Employing an open-source tool to assess astrocyte tridimensional structure.
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.
['Tavares|Gabriela|G|', 'Martins|Manuella|M|', 'Correia|Joana Sofia|JS|', 'Sardinha|Vanessa Morais|VM|', 'Guerra-Gomes|Sónia|S|', 'das Neves|Sofia Pereira|SP|', 'Marques|Fernanda|F|', 'Sousa|Nuno|N|', 'Oliveira|João Filipe|JF|http://orcid.org/0000-0002-1005-2328']
D000818:Animals D001253:Astrocytes D001921:Brain D005904:Glial Fibrillary Acidic Protein D021621:Imaging, Three-Dimensional D008810:Mice, Inbred C57BL D018613:Microscopy, Confocal D017208:Rats, Wistar D012984:Software
Astrocyte Structure Reconstruction GFAP Immunofluorescence