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. 2009 Jul 13;186(1):85–97. doi: 10.1083/jcb.200901084

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© 2009 Rex et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 2. — 3D reconstruction analysis of immunofluorescence. (A) Deconvolution photomicrograph from proximal CA1 str. radiatum shows total cofilin⁺ elements. (B) Images show localization of pCofilin⁺ (left) and PSD95⁺ (middle) elements in the same field (the arrows indicate overlapping structures). pCofilin⁺ elements were of similar size and shape as total cofilin⁺ elements (but less numerous). (C) Photomicrographs (Fluor) and volumetric reconstructions (3D) of a single synapse containing pCofilin ir and PSD95 ir displayed in 30° clockwise turns (left to right). (D) The plot shows numbers of immunolabeled spines (mean ± SEM) in the field of physiological recording for slices receiving control stimulation (con; open bars) or TBS (closed bars). Counts were made from 3D reconstructions. The plot shows separate quantification of total PSD95⁺, double-labeled cofilin (cofilin⁺/PSD95⁺), and double-labeled pCofilin (pCofilin⁺/PSD95⁺) elements. The latter comparison was replicated using the Volocity measurement module. PSD95⁺ elements associated with pCofilin ir represented <5% of the total PSD95⁺ puncta in control slices. Slices receiving TBS showed approximately threefold more pCofilin⁺/PSD95⁺ colabeled elements versus controls (**, P < 0.01). Bars: (A and B) 5 µm; (C) 0.5 µm.