PUBLICATION
Electroporation-based methods for in vivo, whole mount and primary culture analysis of zebrafish brain development
- Authors
- Hendricks, M., and Jesuthasan, S.
- ID
- ZDB-PUB-070330-6
- Date
- 2007
- Source
- Neural Development 2(1): 6 (Journal)
- Registered Authors
- Hendricks, Michael, Jesuthasan, Suresh
- Keywords
- none
- MeSH Terms
-
- Animals
- Brain/embryology*
- Brain/growth & development
- Brain/surgery*
- Cell Culture Techniques
- Cells, Cultured
- Electroporation/instrumentation
- Electroporation/methods*
- Embryo, Nonmammalian/cytology
- Embryo, Nonmammalian/embryology
- Embryo, Nonmammalian/surgery
- Microinjections/instrumentation
- Microinjections/methods
- Microscopy, Video/methods
- Molecular Biology/instrumentation
- Molecular Biology/methods
- Nucleic Acids/pharmacology
- Time Factors
- Transfection/instrumentation
- Transfection/methods
- Zebrafish/embryology*
- Zebrafish/growth & development
- Zebrafish/surgery*
- PubMed
- 17359546 Full text @ Neural Dev.
Citation
Hendricks, M., and Jesuthasan, S. (2007) Electroporation-based methods for in vivo, whole mount and primary culture analysis of zebrafish brain development. Neural Development. 2(1):6.
Abstract
BACKGROUND: Electroporation is a technique for the introduction of nucleic acids and other macromolecules into cells. In chick embryos it has been a particularly powerful technique for the spatial and temporal control of gene expression in developmental studies. Electroporation methods have also been reported for Xenopus, zebrafish, and mouse. RESULTS: We present a new protocol for zebrafish brain electroporation. Using a simple set-up with fixed spaced electrodes and microinjection equipment, it is possible to electroporate 50 to 100 embryos in 1 hour with no lethality and consistently high levels of transgene expression in numerous cells. Transfected cells in the zebrafish brain are amenable to in vivo time lapse imaging. Explants containing transfected neurons can be cultured for in vitro analysis. We also present a simple enzymatic method to isolate whole brains from fixed zebrafish for immunocytochemistry. CONCLUSION: Building on previously described methods, we have optimized several parameters to allow for highly efficient unilateral or bilateral transgenesis of a large number of cells in the zebrafish brain. This method is simple and provides consistently high levels of transgenesis for large numbers of embryos.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping