title: Employing an open-source tool to assess astrocyte tridimensional structure.
abstract: 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.
authors.affiliation: 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.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Tavares
authors.affiliation: 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.
authors.forename: Manuella
authors.initials: M
authors.lastname: Martins
authors.affiliation: 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.
authors.forename: Joana Sofia
authors.initials: JS
authors.lastname: Correia
authors.affiliation: 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.
authors.forename: Vanessa Morais
authors.initials: VM
authors.lastname: Sardinha
authors.affiliation: 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.
authors.forename: Sónia
authors.initials: S
authors.lastname: Guerra-Gomes
authors.affiliation: 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.
authors.forename: Sofia Pereira
authors.initials: SP
authors.lastname: das Neves
authors.affiliation: 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.
authors.forename: Fernanda
authors.initials: F
authors.lastname: Marques
authors.affiliation: 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.
authors.forename: Nuno
authors.initials: N
authors.lastname: Sousa
authors.affiliation: Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal. joaooliveira@ecsaude.uminho.pt.|ICVS/3B's, PT Government Associate Laboratory, Braga, Guimarães, Portugal. joaooliveira@ecsaude.uminho.pt.|DIGARC, Polytechnic Institute of Cávado and Ave, 4750-810, Barcelos, Portugal. joaooliveira@ecsaude.uminho.pt.
authors.forename: João Filipe
authors.identifier: http://orcid.org/0000-0002-1005-2328
authors.initials: JF
authors.lastname: Oliveira
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology* / Q000378:metabolism; D001921:Brain / Q000166:cytology*; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D021621:Imaging, Three-Dimensional; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D017208:Rats, Wistar; D012984:Software
chemical_list: C000614142:Gfap protein, rat; D005904:Glial Fibrillary Acidic Protein; C528900:glial fibrillary astrocytic protein, mouse
keywords: Astrocyte; GFAP; Immunofluorescence; Reconstruction; Structure