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Open Access Green as soon as Postprint is submitted to ZB.
Fluorescence-activated cell sorting-based isolation and characterization of neural stem cells from the adult zebrafish telencephalon.
In: Astrocytes. Berlin [u.a.]: Springer, 2019. 49-66 (Methods Mol. Biol. ; 1938)
Adult mammalian brain, including humans, has rather limited addition of new neurons and poor regenerative capacity. In contrast, neural stem cells (NSC) with glial identity and neurogenesis are highly abundant throughout the adult zebrafish brain. Importantly, the activation of NSC and production of new neurons in response to injuries lead to the brain regeneration in zebrafish brain. Therefore, understanding of the molecular pathways regulating NSC behavior in response to injury is crucial in order to set the basis for experimental modification of these pathways in glial cells after injury in the mammalian brain and to elicit neuronal regeneration. Here, we describe the procedure that we successfully used to prospectively isolate NSCs from adult zebrafish telencephalon, extract RNA, and prepare cDNA libraries for next generation sequencing (NGS) and full transcriptome analysis as the first step toward understanding regulatory mechanisms leading to restorative neurogenesis in zebrafish. Moreover, we describe an alternative approach to analyze antigenic properties of NSC in the adult zebrafish brain using intracellular fluorescence activated cell sorting (FACS). We employ this method to analyze the number of proliferating NSCs positive for proliferating cell nuclear antigen (PCNA) in the prospectively isolated population of stem cells.
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Publication type
Article: Edited volume or book chapter
Keywords
Intracellular Facs ; Neural Stem Cell Purification ; Neural Stem Cells ; Zebrafish
ISSN (print) / ISBN
1064-3745
e-ISSN
1940-6029
Book Volume Title
Astrocytes
Journal
Methods in Molecular Biology
Quellenangaben
Volume: 1938,
Pages: 49-66
Publisher
Springer
Publishing Place
Berlin [u.a.]
Non-patent literature
Publications
Reviewing status
Peer reviewed
Institute(s)
Institute of Stem Cell Research (ISF)