3D quantitative analyses of angiogenic sprout growth dynamics

• Published in: Developmental dynamics 2013-05, Vol.242 (5), p.518-526
• Main Authors: Shirinifard, Abbas, McCollum, Catherine W., Bolin, Maria Bondesson, Gustafsson, Jan‐Åke, Glazier, James A., Clendenon, Sherry G.
• Format: Article
• Language: English
• Subjects: analysis; angiogenesis; Animals; Animals, Genetically Modified; Arsenic - pharmacology; Arsenic - toxicity; Body Patterning - drug effects; Body Patterning - physiology; Cell Movement - physiology; Cell Tracking - methods; Danio rerio; dynamic; Embryo, Nonmammalian - blood supply; Embryo, Nonmammalian - drug effects; Environmental Pollutants - pharmacology; Environmental Pollutants - toxicity; Imaging, Three-Dimensional - methods; intersegmental vessel; Kinetics; Microscopy, Fluorescence; Motion; Neovascularization, Physiologic - drug effects; Neovascularization, Physiologic - physiology; quantitative; Time Factors; Time-Lapse Imaging - methods; trajectory; zebrafish; Zebrafish - embryology; Zebrafish - genetics
• ISSN: 1058-8388; 1097-0177
• DOI: 10.1002/dvdy.23946

Background: Zebrafish intersegmental vessel (ISV) growth is widely used to study angiogenesis and to screen drugs and toxins that perturb angiogenesis. Most current ISV growth assays observe the presence or absence of ISVs or perturbation of ISV morphology but do not measure growth dynamics. We have developed a four‐dimensional (4D, space plus time) quantitative analysis of angiogenic sprout growth dynamics for characterization of both normal and perturbed growth. Results: We tracked the positions of the ISV base and tip for each ISV sprout in 4D. Despite immobilization, zebrafish embryos translocated globally and non‐uniformly during development. We used displacement of the ISV base and the angle between the ISV and the dorsal aorta to correct for displacement and rotation during development. From corrected tip cell coordinates, we computed average ISV trajectories. We fitted a quadratic curve to the average ISV trajectories to produce a canonical ISV trajectory for each experimental group, arsenic treated and untreated. From the canonical ISV trajectories, we computed curvature, average directed migration speed and directionality. Canonical trajectories from treated (arsenic exposed) and untreated groups differed in curvature, average directed migration speed and angle between the ISV and dorsal aorta. Conclusions: 4D analysis of angiogenic sprout growth dynamics: (1) Allows quantitative assessment of ISV growth dynamics and perturbation, and (2) provides critical inputs for computational models of angiogenesis. Developmental Dynamics 242:508–516, 2013. © 2013 Wiley Periodicals, Inc. Key Findings: New technique for quantitative analyses of 3D angiogenic sprout growth dynamics. Useful for analysis of perturbations due to genetic manipulations and exposure to drugs and environmental toxins. Using this new technique we showed that angiogenic sprouts in arsenic treated zebrafish had lower directed migration speed and decreased directional persistence than controls.