Fig. 5

CXCL1 is synaptotoxic. Mouse neurons, transfected at 5-7DIV with GFP, were treated with 40 nM recombinant mouse CXCL1 or vehicle for 24 h to determine if CXCL1 is damaging to neurons. In some cases, neurons were pre-treated for 20 min with 40 ng/ml SB225002, an antagonist of the CXCL1 receptor, CXCR2. Neurons were imaged at 14DIV. A Representative images of dendrites in CXCL1 and SB225002 treated conditions (n = 3, three independent experiments each with three technical replicates) Scale bar = 10 μm. B Quantification of spine density per μm shows reduced spine numbers upon exposure to CXCL1 relative to controls, that was rescued when neurons were pre-treated with the CXCR2 antagonist (n = 6, six independent experiments each with three technical replicates). Data was analysed by ordinary one-way ANOVA and Tukey’s multiple comparison post-hoc test. Measures of neuron health across three wells per experimental repeat showed C reduced total neurite length, D fewer nodes, and E fewer roots on exposure of neurons to CXCL1 that is prevented by pre-treatment with SB225002, demonstrating impairments in neuron health as a result of CXCL1 (n = 3, three independent experiments each with three technical replicates). Data was analysed by ordinary one-way ANOVA and Tukey’s multiple comparison post-hoc test. Data on graphs is mean ± SEM and is shown relative to control conditions. *p < 0.05