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. 2018 Sep 7;8:13429. doi: 10.1038/s41598-018-31759-x

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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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Time-course of corticostriatal network formation. (A) Time recording of neurite outgrowth in the synaptic chamber using lentiviral expression of GFP and mCherry in cortical and striatal neurons, respectively. Images show the growth of cortical axons and striatal dendrites in the synaptic chamber from 3 to 10 days in vitro (DIV). Scale bars, 10 μm. (B,C) Time recording of the formation of corticostriatal connections in the synaptic chamber using selective markers for the presynaptic (synaptophysin, SYP) and postsynaptic (PSD95) compartments. (B) Images show SYP/PSD95 colocalization at DIV 7 and DIV 21. Insets show a 10x magnification of boxes in the left panel. Scale bars, 10 μm (insets, 2 μm). (C) Quantification of the number of SYP/PSD95 adjacent spots per 100 μm branches shows the progressive formation of synaptic contacts until DIV 14 (F_4,112 = 58.3, p < 0.0001; n = 23) and stabilization afterwards (DIV 14 vs DIV 21, n.s). The bar graph recapitulates data obtained at early (DIV 4) and late (DIV 14) stages of neuronal culture. ***p < 0.0001.