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. 2018 Nov 9;8:16644. doi: 10.1038/s41598-018-34523-3

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© The Author(s) 2018

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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MPS VII neuronal activity and MPS VII neurospheroids calcium (Ca⁺⁺) imaging analysis. (A) Ca⁺⁺ release in control and MPS VII neurons, upon KCl induction and reduction of Ca⁺⁺ signaling upon inhibition with nimodipine. (B) FM-1–43 fluorescence decay in control (black) and MPS VII (grey) neurospheroids at day 28. Data are mean ± SD of 3 independent cultures. (C) Percentage of cells per number of spontaneous Ca⁺⁺ events in 300 seconds for control (black) and MPS VII (cl. 8 grey; cl. 13 light grey) neurospheroids. (D) Visual representations of neural networks from control and MPS VII neurospheroids reconstructed using FluoroSNNAP; circles show the position of cells in culture, yellow circles represent functionally connected nodes and red lines represent the functional connectivity of pair-wise neurons. (E) Ca⁺⁺ events peak amplitude, rise time and fall time in control (black) and MPS VII (cl. 8 grey; cl. 13 light grey) neurospheroids. Network properties: Connectivity index (F) and Global synchronization index (G) in control (black) and MPS VII (cl. 8 grey; cl. 13 light grey) neurospheroids. Data are mean ± SD of 3 (control) and 4 (MPS VII; 2 with MPS VII-cl.8 and 2 with MPS VII-cl.13) independent cultures. Asterisks indicate significant difference: *p < 0.05, ***p < 0.01, ***p < 0.001.