Abstract
Growth cones are complex, motile structures at the tip of an outgrowing neurite. They often exhibit a high density of filopodia (thin actin bundles), which complicates the unbiased quantification of their morphologies by software. Contemporary image processing methods require extensive tuning of segmentation parameters, require significant manual curation, and are often not sufficiently adaptable to capture morphology changes associated with switches in regulatory signals. To overcome these limitations, we developed Growth Cone Analyzer (GCA). GCA is designed to quantify growth cone morphodynamics from time-lapse sequences imaged both in vitro and in vivo, but is sufficiently generic that it may be applied to nonneuronal cellular structures. We demonstrate the adaptability of GCA through the analysis of growth cone morphological variation and its relation to motility in both an unperturbed system and in the context of modified Rho GTPase signaling. We find that perturbations inducing similar changes in neurite length exhibit underappreciated phenotypic nuance at the scale of the growth cone.
© 2018 Bagonis et al.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Cell Line, Tumor
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Cell Movement
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Cell Shape / genetics
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Gene Expression Regulation
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Genetic Heterogeneity
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Growth Cones / metabolism
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Growth Cones / ultrastructure*
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Guanine Nucleotide Exchange Factors / deficiency
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Guanine Nucleotide Exchange Factors / genetics
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Image Processing, Computer-Assisted / statistics & numerical data*
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Mice
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Molecular Imaging / methods
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Molecular Imaging / standards*
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Neurons / metabolism
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Neurons / ultrastructure*
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Neuropeptides / deficiency
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Neuropeptides / genetics
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Phosphoproteins / deficiency
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Phosphoproteins / genetics
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Protein Serine-Threonine Kinases / deficiency
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Protein Serine-Threonine Kinases / genetics
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Pseudopodia / metabolism
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Pseudopodia / ultrastructure
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Rho Guanine Nucleotide Exchange Factors / deficiency
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Rho Guanine Nucleotide Exchange Factors / genetics
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Signal Transduction
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Software*
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Time-Lapse Imaging / methods
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Time-Lapse Imaging / standards*
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cdc42 GTP-Binding Protein / deficiency
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cdc42 GTP-Binding Protein / genetics
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rac1 GTP-Binding Protein / deficiency
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rac1 GTP-Binding Protein / genetics
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rho GTP-Binding Proteins / deficiency
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rho GTP-Binding Proteins / genetics*
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rhoA GTP-Binding Protein
Substances
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Arhgef7 protein, mouse
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Cdc42 protein, mouse
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Guanine Nucleotide Exchange Factors
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Neuropeptides
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Phosphoproteins
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Rac1 protein, mouse
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Rho Guanine Nucleotide Exchange Factors
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Trio protein, mouse
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Protein Serine-Threonine Kinases
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RhoA protein, mouse
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cdc42 GTP-Binding Protein
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rac1 GTP-Binding Protein
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rho GTP-Binding Proteins
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rhoA GTP-Binding Protein