Profiling the transcriptome of excitatory neuronal nuclei during pilocarpine-induced seizure
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PROVIDER: PRJNA845771 | ENA |
REPOSITORIES: ENA
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Project description:The initial seizure-induced events in neuronal nuclei can lead to long-term changes in gene expression and cellular responses that likely contribute toward epileptogenesis. Using a transgenic mouse model to specifically isolate excitatory neuronal nuclei, we profiled the seizure-induced nuclear transcriptome by RNA-seq, as well as proteome via tandem mass tag mass spectrometry (available at ProteomeXchange via PRIDE; Accession ID: PXD030637). In the transcriptome, we observed robust induction of genes involved in multiple processes, including MAPK signalling, and downregulation of zinc finger genes harbouring Krüppel-associated box (KRAB) domains, which were strongly enriched for binding sites of the Thap11/Ronin transcriptional repressor. Additionally, we characterised gene expression in introns, which provided further insights into seizure-driven transcriptional regulation and dynamics, including repression of transcription mediated by zinc finger transcription factors. Our results provide a detailed snapshot of nuclear events induced by seizure activity and demonstrate a robust method for cell-type-specific nuclear profiling that can be applied to other cell types and models.
2023-03-31 | GSE205454 | GEO
Project description:Prolonged seizures can disrupt stem cell behavior in the adult hippocampus, an important brain structure for spatial memory. Here, using a mouse model of pilocarpine-induced status epilepticus (SE), we characterized spatiotemporal expression of Lin28a mRNA and proteins after SE. Unlike Lin28a transcripts, induction of LIN28A protein after SE was detected mainly in the subgranular zone, where immunoreactivity was found in progenitors, neuroblasts, and immature and mature granule neurons. To investigate roles of LIN28A in epilepsy, we generated Nestin-Cre::Lin28aloxP/loxP (cKO) and Nestin-Cre::Lin28a+/+ (WT) mice to block LIN28A upregulation in all neuronal lineages after acute seizure. Adult-generated neuron- and hippocampus-associated cognitive impairments were absent in epileptic LIN28A cKO mice as evaluated by pattern separation and contextual fear conditioning tests, respectively, while sham-manipulated WT and cKO animals showed comparable memory function. Moreover, numbers of hilar PROX1-expressing ectopic granule cells (EGCs), together with PROX1/NEUN+ mature EGCs, were significantly reduced in epileptic cKO mice. Transcriptomic analysis and immunohistochemical validation at 3 d post-pilocarpine provided potential LIN28A downstream targets such as serotonin receptor 4. Collectively, our findings indicate that LIN28A is a novel target for regulation of newborn neuron-associated memory dysfunction in epilepsy by modulating seizure-induced aberrant neurogenesis.
2024-01-01 | GSE246519 | GEO
Project description:Temporal changes in mice hippocampus transcriptome after pilocarpine-induced seizure
| PRJNA815649 | ENA
Project description:LC-MS/MS-based proteomics method was used to study activity-dependent changes in the nuclear proteome and transcriptome of forebrain excitatory neurons during Pilocarpine-induced seizures.
2023-08-14 | PXD030637 | Pride
Project description:We performed comprehensive DNA methylation analysis in neuronal and non-neuronal nuclei obtained from the human prefrontal cortex. This revealed that neuronal and non-neuronal nuclei manifest qualitatively and quantitatively distinctive DNA methylation patterns.
2011-02-21 | GSE15014 | GEO
Project description:Neuropsychiatric consequences of poorly controlled seizures that begin in childhood can be devastating. School failure or behavioral difficulty in a child with epilepsy is common and can become the focus of concern for families. Current antiepileptic drugs compound problems with their CNS side effects; effective therapy is currently limited as little is known about the cellular and molecular changes caused by seizures in the developing brain. This study will investigate transcriptional regulation induced by early-life seizures and explore alternative nonpharmacological therapeutic strategies in reversing damages of early-life seizures. We will study the therapeutic efficacy of environmental enrichment in reducing seizure-induced neuronal injury and in modifying gene expression alterations. We will explore molecular mechanisms underlying the beneficial effects of enriched environment and examine how different genes act in concert to influence the outcome of seizure-induced damage. To test the effect of environmental enrichment in modifying KA seizure-induced alterations in gene expression. We hypothesize that environmental enrichment enhances plastic, homeostatic response of immature brain to excessive, dysregulating seizure activity and protect against maladaptive response of developing animals to seizures. After inducing seizures by systemic injection of KA (8 mg/kg) or PBS injections (control) at P20, we will randomly assign P21 male Long Evans rats to 4 groups: enrichment housing (control-enriched; SE-enriched) or to standard vivarium cages (control; SE). Sixteen animals (8 control-enriched; 8 SE-enriched) will be housed as a group in a plastic rectangular cage measuring 115cm x 80 cm containing a running wheel, tunnels, rubber balls, a maze, a mirror, and a clock with a cyclist pendulum. Control and KA will be housed individually in a standard cage. Four litters will be used for each run of experiment and the experiment will be repeated once to obtain 16 animals per group. At P30, 240 h after KA or PBS, animals will be deeply anesthetized with isoflurane, decapitated for total RNA preparation (half brain for each, n = 12 per group). Total RNA will be isolated from each hippocampus individually, and equal amounts of RNA from 4 hippocampi will be pooled for each Genechip. Three independent hybridizations will be performed per condition (total of 12 Genechip Rat Expression Set 230). Keywords: dose response
2004-10-14 | GSE1833 | GEO
Project description:Benzodiazepine (BZ) drugs treat seizures, anxiety, insomnia, and alcohol withdrawal by potentiating γ2 subunit containing GABA type A receptors (GABAARs). BZ clinical use is hampered by tolerance and withdrawal symptoms, which include heightened seizure susceptibility, panic, and sleep disturbances. Here, we undergo a comprehensive investigation of inhibitory GABAergic and excitatory glutamatergic plasticity in mice tolerant to benzodiazepine sedation. Using quantitative proteomics approaches, we reveal cortex neuroadaptations of key pro-excitatory mediators and synaptic plasticity pathways, highlighted by Ca2+/calmodulin-dependent protein kinase II (CAMKII), MAPK, and PKC signaling.
