Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

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

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 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/.

PMC Copyright notice

Fig. 2 — a Neurons expressing membrane-tagged GFP, to emphasize morphology (pink highlight), and stained with Alexa-Fluor-647-phalloidin to monitor F-actin (reverse grayscale). Neurons fixed after 5 min incubation with vehicle (left) or with 50 μM NMDA (right). Insets below show enlarged regions boxed in. Scale bars, 8 µm (above); 2.8 µm (below). b Time course of actinification; n = 4 independent experiments for 0 and 5 min; n = 2 independent experiments for all the other time points. c Examples of two representative time courses where either mApple-F-tractin or Lifeact-mRFP intensity were monitored over time in an individual dendrite shaft (green line) vs the adjacent spines (magenta line) during NMDA-induced actinification. Plotted lines represent mean ± SEM from regions of interest (ROIs) taken across different portions of the same dendrite and different spines of a single neuron. d Time-lapse montage of a dendritic region expressing Lifeact-mRFP at selected time points before and after exposure to NMDA. Scale bar, 5 µm. Blue arrows: examples of spine F-actin decreasing as the F-actin signal in the shaft increases. e Hippocampal cultures incubated for 30 min in vehicle or NMDA, prior to processing for platinum replica electron microscopy (PREM). A neuron representative of each condition is shown at left; yellow arrows: cell soma; yellow boxes: two regions of the dendrite that are displayed at 70× higher magnification in the middle panels, and at a further 3× higher magnification in the right panels. Actin filaments are highlighted (cyan). In control (vehicle), numerous clusters of short, branched actin filaments are observed in presumptive dendritic spines, with occasional actin filaments within the dendrite shaft. After NMDA spine-like protrusions substantially decreased, and numerous long, unbranched filaments were detected within the dendritic shaft (cyan labeled structures within boxed regions). Scale bars, 20 µm for left panels, 500 nm for middle panels, and 200 nm for right panels. All graphs: data presented as mean ± SEM, and source data are provided as source data file.