Cargando…
Differentiation-Dependent Motility-Responses of Developing Neural Progenitors to Optogenetic Stimulation
During neural tissue genesis, neural stem/progenitor cells are exposed to bioelectric stimuli well before synaptogenesis and neural circuit formation. Fluctuations in the electrochemical potential in the vicinity of developing cells influence the genesis, migration and maturation of neuronal precurs...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2017
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5742229/ https://www.ncbi.nlm.nih.gov/pubmed/29311832 http://dx.doi.org/10.3389/fncel.2017.00401 |
_version_ | 1783288336720330752 |
---|---|
author | Köhidi, Tímea Jády, Attila G. Markó, Károly Papp, Noémi Andrási, Tibor Környei, Zsuzsanna Madarász, Emília |
author_facet | Köhidi, Tímea Jády, Attila G. Markó, Károly Papp, Noémi Andrási, Tibor Környei, Zsuzsanna Madarász, Emília |
author_sort | Köhidi, Tímea |
collection | PubMed |
description | During neural tissue genesis, neural stem/progenitor cells are exposed to bioelectric stimuli well before synaptogenesis and neural circuit formation. Fluctuations in the electrochemical potential in the vicinity of developing cells influence the genesis, migration and maturation of neuronal precursors. The complexity of the in vivo environment and the coexistence of various progenitor populations hinder the understanding of the significance of ionic/bioelectric stimuli in the early phases of neuronal differentiation. Using optogenetic stimulation, we investigated the in vitro motility responses of radial glia-like neural stem/progenitor populations to ionic stimuli. Radial glia-like neural stem cells were isolated from CAG(loxp)Stop(loxp)ChR2(H134)-eYFP transgenic mouse embryos. After transfection with Cre-recombinase, ChR2(channelrhodopsin-2)-expressing and non-expressing cells were separated by eYFP fluorescence. Expression of light-gated ion channels were checked by patch clamp and fluorescence intensity assays. Neurogenesis by ChR2-expressing and non-expressing cells was induced by withdrawal of EGF from the medium. Cells in different (stem cell, migrating progenitor and maturing precursor) stages of development were illuminated with laser light (λ = 488 nm; 1.3 mW/mm(2); 300 ms) in every 5 min for 12 h. The displacement of the cells was analyzed on images taken at the end of each light pulse. Results demonstrated that the migratory activity decreased with the advancement of neuronal differentiation regardless of stimulation. Light-sensitive cells, however, responded on a differentiation-dependent way. In non-differentiated ChR2-expressing stem cell populations, the motility did not change significantly in response to light-stimulation. The displacement activity of migrating progenitors was enhanced, while the motility of differentiating neuronal precursors was markedly reduced by illumination. |
format | Online Article Text |
id | pubmed-5742229 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-57422292018-01-08 Differentiation-Dependent Motility-Responses of Developing Neural Progenitors to Optogenetic Stimulation Köhidi, Tímea Jády, Attila G. Markó, Károly Papp, Noémi Andrási, Tibor Környei, Zsuzsanna Madarász, Emília Front Cell Neurosci Neuroscience During neural tissue genesis, neural stem/progenitor cells are exposed to bioelectric stimuli well before synaptogenesis and neural circuit formation. Fluctuations in the electrochemical potential in the vicinity of developing cells influence the genesis, migration and maturation of neuronal precursors. The complexity of the in vivo environment and the coexistence of various progenitor populations hinder the understanding of the significance of ionic/bioelectric stimuli in the early phases of neuronal differentiation. Using optogenetic stimulation, we investigated the in vitro motility responses of radial glia-like neural stem/progenitor populations to ionic stimuli. Radial glia-like neural stem cells were isolated from CAG(loxp)Stop(loxp)ChR2(H134)-eYFP transgenic mouse embryos. After transfection with Cre-recombinase, ChR2(channelrhodopsin-2)-expressing and non-expressing cells were separated by eYFP fluorescence. Expression of light-gated ion channels were checked by patch clamp and fluorescence intensity assays. Neurogenesis by ChR2-expressing and non-expressing cells was induced by withdrawal of EGF from the medium. Cells in different (stem cell, migrating progenitor and maturing precursor) stages of development were illuminated with laser light (λ = 488 nm; 1.3 mW/mm(2); 300 ms) in every 5 min for 12 h. The displacement of the cells was analyzed on images taken at the end of each light pulse. Results demonstrated that the migratory activity decreased with the advancement of neuronal differentiation regardless of stimulation. Light-sensitive cells, however, responded on a differentiation-dependent way. In non-differentiated ChR2-expressing stem cell populations, the motility did not change significantly in response to light-stimulation. The displacement activity of migrating progenitors was enhanced, while the motility of differentiating neuronal precursors was markedly reduced by illumination. Frontiers Media S.A. 2017-12-19 /pmc/articles/PMC5742229/ /pubmed/29311832 http://dx.doi.org/10.3389/fncel.2017.00401 Text en Copyright © 2017 Köhidi, Jády, Markó, Papp, Andrási, Környei and Madarász. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Neuroscience Köhidi, Tímea Jády, Attila G. Markó, Károly Papp, Noémi Andrási, Tibor Környei, Zsuzsanna Madarász, Emília Differentiation-Dependent Motility-Responses of Developing Neural Progenitors to Optogenetic Stimulation |
title | Differentiation-Dependent Motility-Responses of Developing Neural Progenitors to Optogenetic Stimulation |
title_full | Differentiation-Dependent Motility-Responses of Developing Neural Progenitors to Optogenetic Stimulation |
title_fullStr | Differentiation-Dependent Motility-Responses of Developing Neural Progenitors to Optogenetic Stimulation |
title_full_unstemmed | Differentiation-Dependent Motility-Responses of Developing Neural Progenitors to Optogenetic Stimulation |
title_short | Differentiation-Dependent Motility-Responses of Developing Neural Progenitors to Optogenetic Stimulation |
title_sort | differentiation-dependent motility-responses of developing neural progenitors to optogenetic stimulation |
topic | Neuroscience |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5742229/ https://www.ncbi.nlm.nih.gov/pubmed/29311832 http://dx.doi.org/10.3389/fncel.2017.00401 |
work_keys_str_mv | AT kohiditimea differentiationdependentmotilityresponsesofdevelopingneuralprogenitorstooptogeneticstimulation AT jadyattilag differentiationdependentmotilityresponsesofdevelopingneuralprogenitorstooptogeneticstimulation AT markokaroly differentiationdependentmotilityresponsesofdevelopingneuralprogenitorstooptogeneticstimulation AT pappnoemi differentiationdependentmotilityresponsesofdevelopingneuralprogenitorstooptogeneticstimulation AT andrasitibor differentiationdependentmotilityresponsesofdevelopingneuralprogenitorstooptogeneticstimulation AT kornyeizsuzsanna differentiationdependentmotilityresponsesofdevelopingneuralprogenitorstooptogeneticstimulation AT madaraszemilia differentiationdependentmotilityresponsesofdevelopingneuralprogenitorstooptogeneticstimulation |