PUBLICATION

Live imaging of the zebrafish embryonic brain by confocal microscopy

Authors
Graeden, E., and Sive, H.
ID
ZDB-PUB-090407-10
Date
2009
Source
Journal of visualized experiments : JoVE   (26): 1217 (Journal)
Registered Authors
Sive, Hazel
Keywords
none
MeSH Terms
  • Animals
  • Brain/embryology*
  • Microscopy, Confocal/methods*
  • Zebrafish/embryology*
PubMed
19339963 Full text @ J. Vis. Exp.
Abstract
In this video, we demonstrate the method our lab has developed to analyze the cell shape changes and rearrangements required to bend and fold the developing zebrafish brain (Gutzman et al, 2008). Such analysis affords a new understanding of the underlying cell biology required for development of the 3D structure of the vertebrate brain, and significantly increases our ability to study neural tube morphogenesis. The embryonic zebrafish brain is shaped beginning at 18 hours post fertilization (hpf) as the ventricles within the neuroepithelium inflate. By 24 hpf, the initial steps of neural tube morphogenesis are complete. Using the method described here, embryos at the one cell stage are injected with mRNA encoding membrane-targeted green fluorescent protein (memGFP). After injection and incubation, the embryo, now between 18 and 24 hpf, is mounted, inverted, in agarose and imaged by confocal microscopy. Notably, the zebrafish embryo is transparent making it an ideal system for fluorescent imaging. While our analyses have focused on the midbrain-hindbrain boundary and the hindbrain, this method could be extended for analysis of any region in the zebrafish to a depth of 80-100 microm.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping