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Region specific knock-out reveals distinct roles of chromatin modifiers in adult neurogenic niches
Histone methyltransferases (HMTs) are present in heterogeneous cell populations within the adult brain including neurogenic niches. Yet the question remains whether loss of HMTs and the resulting changes in histone methylation alter cell fate in a region-specific manner. We utilized stereotaxic inje...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Taylor & Francis
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5914887/ https://www.ncbi.nlm.nih.gov/pubmed/29433384 http://dx.doi.org/10.1080/15384101.2018.1426417 |
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author | Rhodes, Christopher T. Zunino, Giulia Huang, Shu-Wei Angela Cardona, Sandra M. Cardona, Astrid E. Berger, Mitchel S. Lemmon, Vance P. Lin, Chin-Hsing Annie |
author_facet | Rhodes, Christopher T. Zunino, Giulia Huang, Shu-Wei Angela Cardona, Sandra M. Cardona, Astrid E. Berger, Mitchel S. Lemmon, Vance P. Lin, Chin-Hsing Annie |
author_sort | Rhodes, Christopher T. |
collection | PubMed |
description | Histone methyltransferases (HMTs) are present in heterogeneous cell populations within the adult brain including neurogenic niches. Yet the question remains whether loss of HMTs and the resulting changes in histone methylation alter cell fate in a region-specific manner. We utilized stereotaxic injection of Cre recombinant protein into the adult neurogenic niches, the subventricular zone (SVZ) adjacent to the lateral ventricle and the subgranular zone (SGZ) of the dentate gyrus. We confirmed that Cre protein was enzymatically active in vivo and recombination events were restricted to the vicinity of injection areas. In this study, we focus on using Cre mediated recombination in mice harboring floxed HMT: enhancer of zeste homolog 2 (EZH2) or suppressor of variegation homolog (Suv4-20h). Injectable Cre protein successfully knocked out either EZH2 or Suv4-20h, allowing assessment of long-term effects in a region-specific fashion. We performed meso-scale imaging and flow cytometry for phenotype analysis and unbiased quantification. We demonstrated that regional loss of EZH2 affects the differentiation paradigm of neural stem progenitor cells as well as the maintenance of stem cell population. We further demonstrated that regional loss of Suv4-20h influences the cell cycle but does not affect stem cell differentiation patterns. Therefore, Cre protein mediated knock-out a given HMT unravel their distinguishable and important roles in adult neurogenic niches. This Cre protein-based approach offers tightly-controlled knockouts in multiple cell types simultaneously for studying diverse regulatory mechanisms and is optimal for region-specific manipulation within complex, heterogeneous brain architectures. |
format | Online Article Text |
id | pubmed-5914887 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Taylor & Francis |
record_format | MEDLINE/PubMed |
spelling | pubmed-59148872018-04-27 Region specific knock-out reveals distinct roles of chromatin modifiers in adult neurogenic niches Rhodes, Christopher T. Zunino, Giulia Huang, Shu-Wei Angela Cardona, Sandra M. Cardona, Astrid E. Berger, Mitchel S. Lemmon, Vance P. Lin, Chin-Hsing Annie Cell Cycle Report Histone methyltransferases (HMTs) are present in heterogeneous cell populations within the adult brain including neurogenic niches. Yet the question remains whether loss of HMTs and the resulting changes in histone methylation alter cell fate in a region-specific manner. We utilized stereotaxic injection of Cre recombinant protein into the adult neurogenic niches, the subventricular zone (SVZ) adjacent to the lateral ventricle and the subgranular zone (SGZ) of the dentate gyrus. We confirmed that Cre protein was enzymatically active in vivo and recombination events were restricted to the vicinity of injection areas. In this study, we focus on using Cre mediated recombination in mice harboring floxed HMT: enhancer of zeste homolog 2 (EZH2) or suppressor of variegation homolog (Suv4-20h). Injectable Cre protein successfully knocked out either EZH2 or Suv4-20h, allowing assessment of long-term effects in a region-specific fashion. We performed meso-scale imaging and flow cytometry for phenotype analysis and unbiased quantification. We demonstrated that regional loss of EZH2 affects the differentiation paradigm of neural stem progenitor cells as well as the maintenance of stem cell population. We further demonstrated that regional loss of Suv4-20h influences the cell cycle but does not affect stem cell differentiation patterns. Therefore, Cre protein mediated knock-out a given HMT unravel their distinguishable and important roles in adult neurogenic niches. This Cre protein-based approach offers tightly-controlled knockouts in multiple cell types simultaneously for studying diverse regulatory mechanisms and is optimal for region-specific manipulation within complex, heterogeneous brain architectures. Taylor & Francis 2018-02-19 /pmc/articles/PMC5914887/ /pubmed/29433384 http://dx.doi.org/10.1080/15384101.2018.1426417 Text en © 2018 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way. |
spellingShingle | Report Rhodes, Christopher T. Zunino, Giulia Huang, Shu-Wei Angela Cardona, Sandra M. Cardona, Astrid E. Berger, Mitchel S. Lemmon, Vance P. Lin, Chin-Hsing Annie Region specific knock-out reveals distinct roles of chromatin modifiers in adult neurogenic niches |
title | Region specific knock-out reveals distinct roles of chromatin modifiers in adult neurogenic niches |
title_full | Region specific knock-out reveals distinct roles of chromatin modifiers in adult neurogenic niches |
title_fullStr | Region specific knock-out reveals distinct roles of chromatin modifiers in adult neurogenic niches |
title_full_unstemmed | Region specific knock-out reveals distinct roles of chromatin modifiers in adult neurogenic niches |
title_short | Region specific knock-out reveals distinct roles of chromatin modifiers in adult neurogenic niches |
title_sort | region specific knock-out reveals distinct roles of chromatin modifiers in adult neurogenic niches |
topic | Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5914887/ https://www.ncbi.nlm.nih.gov/pubmed/29433384 http://dx.doi.org/10.1080/15384101.2018.1426417 |
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