Project description:This SuperSeries is composed of the following subset Series:; GSE11256: KCl depolarization-regulated genes in mouse cortical neurons; GSE11258: Npas4-regulated genes in mouse hippocampal neurons Experiment Overall Design: Refer to individual Series
Project description:5’ ExoSeq of total RNA (rRNA & signal recognition particle RNA depleted) from mouse cortical neurons before and after membrane depolarization by potassium chloride (KCl).
Project description:Activation of neurons is one of the fundamental events for the functioning of nervous system. Neuronal activation relays information to next neurons. On the other hand, the activated neurons themselves are also influenced by neuronal activation. Depending on the type and condition of neuronal activation, these activated neurons change their gene expressions, thereby being able to process information more or less efficiently. We applied the microarray technology to identify hither-to-uncharacterized as activity-dependent genes. Especially, we screened the transcription factors, because early changes in the transcription factors should result in alterations of gene expression profiles and subsequent neuronal properties. Experiment Overall Design: Rat primary cortical neurons with or without KCl treatment were selected for RNA extraction and hybridization on Affymetrix microarrays. To identify the genes whose expression was induced by depolarization, we first compared gene expression profiles in control vs. 4 hr after KCl (25 mM) treated cortical neurons using Affymetrix Genechips specified for neurobiology. All four hybridizations were analyzed for correlation accuracy between the replicates of the same treatments .Control replicates (control 1, 2) KCl-treated replicates (KCl 1, 2)
Project description:RNA-seq of total RNA from Rad21lox/lox;NexCre/+ and Rad21+/+;NexCre/+ dissociated cortical neurons at Day In Vitro (DIV10) without stimulation or after treatment with TTX/D-AP5 and 1h or 6h of KCl depolarization.
Project description:5C-seq from Rad21lox/lox;NexCre/+ and Rad21+/+;NexCre/+ dissociated cortical neurons at Day In Vitro (DIV10) without stimulation or after treatment with TTX/D-AP5 and 1h of KCl depolarization. And, 5C of and doxycycline treated Rad21-TEV primary neurons transduced with NLS-TEV
Project description:To examine potential differences in activity-dependent gene expression, we analyzed mRNA expression in cultured neurons isolated from Wild-type vs MeCP2 S421A mice at 0 (unstimulated), 1 or 6 hours after membrane depolarization by exposure to high extracellular KCl (55mM) All mice were male littermates from one of three litters. We isolated RNA from dissociated cortical cultures (E16+7DIV) isolated from Wild-Type or MeCP2 S421A knock-in mice littermates. Cells were either left unstimulated or depolarized for 1 or 6 hours by addition of 55mM KCl to the media. mRNA expression was analyzed using the Affymetrix GeneChip Mouse Expression Set 430 2.0 microarray platform.
Project description:Learning and memory require activity-induced changes in dendritic translation, but which messenger RNAs (mRNAs) are involved and how they are regulated are unclear. Here, to monitor how depolarization impacts local dendritic biology, we employed a dendritically-targeted proximity labeling approach, followed by cross-linking immunoprecipitation (CLIP), ribosome profiling, and mass spectrometry. Depolarization of primary cortical neurons with KCl or the glutamate agonist DHPG causes rapid reprogramming of dendritic protein expression, where changes in dendritic mRNAs and proteins are weakly correlated. For a subset of pre-localized messages, depolarization increases translation of upstream open reading frames (uORFs) and their downstream coding sequences, enabling localized production of proteins involved in long term potentiation, cell signaling, and energy metabolism. This activity-dependent translation is accompanied by the phosphorylation and recruitment of the non-canonical translation initiation factor eIF4G2, and the translated uORFs are sufficient to confer depolarization-induced, eIF4G2-dependent translational control. These studies uncover an unanticipated mechanism by which activity-dependent uORF translational control by eIF4G2 couples activity to local dendritic remodeling.
Project description:RNA-seq of human and mouse stem-cell derived neurons and mouse primary cortical neurons after KCl-induced membrane depolarization against controls.
