Growth cone repulsion to Netrin-1 depends on lipid raft microdomains enriched in UNC5 receptors
- PDF / 11,576,242 Bytes
- 24 Pages / 595.276 x 790.866 pts Page_size
- 16 Downloads / 179 Views
Cellular and Molecular Life Sciences
ORIGINAL ARTICLE
Growth cone repulsion to Netrin‑1 depends on lipid raft microdomains enriched in UNC5 receptors Marc Hernaiz‑Llorens1,2 · Cristina Roselló‑Busquets1,2 · Nela Durisic5 · Adam Filip1,7 · Fausto Ulloa1,2 · Ramón Martínez‑Mármol1,6 · Eduardo Soriano1,2,3,4 Received: 8 May 2020 / Revised: 8 September 2020 / Accepted: 30 September 2020 © The Author(s) 2020
Abstract 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. Keywords UNC5 · Netrin-1 · Cerebellar EGL neurons · Axonal repulsion · Lipid raft microdomain · Single particle tracking
Introduction The establishment of proper neuronal connections relies on the ability of axons to locate and reach their target during neural development. Axons must elongate to find and contact their partners, which can be very distant from the Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00018-020-03663-z) contains supplementary material, which is available to authorized users.
soma of the neuron. Thus, axons need specific extracellular cues to properly navigate towards their final destination. The leading edge of the axon is known as the growth cone, which is the major motile structure that senses extracellular signals. Netrins are guidance cues involved in a variety of cellular processes such as cell survival [1], cell migration [2] and neuronal axon guidance [3]. In vertebrates, four netrins (Netrin1-4) have been identified [4], and are recognized by four different families of receptors
* Ramón Martínez‑Mármol [email protected]
4
Institució Catalana de Recerca I Estudis Avançats (ICREA), 08010 Barcelona, Spain
* Eduardo Soriano [email protected]
5
Queensland Brain Institute (QBI), The University of Queensland, St Lucia Campus, Brisbane, QLD 4072, Australia
6
Clem Jones Centre for Ageing Dementia Research (CJCADR), Queensland Brain Institute (QBI), Th
Data Loading...
