PUBLICATION

Heart Dissection in Larval, Juvenile and Adult Zebrafish, Danio rerio

Authors
Singleman, C., and Holtzman, N.G.
ID
ZDB-PUB-111026-5
Date
2011
Source
Journal of visualized experiments : JoVE : (Movie)
Registered Authors
Holtzman, Nathalia Glickman
Keywords
none
MeSH Terms
  • Animals
  • Cardiac Surgical Procedures/methods*
  • Dissection/methods*
  • Female
  • Heart/anatomy & histology*
  • Heart/growth & development
  • Male
  • Zebrafish/anatomy & histology*
  • Zebrafish/embryology
  • Zebrafish/growth & development
  • Zebrafish/surgery*
PubMed
21989462 Full text @ J. Vis. Exp.
Citation
Singleman, C., and Holtzman, N.G. (2011) Heart Dissection in Larval, Juvenile and Adult Zebrafish, Danio rerio. Journal of visualized experiments : JoVE. .
Abstract
Zebrafish have become a beneficial and practical model organism for the study of embryonic heart development (see recent reviews1-6), however, work examining post-embryonic through adult cardiac development has been limited7-10. Examining the changing morphology of the maturing and aging heart are restricted by the lack of techniques available for staging and isolating juvenile and adult hearts. In order to analyze heart development over the fish's lifespan, we dissect zebrafish hearts at numerous stages and photograph them for further analysis11. The morphological features of the heart can easily be quantified and individual hearts can be further analyzed by a host of standard methods. Zebrafish grow at variable rates and maturation correlates better with fish size than age, thus, post-fixation, we photograph and measure fish length as a gauge of fish maturation. This protocol explains two distinct, size dependent dissection techniques for zebrafish, ranging from larvae 3.5mm standard length (SL) with hearts of 100µm ventricle length (VL), to adults, with SL of 30mm and VL 1mm or larger. Larval and adult fish have quite distinct body and organ morphology. Larvae are not only significantly smaller, they have less pigment and each organ is visually very difficult to identify. For this reason, we use distinct dissection techniques.

We used pre-dissection fixation procedures, as we discovered that hearts dissected directly after euthanization have a more variable morphology, with very loose and balloon like atria compared with hearts removed following fixation. The fish fixed prior to dissection, retain in vivo morphology and chamber position (data not shown). In addition, for demonstration purposes, we take advantage of the heart (myocardial) specific GFP transgenic Tg(myl7:GFP)twu34 (12), which allows us to visualize the entire heart and is particularly useful at early stages in development when the cardiac morphology is less distinct from surrounding tissues. Dissection of the heart makes further analysis of the cell and molecular biology underlying heart development and maturation using in situ hybridization, immunohistochemistry, RNA extraction or other analytical methods easier in post-embryonic zebrafish. This protocol will provide a valuable technique for the study of cardiac development maturation and aging.

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