Growth cone repulsion to Netrin-1 depends on lipid raft microdomains enriched in UNC5 receptors.
Marc Hernaiz-Llorens, Cristina Roselló-Busquets, Nela Durisic, Adam Filip, Fausto Ulloa, Ramón Martínez-Mármol, Eduardo Soriano
Author Information
Marc Hernaiz-Llorens: Department of Cell Biology, Physiology and Immunology, Faculty of Biology and Institute of Neurosciences, University of Barcelona, 08028, Barcelona, Spain. ORCID
Cristina Roselló-Busquets: Department of Cell Biology, Physiology and Immunology, Faculty of Biology and Institute of Neurosciences, University of Barcelona, 08028, Barcelona, Spain.
Nela Durisic: Queensland Brain Institute (QBI), The University of Queensland, St Lucia Campus, Brisbane, QLD, 4072, Australia.
Adam Filip: Department of Cell Biology, Physiology and Immunology, Faculty of Biology and Institute of Neurosciences, University of Barcelona, 08028, Barcelona, Spain.
Fausto Ulloa: Department of Cell Biology, Physiology and Immunology, Faculty of Biology and Institute of Neurosciences, University of Barcelona, 08028, Barcelona, Spain.
Ramón Martínez-Mármol: Department of Cell Biology, Physiology and Immunology, Faculty of Biology and Institute of Neurosciences, University of Barcelona, 08028, Barcelona, Spain. r.martinezmarmol@uq.edu.au. ORCID
Eduardo Soriano: Department of Cell Biology, Physiology and Immunology, Faculty of Biology and Institute of Neurosciences, University of Barcelona, 08028, Barcelona, Spain. esoriano@ub.edu.
During brain development, Uncoordinated locomotion 5 (UNC5) receptors control axonal extension through their sensing of the guidance molecule Netrin-1. The correct positioning of receptors into cholesterol-enriched membrane raft microdomains is crucial for the efficient transduction of the recognized signals. However, whether such microdomains are required for the appropriate axonal guidance mediated by UNC5 receptors remains unknown. Here, we combine the use of confocal microscopy, live-cell FRAP analysis and single-particle tracking PALM to characterize the distribution of UNC5 receptors into raft microdomains, revealing differences in their membrane mobility properties. Using pharmacological and genetic approaches in primary neuronal cultures and brain cerebellar explants we further demonstrate that disrupting raft microdomains inhibits the chemorepulsive response of growth cones and axons against Netrin-1. Together, our findings indicate that the distribution of all UNC5 receptors into cholesterol-enriched raft microdomains is heterogeneous and that the specific localization has functional consequences for the axonal chemorepulsion against Netrin-1.
