PUBLICATION

Intravital imaging of hair-cell development and regeneration in the zebrafish

Authors
Pinto-Teixeira, F., Muzzopappa, M., Swoger, J., Mineo, A., Sharpe, J., and López-Schier, H.
ID
ZDB-PUB-131119-3
Date
2013
Source
Frontiers in neuroscience   7: 33 (Journal)
Registered Authors
Lopez-Schier, Hernan, Mineo, Alessandro, Muzzopappa, Mariana
Keywords
hair cells, auditory, zebrafish, intravital fluorescence microscopy, regeneration, lateral line system, development
MeSH Terms
none
PubMed
24130521 Full text @ Front. Neurosci.
Abstract

Direct videomicroscopic visualization of organ formation and regeneration in toto is a powerful strategy to study cellular processes that often cannot be replicated in vitro. Intravital imaging aims at quantifying changes in tissue architecture or subcellular organization over time during organ development, regeneration or degeneration. A general feature of this approach is its reliance on the optical isolation of defined cell types in the whole animals by transgenic expression of fluorescent markers. Here we describe a simple and robust method to analyze sensory hair-cell development and regeneration in the zebrafish lateral line by high-resolution intravital imaging using laser-scanning confocal microscopy (LSCM) and selective plane illumination microscopy (SPIM). The main advantage of studying hair-cell regeneration in the lateral line is that it occurs throughout the life of the animal, which allows its study in the most natural context. We detail protocols to achieve continuous videomicroscopy for up to 68 hours, enabling direct observation of cellular behavior, which can provide a sensitive assay for the quantitative classification of cellular phenotypes and cell-lineage reconstruction. Modifications to this protocol should facilitate pharmacogenetic assays to identify or validate otoprotective or reparative drugs for future clinical strategies aimed at preserving aural function in humans.

Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping