PUBLICATION

Zebrafish brain ventricle injection

Authors
Gutzman, J.H., and Sive, H.
ID
ZDB-PUB-090413-6
Date
2009
Source
Journal of visualized experiments : JoVE   (26): (Journal)
Registered Authors
Gutzman, Jennifer, Sive, Hazel
Keywords
none
MeSH Terms
  • Animals
  • Brain/embryology*
  • Cerebral Ventricles/embryology*
  • Fluorescent Dyes/administration & dosage*
  • Injections, Intraventricular/methods
  • Microscopy, Fluorescence/methods*
  • Zebrafish/embryology*
PubMed
19352312 Full text @ J. Vis. Exp.
Abstract
Proper brain ventricle formation during embryonic brain development is required for normal brain function. Brain ventricles are the highly conserved cavities within the brain that are filled with cerebrospinal fluid. In zebrafish, after neural tube formation, the neuroepithelium undergoes a series of constrictions and folds while it fills with fluid resulting in brain ventricle formation. In order to understand the process of ventricle formation, and the neuroepithelial shape changes that occur at the same time, we needed a way to visualize the ventricle space in comparison to the brain tissue. However, the nature of transparent zebrafish embryos makes it difficult to differentiate the tissue from the ventricle space. Therefore, we developed a brain ventricle injection technique where the ventricle space is filled with a fluorescent dye and imaged by brightfield and fluorescent microscopy. The brightfield and the fluorescent images are then processed and superimposed in Photoshop. This technique allows for visualization of the ventricle space with the fluorescent dye, in comparison to the shape of the neuroepithelium in the brightfield image. Brain ventricle injection in zebrafish can be employed from 18 hours post fertilization through early larval stages. We have used this technique extensively in our studies of brain ventricle formation and morphogenesis as well as in characterizing brain morphogenesis mutants (1-3).
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping