PUBLICATION
Isolation and Culture of Adult Zebrafish Brain-derived Neurospheres
- Authors
- Lopez-Ramirez, M.A., Calvo, C.F., Ristori, E., Thomas, J.L., Nicoli, S.
- ID
- ZDB-PUB-160312-3
- Date
- 2016
- Source
- Journal of visualized experiments : JoVE (108): 53617 (Journal)
- Registered Authors
- Ristori, Emma
- Keywords
- none
- MeSH Terms
-
- Adult Stem Cells/cytology*
- Animals
- Brain/cytology*
- Brain/physiology
- Cell Culture Techniques/methods
- Cell Differentiation/physiology
- Cell Proliferation/physiology
- Cerebellum/cytology
- Multipotent Stem Cells/cytology*
- Neural Stem Cells/cytology*
- Neurogenesis/physiology
- Regeneration/physiology
- Telencephalon/cytology
- Zebrafish
- PubMed
- 26967835 Full text @ J. Vis. Exp.
Citation
Lopez-Ramirez, M.A., Calvo, C.F., Ristori, E., Thomas, J.L., Nicoli, S. (2016) Isolation and Culture of Adult Zebrafish Brain-derived Neurospheres. Journal of visualized experiments : JoVE. (108):53617.
Abstract
The zebrafish is a highly relevant model organism for understanding the cellular and molecular mechanisms involved in neurogenesis and brain regeneration in vertebrates. However, an in-depth analysis of the molecular mechanisms underlying zebrafish adult neurogenesis has been limited due to the lack of a reliable protocol for isolating and culturing neural adult stem/progenitor cells. Here we provide a reproducible method to examine adult neurogenesis using a neurosphere assay derived from zebrafish whole brain or from the telencephalon, tectum and cerebellum regions of the adult zebrafish brain. The protocol involves, first the microdissection of zebrafish adult brain, then single cell dissociation and isolation of self-renewing multipotent neural stem/progenitor cells. The entire procedure takes eight days. Additionally, we describe how to manipulate gene expression in zebrafish neurospheres, which will be particularly useful to test the role of specific signaling pathways during adult neural stem/progenitor cell proliferation and differentiation in zebrafish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping