Top

Published in:

Open Access 01-05-2017 | Methods Paper

Employing an open-source tool to assess astrocyte tridimensional structure

Authors: Gabriela Tavares, Manuella Martins, Joana Sofia Correia, Vanessa Morais Sardinha, Sónia Guerra-Gomes, Sofia Pereira das Neves, Fernanda Marques, Nuno Sousa, João Filipe Oliveira

Published in: Brain Structure and Function | Issue 4/2017

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.

Araque A, Carmignoto G, Haydon PG et al (2014) Gliotransmitters travel in time and space. Neuron 81:728–739. doi:10.1016/j.neuron.2014.02.007 CrossRefPubMedPubMedCentral

Burda JE, Bernstein AM, Sofroniew MV (2016) Astrocyte roles in traumatic brain injury. Exp Neurol 275(3):305–315. doi:10.1016/j.expneurol.2015.03.020 CrossRefPubMed

Bushong EA, Martone ME, Jones YZ, Ellisman MH (2002) Protoplasmic astrocytes in CA1 stratum radiatum occupy separate anatomical domains. J Neurosci 22:183–192PubMed

Donohue DE, Ascoli GA (2011) Automated reconstruction of neuronal morphology: an overview. Brain Res Rev 67:94–102. doi:10.1016/j.brainresrev.2010.11.003 CrossRefPubMed

Doyle JP, Dougherty JD, Heiman M et al (2008) Application of a translational profiling approach for the comparative analysis of CNS cell types. Cell 135:749–762. doi:10.1016/j.cell.2008.10.029 CrossRefPubMedPubMedCentral

Ghézali G, Dallérac G, Rouach N (2016) Perisynaptic astroglial processes: dynamic processors of neuronal information. Brain Struct Funct 221:2427–2442. doi:10.1007/s00429-015-1070-3 CrossRefPubMed

Haber M, Zhou L, Murai KK (2006) Cooperative astrocyte and dendritic spine dynamics at hippocampal excitatory synapses. J Neurosci 26:8881–8891. doi:10.1523/JNEUROSCI.1302-06.2006 CrossRefPubMed

Heller JP, Rusakov DA (2015) Morphological plasticity of astroglia: understanding synaptic microenvironment: morphological plasticity of astroglia. Glia 63:2133–2151. doi:10.1002/glia.22821 CrossRefPubMedPubMedCentral

Hol EM, Pekny M (2015) Glial fibrillary acidic protein (GFAP) and the astrocyte intermediate filament system in diseases of the central nervous system. Curr Opin Cell Biol 32:121–130. doi:10.1016/j.ceb.2015.02.004 CrossRefPubMed

Jinno S (2011) Regional and laminar differences in antigen profiles and spatial distributions of astrocytes in the mouse hippocampus, with reference to aging. Neuroscience 180:41–52. doi:10.1016/j.neuroscience.2011.02.013 CrossRefPubMed

Khakh BS, Sofroniew MV (2015) Diversity of astrocyte functions and phenotypes in neural circuits. Nat Neurosci 18:942–952. doi:10.1038/nn.4043 CrossRefPubMedPubMedCentral

Kulkarni PM, Barton E, Savelonas M et al (2015) Quantitative 3-D analysis of GFAP labeled astrocytes from fluorescence confocal images. J Neurosci Methods 246:38–51. doi:10.1016/j.jneumeth.2015.02.014 CrossRefPubMed

Lima A, Sardinha VM, Oliveira AF et al (2014) Astrocyte pathology in the prefrontal cortex impairs the cognitive function of rats. Mol Psychiatry 19:834–841. doi:10.1038/mp.2013.182 CrossRefPubMed

Longair MH, Baker DA, Armstrong JD (2011) Simple Neurite Tracer: open source software for reconstruction, visualization and analysis of neuronal processes. Bioinformatics 27:2453–2454. doi:10.1093/bioinformatics/btr390 CrossRefPubMed

Matyash V, Kettenmann H (2010) Heterogeneity in astrocyte morphology and physiology. Brain Res Rev 63:2–10. doi:10.1016/j.brainresrev.2009.12.001 CrossRefPubMed

Medvedev N, Popov V, Henneberger C et al (2014) Glia selectively approach synapses on thin dendritic spines. Phil Trans R Soc B 369:20140047. doi:10.1098/rstb.2014.0047 CrossRefPubMedPubMedCentral

Myatt D, Hadlington T, Ascoli G, Nasuto S (2012) Neuromantic—from semi-manual to semi-automatic reconstruction of neuron morphology. Front Neuroinformatics 6:4. doi:10.3389/fninf.2012.00004 CrossRef

Nash B, Thomson CE, Linington C et al (2011) Functional duality of astrocytes in myelination. J Neurosci 31:13028–13038. doi:10.1523/JNEUROSCI.1449-11.2011 CrossRefPubMed

Oberheim NA, Takano T, Han X et al (2009) Uniquely hominid features of adult human astrocytes. J Neurosci 29:3276–3287. doi:10.1523/JNEUROSCI.4707-08.2009 CrossRefPubMedPubMedCentral

Oberheim NA, Goldman SA, Nedergaard M (2012) Heterogeneity of astrocytic form and function. Methods Mol Biol Clifton NJ 814:23–45. doi:10.1007/978-1-61779-452-0_3 CrossRef

Oliveira JF, Sardinha VM, Guerra-Gomes S et al (2015) Do stars govern our actions? Astrocyte involvement in rodent behavior. Trends Neurosci 38:535–549. doi:10.1016/j.tins.2015.07.006 CrossRefPubMed

Parekh R, Ascoli GA (2013) Neuronal morphology goes digital: a research hub for cellular and system neuroscience. Neuron 77:1017–1038. doi:10.1016/j.neuron.2013.03.008 CrossRefPubMedPubMedCentral

Parpura V, Heneka MT, Montana V et al (2012) Glial cells in (patho)physiology. J Neurochem 121:4–27. doi:10.1111/j.1471-4159.2012.07664.x CrossRefPubMedPubMedCentral

Paxinos VG, Watson C (2005) The rat brain in stereotaxic coordinates, 5th edn. Academic Press, New York

Perea G, Navarrete M, Araque A (2009) Tripartite synapses: astrocytes process and control synaptic information. Trends Neurosci 32:421–431. doi:10.1016/j.tins.2009.05.001 CrossRefPubMed

Perez-Alvarez A, Navarrete M, Covelo A et al (2014) Structural and functional plasticity of astrocyte processes and dendritic spine interactions. J Neurosci 34:12738–12744. doi:10.1523/JNEUROSCI.2401-14.2014 CrossRefPubMed

Pool M, Thiemann J, Bar-Or A, Fournier AE (2008) NeuriteTracer: a novel ImageJ plugin for automated quantification of neurite outgrowth. J Neurosci Methods 168:134–139. doi:10.1016/j.jneumeth.2007.08.029 CrossRefPubMed

Reichenbach A, Derouiche A, Kirchhoff F (2010) Morphology and dynamics of perisynaptic glia. Brain Res Rev 63:11–25. doi:10.1016/j.brainresrev.2010.02.003 CrossRefPubMed

Rodríguez-Arellano JJ, Parpura V, Zorec R, Verkhratsky A (2016) Astrocytes in physiological aging and Alzheimer’s disease. Neuroscience 323:170–182. doi:10.1016/j.neuroscience.2015.01.007 CrossRefPubMed

Rusakov DA, Bard L, Stewart MG, Henneberger C (2014) Diversity of astroglial functions alludes to subcellular specialisation. Trends Neurosci 37:228–242. doi:10.1016/j.tins.2014.02.008 CrossRefPubMed

Saur L, Baptista PPA, de Senna PN et al (2014) Physical exercise increases GFAP expression and induces morphological changes in hippocampal astrocytes. Brain Struct Funct 219:293–302. doi:10.1007/s00429-012-0500-8 CrossRefPubMed

Sofroniew MV (2015) Astrogliosis. Cold Spring Harb Perspect Biol 7:a020420. doi:10.1101/cshperspect.a020420 CrossRefPubMedCentral

Volterra A, Liaudet N, Savtchouk I (2014) Astrocyte Ca²⁺ signalling: an unexpected complexity. Nat Rev Neurosci 15:327–335. doi:10.1038/nrn3725 CrossRefPubMed

Wang D, Bordey A (2008) The astrocyte odyssey. Prog Neurobiol 86:342–367. doi:10.1016/j.pneurobio.2008.09.015 PubMedPubMedCentral

Wilhelmsson U, Bushong EA, Price DL et al (2006) Redefining the concept of reactive astrocytes as cells that remain within their unique domains upon reaction to injury. Proc Natl Acad Sci 103:17513–17518. doi:10.1073/pnas.0602841103 CrossRefPubMedPubMedCentral

Yang Z, Wang KKW (2015) Glial fibrillary acidic protein: from intermediate filament assembly and gliosis to neurobiomarker. Trends Neurosci 38:364–374. doi:10.1016/j.tins.2015.04.003 CrossRefPubMedPubMedCentral

Title: Employing an open-source tool to assess astrocyte tridimensional structure
Authors: Gabriela Tavares
Manuella Martins
Joana Sofia Correia
Vanessa Morais Sardinha
Sónia Guerra-Gomes
Sofia Pereira das Neves
Fernanda Marques
Nuno Sousa
João Filipe Oliveira
Publication date: 01-05-2017
Publisher: Springer Berlin Heidelberg
Published in: Brain Structure and Function / Issue 4/2017
Print ISSN: 1863-2653
Electronic ISSN: 1863-2661
DOI: https://doi.org/10.1007/s00429-016-1316-8

At a glance: The STEP trials

Springer Medicine

Employing an open-source tool to assess astrocyte tridimensional structure

Abstract

At a glance: The STEP trials

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 4/2017

Cerebello-cerebral connectivity in the developing brain

Local morphology informs location of activation during navigation within the parahippocampal region of the human brain

Regional cerebellar volumetric correlates of manual motor and cognitive function

Temporal stability of reference frames in monkey area V6A during a reaching task in 3D space

Neurochemical evidence supporting dopamine D1–D2 receptor heteromers in the striatum of the long-tailed macaque: changes following dopaminergic manipulation

White matter properties differ in 6-year old Readers and Pre-readers