title: Computer vision profiling of neurite outgrowth dynamics reveals spatiotemporal modularity of Rho GTPase signaling.
abstract: Rho guanosine triphosphatases (GTPases) control the cytoskeletal dynamics that power neurite outgrowth. This process consists of dynamic neurite initiation, elongation, retraction, and branching cycles that are likely to be regulated by specific spatiotemporal signaling networks, which cannot be resolved with static, steady-state assays. We present NeuriteTracker, a computer-vision approach to automatically segment and track neuronal morphodynamics in time-lapse datasets. Feature extraction then quantifies dynamic neurite outgrowth phenotypes. We identify a set of stereotypic neurite outgrowth morphodynamic behaviors in a cultured neuronal cell system. Systematic RNA interference perturbation of a Rho GTPase interactome consisting of 219 proteins reveals a limited set of morphodynamic phenotypes. As proof of concept, we show that loss of function of two distinct RhoA-specific GTPase-activating proteins (GAPs) leads to opposite neurite outgrowth phenotypes. Imaging of RhoA activation dynamics indicates that both GAPs regulate different spatiotemporal Rho GTPase pools, with distinct functions. Our results provide a starting point to dissect spatiotemporal Rho GTPase signaling networks that regulate neurite outgrowth.
authors.affiliation: Department of Biomedicine, University of Basel, 4058 Basel, Switzerland.
authors.forename: Ludovico
authors.initials: L
authors.lastname: Fusco
authors.affiliation: Institut Dalla Molle d'Intelligence Artificielle Perceptive (IDIAP Research Institute), 1920 Martigny, Switzerland.
authors.forename: Riwal
authors.initials: R
authors.lastname: Lefort
authors.affiliation: Computer Vision Laboratory, École Polytechnique Fédérale de Lausanne, 1015 Lausanne, Switzerland.
authors.forename: Kevin
authors.initials: K
authors.lastname: Smith
authors.affiliation: Computer Vision Laboratory, École Polytechnique Fédérale de Lausanne, 1015 Lausanne, Switzerland.
authors.forename: Fethallah
authors.initials: F
authors.lastname: Benmansour
authors.affiliation: Computer Vision Laboratory, École Polytechnique Fédérale de Lausanne, 1015 Lausanne, Switzerland.
authors.forename: German
authors.initials: G
authors.lastname: Gonzalez
authors.affiliation: Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule, 4058 Basel, Switzerland.
authors.forename: Caterina
authors.initials: C
authors.lastname: Barillari
authors.affiliation: Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule, 4058 Basel, Switzerland.
authors.forename: Bernd
authors.initials: B
authors.lastname: Rinn
authors.affiliation: Institut Dalla Molle d'Intelligence Artificielle Perceptive (IDIAP Research Institute), 1920 Martigny, Switzerland.
authors.forename: Francois
authors.initials: F
authors.lastname: Fleuret
authors.affiliation: Computer Vision Laboratory, École Polytechnique Fédérale de Lausanne, 1015 Lausanne, Switzerland.
authors.forename: Pascal
authors.initials: P
authors.lastname: Fua
authors.affiliation: Department of Biomedicine, University of Basel, 4058 Basel, Switzerland olivier.pertz@izb.unibe.ch.
authors.forename: Olivier
authors.initials: O
authors.lastname: Pertz
mesh_terms: D000818:Animals; D051379:Mice; D016501:Neurites / Q000201:enzymology* / Q000378:metabolism; D010641:Phenotype; D015398:Signal Transduction; D062211:Spatio-Temporal Analysis; D014407:Tumor Cells, Cultured; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
chemical_list: D020742:rhoA GTP-Binding Protein