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Automated and manual patch clamp data of human induced pluripotent stem cell-derived dopaminergic neurons

Human induced pluripotent stem cells can be differentiated into dopaminergic neurons (Dopa.4U). Dopa.4U neurons expressed voltage-gated Na(V) and K(V) channels and showed neuron-like spontaneous electrical activity. In automated patch clamp measurements with suspended Dopa.4U neurons, delayed rectif...

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Autores principales: Franz, Denise, Olsen, Hervør Lykke, Klink, Oliver, Gimsa, Jan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5404656/
https://www.ncbi.nlm.nih.gov/pubmed/28440808
http://dx.doi.org/10.1038/sdata.2017.56
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author Franz, Denise
Olsen, Hervør Lykke
Klink, Oliver
Gimsa, Jan
author_facet Franz, Denise
Olsen, Hervør Lykke
Klink, Oliver
Gimsa, Jan
author_sort Franz, Denise
collection PubMed
description Human induced pluripotent stem cells can be differentiated into dopaminergic neurons (Dopa.4U). Dopa.4U neurons expressed voltage-gated Na(V) and K(V) channels and showed neuron-like spontaneous electrical activity. In automated patch clamp measurements with suspended Dopa.4U neurons, delayed rectifier K(+) current (delayed K(V)) and rapidly inactivating A-type K(+) current (fast K(V)) were identified. Examination of the fast K(V) current with inhibitors yielded IC(50) values of 0.4 mM (4-aminopyridine) and 0.1 mM (tetraethylammonium). In manual patch clamp measurements with adherent Dopa.4U neurons, fast K(V) current could not be detected, while the delayed K(V) current showed an IC(50) of 2 mM for 4-aminopyridine. The Na(V) channels in adherent and suspended Dopa.4U neurons showed IC(50) values for tetrodotoxin of 27 and 2.9 nM, respectively. GABA-induced currents that could be observed in adherent Dopa.4U neurons could not be detected in suspended cells. Application of current pulses induced action potentials in approx. 70 % of the cells. Our results proved the feasibility of automated electrophysiological characterization of neuronal cells.
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spelling pubmed-54046562017-05-01 Automated and manual patch clamp data of human induced pluripotent stem cell-derived dopaminergic neurons Franz, Denise Olsen, Hervør Lykke Klink, Oliver Gimsa, Jan Sci Data Data Descriptor Human induced pluripotent stem cells can be differentiated into dopaminergic neurons (Dopa.4U). Dopa.4U neurons expressed voltage-gated Na(V) and K(V) channels and showed neuron-like spontaneous electrical activity. In automated patch clamp measurements with suspended Dopa.4U neurons, delayed rectifier K(+) current (delayed K(V)) and rapidly inactivating A-type K(+) current (fast K(V)) were identified. Examination of the fast K(V) current with inhibitors yielded IC(50) values of 0.4 mM (4-aminopyridine) and 0.1 mM (tetraethylammonium). In manual patch clamp measurements with adherent Dopa.4U neurons, fast K(V) current could not be detected, while the delayed K(V) current showed an IC(50) of 2 mM for 4-aminopyridine. The Na(V) channels in adherent and suspended Dopa.4U neurons showed IC(50) values for tetrodotoxin of 27 and 2.9 nM, respectively. GABA-induced currents that could be observed in adherent Dopa.4U neurons could not be detected in suspended cells. Application of current pulses induced action potentials in approx. 70 % of the cells. Our results proved the feasibility of automated electrophysiological characterization of neuronal cells. Nature Publishing Group 2017-04-25 /pmc/articles/PMC5404656/ /pubmed/28440808 http://dx.doi.org/10.1038/sdata.2017.56 Text en Copyright © 2017, The Author(s) http://creativecommons.org/licenses/by/4.0 This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0 Metadata associated with this Data Descriptor is available at http://www.nature.com/sdata/ and is released under the CC0 waiver to maximize reuse.
spellingShingle Data Descriptor
Franz, Denise
Olsen, Hervør Lykke
Klink, Oliver
Gimsa, Jan
Automated and manual patch clamp data of human induced pluripotent stem cell-derived dopaminergic neurons
title Automated and manual patch clamp data of human induced pluripotent stem cell-derived dopaminergic neurons
title_full Automated and manual patch clamp data of human induced pluripotent stem cell-derived dopaminergic neurons
title_fullStr Automated and manual patch clamp data of human induced pluripotent stem cell-derived dopaminergic neurons
title_full_unstemmed Automated and manual patch clamp data of human induced pluripotent stem cell-derived dopaminergic neurons
title_short Automated and manual patch clamp data of human induced pluripotent stem cell-derived dopaminergic neurons
title_sort automated and manual patch clamp data of human induced pluripotent stem cell-derived dopaminergic neurons
topic Data Descriptor
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5404656/
https://www.ncbi.nlm.nih.gov/pubmed/28440808
http://dx.doi.org/10.1038/sdata.2017.56
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