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Fig. 3 | Acta Neuropathologica Communications

Fig. 3

From: Age-dependent changes in synaptic plasticity enhance tau oligomerization in the mouse hippocampus

Fig. 3

The ALP in aged neurons contributes not only to LTD but also to the formation of tau oligomers. a LFS (1800 pulses, 1 Hz) reduced type I LC3 and increased type II LC3 mainly in the ipsilateral side of the aged hippocampus (immunoblot inset), resulting in a significant difference in the type II/type I LC3 ratio between the stimulated (ipsilateral, I) and contralateral (control) side (C) of the hippocampus (graph; n = 4, p = 0.0116, paired t-test). b Immunoprecipitation of LFS-induced tau oligomers by T22 indicated the presence of GluA2 (top blot) and NDP52 (middle blot) in addition to tau (bottom blot) in the molecular complexes obtained from the stimulated side (I) after LFS. Within the control (contralateral, C) side, the molecular complex was barely detected. These tendencies were confirmed in three independent experiments. c Intracerebral ventricle application of 3MA (1 μl/min of a 10 mM solution for 3 min) was carried out from 15 min before LFS (1800 pulses, 1 Hz), preventing LFS-induced tau oligomerization. The left panel shows the experimental procedure. The right panel shows an example of western blot analysis of sarkosyl-soluble (SS) and sarkosyl-insoluble (SI) fractions using A0024 tau antibody. d When Bafilomycin was applied (1 μl/min of a 10 μM solution for 3 min) from 15 min after the start of LFS (after 900 of 1800 pulses), LFS-induced tau oligomerization was not prevented, but 60- to 64-kDa hyperphosphorylated tau was formed. e Summary of normalized-tau levels in SI fractions from ipsilateral and contralateral hippocampi, which were obtained after LFS plus 3MA or Bafilomycin application (n = 5 for each group). Data are shown as the mean ± SEM. #p < 0.05 for a one-sample t-test against a theoretical value of ‘1’. f An electron microscopy image showing the morphology of immunogold-labeled tau oligomers formed by LFS under Bafilomycin treatment. Bar: 10 nm. In all experiments described here, aged wild-type mice (20–24 months old) were used

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