Cargando…
Serotonin-specific neurons differentiated from human iPSCs form distinct subtypes with synaptic protein assembly
Human induced pluripotent stem cells (hiPSCs) have revolutionized the generation of experimental disease models, but the development of protocols for the differentiation of functionally active neuronal subtypes with defined specification is still in its infancy. While dysfunction of the brain seroto...
Autores principales: | , , , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Vienna
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7914246/ https://www.ncbi.nlm.nih.gov/pubmed/33560471 http://dx.doi.org/10.1007/s00702-021-02303-5 |
_version_ | 1783656972759859200 |
---|---|
author | Jansch, Charline Ziegler, Georg C. Forero, Andrea Gredy, Sina Wäldchen, Sina Vitale, Maria Rosaria Svirin, Evgeniy Zöller, Johanna E. M. Waider, Jonas Günther, Katharina Edenhofer, Frank Sauer, Markus Wischmeyer, Erhard Lesch, Klaus-Peter |
author_facet | Jansch, Charline Ziegler, Georg C. Forero, Andrea Gredy, Sina Wäldchen, Sina Vitale, Maria Rosaria Svirin, Evgeniy Zöller, Johanna E. M. Waider, Jonas Günther, Katharina Edenhofer, Frank Sauer, Markus Wischmeyer, Erhard Lesch, Klaus-Peter |
author_sort | Jansch, Charline |
collection | PubMed |
description | Human induced pluripotent stem cells (hiPSCs) have revolutionized the generation of experimental disease models, but the development of protocols for the differentiation of functionally active neuronal subtypes with defined specification is still in its infancy. While dysfunction of the brain serotonin (5-HT) system has been implicated in the etiology of various neuropsychiatric disorders, investigation of functional human 5-HT specific neurons in vitro has been restricted by technical limitations. We describe an efficient generation of functionally active neurons from hiPSCs displaying 5-HT specification by modification of a previously reported protocol. Furthermore, 5-HT specific neurons were characterized using high-end fluorescence imaging including super-resolution microscopy in combination with electrophysiological techniques. Differentiated hiPSCs synthesize 5-HT, express specific markers, such as tryptophan hydroxylase 2 and 5-HT transporter, and exhibit an electrophysiological signature characteristic of serotonergic neurons, with spontaneous rhythmic activities, broad action potentials and large afterhyperpolarization potentials. 5-HT specific neurons form synapses reflected by the expression of pre- and postsynaptic proteins, such as Bassoon and Homer. The distribution pattern of Bassoon, a marker of the active zone along the soma and extensions of neurons, indicates functionality via volume transmission. Among the high percentage of 5-HT specific neurons (~ 42%), a subpopulation of CDH13 + cells presumably designates dorsal raphe neurons. hiPSC-derived 5-HT specific neuronal cell cultures reflect the heterogeneous nature of dorsal and median raphe nuclei and may facilitate examining the association of serotonergic neuron subpopulations with neuropsychiatric disorders. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00702-021-02303-5. |
format | Online Article Text |
id | pubmed-7914246 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Springer Vienna |
record_format | MEDLINE/PubMed |
spelling | pubmed-79142462021-03-15 Serotonin-specific neurons differentiated from human iPSCs form distinct subtypes with synaptic protein assembly Jansch, Charline Ziegler, Georg C. Forero, Andrea Gredy, Sina Wäldchen, Sina Vitale, Maria Rosaria Svirin, Evgeniy Zöller, Johanna E. M. Waider, Jonas Günther, Katharina Edenhofer, Frank Sauer, Markus Wischmeyer, Erhard Lesch, Klaus-Peter J Neural Transm (Vienna) Psychiatry and Preclinical Psychiatric Studies - Original Article Human induced pluripotent stem cells (hiPSCs) have revolutionized the generation of experimental disease models, but the development of protocols for the differentiation of functionally active neuronal subtypes with defined specification is still in its infancy. While dysfunction of the brain serotonin (5-HT) system has been implicated in the etiology of various neuropsychiatric disorders, investigation of functional human 5-HT specific neurons in vitro has been restricted by technical limitations. We describe an efficient generation of functionally active neurons from hiPSCs displaying 5-HT specification by modification of a previously reported protocol. Furthermore, 5-HT specific neurons were characterized using high-end fluorescence imaging including super-resolution microscopy in combination with electrophysiological techniques. Differentiated hiPSCs synthesize 5-HT, express specific markers, such as tryptophan hydroxylase 2 and 5-HT transporter, and exhibit an electrophysiological signature characteristic of serotonergic neurons, with spontaneous rhythmic activities, broad action potentials and large afterhyperpolarization potentials. 5-HT specific neurons form synapses reflected by the expression of pre- and postsynaptic proteins, such as Bassoon and Homer. The distribution pattern of Bassoon, a marker of the active zone along the soma and extensions of neurons, indicates functionality via volume transmission. Among the high percentage of 5-HT specific neurons (~ 42%), a subpopulation of CDH13 + cells presumably designates dorsal raphe neurons. hiPSC-derived 5-HT specific neuronal cell cultures reflect the heterogeneous nature of dorsal and median raphe nuclei and may facilitate examining the association of serotonergic neuron subpopulations with neuropsychiatric disorders. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00702-021-02303-5. Springer Vienna 2021-02-09 2021 /pmc/articles/PMC7914246/ /pubmed/33560471 http://dx.doi.org/10.1007/s00702-021-02303-5 Text en © The Author(s) 2021 Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Psychiatry and Preclinical Psychiatric Studies - Original Article Jansch, Charline Ziegler, Georg C. Forero, Andrea Gredy, Sina Wäldchen, Sina Vitale, Maria Rosaria Svirin, Evgeniy Zöller, Johanna E. M. Waider, Jonas Günther, Katharina Edenhofer, Frank Sauer, Markus Wischmeyer, Erhard Lesch, Klaus-Peter Serotonin-specific neurons differentiated from human iPSCs form distinct subtypes with synaptic protein assembly |
title | Serotonin-specific neurons differentiated from human iPSCs form distinct subtypes with synaptic protein assembly |
title_full | Serotonin-specific neurons differentiated from human iPSCs form distinct subtypes with synaptic protein assembly |
title_fullStr | Serotonin-specific neurons differentiated from human iPSCs form distinct subtypes with synaptic protein assembly |
title_full_unstemmed | Serotonin-specific neurons differentiated from human iPSCs form distinct subtypes with synaptic protein assembly |
title_short | Serotonin-specific neurons differentiated from human iPSCs form distinct subtypes with synaptic protein assembly |
title_sort | serotonin-specific neurons differentiated from human ipscs form distinct subtypes with synaptic protein assembly |
topic | Psychiatry and Preclinical Psychiatric Studies - Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7914246/ https://www.ncbi.nlm.nih.gov/pubmed/33560471 http://dx.doi.org/10.1007/s00702-021-02303-5 |
work_keys_str_mv | AT janschcharline serotoninspecificneuronsdifferentiatedfromhumanipscsformdistinctsubtypeswithsynapticproteinassembly AT zieglergeorgc serotoninspecificneuronsdifferentiatedfromhumanipscsformdistinctsubtypeswithsynapticproteinassembly AT foreroandrea serotoninspecificneuronsdifferentiatedfromhumanipscsformdistinctsubtypeswithsynapticproteinassembly AT gredysina serotoninspecificneuronsdifferentiatedfromhumanipscsformdistinctsubtypeswithsynapticproteinassembly AT waldchensina serotoninspecificneuronsdifferentiatedfromhumanipscsformdistinctsubtypeswithsynapticproteinassembly AT vitalemariarosaria serotoninspecificneuronsdifferentiatedfromhumanipscsformdistinctsubtypeswithsynapticproteinassembly AT svirinevgeniy serotoninspecificneuronsdifferentiatedfromhumanipscsformdistinctsubtypeswithsynapticproteinassembly AT zollerjohannaem serotoninspecificneuronsdifferentiatedfromhumanipscsformdistinctsubtypeswithsynapticproteinassembly AT waiderjonas serotoninspecificneuronsdifferentiatedfromhumanipscsformdistinctsubtypeswithsynapticproteinassembly AT guntherkatharina serotoninspecificneuronsdifferentiatedfromhumanipscsformdistinctsubtypeswithsynapticproteinassembly AT edenhoferfrank serotoninspecificneuronsdifferentiatedfromhumanipscsformdistinctsubtypeswithsynapticproteinassembly AT sauermarkus serotoninspecificneuronsdifferentiatedfromhumanipscsformdistinctsubtypeswithsynapticproteinassembly AT wischmeyererhard serotoninspecificneuronsdifferentiatedfromhumanipscsformdistinctsubtypeswithsynapticproteinassembly AT leschklauspeter serotoninspecificneuronsdifferentiatedfromhumanipscsformdistinctsubtypeswithsynapticproteinassembly |