PUBLICATION
Chapter 1 - Analyzing retinal axon guidance in zebrafish
- Authors
- Poulain, F.E., Gaynes, J.A., Hörndli, C.S., Law, M.Y., and Chien, C.B.
- ID
- ZDB-PUB-101201-24
- Date
- 2010
- Source
- Methods in cell biology 100: 2-26 (Chapter)
- Registered Authors
- Chien, Chi-Bin, Gaynes, John, Hörndli, Coni Stacher, Law, Mei-Yee, Poulain, Fabienne
- Keywords
- none
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Axons*
- Electroporation
- Embryo, Nonmammalian/metabolism
- Neurobiology/methods
- Neurogenesis*
- Retina/cytology
- Retinal Ganglion Cells/cytology*
- Retinal Ganglion Cells/metabolism
- Time-Lapse Imaging
- Zebrafish*
- PubMed
- 21111212 Full text @ Meth. Cell. Biol.
Citation
Poulain, F.E., Gaynes, J.A., Hörndli, C.S., Law, M.Y., and Chien, C.B. (2010) Chapter 1 - Analyzing retinal axon guidance in zebrafish. Methods in cell biology. 100:2-26.
Abstract
How neuronal connections are established during development is one of the most fascinating questions in the field of neurobiology. The zebrafish retinotectal system offers distinct advantages for studying axon guidance in an in vivo context. Its accessibility and the larva's transparency not only allow its direct visualization, but also facilitate experimental manipulations to address the mechanisms of its development. Here we describe methods for labeling and visualizing retinal axons in vivo, including transient expression of DNA constructs, injection of lipophilic dyes, and time-lapse imaging. We describe in detail the available transgenic lines for marking retinal ganglion cells (RGCs); a protocol for very precise lipophilic dye labeling; and a protocol for single cell electroporation of RGCs. We then describe several approaches for perturbing the retinotectal system, including morpholino or DNA injection; localized heat shock to induce misexpression of genes; a comprehensive list of known retinotectal mutants; and a detailed protocol for RGC transplants to test cell autonomy. These methods not only provide new ways for examining how retinal axons are guided by their environment, but also can be used to study other axonal tracts in the living embryo.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping