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Computer vision profiling of neurite outgrowth dynamics reveals spatio-temporal modularity of Rho GTPase signaling
École Polytechnique Fédérale de Lausanne. (Smith)ORCID iD: 0000-0002-6163-191X
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2016 (English)In: Journal of Cell Biology, ISSN 0021-9525, E-ISSN 1540-8140, Vol. 212, no 1, 91-111 p.Article in journal (Refereed) Published
Abstract [en]

Rho guanosine triphosphatases (GTPases) control the cytoskeletal dynamics that power neurite outgrowth. This process consists of dynamic neuriteinitiation, 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 Neurite-Tracker, 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.

Place, publisher, year, edition, pages
Rockefeller University Press, 2016. Vol. 212, no 1, 91-111 p.
National Category
Cell Biology
URN: urn:nbn:se:kth:diva-181938DOI: 10.1083/jcb.201506018ISI: 000370486100010OAI: diva2:901994

QC 20160224. QC 20160319

Available from: 2016-02-09 Created: 2016-02-09 Last updated: 2016-03-19Bibliographically approved

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