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A robust ex vivo experimental platform for molecular-genetic dissection of adult human neocortical cell types and circuits
The powerful suite of available genetic tools is driving tremendous progress in understanding mouse brain cell types and circuits. However, the degree of conservation in human remains largely unknown in large part due to the lack of such tools and healthy tissue preparations. To close this gap, we d...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5976666/ https://www.ncbi.nlm.nih.gov/pubmed/29849137 http://dx.doi.org/10.1038/s41598-018-26803-9 |
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author | Ting, Jonathan T. Kalmbach, Brian Chong, Peter de Frates, Rebecca Keene, C. Dirk Gwinn, Ryder P. Cobbs, Charles Ko, Andrew L. Ojemann, Jeffrey G. Ellenbogen, Richard G. Koch, Christof Lein, Ed |
author_facet | Ting, Jonathan T. Kalmbach, Brian Chong, Peter de Frates, Rebecca Keene, C. Dirk Gwinn, Ryder P. Cobbs, Charles Ko, Andrew L. Ojemann, Jeffrey G. Ellenbogen, Richard G. Koch, Christof Lein, Ed |
author_sort | Ting, Jonathan T. |
collection | PubMed |
description | The powerful suite of available genetic tools is driving tremendous progress in understanding mouse brain cell types and circuits. However, the degree of conservation in human remains largely unknown in large part due to the lack of such tools and healthy tissue preparations. To close this gap, we describe a robust and stable adult human neurosurgically-derived ex vivo acute and cultured neocortical brain slice system optimized for rapid molecular-genetic manipulation. Surprisingly, acute human brain slices exhibited exceptional viability, and neuronal intrinsic membrane properties could be assayed for at least three days. Maintaining adult human slices in culture under sterile conditions further enabled the application of viral tools to drive rapid expression of exogenous transgenes. Widespread neuron-specific labeling was achieved as early as two days post infection with HSV-1 vectors, with virally-transduced neurons exhibiting membrane properties largely comparable to uninfected neurons over this short timeframe. Finally, we demonstrate the suitability of this culture paradigm for optical manipulation and monitoring of neuronal activity using genetically encoded probes, opening a path for applying modern molecular-genetic tools to study human brain circuit function. |
format | Online Article Text |
id | pubmed-5976666 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-59766662018-05-31 A robust ex vivo experimental platform for molecular-genetic dissection of adult human neocortical cell types and circuits Ting, Jonathan T. Kalmbach, Brian Chong, Peter de Frates, Rebecca Keene, C. Dirk Gwinn, Ryder P. Cobbs, Charles Ko, Andrew L. Ojemann, Jeffrey G. Ellenbogen, Richard G. Koch, Christof Lein, Ed Sci Rep Article The powerful suite of available genetic tools is driving tremendous progress in understanding mouse brain cell types and circuits. However, the degree of conservation in human remains largely unknown in large part due to the lack of such tools and healthy tissue preparations. To close this gap, we describe a robust and stable adult human neurosurgically-derived ex vivo acute and cultured neocortical brain slice system optimized for rapid molecular-genetic manipulation. Surprisingly, acute human brain slices exhibited exceptional viability, and neuronal intrinsic membrane properties could be assayed for at least three days. Maintaining adult human slices in culture under sterile conditions further enabled the application of viral tools to drive rapid expression of exogenous transgenes. Widespread neuron-specific labeling was achieved as early as two days post infection with HSV-1 vectors, with virally-transduced neurons exhibiting membrane properties largely comparable to uninfected neurons over this short timeframe. Finally, we demonstrate the suitability of this culture paradigm for optical manipulation and monitoring of neuronal activity using genetically encoded probes, opening a path for applying modern molecular-genetic tools to study human brain circuit function. Nature Publishing Group UK 2018-05-30 /pmc/articles/PMC5976666/ /pubmed/29849137 http://dx.doi.org/10.1038/s41598-018-26803-9 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Ting, Jonathan T. Kalmbach, Brian Chong, Peter de Frates, Rebecca Keene, C. Dirk Gwinn, Ryder P. Cobbs, Charles Ko, Andrew L. Ojemann, Jeffrey G. Ellenbogen, Richard G. Koch, Christof Lein, Ed A robust ex vivo experimental platform for molecular-genetic dissection of adult human neocortical cell types and circuits |
title | A robust ex vivo experimental platform for molecular-genetic dissection of adult human neocortical cell types and circuits |
title_full | A robust ex vivo experimental platform for molecular-genetic dissection of adult human neocortical cell types and circuits |
title_fullStr | A robust ex vivo experimental platform for molecular-genetic dissection of adult human neocortical cell types and circuits |
title_full_unstemmed | A robust ex vivo experimental platform for molecular-genetic dissection of adult human neocortical cell types and circuits |
title_short | A robust ex vivo experimental platform for molecular-genetic dissection of adult human neocortical cell types and circuits |
title_sort | robust ex vivo experimental platform for molecular-genetic dissection of adult human neocortical cell types and circuits |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5976666/ https://www.ncbi.nlm.nih.gov/pubmed/29849137 http://dx.doi.org/10.1038/s41598-018-26803-9 |
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