PUBLICATION

High resolution imaging of vascular function in zebrafish

Authors
Watkins, S.C., Maniar, S., Mosher, M., Roman, B.L., Tsang, M., and St Croix, C.M.
ID
ZDB-PUB-120907-21
Date
2012
Source
PLoS One   7(8): e44018 (Journal)
Registered Authors
Roman, Beth, Tsang, Michael
Keywords
none
MeSH Terms
  • Animals
  • Blood Circulation/drug effects
  • Blood Vessels/cytology
  • Blood Vessels/drug effects
  • Blood Vessels/physiology*
  • Embryo, Nonmammalian/blood supply
  • Endothelium, Vascular/drug effects
  • Endothelium, Vascular/metabolism
  • Erythrocytes/cytology
  • Fertilization
  • Imaging, Three-Dimensional
  • Larva/physiology
  • Molecular Imaging/methods*
  • Thromboxane A2/analogs & derivatives
  • Thromboxane A2/pharmacology
  • Time Factors
  • Zebrafish*/blood
  • Zebrafish*/embryology
  • Zebrafish*/physiology
PubMed
22952858 Full text @ PLoS One
Abstract

Rationale

The role of the endothelium in the pathogenesis of cardiovascular disease is an emerging field of study, necessitating the development of appropriate model systems and methodologies to investigate the multifaceted nature of endothelial dysfunction including disturbed barrier function and impaired vascular reactivity.

Objective

We aimed to develop and test an optimized high-speed imaging platform to obtain quantitative real-time measures of blood flow, vessel diameter and endothelial barrier function in order to assess vascular function in live vertebrate models.

Methods and Results

We used a combination of cutting-edge optical imaging techniques, including high-speed, camera-based imaging (up to 1000 frames/second), and 3D confocal methods to collect real time metrics of vascular performance and assess the dynamic response to the thromboxane A2 (TXA2) analogue, U-46619 (1 ┬ÁM), in transgenic zebrafish larvae. Data obtained in 3 and 5 day post-fertilization larvae show that these methods are capable of imaging blood flow in a large (1 mm) segment of the vessel of interest over many cardiac cycles, with sufficient speed and sensitivity such that the trajectories of individual erythrocytes can be resolved in real time. Further, we are able to map changes in the three dimensional sizes of vessels and assess barrier function by visualizing the continuity of the endothelial layer combined with measurements of extravasation of fluorescent microspheres.

Conclusions

We propose that this system-based microscopic approach can be used to combine measures of physiologic function with molecular behavior in zebrafish models of human vascular disease.

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