehensive proteome analysis of isolated mitochondria from primary neurons and astrocytes, to be only partially and at late stages regulated during the process. To improve this, we used dCas9 technology to induce neuron-specific mitochondrial proteins early during reprogramming. This resulted not only in increased conversion efficiency, but also in faster neuronal generation. Taken together, reprogramming mitochondria in a cell type-specific manner has powerful effects on astrocyte-to-neuron conversion, suggesting mitochondria to be a driving force in this process.
2021-03-29 | PXD014886 | Pride
Project description:SORL1 is implicated in the pathogenesis of Alzheimer’s disease (AD) through genetic studies. To interrogate the role(s) of SORL1 in human brain cells, SORL1 null iPSCs are differentiated to neuron, astrocyte, microglial, and endothelial cell fates. Loss of SORL1 leads to alterations in both overlapping and distinct pathways across cell types, with the greatest effects in neurons and astrocytes. SORL1 loss induces a neuron-specific reduction in APOE and CLU and altered lipid profiles. Enhancement of retromer-mediated trafficking rescues tau phenotypes observed in SORL1 null neurons but does not rescue APOE levels. Pathway analyses implicate TGF-β/SMAD signaling in SORL1 function, and modulating SMAD signaling in neurons alters APOE RNA levels in a SORL1-dependent manner. Analyses of iPSCs derived from a large cohort reveal a neuron-specific association between SORL1, APOE, and CLU levels, a finding validated in post-mortem brain. These studies provide a mechanistic link between strong genetic risk factors for AD.
2023-10-24 | PXD044093 | Pride
Project description:Neurons induce a dramatic transformation in developing astrocytes, causing them to develop a complex stellate morphology resembling their appearance in vivo. However, the transcriptional changes that accompany this transformation are not known, nor are the signalling mechanisms responsible. Similarly, whether synaptic activity controls astrocytic gene expression and whether this leads to altered astrocytic function is unclear. This experiment seeks to investigate this non-cell-autonomously regulated gene expression by co-culturing astrocytes and neurons derived from closely related species (mouse and rat), and separating RNA-seq reads derived from each cell type in silico, thus shedding light on the signalling mechanisms underlying neuron-to-astrocyte communication and the functional consequences for astrocytes.
2017-05-02 | E-MTAB-5514 | biostudies-arrayexpress
Project description:The dynamics of microglial activity within neuron-astrocyte-microglia tri-cultures derived from human induced pluripotent stem cells (iPSCs) present a complex interplay and offer an opportunity to obtain new insights into neuron-glia interactions. Iron-laden microglia, correlating with functional changes, represent a key pathological feature of Alzheimer's disease (AD). This study characterized the cellular crosstalk and transcriptional states of microglia in tri-cultures. Complement C3 can be detected in culture media when microglia are cocultured with neurons, and the addition of astrocytes in the coculture led to an increased amount of C3, indicating that the impact of glial interactions can be evaluated in this model system. We compared microglial gene expression profiles comprehensively in monoculture, coculture, and tri-culture settings. Single-cell RNA sequencing (scRNA-seq) revealed various microglial states with gene expression changes associated with endocytosis and neuron-related functions in tri-culture settings, suggesting that microglial behavior is profoundly impacted by the presence of neurons and astrocytes. We assessed microglial responses to iron overload combined with the ferroptosis inducer RSL3 (a GPX4 inhibitor) in tri-cultures. Microglial cell death was accompanied by ferritin heavy-chain expression, indicating microglia ferroptosis. scRNA-seq analyses highlighted alterations in pathways related to ferroptosis, stress response, and autophagy, indicating substantial shifts in microglial profiles upon iron perturbation. These findings underscore the necessity of using tri-cultures as a model to capture certain degrees of complex cellular interactions occurring in vivo. These results offer critical insights for establishing in vitro models for therapeutic development of neurodegenerative diseases, including AD.
2026-03-31 | GSE325931 | GEO
Project description:Bulk RNA sequencing was performed to compare mRNA expression profiles across six experimental conditions: monoculture of primary dorsal root ganglion (DRG) neurons isolated from C57BL/6J mice; conditioned medium from EO771 cells applied to DRG neurons; and direct co-culture of EO771 and DRG neurons followed by fluorescence-activated cell sorting (FACS) to DRG neuron population. The aim was to identify transcriptional programs regulated by tumor–neuron interactions, including acute gene expression changes, intercellular signaling pathways, and broader pathway-level alterations. The dataset is intended for differential expression analysis, pathway enrichment, and hypothesis generation for downstream functional studies.
2026-05-15 | GSE330238 | GEO