PUBLICATION

Studying Diabetes Through the Eyes of a Fish: Microdissection, Visualization, and Analysis of the Adult tg(fli:EGFP) Zebrafish Retinal Vasculature

Authors
Wiggenhauser, L.M., Kohl, K., Dietrich, N., Hammes, H.P., Kroll, J.
ID
ZDB-PUB-180105-6
Date
2017
Source
Journal of visualized experiments : JoVE   (130): (Journal)
Registered Authors
Kroll, Jens
Keywords
protocol; retinal vasculature; neoangiogenesis;
MeSH Terms
  • Animals
  • Diabetes Mellitus, Experimental/diagnosis*
  • Diabetes Mellitus, Experimental/physiopathology*
  • Disease Models, Animal
  • Humans
  • Microdissection/methods*
  • Retina/pathology*
  • Retinal Neovascularization/pathology*
  • Zebrafish
PubMed
29364210 Full text @ J. Vis. Exp.
Abstract
Diabetic retinopathy is the leading cause of blindness among middle-aged adults. The rising prevalence of diabetes worldwide will make the prevention of diabetic microvascular complications one of the key research fields of the next decades. Specialized, targeted therapy and novel therapeutic drugs are needed to manage the increasing number of patients at risk of vision-loss. The zebrafish is an established animal model for developmental research questions with increasing relevance for modeling metabolic multifactorial disease processes. The advantages of the species allow for optimal visualization and high throughput drug screening approaches, combined with the strong ability to knock out genes of interest. Here, we describe a protocol which will allow easy analysis of the adult tg(fli:EGFP) zebrafish retinal vasculature as a fast read-out in settings of long-term vascular pathologies linked to neoangiogenesis or vessel damage. This is achieved via dissection of the zebrafish retina and whole-mounting of the tissue. Visualization of the exposed vessels is then achieved via confocal microscopy of the green EGFP reporter expressed in the adult retinal vasculature. Correct handling of the tissue will lead to better outcomes and less internal vessel breakage to assure the visualization of the unaltered vascular structure. The method can be utilized in zebrafish models of retinal vasculopathy linked to changes in the vessel architecture as well as neoangiogenesis.
Genes / Markers
Figures
Show all Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping