Granule neuron differentiation status and substrate modulate the Netrin-1 signaling response in the cerebellum

Abstract

During development, granule neuron progenitors often have to migrate over long distances from their germinal zone to their final location. They navigate by integrating chemorepulsive or chemoattractive guidance cues, which sometimes come from the germinal zone through guidance molecules. However, it is unclear how neurons, upon differentiation, modulate their response to a given signal. In this study, we focus on the guidance-signaling protein Netrin-1 (Ntn1) and its role in mouse cerebellar development. Using FACS sorting to isolate the granule neurons of Atoh1-GFP transgenic mice cerebella based on their differentiation status, we ran an in vitro granule neurons display different migration behaviors in response to Ntn1 depending on their differentiation status and the surrounding substrate. We show that gradient assay to track the netrin-driven migration patterns of different stages of granule neurons based on their GFP fluorescence on two substrates: laminin and vitronectin.