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Altered synaptic connectivity in an in vitro human model of STXBP1 encephalopathy

Early infantile developmental and epileptic encephalopathies are devastating conditions, generally of genetic origin, but the pathological mechanisms often remain obscure. A major obstacle in this field of research is the difficulty of studying cortical brain development in humans, at the relevant t...

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Autores principales: McLeod, Faye, Dimtsi, Anna, Marshall, Amy C, Lewis-Smith, David, Thomas, Rhys, Clowry, Gavin J, Trevelyan, Andrew J
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9976961/
https://www.ncbi.nlm.nih.gov/pubmed/36315647
http://dx.doi.org/10.1093/brain/awac396
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author McLeod, Faye
Dimtsi, Anna
Marshall, Amy C
Lewis-Smith, David
Thomas, Rhys
Clowry, Gavin J
Trevelyan, Andrew J
author_facet McLeod, Faye
Dimtsi, Anna
Marshall, Amy C
Lewis-Smith, David
Thomas, Rhys
Clowry, Gavin J
Trevelyan, Andrew J
author_sort McLeod, Faye
collection PubMed
description Early infantile developmental and epileptic encephalopathies are devastating conditions, generally of genetic origin, but the pathological mechanisms often remain obscure. A major obstacle in this field of research is the difficulty of studying cortical brain development in humans, at the relevant time period in utero. To address this, we established an in vitro assay to study the impact of gene variants on the developing human brain by using living organotypic cultures of the human subplate and neighbouring cortical regions, prepared from ethically sourced, 14–17 post-conception week brain tissue (www.hdbr.org). We were able to maintain cultures for several months, during which time the gross anatomical structures of the cortical plate, subplate and marginal zone persisted, while neurons continued to develop morphologically and form new synaptic networks. This preparation thus permits the study of genetic manipulations and their downstream effects on an intact developing human cortical network. We focused on STXBP1 haploinsufficiency, which is among the most common genetic causes of developmental and epileptic encephalopathy. This was induced using shRNA interference, leading to impaired synaptic function and a reduced density of glutamatergic synapses. We thereby provide a critical proof-of-principle for how to study the impact of any gene of interest on the development of the human cortex.
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spelling pubmed-99769612023-03-02 Altered synaptic connectivity in an in vitro human model of STXBP1 encephalopathy McLeod, Faye Dimtsi, Anna Marshall, Amy C Lewis-Smith, David Thomas, Rhys Clowry, Gavin J Trevelyan, Andrew J Brain Report Early infantile developmental and epileptic encephalopathies are devastating conditions, generally of genetic origin, but the pathological mechanisms often remain obscure. A major obstacle in this field of research is the difficulty of studying cortical brain development in humans, at the relevant time period in utero. To address this, we established an in vitro assay to study the impact of gene variants on the developing human brain by using living organotypic cultures of the human subplate and neighbouring cortical regions, prepared from ethically sourced, 14–17 post-conception week brain tissue (www.hdbr.org). We were able to maintain cultures for several months, during which time the gross anatomical structures of the cortical plate, subplate and marginal zone persisted, while neurons continued to develop morphologically and form new synaptic networks. This preparation thus permits the study of genetic manipulations and their downstream effects on an intact developing human cortical network. We focused on STXBP1 haploinsufficiency, which is among the most common genetic causes of developmental and epileptic encephalopathy. This was induced using shRNA interference, leading to impaired synaptic function and a reduced density of glutamatergic synapses. We thereby provide a critical proof-of-principle for how to study the impact of any gene of interest on the development of the human cortex. Oxford University Press 2022-10-31 /pmc/articles/PMC9976961/ /pubmed/36315647 http://dx.doi.org/10.1093/brain/awac396 Text en © The Author(s) 2022. Published by Oxford University Press on behalf of the Guarantors of Brain. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Report
McLeod, Faye
Dimtsi, Anna
Marshall, Amy C
Lewis-Smith, David
Thomas, Rhys
Clowry, Gavin J
Trevelyan, Andrew J
Altered synaptic connectivity in an in vitro human model of STXBP1 encephalopathy
title Altered synaptic connectivity in an in vitro human model of STXBP1 encephalopathy
title_full Altered synaptic connectivity in an in vitro human model of STXBP1 encephalopathy
title_fullStr Altered synaptic connectivity in an in vitro human model of STXBP1 encephalopathy
title_full_unstemmed Altered synaptic connectivity in an in vitro human model of STXBP1 encephalopathy
title_short Altered synaptic connectivity in an in vitro human model of STXBP1 encephalopathy
title_sort altered synaptic connectivity in an in vitro human model of stxbp1 encephalopathy
topic Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9976961/
https://www.ncbi.nlm.nih.gov/pubmed/36315647
http://dx.doi.org/10.1093/brain/awac396
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