Astrocytes pp 49-66 | Cite as

Fluorescence-Activated Cell Sorting-Based Isolation and Characterization of Neural Stem Cells from the Adult Zebrafish Telencephalon

  • Rossella Di GiaimoEmail author
  • Sven Aschenbroich
  • Jovica Ninkovic
Part of the Methods in Molecular Biology book series (MIMB, volume 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.

Key words

Neural stem cells Zebrafish Neural stem cell purification Intracellular FACS 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Rossella Di Giaimo
    • 1
    Email author
  • Sven Aschenbroich
    • 2
    • 3
    • 4
  • Jovica Ninkovic
    • 2
    • 3
    • 4
  1. 1.Department of BiologyUniversity of Naples Federico II, Complesso Universitario Monte S. AngeloNaplesItaly
  2. 2.Institute of Stem Cell Research, Helmholtz Center MunichMunichGermany
  3. 3.Physiological Genomics, Biomedical CenterUniversity of MunichMunichGermany
  4. 4.Institute of Cell Biology, Biomedical CenterUniversity of MunichMunichGermany

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