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High Resolution Imaging of Vascular Function in Zebrafish

Watkins, SC and Maniar, S and Mosher, M and Roman, BL and Tsang, M and St Croix, CM (2012) High Resolution Imaging of Vascular Function in Zebrafish. PLoS ONE, 7 (8).

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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 μM), 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. © 2012 Watkins et al.


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Watkins, SCsimon.watkins@pitt.eduSWATKINS
Maniar, S
Mosher, M
Roman, BLromanb@pitt.eduROMANB
Tsang, Mtsang@pitt.eduTSANG
St Croix, CMclaudette.stcroix@pitt.eduCLS13
Contributors:
ContributionContributors NameEmailPitt UsernameORCID
EditorLuttun, AernoutUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Date: 30 August 2012
Date Type: Publication
Journal or Publication Title: PLoS ONE
Volume: 7
Number: 8
DOI or Unique Handle: 10.1371/journal.pone.0044018
Schools and Programs: Graduate School of Public Health > Environmental and Occupational Health
Dietrich School of Arts and Sciences > Biological Sciences
School of Medicine > Cell Biology
School of Medicine > Developmental Biology
Refereed: Yes
Other ID: NLM PMC3431338
PubMed Central ID: PMC3431338
PubMed ID: 22952858
Date Deposited: 19 Oct 2012 21:20
Last Modified: 02 Feb 2019 14:56
URI: http://d-scholarship.pitt.edu/id/eprint/15898

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