Unknown,Transcriptomics,Genomics,Proteomics

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RNA-seq profiling of three distinct cell populations in a murine model of epilepsia with and without drug treatment

ABSTRACT: This experiment aimed at identifying transcriptome-wide differences in a murine model of epilepsia after treatment with soticlestat. We used three different cell populations (neurons, microglia, and astrocytes) from soticlestat-treated mice, mice where the epileptic state had been induced, and sham mice as control. We compared the differences within each group (treatment / status epilepticus versus sham) and across groups (treatment - status epilepticus) to identify genes involved in the disease, and on the other hand, genes involved in the therapeutic effect of the drug.

INSTRUMENT(S): NextSeq 500

ORGANISM(S): Mus musculus

SUBMITTER: Luca Beltrame

PROVIDER: E-MTAB-9943 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:All established protocols for differentiation of mouse and human pluripotent stem cells into specific neural subpopulations generate a considerable cellular heterogeneity that hampers experimental and clinical progress. In order to obtain a homogenous population of neuronal precursor cells and to streamline the differentiation of embryonic stem cells (ESCs), we assessed PSA-NCAM, a surface glycoprotein that is specifically expressed on immature neurons. We developed an optimized strategy for magnetic isolation of PSA-NCAM positive neuronal precursors from differentiated ESC cultures and characterized their neuronal differentiation potential in vitro. PSA-NCAM enrichment at an early step of neural differentiation increased the number of ES cell derived neurons and reduced cellular diversity. Gene expression analysis revealed that mainly genes involved in neuronal activity were over-represented after purification. The in vivo potential of in vitro derived PSA-NCAM+ enriched precursors was functionally characterized by grafting into the forebrain of adult mice. Analysis for several neuronal and glia markers at 10 or 40 days post graft showed a distinct differentiation pattern. While unsorted control cells gave rise to a mixed population composed of immature precursors, early postmitotic neurons or glial cells, the majority of PSA-NCAM+ enriched cells differentiated into NeuN positive neurons. Furthermore, when in contact with the rostral migratory stream, higher numbers of cells integrated into the stream and migrated towards the olfactory bulb when the PSA-NCAM enriched population was grafted. Thus, enrichment of neuronal precursors based on PSA-NCAM expression represents a general and straightforward approach to narrow cellular heterogeneity during neuronal differentiation of pluripotent cells. Two conditions (step 4, step 5), each represented by three biological replicates of control and enriched cells (Cy5); mESC was used as common reference (Cy3)

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RNA-seq profiling of three distinct cell populations in a murine model of epilepsia with and without drug treatment

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