PUBLICATION
Isolation and Culture of Primary Embryonic Zebrafish Neural Tissue
- Authors
- Patel, B.B., Clark, K.L., Kozik, E.M., Dash, L., Kuhlman, J.A., Sakaguchi, D.S.
- ID
- ZDB-PUB-190902-24
- Date
- 2019
- Source
- Journal of Neuroscience Methods 328: 108419 (Journal)
- Registered Authors
- Kuhlman, Julie
- Keywords
- Primary neuron culture, cell migration, time-lapse microscopy, zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Embryo, Nonmammalian
- Neuroglia*/cytology
- Neuroglia*/physiology
- Neurons*/cytology
- Neurons*/physiology
- Neurosciences/methods*
- Primary Cell Culture/methods*
- Zebrafish*
- PubMed
- 31472190 Full text @ J. Neurosci. Methods
Citation
Patel, B.B., Clark, K.L., Kozik, E.M., Dash, L., Kuhlman, J.A., Sakaguchi, D.S. (2019) Isolation and Culture of Primary Embryonic Zebrafish Neural Tissue. Journal of Neuroscience Methods. 328:108419.
Abstract
Background Primary cell culture is a valuable tool to utilize in parallel with in vivo studies in order to maximize our understanding of the mechanisms surrounding neurogenesis and central nervous system (CNS) regeneration and plasticity. The zebrafish is an important model for biomedical research and primary neural cells are readily obtainable from their embryonic stages via tissue dissociation. Further, transgenic reporter lines with cell type-specific expression allows for observation of distinct cell populations within the dissociated tissue.
New method Here, we define an efficient method for ex vivo quantification and characterization of neuronal and glial tissue dissociated from embryonic zebrafish.
Results Zebrafish brain dissociated cells have been documented to survive in culture for at least 9 days in vitro (div). Anti-HuC/D and anti-Acetylated Tubulin antibodies were used to identify neurons in culture; at 3 div approximately 48% of cells were HuC/D positive and 85% expressed serotonin, suggesting our protocol can efficiently isolate neurons from whole embryonic zebrafish brains. Live time-lapse imaging was also carried out to analyze cell migration in vitro.
Comparison with existing methods Primary cultures of zebrafish neural cells typically have low rates of survivability in vitro. We have developed a culture system that has long term cell viability, enabling direct analysis of cell-cell and cell-extracellular matrix interactions.
Conclusions These results demonstrate a practical method for isolating, dissociating and culturing of embryonic zebrafish neural tissue. This approach could further be utilized to better understand zebrafish regeneration in vitro.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping