Cargando…
Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line
BACKGROUND: Satellite cells are residential muscle stem cells that express a paired box protein, PAX7. RESULTS: Here, we report a knock-in mouse line expressing a PAX7-enhanced yellow fluorescent protein (YFP) fusion protein that enables visualization of PAX7 protein dynamics in living satellite cel...
Autores principales: | , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6108100/ https://www.ncbi.nlm.nih.gov/pubmed/30139390 http://dx.doi.org/10.1186/s13395-018-0174-x |
_version_ | 1783350084931420160 |
---|---|
author | Kitajima, Yasuo Ono, Yusuke |
author_facet | Kitajima, Yasuo Ono, Yusuke |
author_sort | Kitajima, Yasuo |
collection | PubMed |
description | BACKGROUND: Satellite cells are residential muscle stem cells that express a paired box protein, PAX7. RESULTS: Here, we report a knock-in mouse line expressing a PAX7-enhanced yellow fluorescent protein (YFP) fusion protein that enables visualization of PAX7 protein dynamics in living satellite cells through YFP fluorescence. The YFP fluorescence signals in Pax7-YFP knock-in mice clearly recapitulated the endogenous expression of PAX7 protein in satellite cells. YFP+ satellite cells were efficiently isolated from muscle tissues by fluorescence-activated cell sorting. Homozygous Pax7-YFP knock-in mice (Pax7(YFP/YFP)) were viable, grew and regenerated muscle normally, and Pax7(YFP/YFP) mouse-derived satellite cells proliferated, differentiated, and self-renewed as efficiently as those from wild-type (Pax7(+/+)) mice. CONCLUSIONS: Taken together, our Pax7-YFP mouse line is a useful tool to aid the development of stem-cell-based therapies for muscle diseases. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13395-018-0174-x) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6108100 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-61081002018-08-28 Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line Kitajima, Yasuo Ono, Yusuke Skelet Muscle Methodology BACKGROUND: Satellite cells are residential muscle stem cells that express a paired box protein, PAX7. RESULTS: Here, we report a knock-in mouse line expressing a PAX7-enhanced yellow fluorescent protein (YFP) fusion protein that enables visualization of PAX7 protein dynamics in living satellite cells through YFP fluorescence. The YFP fluorescence signals in Pax7-YFP knock-in mice clearly recapitulated the endogenous expression of PAX7 protein in satellite cells. YFP+ satellite cells were efficiently isolated from muscle tissues by fluorescence-activated cell sorting. Homozygous Pax7-YFP knock-in mice (Pax7(YFP/YFP)) were viable, grew and regenerated muscle normally, and Pax7(YFP/YFP) mouse-derived satellite cells proliferated, differentiated, and self-renewed as efficiently as those from wild-type (Pax7(+/+)) mice. CONCLUSIONS: Taken together, our Pax7-YFP mouse line is a useful tool to aid the development of stem-cell-based therapies for muscle diseases. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13395-018-0174-x) contains supplementary material, which is available to authorized users. BioMed Central 2018-08-24 /pmc/articles/PMC6108100/ /pubmed/30139390 http://dx.doi.org/10.1186/s13395-018-0174-x Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Methodology Kitajima, Yasuo Ono, Yusuke Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line |
title | Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line |
title_full | Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line |
title_fullStr | Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line |
title_full_unstemmed | Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line |
title_short | Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line |
title_sort | visualization of pax7 protein dynamics in muscle satellite cells in a yfp knock-in-mouse line |
topic | Methodology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6108100/ https://www.ncbi.nlm.nih.gov/pubmed/30139390 http://dx.doi.org/10.1186/s13395-018-0174-x |
work_keys_str_mv | AT kitajimayasuo visualizationofpax7proteindynamicsinmusclesatellitecellsinayfpknockinmouseline AT onoyusuke visualizationofpax7proteindynamicsinmusclesatellitecellsinayfpknockinmouseline |