Journal: The Journal of Neuroscience

Article Title: Memory-Related Synaptic Plasticity Is Sexually Dimorphic in Rodent Hippocampus

doi: 10.1523/JNEUROSCI.0801-18.2018

Figure Lengend Snippet: ERα is present at higher concentrations in female than in male CA1 synapses. A, Top left, Deconvolved two-photon microscopic images of immunofluorescent labeling were used to construct a 3D montage of the CA1 sample field (shown); one can see that a subpopulation of PSD-95-IR contacts (green) also contains ERα immunoreactivity (double labeling appears yellow). Scale bar, 2 μm. Top right, Image of a single double-labeled PSD shows the spatial relationship of areas occupied by PSD-95 (green) and ERα (red) immunoreactivities and the extent of overlap (merge, yellow). Scale bar, 0.1 μm. Bottom, Image shows the montage from the same z-stack illustrated in the top left but with the top of that panel rotated away from the viewer to show double labeling of the same puncta (arrows) from a different 3D viewpoint. B, The density frequency distribution for ERα-IR (colocalized with PSD-95) shows a greater rightward skew in females relative to males (p < 0.0001, F(19,646) = 17.28; males, n = 12; female, n = 24). C, Bar graph shows the percentage of double-labeled synapses with high concentrations of ERα immunolabeling (density units of ≥90) normalized to the mean male value shows that there were far more dense ERα-IR synapses in females than in males (***p = 0.0001, t(34) = 4.40). D, E, Density frequency distributions for all synapse-sized clusters of ERβ (D) and GPER1 (E) immunoreactivities colocalized with PSD-95 in the CA1 SR sample field (n = 12/group, females in diestrus). For ERβ-IR (D), there was a significant interaction between sex and immunolabeling density (p = 0.0005, F(21,462) = 2.4) because of a slightly greater proportion of synapses with low-density ERβ-IR (density units 83–88) in males than in females. There was no effect of sex on the numbers of densely ERβ-IR synapses (density units, ≥90; p = 0.12; t(22) = 1.6). For GPER1-IR, there were no group differences (GPER1: p = 0.89; F(20,440) = 0.33). F, Deconvolved epifluorescence images shows that ERβ and GPER1 (red) are both localized to synapse-sized puncta in CA1 SR and that some of those are colocalized with PSD-95 (green; doubles appear yellow). Scale bar, 2 μm.

Article Snippet: Stacks were deconvolved using AutoQuant version 2.2.1 (Media Cybernetics) and constructed into a 3D montage using Imaris (Bitplane; Lauterborn et al., 2016 ).

Techniques: Labeling, Construct, Immunolabeling