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. 2023 Jan 4;55(1):108–119. doi: 10.1038/s12276-022-00918-6

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© The Author(s) 2023

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 Schematic overview of nArgBP2 KD in developing and mature neurons. b Representative images of spine morphologies and the proportions of mushroom-shaped, stubby, and thin spines in neurons expressing control or shRNA-nArgBP2 in developing neurons and mature neurons. Developing neurons were transfected at DIV 9 and fixed at DIV 16, whereas mature neurons were transfected at DIV 16 and fixed at DIV 21. Scale bar: 2 μm. The proportion of each is shown above the bar. c Representative time-lapse images of spines in control and nArgBP2 KD neurons during cLTP induction. Neurons at DIV 16 were transfected with shRNA-nArgBP2 alone or combined with EGFP-nArgBP2-res and were imaged at DIV 21. Note the gradual enlargement of the spine heads in control and KD + EGFP-nArgBP2-res neurons but not in KD neurons. The yellow arrowheads indicate the spine heads. Scale bars: 2 μm. d The relative size of spine heads in control and nArgBP2 KD neurons after cLTP, normalized to the initial value. n = 9 for control, 11 for KD, and 7 for KD + EGFP-nArgBP2-res (one-way ANOVA followed by Tukey’s HSD test). *p < 0.05; **p < 0.01. Error bars indicate the means ± s.e.m.s. e Schematic diagram of the process of 3D geometry analysis of dendritic spines using DXplorer. The left panel shows the acquisition of 3D-SIM images and automatic detection of dendritic spines. 3D geometrical features were extracted and calculated. The datasets were further analyzed and classified by dimensionality reduction and machine-learning-based automatic classification. f Representative morphological features extracted from the 3D mesh of dendritic spines. S, Surface; V, Volume; L, length of spine; hMax, the maximum diameter of the spine head; nMax, the maximum diameter of the spine neck; CS, the centroid of the spine. g Normalized differences in morphological features of spines between pre-and post cLTP in control and nArgBP2 KD neurons. HNR, the ratio of head to the neck (HNR = hMax/nMax); n = 39 for control, 34 for KD. *p < 0.05; **p < 0.01; ***p < 0.001. Student’s t-test; error bars indicate the mean ± s.d.