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. 2022 Oct 13;13:6037. doi: 10.1038/s41467-022-33268-y

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Cultured rat hippocampal neurons stained using Alexa-Fluor-647-phalloidin in the absence or presence of oxygen and glucose deprivation (OGD). Selected inverted grayscale somatodendritic regions are highlighted in pink. Note: in control F-actin is concentrated in dendritic spines (dark puncta), while OGD induces a decrease in spine F-actin and an accumulation of linear bundles of F-actin within the soma and dendrite of many neurons (actinification). Scale bar, 5 µm. b Numbered insets correspond to blue boxed regions in a. Width blue boxes = 17 µm. c Quantification of the fraction of actinified neurons in control vs OGD or OD (oxygen deprivation) for the indicated durations; the NMDA receptor antagonist APV prevented actinification seen with 6 h OGD. n = 4 independent experiments; Kruskal–Wallis one-way ANOVA (p = 0.0066), with Dunn’s post hoc multiple comparison analysis (p = 0.0421 OGD 0 m vs 6 h; p = 0.0149 OGD 6 h vs APV). d Coronal brain section of a Thy1-YFP (green) expressing mouse stained for the neuronal cell body marker NeuN (magenta) 6 h post stroke. Regions marked “1” and “2”: areas from which the corresponding enlarged images in e are shown (below). Scale bar, 350 µm. Right: Higher magnification of the individual fluorescent labels of the ischemic area. e Alexa-Fluor-647-Phalloidin staining of neurons within somatosensory cortex within (2) infarcted region (middle, right panels at different magnification), or (1) corresponding region contralateral to the stroke (left single panel). Blue arrows: accumulation of linear actin bundles within the somatodendritic region of YFP-positive neurons. F-actin staining within the control, contralateral region (left) exhibits the punctate pattern of dendritic spines. Scale bars, 5 µm (left and middle), 8 µm (right). f Quantification of the fraction of YFP-positive layer 5/6 neurons exhibiting actinification under the following conditions and regions: sham-operated mice (somatosensory cortex); or stroke-induced mice within the contralateral cortex, ipsilateral cortex, the ischemic core, or the penumbra.; n = 3 biologically independent animals each, sham vs stroke; one-way ANOVA (p = 0.0001), with Tukey’s post hoc multiple comparison analysis (p = 0.0001 sham vs core; p = 0.0004 core vs pen). All graphs: data presented as mean ± SEM. Source data are provided as source data file.