2023-10-24 | PXD041332 | Pride
Project description:Neurodegenerative brain disorders become more common in the aged. Most of these disorders are associated with or caused by selective death of certain neuronal subpopulations. The mechanisms underlying the differential vulnerability of certain neuronal populations are still largely unexplored and few neuroprotective treatments are available to date. Elucidation of these mechanisms may lead to a greater understanding of the pathogenesis and treatment of neurodegenerative diseases. Moreover, preconditioning by a short seizure confers neuroprotection following a subsequent prolonged seizure. Our goal is to identify pathways that confer vulnerability and resistance to neurotoxic conditions by comparing the basal and preconditioned gene expression profiles of three differentially vulnerable hippocampal neuron populations. Hippocampal CA1 and CA3 pyramidal neurons are highly susceptible to seizures and ischemia, whereas dentate gyrus granule cells are relatively resistant. A brief preconditioning seizure confers protection to the pyramidal cells. We will first determine gene expression profiles of untreated rat CA1 and CA3 pyramidal cells, and dentate granule cells, using laser capture microscopy to obtain region-specific neuronal mRNA. We will then determine the effect of a brief preconditioning seizure, which is neuroprotective in CA1 and CA3, on these expression profiles. We hypothesize that common molecular mechanisms exist in neurons that determine their susceptibility to seizure-induced injury. Intrinsic differences in gene expression exist between hippocampal glutamatergic CA1 and CA3 pyramidal neurons, on the one hand, and dentate granule cells on the other, which contribute to the greater susceptibility of pyramidal neurons to degeneration in experimental stroke and epilepsy. We specifically hypothesize that differences in basal energy metabolism genes may confer differential susceptibility to neurodegeneration produced by seizures and ischemia. Anesthetized animals will be sacrificed by decapitation, and frozen 10 micron sections will be lightly stained with cresyl violet to identify cell layers in the hippocampus. Approximately 1000 neurons from each of the three cell layers will be isolated by LCM. Poly-A RNA will be amplified using a modified Eberwine protocol. The quality of our aRNA will be evaluated by quantitative RT-PCR of GluR6 and KA2 mRNA levels before we send the samples to the Center for labeling and hybridization to Affymetrix rat 230A arrays. We will provide a one-round amplification cDNA product to the center for labeling and hybridization. This protocol is identical to a previously approved study by Jim Greene in our laboratory. Keywords: other
2004-11-24 | GSE1994 | GEO
Project description:In the cerebellum, the excitatory neurons of the three cerebellar nuclei (eCN) form the primary output for the circuit. The medial eCN (eCNm) were recently divided into molecularly defined subdomains in the adult, however how they are established during development is not known. We define molecular subdomains of the eCNm using scRNA-seq and spatial expression analysis during embryogenesis. Our analysis shows that the eCNm are transcriptionally divergent from the rest of the eCN by E14.5. We previously showed that conditional loss of the homeobox genesEn1andEn2leads to death of a subset of embryonic eCNm. We demonstrate that loss ofEn1/2in embryonic eCNm results in cell death of a specific posterior eCNm molecular subdomain and loss of TBR2 expression in an anterior subdomain, as well as reduced synaptic gene expression. We further reveal a similar function for EN1/2 in mediating TBR2 expression, neuron differentiation and survival in the two other cerebellar excitatory neuron types. Thus, our work defines embryonic eCNm molecular diversity and reveals conserved roles for EN1/2 in the cerebellar excitatory neuron lineage.
2024-07-09 | GSE246894 | GEO
Project description:We mapped out temporal changes in mice hippocampus transcriptome as mRNA-seq and microRNA-seq after pilocarpine-induced SE to understand better molecular and cellular pathways that lead to neurodegenerative and neuroprotective processes after SE.
2023-12-31 | GSE198498 | GEO