@article{033a8fa037294f04b9fd1882c5966efd,
title = "Region specific knock-out reveals distinct roles of chromatin modifiers in adult neurogenic niches",
abstract = "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.",
keywords = "enhancer of zeste homolog 2 (EZH2), subgranular zone, subventricular zone, suppressor of variegation homolog (Suv4-20h)",
author = "Rhodes, {Christopher T.} and Giulia Zunino and Huang, {Shu Wei Angela} and Cardona, {Sandra M.} and Cardona, {Astrid E.} and Berger, {Mitchel S.} and Lemmon, {Vance P.} and Lin, {Chin Hsing Annie}",
note = "Funding Information: HHS | NIH | National Institute of General Medical Sciences (NIGMS) [grant number NIH/NIGMS SC3GM112543] We thank Drs. Alexander Tarakhovsky and Gunnar Schotta for generously providing conditional Ezh2 and Suv4-20h mouse lines, respectively. Authors are grateful to Dr. Fred Gage for his insightful comments and technical support. We also thank Drs. Arturo Alvarez-Buylla, Douglas Grow, ChiungYu Hung, and Sebastian Jessberger for technical advice. Stereotaxic injections, confocal imaging, and flow cytometry/imaging flow were done in the Neuroscience Institute Opto-excitability Core Facility, the Imaging Core, and the Immune Defense Core at UTSA, respectively. Light Sheet Fluorescent Microscopy was conducted in The Miami Project to Cure Paralysis Image Analysis Core. CTR and SAH were supported by NIH/NIGMS SC3GM112543. SMC and AEC were supported by NIH/NIGMS SC1GM095426 and NIH/NINDS NS078501. GZ and VPL were supported by The Walter G. Ross Foundation and NIH/NICHD HD057632. This project was supported by NIH/NIGMS SC3GM112543 and TRAC award to CAL. Funding Information: We thank Drs. Alexander Tarakhovsky and Gunnar Schotta for generously providing conditional Ezh2 and Suv4-20h mouse lines, respectively. Authors are grateful to Dr. Fred Gage for his insightful comments and technical support. We also thank Drs. Arturo Alvarez-Buylla, Douglas Grow, ChiungYu Hung, and Sebastian Jessberger for technical advice. Stereotaxic injections, confocal imaging, and flow cytometry/imaging flow were done in the Neuroscience Institute Opto-excitability Core Facility, the Imaging Core, and the Immune Defense Core at UTSA, respectively. Light Sheet Fluorescent Microscopy was conducted in The Miami Project to Cure Paralysis Image Analysis Core. CTR and SAH were supported by NIH/NIGMS SC3GM112543. SMC and AEC were supported by NIH/NIGMS SC1GM095426 and NIH/NINDS NS078501. GZ and VPL were supported by The Walter G. Ross Foundation and NIH/NICHD HD057632. This project was supported by NIH/NIGMS SC3GM112543 and TRAC award to CAL. Publisher Copyright: {\textcopyright} 2018 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.",
year = "2018",
month = feb,
day = "1",
doi = "10.1080/15384101.2018.1426417",
language = "English (US)",
volume = "17",
pages = "377--389",
journal = "Cell Cycle",
issn = "1538-4101",
publisher = "Landes Bioscience",
number = "3",
}