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. 2019 Oct 16;10:4694. doi: 10.1038/s41467-019-12681-w

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© The Author(s) 2019

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. 4 — Imaging the orientation of short actin filaments. a Dynamic imaging of the myosin-driven movement of phalloidin-labeled actin filament. The white box contains the trajectory of a short actin filament. b Magnified view of the yellow boxed region in a. The dipole orientations of the actin filaments change as they move. c Time-lapse orientation and position of the fragmented actin in b. d 2D-pSIM imaging of the actin filaments in hippocampal neurons, which clearly distinguishes between the continuous long actin filaments in the dendrite and the region of discrete actin ring structure in the axon. e Another illustration of the actin ring structure in the axon. f, g Magnified views of the boxed regions in d, which compare the results of PM and pSIM imaging. h The intensity profile of the line indicated in e, whose Fourier transform (i) shows a 184 nm periodicity, consistent with previously reported results. j, k The actin ring structure is critical to the membrane-associated periodic skeleton (MPS) in neurons. The previous model assumes an end-to-end organization for the adducin-capped actin filaments. However, pSIM reveals that the orientation of the short actin filaments is parallel to the axon shaft, supporting a side-by-side organization for the actin ring structures. Scale bars: a 2 μm and d–g 1 μm