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

. 2020 Feb 12;7(1):ENEURO.0303-19.2019. doi: 10.1523/ENEURO.0303-19.2019

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

Copyright © 2020 Hollos et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

PMC Copyright notice

Figure 3. — Photoactivation of the LOV2-JBD inhibitor rapidly reduces actin motility in the peripheral domain of the spine. A, Time-lapse sequences from 16-d hippocampal neurons expressing mCherry-actin and LOV2-JBD variants. ROIs (blue squares) encompassing spine-heads were photostimulated for 1 s at 36 s using 0.4 mW of 458-nm light. Spine-head motility was reduced following light only in cells expressing LOV2-JBD. Additional examples are in Extended Data Figure 3-1A. B, Arithmetic difference projections of mCherry-actin before (pre) and after (post) 458-nm illumination of the spine (blue box). LOV2-JBD immobilized mCherry-actin in photoactivated spines only. Dendritic shaft mCherry-actin was unchanged. C, Quantitative data on mCherry actin motility. Mean data ± SEM are shown. Spine numbers are indicated on the bars. D, mCherry-actin motility changes are plotted according to spine head diameter:neck length ratio. Data from spines with head diameter:neck length ratios corresponding to “mushroom” spines is in Extended Data Figure 3-1B. E, Time-lapse of spine-head mCherry-actin motility shows LOV2-JBD immobilizes actin motility within 6 s of photoactivation. F, Light alone does not alter actin dynamics. For E-F, more than or equal to four spines were measured per cell. G, Color-coded time projections provide spatial information on mCherry-actin motility over 90-s recording. Photoactivation is at 0 s. Images are acquired at 6-s intervals and coded with a unique hue (LUT). Mixed color (white-ish) indicates continued motility over time. Blue indicates no further movement after that time point. LOV2-JBD-expressing neurons display reduced motility following light. H, Higher resolution maximum projections of temporally color-coded dendritic spines are shown from 10-s time lapses acquired at 1-s intervals. These show mCherry-actin is immobilized in the spine periphery. Additional examples with actin footprints and movies generated from the time lapses are in Extended Data Figure 3-1C–E. I, The effect of latrunculin and phalloidin on dendritic spine motility were tested. Arithmetic projection images before and after 458-nm light indicate effect on actin dynamics. J, Quantitative data from I indicate that latrunculin reduces actin motility. Measurements are from ≥10 cells and multiple spines from separate experiments. Mean data ± SEM are shown. K, Temporal color coding of spines from neurons treated with latrunculin or phalloidin are shown.