PUBLICATION

Laser-scanning velocimetry: A confocal microscopy method for quantitative measurement of cardiovascular performance in zebrafish embryos and larvae

Authors
Malone, M.H., Sciaky, N., Stalheim, L., Hahn, K.M., Linney, E., and Johnson, G.L.
ID
ZDB-PUB-070726-6
Date
2007
Source
BMC Biotechnology   7(1): 40 (Journal)
Registered Authors
Linney, Elwood
Keywords
none
MeSH Terms
  • Aminobenzoates/pharmacology
  • Animals
  • Aorta/drug effects
  • Aorta/embryology
  • Aorta/physiology
  • Blood Flow Velocity/drug effects
  • Blotting, Western
  • Embryo, Nonmammalian/drug effects
  • Embryo, Nonmammalian/metabolism
  • Embryo, Nonmammalian/physiology
  • Gene Expression Regulation, Developmental
  • Genetic Complementation Test
  • Heart/embryology
  • Heart/physiology*
  • Larva/cytology
  • Larva/genetics
  • Larva/physiology
  • Microfilament Proteins/genetics
  • Microfilament Proteins/metabolism
  • Microfilament Proteins/physiology
  • Microinjections
  • Microscopy, Confocal/methods*
  • Morpholines/pharmacology
  • Mutation
  • Myocardium/cytology
  • Myocardium/metabolism
  • RNA, Messenger/administration & dosage
  • RNA, Messenger/genetics
  • RNA, Messenger/metabolism
  • Reproducibility of Results
  • Zebrafish/embryology
  • Zebrafish/physiology*
PubMed
17623073 Full text @ BMC Biotechnol.
Abstract
BACKGROUND: The zebrafish Danio rerio is an important model system for drug discovery and to study cardiovascular development. Using a laser-scanning confocal microscope, we have developed a non-invasive method of measuring cardiac performance in zebrafish embryos and larvae that obtains cardiovascular parameters similar to those obtained using Doppler echocardiography in mammals. A laser scan line placed parallel to the path of blood in the dorsal aorta measures blood cell velocity, from which cardiac output and indices of vascular resistance and contractility are calculated. RESULTS: This technique, called laser-scanning velocimetry, was used to quantify the effects of pharmacological, developmental, and genetic modifiers of cardiac function. Laser-scanning velocimetry was applied to analyze the cardiovascular effects of morpholino knockdown of osmosensing scaffold for MEKK3 (OSM), which when mutated causes the human vascular disease cerebral cavernous malformations. OSM-deficient embryos had a constricted aortic arch and markedly increased peak cell velocity, a characteristic indicator of aortic stenosis. CONCLUSIONS: These data validate laser-scanning velocimetry as a quantitative tool to measure cardiovascular performance for pharmacological and genetic analysis in zebrafish, which requires no specialized equipment other than a laser-scanning confocal microscope.
Genes / Markers
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Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
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Mapping