Cargando…
Regulation of Progenitor Cell Proliferation and Neuronal Differentiation in Enteric Nervous System Neurospheres
Enteric nervous system (ENS) progenitor cells isolated from mouse and human bowel can be cultured in vitro as neurospheres which are aggregates of the proliferating progenitor cells, together with neurons and glial cells derived from them. To investigate the factors regulating progenitor cell prolif...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3553067/ https://www.ncbi.nlm.nih.gov/pubmed/23372773 http://dx.doi.org/10.1371/journal.pone.0054809 |
_version_ | 1782256777141682176 |
---|---|
author | Theocharatos, Sokratis Wilkinson, David J. Darling, Sarah Wilm, Bettina Kenny, Simon E. Edgar, David |
author_facet | Theocharatos, Sokratis Wilkinson, David J. Darling, Sarah Wilm, Bettina Kenny, Simon E. Edgar, David |
author_sort | Theocharatos, Sokratis |
collection | PubMed |
description | Enteric nervous system (ENS) progenitor cells isolated from mouse and human bowel can be cultured in vitro as neurospheres which are aggregates of the proliferating progenitor cells, together with neurons and glial cells derived from them. To investigate the factors regulating progenitor cell proliferation and differentiation, we first characterised cell proliferation in mouse ENS neurospheres by pulse chase experiments using thymidine analogs. We demonstrate rapid and continuous cell proliferation near the neurosphere periphery, after which postmitotic cells move away from the periphery to become distributed throughout the neurosphere. While many proliferating cells expressed glial markers, expression of the neuronal markers β-tubulin III (Tuj1) and nitric oxide synthase was detected in increasing numbers of post-mitotic cells after a delay of several days. Treatment of both mouse and human neurospheres with the γ-secretase inhibitor N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT) reduced expression of the transcription factors Hes1 and Hes5, demonstrating inhibition of Notch signaling. DAPT treatment also inhibited progenitor cell proliferation and increased the numbers of differentiating neurons expressing Tuj1 and nitric oxide synthase. To confirm that the cellular effects of DAPT treatment were due to inhibition of Notch signaling, siRNA knockdown of RBPjκ, a key component of the canonical Notch signaling pathway, was demonstrated both to reduce proliferation and to increase neuronal differentiation in neurosphere cells. These observations indicate that Notch signaling promotes progenitor cell proliferation and inhibits neuronal differentiation in ENS neurospheres. |
format | Online Article Text |
id | pubmed-3553067 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-35530672013-01-31 Regulation of Progenitor Cell Proliferation and Neuronal Differentiation in Enteric Nervous System Neurospheres Theocharatos, Sokratis Wilkinson, David J. Darling, Sarah Wilm, Bettina Kenny, Simon E. Edgar, David PLoS One Research Article Enteric nervous system (ENS) progenitor cells isolated from mouse and human bowel can be cultured in vitro as neurospheres which are aggregates of the proliferating progenitor cells, together with neurons and glial cells derived from them. To investigate the factors regulating progenitor cell proliferation and differentiation, we first characterised cell proliferation in mouse ENS neurospheres by pulse chase experiments using thymidine analogs. We demonstrate rapid and continuous cell proliferation near the neurosphere periphery, after which postmitotic cells move away from the periphery to become distributed throughout the neurosphere. While many proliferating cells expressed glial markers, expression of the neuronal markers β-tubulin III (Tuj1) and nitric oxide synthase was detected in increasing numbers of post-mitotic cells after a delay of several days. Treatment of both mouse and human neurospheres with the γ-secretase inhibitor N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT) reduced expression of the transcription factors Hes1 and Hes5, demonstrating inhibition of Notch signaling. DAPT treatment also inhibited progenitor cell proliferation and increased the numbers of differentiating neurons expressing Tuj1 and nitric oxide synthase. To confirm that the cellular effects of DAPT treatment were due to inhibition of Notch signaling, siRNA knockdown of RBPjκ, a key component of the canonical Notch signaling pathway, was demonstrated both to reduce proliferation and to increase neuronal differentiation in neurosphere cells. These observations indicate that Notch signaling promotes progenitor cell proliferation and inhibits neuronal differentiation in ENS neurospheres. Public Library of Science 2013-01-23 /pmc/articles/PMC3553067/ /pubmed/23372773 http://dx.doi.org/10.1371/journal.pone.0054809 Text en © 2013 Theocharatos et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Theocharatos, Sokratis Wilkinson, David J. Darling, Sarah Wilm, Bettina Kenny, Simon E. Edgar, David Regulation of Progenitor Cell Proliferation and Neuronal Differentiation in Enteric Nervous System Neurospheres |
title | Regulation of Progenitor Cell Proliferation and Neuronal Differentiation in Enteric Nervous System Neurospheres |
title_full | Regulation of Progenitor Cell Proliferation and Neuronal Differentiation in Enteric Nervous System Neurospheres |
title_fullStr | Regulation of Progenitor Cell Proliferation and Neuronal Differentiation in Enteric Nervous System Neurospheres |
title_full_unstemmed | Regulation of Progenitor Cell Proliferation and Neuronal Differentiation in Enteric Nervous System Neurospheres |
title_short | Regulation of Progenitor Cell Proliferation and Neuronal Differentiation in Enteric Nervous System Neurospheres |
title_sort | regulation of progenitor cell proliferation and neuronal differentiation in enteric nervous system neurospheres |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3553067/ https://www.ncbi.nlm.nih.gov/pubmed/23372773 http://dx.doi.org/10.1371/journal.pone.0054809 |
work_keys_str_mv | AT theocharatossokratis regulationofprogenitorcellproliferationandneuronaldifferentiationinentericnervoussystemneurospheres AT wilkinsondavidj regulationofprogenitorcellproliferationandneuronaldifferentiationinentericnervoussystemneurospheres AT darlingsarah regulationofprogenitorcellproliferationandneuronaldifferentiationinentericnervoussystemneurospheres AT wilmbettina regulationofprogenitorcellproliferationandneuronaldifferentiationinentericnervoussystemneurospheres AT kennysimone regulationofprogenitorcellproliferationandneuronaldifferentiationinentericnervoussystemneurospheres AT edgardavid regulationofprogenitorcellproliferationandneuronaldifferentiationinentericnervoussystemneurospheres |