Figure 10.

Localization of ERα in hippocampal synapses. (A) – (C) Immunoelectron microscopic analysis of the distribution of ERα within axospinous synapses in the stratum radiatum of the hippocampal slices from adult male rat. (A) Gold particles (arrowheads) were localized in the pre- and postsynaptic regions. (B) In dendritic spines, gold particles were associated with PSD regions. (C) Gold particles were also localized in the nuclei. Pre, presynaptic region; post, postsynaptic region; Scale bar, 200 nm. (D–F) Immunohistochemical staining of ERα in the hippocampal slices from adult male rat [(D): whole hippocampus; (E): DG] and adult male ERαKO mouse [(F): DG]. gcl, Granule cell layer; hl, hilus. Scale bar, 500 μm for (D), and 200 μm for (E,F). (G) Western blot analysis of ERα in male rat hippocampal neurons. Blot of ERα in postsynaptic density (PSD), dendritic raft (D-Raft), and cytoplasm (CYT). From left to middle, blot of PSD fractions with RC-19 IgG (ERα), PSD-95 IgG (PSD-95), and Erk MAP kinase IgG (MAPK). From middle to right, blot of D-Raft with RC-19 (ERα) and flotillin-1 IgG (flotillin-1). At right-most lane, blot of CYT with RC-19 (ERα). The amount of protein applied was 20 μg for each lane, except for left-most PSD lane in which 60 μg protein was applied in order to improve the signal to noise ratio. Modified from Mukai et al. (2007, 2010).