Figure 3
Extrasynaptic NMDAR activation induces hTau-dependent toxicity in primary neuronal cultures and hippocampal slice cultures. (a) Representative confocal images of primary neurons expressing EGFP or EGFP-hTau after synaptic or extrasynaptic activation. (b) Confocal images of EGFP-hTau-expressing neurons after extrasynaptic activation and immunostaining against βIII tubulin. Arrows mark non-infected neurons. (c) Quantification of hTau-dependent toxicity. Shown is the ratio of non-degenerated infected primary neurons (neurons without fragmented or beaded neurites or ballooned morphology) to total infected neurons. (d) Representative western blot with phospho-ERK 1/2 (pERK) and ERK 1/2 antibodies of lysates from primary neurons after synaptic or extrasynaptic activation. (e) Cytotoxicity of hTau in wt hippocampal slice cultures after extrasynaptic activation measured by Cytotox-Glo assay. (f) Western blot showing AT8 phosphorylation of hTau after extrasynaptic activation in slice cultures. (g) Representative dendritic segments from CA1 neurons of wt slice cultures after extrasynaptic activation. (h) Quantification of spine density in wt slices analyzed 1 day after extrasynaptic activation. eSyn, extrasynaptic; Syn, synaptic activation; veh, vehicle; values are shown as mean±S.E.M. with **P<0.01 and ***P<0.001; Mann–Whitney-U-test; n=10–13 (c), n=3 (d); n=8 (e), n=4 (f); n=10 (h) scale bars: 50 μm (a, b), 5 μm (g)
