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. 2022 May 4;10:836746. doi: 10.3389/fcell.2022.836746

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Copyright © 2022 Byeon, Werner, Falter, Davidsen, Snyder, Ong and Yadav.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Phosphorylation of Sept7 alters its localization and dynamics. (A) Hippocampal neurons were transfected with GFP-tagged Sept7-WT, phosphomimetic Sept7-T426D, or phosphomutant Sept7-T426A along with membrane marker myristoylated-tdTomato at DIV9 and then fixed at DIV11. Phosphomimetic GFP-Sept7 (T426D) expression leads to early maturation of dendritic spines compared to Sept7-WT, while phosphomutant Sept7-T426A expressing neurons exhibited only filopodial protrusions. (B) Montage of time lapse images showing DIV11 hippocampal neurons transfected at DIV9 with GFP-tagged wild-type Sept7-WT, phosphomimetic Sept7-T426D, or phosphomutant Sept7-T426A. Phosphomimetic Sept7-T426D is stably localized within the dendritic spine head, phosphomutant Sept7-T426A localizes to the base of the dendritic spine, whereas the wild-type Sept7-WT shows dynamic changes in its localization within protrusions. Scale bar represents 5 µm (green) and 1 µm (montage), respectively. (C) Quantification of GFP intensity in protrusions of DIV11 neurons transfected with GFP-tagged wild-type Sept7-WT, phosphomimetic Sept7-T426D, and phosphomutant Sept7-T426A. Values represent mean intensity, error bars represent SEM and n = 50 protrusions from 10 neurons in each condition. Dunnett’s multiple comparison test, p < 0.0001.