دورية أكاديمية
New insights into the early mechanisms of epileptogenesis in a zebrafish model of Dravet syndrome.
العنوان: | New insights into the early mechanisms of epileptogenesis in a zebrafish model of Dravet syndrome. |
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المؤلفون: | Tiraboschi E; Chemical Neuroscience Group, Center for Molecular Medicine Norway, University of Oslo, Oslo, Norway., Martina S; Integrative Cell Signaling Group, Luxembourg Center for Systems Biomedicine, University of Luxembourg, Esch-sur-Alzette, Luxembourg., van der Ent W; Chemical Neuroscience Group, Center for Molecular Medicine Norway, University of Oslo, Oslo, Norway., Grzyb K; Integrative Cell Signaling Group, Luxembourg Center for Systems Biomedicine, University of Luxembourg, Esch-sur-Alzette, Luxembourg., Gawel K; Chemical Neuroscience Group, Center for Molecular Medicine Norway, University of Oslo, Oslo, Norway.; Department of Experimental and Clinical Pharmacology, Medical University of Lublin, Lublin, Poland., Cordero-Maldonado ML; Integrative Cell Signaling Group, Luxembourg Center for Systems Biomedicine, University of Luxembourg, Esch-sur-Alzette, Luxembourg., Poovathingal SK; Integrative Cell Signaling Group, Luxembourg Center for Systems Biomedicine, University of Luxembourg, Esch-sur-Alzette, Luxembourg., Heintz S; Chemical Neuroscience Group, Center for Molecular Medicine Norway, University of Oslo, Oslo, Norway., Satheesh SV; Molecular Toxicology Group, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway., Brattespe J; Department of Biological Sciences, University of Bergen, Bergen, Norway., Xu J; Department of Biomedicine, University of Bergen, Bergen, Norway., Suster M; allmyhomes, Berlin, Germany., Skupin A; Integrative Cell Signaling Group, Luxembourg Center for Systems Biomedicine, University of Luxembourg, Esch-sur-Alzette, Luxembourg., Esguerra CV; Chemical Neuroscience Group, Center for Molecular Medicine Norway, University of Oslo, Oslo, Norway.; Department of Pharmacy, University of Oslo, Oslo, Norway. |
المصدر: | Epilepsia [Epilepsia] 2020 Mar; Vol. 61 (3), pp. 549-560. Date of Electronic Publication: 2020 Feb 24. |
نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
اللغة: | English |
بيانات الدورية: | Publisher: Blackwell Science Country of Publication: United States NLM ID: 2983306R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1528-1167 (Electronic) Linking ISSN: 00139580 NLM ISO Abbreviation: Epilepsia Subsets: MEDLINE |
أسماء مطبوعة: | Publication: Malden, MA : Blackwell Science Original Publication: Copenhagen : Munskgaard |
مواضيع طبية MeSH: | Brain/*physiopathology , Epilepsies, Myoclonic/*genetics , NAV1.1 Voltage-Gated Sodium Channel/*genetics , Neuronal Plasticity/*genetics , Zebrafish Proteins/*genetics, Animals ; Anticonvulsants/pharmacology ; Brain/drug effects ; Brain/metabolism ; Brain/pathology ; CRISPR-Cas Systems ; Cell Proliferation/drug effects ; Diazepam/pharmacology ; Disease Models, Animal ; Electroencephalography ; Epilepsies, Myoclonic/metabolism ; Epilepsies, Myoclonic/pathology ; Epilepsies, Myoclonic/physiopathology ; Fenfluramine/pharmacology ; GABAergic Neurons/drug effects ; GABAergic Neurons/metabolism ; GABAergic Neurons/pathology ; Gene Expression Profiling ; Gliosis/genetics ; Gliosis/pathology ; Locomotion/drug effects ; Mutation, Missense ; NAV1.1 Voltage-Gated Sodium Channel/metabolism ; Neuronal Plasticity/drug effects ; RNA-Seq ; Real-Time Polymerase Chain Reaction ; Serotonin 5-HT2 Receptor Agonists/pharmacology ; Single-Cell Analysis ; Zebrafish ; Zebrafish Proteins/metabolism |
مستخلص: | Objective: To pinpoint the earliest cellular defects underlying seizure onset (epileptogenic period) during perinatal brain development in a new zebrafish model of Dravet syndrome (DS) and to investigate potential disease-modifying activity of the 5HT Methods: We used CRISPR/Cas9 mutagenesis to introduce a missense mutation, designed to perturb ion transport function in all channel isoforms, into scn1lab, the zebrafish orthologue of SCN1A (encoding voltage-gated sodium channel alpha subunit 1). We performed behavioral analysis and electroencephalographic recordings to measure convulsions and epileptiform discharges, followed by single-cell RNA-Seq, morphometric analysis of transgenic reporter-labeled γ-aminobutyric acidergic (GABAergic) neurons, and pharmacological profiling of mutant larvae. Results: Homozygous mutant (scn1lab mut/mut ) larvae displayed spontaneous seizures with interictal, preictal, and ictal discharges (mean = 7.5 per 20-minute recording; P < .0001; one-way analysis of variance). Drop-Seq analysis revealed a 2:1 shift in the ratio of glutamatergic to GABAergic neurons in scn1lab mut/mut larval brains versus wild type (WT), with dynamic changes in neuronal, glial, and progenitor cell populations. To explore disease pathophysiology further, we quantified dendritic arborization in GABAergic neurons and observed a 40% reduction in arbor number compared to WT (P < .001; n = 15 mutant, n = 16 WT). We postulate that the significant reduction in inhibitory arbors causes an inhibitory to excitatory neurotransmitter imbalance that contributes to seizures and enhanced electrical brain activity in scn1lab mut/mut larvae (high-frequency range), with subsequent GABAergic neuronal loss and astrogliosis. Chronic fenfluramine administration completely restored dendritic arbor numbers to normal in scn1lab mut/mut larvae, whereas similar treatment with the benzodiazepine diazepam attenuated seizures, but was ineffective in restoring neuronal cytoarchitecture. BrdU labeling revealed cell overproliferation in scn1lab mut/mut larval brains that were rescued by fenfluramine but not diazepam. Significance: Our findings provide novel insights into early mechanisms of DS pathogenesis, describe dynamic cell population changes in the scn1lab mut/mut brain, and present first-time evidence for potential disease modification by fenfluramine. (© 2020 The Authors. Epilepsia published by Wiley Periodicals, Inc. on behalf of International League Against Epilepsy.) |
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معلومات مُعتمدة: | International Centre for Molecular Medicine Norway (NCMM) Start up grant; C14/BM/7975668/CaSCAD International National Research Fund of Luxembourg; INTER/DFG/17/11583046 MechEPI (Mechanisms of Epileptogenesis) International National Research Fund of Luxembourg |
فهرسة مساهمة: | Keywords: Dravet syndrome; epileptogenesis; fenfluramine; sodium channel; zebrafish |
المشرفين على المادة: | 0 (Anticonvulsants) 0 (NAV1.1 Voltage-Gated Sodium Channel) 0 (Serotonin 5-HT2 Receptor Agonists) 0 (Zebrafish Proteins) 0 (scn1laa protein, zebrafish) 2DS058H2CF (Fenfluramine) Q3JTX2Q7TU (Diazepam) |
تواريخ الأحداث: | Date Created: 20200226 Date Completed: 20201019 Latest Revision: 20201019 |
رمز التحديث: | 20240628 |
DOI: | 10.1111/epi.16456 |
PMID: | 32096222 |
قاعدة البيانات: | MEDLINE |
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