Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Medical School, Beech Hill Road, Sheffield S10 2RX, UK.
The Bateson Centre, Firth Court, University of Sheffield, Western Bank, Sheffield S10 2TN, UK.
Development. 2022 Feb 1;149(3). doi: 10.1242/dev.199720. Epub 2022 Feb 14.
Zebrafish transgenic lines and light sheet fluorescence microscopy allow in-depth insights into three-dimensional vascular development in vivo. However, quantification of the zebrafish cerebral vasculature in 3D remains highly challenging. Here, we describe and test an image analysis workflow for 3D quantification of the total or regional zebrafish brain vasculature, called zebrafish vasculature quantification (ZVQ). It provides the first landmark- or object-based vascular inter-sample registration of the zebrafish cerebral vasculature, producing population average maps allowing rapid assessment of intra- and inter-group vascular anatomy. ZVQ also extracts a range of quantitative vascular parameters from a user-specified region of interest, including volume, surface area, density, branching points, length, radius and complexity. Application of ZVQ to 13 experimental conditions, including embryonic development, pharmacological manipulations and morpholino-induced gene knockdown, shows that ZVQ is robust, allows extraction of biologically relevant information and quantification of vascular alteration, and can provide novel insights into vascular biology. To allow dissemination, the code for quantification, a graphical user interface and workflow documentation are provided. Together, ZVQ provides the first open-source quantitative approach to assess the 3D cerebrovascular architecture in zebrafish.
斑马鱼转基因系和光片荧光显微镜使我们能够深入了解体内三维血管发育。然而，对斑马鱼大脑血管的三维定量仍然极具挑战性。在这里，我们描述并测试了一种用于三维定量斑马鱼大脑血管的图像分析工作流程，称为斑马鱼血管定量（ZVQ）。它提供了第一个基于标志或对象的斑马鱼大脑血管的样本间血管配准，生成群体平均图谱，可快速评估血管解剖结构的组内和组间差异。ZVQ 还可以从用户指定的感兴趣区域提取一系列定量血管参数，包括体积、表面积、密度、分支点、长度、半径和复杂度。将 ZVQ 应用于 13 种实验条件，包括胚胎发育、药理学处理和 morpholino 诱导的基因敲低，结果表明 ZVQ 具有稳健性，可提取有生物学意义的信息并定量评估血管变化，并为血管生物学提供新的见解。为了便于传播，提供了用于定量的代码、图形用户界面和工作流程文档。总之，ZVQ 提供了评估斑马鱼三维脑血管结构的第一个开源定量方法。
