Abstract
Zebrafish with fluorescently labelled blood vessels provide an excellent model for studying angiogenesis. Most commonly the growth of the intersegmental blood vessels is investigated in response to compounds or manipulation of gene expression and analysed using manual methods, typically scoring the connectivity of these blood vessels to the dorsal longitudinal anastomotic vessel. Such methods are laborious and best suited to time points after the connectivity of these vessels have been established. By contrast, reported image processing-based methods are difficult to implement and often depend on specialist software. This study aimed to develop and evaluate a computational method using the freely available ImageJ software to quantify the development of intersegmental blood vessel formation. This methodology developed allowed rapid analysis of vascular development. The outputs of total vessel length and number of junctions best documented defective vascular development at differing levels of severity and gave comparable results to the frequently used manual approach of calculating percentage connectivity of intersegmental vessels. This ImageJ-based method allowed objective quantitation of vascular network formation in zebrafish enabling a free, straightforward and rapid approach to determine the effect of novel compounds or genetic manipulation of the angiogenic process.
Highlights
The zebrafish (Danio rerio) is a powerful and widely used model organism for the study of vascular development and angiogenesis
Researchers focus on the growth of intersegmental vessels (ISVs) and the formation of the dorsal longitudinal anastomotic vessel (DLAV) in zebrafish, as the ISVs represent the first angiogenic vessels formed in this organism [
A segment as calculated by this method is defined as a set of connected vessels; this is distinct from an intersegmental vessel, which is a vessel that grows between the somites and collectively these are considered to be the first angiogenic vessels to form in this organism [
Summary
Danio rerio) is a powerful and widely used model organism for the study of vascular development and angiogenesis. Features such as the embryo’s rapid and external development, small size, optical transparency, fluorescent tagging of vessels and methods to modify its gene expression all contribute to its utility in studying vessel development [. The widespread use of zebrafish in studying the effects of genes or compounds in vessel development is accompanied by a variety of methods used to analyse images of the zebrafish vasculature. The majority of ISV analysis methods involve manually scoring images of zebrafish embryos, typically counting the number of normal ISVs formed [
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