PUBLICATION
Assessment of Zebrafish Lens Nucleus Localization and Sutural Integrity
- Authors
- Vorontsova, I., Hall, J.E., Schilling, T.F.
- ID
- ZDB-PUB-190521-8
- Date
- 2019
- Source
- Journal of visualized experiments : JoVE (147): (Journal)
- Registered Authors
- Schilling, Tom
- Keywords
- none
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Aquaporins/metabolism
- Embryo, Nonmammalian/anatomy & histology
- Eye Proteins/metabolism
- Larva/anatomy & histology
- Lens, Crystalline/anatomy & histology*
- Zebrafish/anatomy & histology*
- Zebrafish/embryology
- PubMed
- 31107462 Full text @ J. Vis. Exp.
Citation
Vorontsova, I., Hall, J.E., Schilling, T.F. (2019) Assessment of Zebrafish Lens Nucleus Localization and Sutural Integrity. Journal of visualized experiments : JoVE. (147):.
Abstract
The zebrafish is uniquely suited to genetic manipulation and in vivo imaging, making it an increasingly popular model for reverse genetic studies and for generation of transgenics for in vivo imaging. These unique capabilities make the zebrafish an ideal platform to study ocular lens development and physiology. Our recent findings that an Aquaporin-0, Aqp0a, is required for stability of the anterior lens suture, as well as for the shift of the lens nucleus to the lens center with age led us to develop tools especially suited to analyzing the properties of zebrafish lenses. Here we outline detailed methods for lens dissection that can be applied to both larval and adult lenses, to prepare them for histological analysis, immunohistochemistry and imaging. We focus on analysis of lens suture integrity and cortical cell morphology and compare data generated from dissected lenses with data obtained from in vivo imaging of lens morphology made possible by a novel transgenic zebrafish line with a genetically encoded fluorescent marker. Analysis of dissected lenses perpendicular to their optical axis allows quantification of the relative position of the lens nucleus along the anterior-posterior axis. Movement of the lens nucleus from an initial anterior position to the center is required for normal lens optics in adult zebrafish. Thus, a quantitative measure of lens nuclear position directly correlates with its optical properties.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping