Figure 1: Tau localizes to presynaptic terminals and binds to synaptic vesicles via its N-terminal domain.
From: Tau association with synaptic vesicles causes presynaptic dysfunction
(a) Tau and CSP immunolabeling at neuromuscular junctions (NMJs) of Drosophila larvae expressing WT or FTDP-17 pathogenic mutant Tau (R406W, V337M or P301L) under the D42-Gal4 motor neuron driver. Axons (arrowheads) and synaptic boutons (arrows) are indicated. Scale bar, 20 μm. (b) Quantification of fluorescence intensity of Tau within synaptic boutons (SBs) as ratio to the intensity of axonal Tau. One-way ANOVA, **P=0.0030, 0.0019, 0.0041 (R406W, V337M, P301L) n=10 (R406W, V337M, P301L) or 12 (WT) NMJs from 5 to 6 animals. Data present mean±s.e.m. (c,d) Super-resolution structured illumination microscopy analysis of Tau and CSP immunolabeling within SBs under non-treated condition (c) or after depletion of synaptic vesicles in Shits1 mutant background by KCl stimulation at the non-permissive temperature (d). Scale bar, 5 μm. (e,f) Immunoblots of Tau (anti-His tag) and synaptic vesicle (SV) proteins Synaptobrevin (Syb), Synaptotagmin (Syt) and Synapsin (Syn) from sedimentation assay (e) and co-immunoprecipitation (co-IP) using anti-His antibodies (f) assessing recombinant human Tau binding to purified synaptic vesicles. (g) Electron microscopy imaging of recombinant Tau (probed by Ni-NTA-Nanogold) bound to ultrapure synaptic vesicles in vitro. Scale bar, 50 nm. (h–j) Mapping of the vesicle-binding domain of Tau in vitro by co-IP assay. Truncations of the N-terminal (NT), proline-rich (PRD), microtubule-binding (MTB) or C-terminal (CT) domains of Tau were generated as indicated in the schematic (h). Immunoblots of recombinant Tau domain-truncations and Syb (SV marker) from co-IP using anti-His antibodies (i) and quantification of relative Syb intensity (j). One-way ANOVA, ***P=0.002, n=3 independent experiments.
