Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Single-cell RNA-seq of mouse cardiac interstitial cells 3 & 7 days after sham or myocardial infarction injury

ABSTRACT: The aims of the experiment were to profile the cell types in the adult mouse cardiac interstitium (non-myocyte cells) and how they respond to myocardial infarction injury. Adult, male, Pdgfra +/GFP mice were subject to either a myocardial infarction or sham injury, with cells isolated from cardiac ventricles 3 or 7 days following surgery. We obtained scRNA-seq profiles of two cell fractions: total interstitial (non-myocyte) cell population (TIP) and FACS-sorted GFP+/Cd31- cells (GFP).

INSTRUMENT(S): Illumina HiSeq 2500, NextSeq 500

ORGANISM(S): Mus musculus

SUBMITTER: Ralph Patrick

PROVIDER: E-MTAB-7376 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Ischemic stroke is a severe medical condition that leads to neurological impairments, including the loss of sensory, motor, and cognitive functions. Focal cerebral ischemia (FCI) occurs when blood supply to a specific brain region is interrupted, resulting in cell death. Cells expressing neural-glial antigen 2 (NG2) include glial cells, primarily oligodendrocyte precursors, and perivascular cells. In both cell types, the canonical Wnt signaling pathway is active, with its activity increasing after ischemic injury, influencing the tissue’s response to damage. Following ischemia, NG2 glia rapidly proliferate, migrate to the injury site, contribute to glial scar formation alongside astrocytes, and play a role in brain tissue regeneration. Their potential to differentiate into cell types beyond oligodendrocytes has been documented. Perivascular cells, which regulate vascular contraction, serve as an informational bridge between vascular endothelial cells and glial cells, particularly astrocytes. The plasticity of pericytes following brain injury has also been suggested. Previously, we analyzed NG2-expressing cells in Rosa26-tdTomato/Cspg4-CreERT2 mice three days after middle cerebral artery occlusion (MCAO) or sham surgery (CTRL) without Wnt pathway modulation (ArrayExpress accession number: E-MTAB-11967). In this study, we extended our analysis by inhibiting the Wnt signaling pathway, which is crucial for stem cell maintenance, proliferation, and neuronal differentiation. To investigate these processes, we isolated NG2 cells and their derivatives labeled with the red fluorescent protein tdTomato from the cortex of Rosa26-tdTomato/Cspg4-CreERT2 mice three days after MCAO. Mice subjected to sham surgery served as healthy controls. Wnt pathway inhibition was achieved using an allele that enables inducible expression of the Wnt inhibitor Dickkopf1 (Dkk1). Our aim was to determine how the Wnt pathway inhibition affects the distribution of NG2 glial and perivascular cell subtypes, identify their derivatives in the cortex of healthy and ischemic mice, and characterize their abundance and gene expression profiles.

Project description:Ischemic stroke is a severe medical condition that leads to neurological impairments, including the loss of sensory, motor, and cognitive functions. Focal cerebral ischemia (FCI) occurs when blood supply to a specific brain region is interrupted, resulting in cell death. Cells expressing neural-glial antigen 2 (NG2) include glial cells, primarily oligodendrocyte precursors, and perivascular cells. In both cell types, the canonical Wnt signaling pathway is active, with its activity increasing after ischemic injury, influencing the tissue’s response to damage. Following ischemia, NG2 glia rapidly proliferate, migrate to the injury site, contribute to glial scar formation alongside astrocytes, and play a role in brain tissue regeneration. Their potential to differentiate into cell types beyond oligodendrocytes has been documented. Perivascular cells, which regulate vascular contraction, serve as an informational bridge between vascular endothelial cells and glial cells, particularly astrocytes. The plasticity of pericytes following brain injury has also been suggested. Previously, we analyzed NG2-expressing cells in Rosa26-tdTomato/Cspg4-CreERT2 mice three days after middle cerebral artery occlusion (MCAO) or sham surgery (CTRL) without Wnt pathway modulation (ArrayExpress accession number: E-MTAB-11967). In this study, we extended our analysis by constitutively activating the Wnt/β-catenin signaling pathway, which is crucial for stem cell maintenance, proliferation, and neuronal differentiation. To investigate these processes, we isolated NG2 cells and their derivatives labeled with the red fluorescent protein tdTomato from the cortex of Rosa26-tdTomato/Cspg4-CreERT2 mice three days after MCAO. Mice subjected to sham surgery served as healthy controls. Wnt pathway activation was achieved using an allele that enables constitutive stabilization of β-catenin (Ctnnb1 delEx3). Our aim was to determine how the Wnt pathway overactivation affects the distribution of NG2 glial and perivascular cell subtypes, identify their derivatives in the cortex of healthy and ischemic mice, and characterize their abundance and gene expression profiles.

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Single-cell RNA-seq of mouse cardiac interstitial cells 3 & 7 days after sham or myocardial infarction injury

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