title: High resolution imaging of vascular function in zebrafish.
abstract: BACKGROUND
  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.
  
  
  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 A(2) (TXA(2)) 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.
authors.affiliation: Department of Cell Biology, The University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Simon C
authors.initials: SC
authors.lastname: Watkins
authors.forename: Salony
authors.initials: S
authors.lastname: Maniar
authors.forename: Mackenzie
authors.initials: M
authors.lastname: Mosher
authors.forename: Beth L
authors.initials: BL
authors.lastname: Roman
authors.forename: Michael
authors.initials: M
authors.lastname: Tsang
authors.forename: Claudette M
authors.initials: CM
authors.lastname: St Croix
mesh_terms: D000818:Animals; D001775:Blood Circulation / Q000187:drug effects; D001808:Blood Vessels / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D004625:Embryo, Nonmammalian / Q000098:blood supply; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism; D004912:Erythrocytes / Q000166:cytology; D005306:Fertilization; D021621:Imaging, Three-Dimensional; D007814:Larva / Q000502:physiology; D057054:Molecular Imaging / Q000379:methods*; D013928:Thromboxane A2 / Q000031:analogs & derivatives / Q000494:pharmacology; D013997:Time Factors; D015027:Zebrafish / Q000097:blood / Q000196:embryology / Q000502:physiology
chemical_list: D013928:Thromboxane A2