Tavares Gabriela, Martins Manuella, Correia Joana Sofia, Sardinha Vanessa Morais, Guerra-Gomes Sónia, das Neves Sofia Pereira, Marques Fernanda, Sousa Nuno, Oliveira João Filipe
Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal.
ICVS/3B's, PT Government Associate Laboratory, Braga, Guimarães, Portugal.
Brain Struct Funct. 2017 May;222(4):1989-1999. doi: 10.1007/s00429-016-1316-8. Epub 2016 Sep 30.
Astrocytes display important features that allow them to maintain a close dialog with neurons, ultimately impacting brain function. The complex morphological structure of astrocytes is crucial to the role of astrocytes in brain networks. Therefore, assessing morphologic features of astrocytes will help provide insights into their physiological relevance in healthy and pathological conditions. Currently available tools that allow the tridimensional reconstruction of astrocytes present a number of disadvantages, including the need for advanced computational skills and powerful hardware, and are either time-consuming or costly. In this study, we optimized and validated the FIJI-ImageJ, Simple Neurite Tracer (SNT) plugin, an open-source software that aids in the reconstruction of GFAP-stained structure of astrocytes. We describe (1) the loading of confocal microscopy Z-stacks, (2) the selection criteria, (3) the reconstruction process, and (4) the post-reconstruction analysis of morphological features (process length, number, thickness, and arbor complexity). SNT allows the quantification of astrocyte morphometric parameters in a simple, efficient, and semi-automated manner. While SNT is simple to learn, and does not require advanced computational skills, it provides reproducible results, in different brain regions or pathophysiological states.
星形胶质细胞具有重要特征，使其能够与神经元保持密切对话，最终影响脑功能。星形胶质细胞复杂的形态结构对于其在脑网络中的作用至关重要。因此，评估星形胶质细胞的形态特征将有助于深入了解其在健康和病理状态下的生理相关性。目前可用于星形胶质细胞三维重建的工具存在许多缺点，包括需要先进的计算技能和强大的硬件，而且要么耗时要么成本高昂。在本研究中，我们优化并验证了FIJI-ImageJ的简易神经突追踪器（SNT）插件，这是一款有助于重建星形胶质细胞GFAP染色结构的开源软件。我们描述了（1）共聚焦显微镜Z-stack图像的加载，（2）选择标准，（3）重建过程，以及（4）形态特征（突起长度、数量、厚度和分支复杂性）的重建后分析。SNT能够以简单、高效和半自动的方式对星形胶质细胞的形态计量参数进行量化。虽然SNT易于学习，不需要先进的计算技能，但它在不同脑区或病理生理状态下都能提供可重复的结果。
