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A Computational Tool for Quantitative Analysis of Vascular Networks

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      Abstract

      Angiogenesis is the generation of mature vascular networks from pre-existing vessels. Angiogenesis is crucial during the organism' development, for wound healing and for the female reproductive cycle. Several murine experimental systems are well suited for studying developmental and pathological angiogenesis. They include the embryonic hindbrain, the post-natal retina and allantois explants. In these systems vascular networks are visualised by appropriate staining procedures followed by microscopical analysis. Nevertheless, quantitative assessment of angiogenesis is hampered by the lack of readily available, standardized metrics and software analysis tools. Non-automated protocols are being used widely and they are, in general, time - and labour intensive, prone to human error and do not permit computation of complex spatial metrics. We have developed a light-weight, user friendly software, AngioTool, which allows for quick, hands-off and reproducible quantification of vascular networks in microscopic images. AngioTool computes several morphological and spatial parameters including the area covered by a vascular network, the number of vessels, vessel length, vascular density and lacunarity. In addition, AngioTool calculates the so-called “branching index” (branch points / unit area), providing a measurement of the sprouting activity of a specimen of interest. We have validated AngioTool using images of embryonic murine hindbrains, post-natal retinas and allantois explants. AngioTool is open source and can be downloaded free of charge.

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      Most cited references 30

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      Angiogenesis in health and disease.

      Blood vessels constitute the first organ in the embryo and form the largest network in our body but, sadly, are also often deadly. When dysregulated, the formation of new blood vessels contributes to numerous malignant, ischemic, inflammatory, infectious and immune disorders. Molecular insights into these processes are being generated at a rapidly increasing pace, offering new therapeutic opportunities that are currently being evaluated.
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        VEGF guides angiogenic sprouting utilizing endothelial tip cell filopodia

        Vascular endothelial growth factor (VEGF-A) is a major regulator of blood vessel formation and function. It controls several processes in endothelial cells, such as proliferation, survival, and migration, but it is not known how these are coordinately regulated to result in more complex morphogenetic events, such as tubular sprouting, fusion, and network formation. We show here that VEGF-A controls angiogenic sprouting in the early postnatal retina by guiding filopodial extension from specialized endothelial cells situated at the tips of the vascular sprouts. The tip cells respond to VEGF-A only by guided migration; the proliferative response to VEGF-A occurs in the sprout stalks. These two cellular responses are both mediated by agonistic activity of VEGF-A on VEGF receptor 2. Whereas tip cell migration depends on a gradient of VEGF-A, proliferation is regulated by its concentration. Thus, vessel patterning during retinal angiogenesis depends on the balance between two different qualities of the extracellular VEGF-A distribution, which regulate distinct cellular responses in defined populations of endothelial cells.
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          Molecular regulation of angiogenesis and lymphangiogenesis.

          Blood vessels and lymphatic vessels form extensive networks that are essential for the transport of fluids, gases, macromolecules and cells within the large and complex bodies of vertebrates. Both of these vascular structures are lined with endothelial cells that integrate functionally into different organs, acquire tissue-specific specialization and retain plasticity; thereby, they permit growth during tissue repair or in disease settings. The angiogenic growth of blood vessels and lymphatic vessels coordinates several biological processes such as cell proliferation, guided migration, differentiation and cell-cell communication.
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            Author and article information

            Affiliations
            [1 ]Angiogenesis Core Facility, Radiation Oncology Branch, National Institute of Health National Cancer Institute, Gaithersburg, Maryland, United States of America
            [2 ]Inositide Laboratory, The Babraham Institute, Babraham Research Campus, Cambridge, United Kingdom
            [3 ]Advanced Biomedical Computing Center - Information System Program, Science Applications International Corporation-Frederick Inc, Frederick, Maryland, United States of America
            University College London, United Kingdom
            Author notes

            Conceived and designed the experiments: SV LG. Performed the experiments: LG. Analyzed the data: LG EZ SV. Wrote the paper: EZ SV. Wrote the software: EZ CK.

            Contributors
            Role: Editor
            Journal
            PLoS One
            plos
            plosone
            PLoS ONE
            Public Library of Science (San Francisco, USA )
            1932-6203
            2011
            16 November 2011
            : 6
            : 11
            3217985
            22110636
            PONE-D-11-13876
            10.1371/journal.pone.0027385
            (Editor)
            This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.
            Counts
            Pages: 12
            Categories
            Research Article
            Biology
            Anatomy and Physiology
            Cardiovascular System
            Cardiovascular Anatomy
            Computational Biology
            Biological Data Management
            Developmental Biology
            Molecular Development
            Signaling
            Organism Development
            Organogenesis
            Model Organisms
            Animal Models
            Mouse
            Medicine
            Cardiovascular
            Vascular Biology

            Uncategorized

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