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Directed differentiation of human pluripotent stem cells into cortical interneurons

ABSTRACT: Here we demonstrate the highly efficient derivation of human cortical interneurons in a NKX2.1::GFP hESC reporter line. Manipulating the timing of SHH activation yields three distinct GFP+ populations with specific transcriptional profiles, neurotransmitter phenotypes and migratory behaviors. Total RNA was isolated at day 18 of differentiation from FACS sorted NKX2.1::GFP+ cells from three varying differentiation conditions and a “No SHH” condition (no sonic hedgehog treatment) that did not contain any GFP-expressing cells (Trizol, Sigma). Samples for each group in triplicate were processed for Illumina bead arrays (Illumina HT-12) by the MSKCC genomics core facility according to the specifications of the manufacturer.

ORGANISM(S): Homo sapiens

SUBMITTER: Jeffrey Zhao

PROVIDER: E-GEOD-46098 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Directed differentiation of human pluripotent stem cells into cortical interneurons

Project description:Here we demonstrate the highly efficient derivation of human cortical interneurons in a NKX2.1::GFP hESC reporter line. Manipulating the timing of SHH activation yields three distinct GFP+ populations with specific transcriptional profiles, neurotransmitter phenotypes and migratory behaviors.

2013-12-31 | GSE46098 | GEO

Sonic hedgehog Activation Restores Interneurons and Cognitive Function in Newborns with Intraventricular Hemorrhage

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Project description:Unilateral Traumatic Brain Injury (TBI) causes functional disturbances of the neuronal networks spreading even to the undamaged cortical hemisphere. The phenomenon, referred to as transhemispheric diaschisis, is suggested to be mediated by an imbalance of the strength of glutamatergic, excitatory vs. GABAergic, inhibitory neurotransmission. Here we present evidence that a switch in expression of α Subunits of pore-forming L-Type voltage-gated calcium channels (VGCC), by an expression of CaV1.3 and simultaneous ablation of CaV1.2 in GABAergic interneurons could balance early cortical disturbances manifested as contralateral hyperexcitability in the early phase after TBI. The switch of the VGCC alpha Subunits in GABAergic interneurons was detected using the GAD67-GFP (Glutamate Decarboxylase 67 – Green Fluorescent Protein) Knock-in mouse line. Mice received a TBI with a Controlled Cortical Impact (CCI) to the primary motor and somatosensory cortex at postnatal day 19-21 under anesthesia in vivo. Single GFP+ interneurons located in the undamaged, contralateral cortex were isolated by Fluorescence-Activated Cell Sorting (FACS) and further analyzed by Mass Spectrometry (MS). The switch was associated with an increased excitability of Somatostatin (SST) interneurons and extracellular network activity in acute brain slices in Microelectrode Array (MEA) recordings, which could be restored in presence of isradipine (100 nM), which selectively blocks CaV1.3-containing VGCCs. These data suggest that a switch in alpha.subunits of VGCCs expressed on SST-positive interneurons stabilizes early hyperactivity of the contralateral cortical network at 72 h after TBI, thereby promoting an adaptive mechanism to counterbalance post-traumatic hyperexcitabilty that might lead to epileptogenesis.

2022-01-03 | PXD023551 | Pride

Directed differentiation of oligodendrocytes from human embryonic stem cells

Project description:Here we compare the transcriptional profile of the undifferentiated hESC with the anterior oligodendroctes derived from the hESC using our new differentiation protocol. Total RNA was isolated from undifferentiated hES cells and from hES derived oligodendrocytes, 70 days old and FACS sorted for O4 marker. Samples for each group in triplicate were processed for Illumina bead arrays (Illumina HT-12) by the MSKCC genomics core facility according to the specifications of the manufacturer.

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Transcription profiling by array of human NKX2.1GFP/w spin embryoid bodies differentiated and then FACS sorted based on FORSE-1, NKX2.1-GFP and PDGFR expression

Project description:The gene targeted NKX2.1GFP/w hESC line was differentiated as spin EBs in BPEL medium supplemented with FGF2 and retinoic acid to generate NKX2.1-GFP+ neural cells. NKX2.1-GFP+ cells were FACS sorted and further differentiated in serum free BEL medium supplemented with FGF2 (20 ng/ml) and EGF1 (20 ng/ml) on laminin-coated flasks. After ~ 90 days in culture, cells were FACS sorted based on FORSE-1 and NKX2.1-GFP expression, with the FORSE-1-GFP- fraction further fractionated on the basis of PDGFR? expression. Sorted fractions were subjected to Illumina microarray processing.

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Project description:To determine how Hedgehog signaling regulates salivary function, we performed single cell RNA sequencing (scRNA-seq) of adult mouse submandibular glands (SMGs) harvested 7 days after intra-SMG delivery of adenoviral vectors (Ad) carrying rat Sonic Hedgehog (Shh) or control GFP gene.

2023-04-10 | GSE148541 | GEO

NOTCH, ERK and SHH signaling respectively control the fate determination of cortical astrocytes, oligodendrocytes, and olfactory bulb interneurons

Project description:During cortical development, radial glial cells, known as neural stem cells, initially generate a large number of cortical glutamatergic pyramidal neurons through a process called neurogenesis. This is followed by the generation of a diversity of cortical astrocytes, oligodendrocytes, and olfactory bulb interneurons, known as gliogenesis. However, the molecular mechanisms underlying the switch from cortical neurogenesis to gliogenesis, and the subsequent fate determination of cortical astrocytes, oligodendrocytes, and olfactory bulb interneurons, remain unclear. Here, we report that extracellular signal-regulated kinase (ERK) signaling plays a fundamental role in promoting cortical gliogenesis and the generation of cortical glial progenitors. Additionally, SHH-SMO-GLI activator signaling has an auxiliary function to ERK during these processes. We further demonstrate that NOTCH signaling is absolutely required for the fate determination of astrocytes, while ERK signaling plays a prominent role in oligodendrocyte fate specification, and SHH signaling is crucial for the fate determination of olfactory bulb interneurons from cortical progenitors. We provide evidence suggesting that this mechanism is conserved in both mice and humans. Finally, we propose a unifying principle of mammalian cortical gliogenesis.

2024-08-23 | GSE274547 | GEO

NOTCH, ERK and SHH signaling respectively control the fate determination of cortical astrocytes, oligodendrocytes, and olfactory bulb interneurons [CUT&Tag]

2024-08-21 | GSE275004 | GEO

Chronic stress causes striatal disinhibition mediated by SOM-interneurons

Project description:Chronic stress is a major triggering factor for neuropsychiatric disorders including obsessive-compulsive disorder (OCD), a mental health condition characterized by motor stereotypies and striatal overactivation. However, the mechanisms at the cell- and microcircuit-level through which stress triggers motor symptoms is currently unknown. Here, we report that chronic stress (CS) in mice alters dorsomedial striatum (DMS) function, by affecting GABAergic interneuron populations and somatostatin-positive (SOM) interneurons in particular. Specifically, we show that CS impairs communication between SOM interneurons and medium spiny neurons, promoting striatal overactivation / disinhibition and increased motor output. Using probabilistic machine learning for analyzing animal behavior we further demonstrate that in vivo chemogenetic manipulation of SOM interneurons in DMS modulates motor phenotypes in stressed mice. Altogether, we propose a causal link between dysfunction of striatal SOM interneurons and motor symptoms in stress-related neuropsychiatric disorders.

2022-12-13 | PXD031193 | Pride

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