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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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Progressive acceleration of axonal vesicular flow during network maturation. (A) Kymograph analyses obtained at different stages of network maturation (early, mid, late) show progressive increase in axonal transport of dense core vesicles. Number in brackets indicates DIV. Anterograde and retrograde movements, pauses and static vesicles are shown. (B) Increased segmental velocity (Antero, F_3,201 = 121.3, p < 0.0001; Retro, F_3,201 = 11.5, p < 0.0001; n = 52) and (C) increased number of vesicles (Antero, F_3,201 = 105.9, p < 0.0001; Retro, F_3,201 = 14.7, p < 0.0001) translate into (D) a redistribution of the pool of motile vesicles (Antero, Χ² = 452.8, p < 0.0001; Retro, Χ² = 26.26, p < 0.0001). (E) The pausing time of motile vesicles rapidly decreases to a minimum at mid stages (F_3,201 = 42.1, p < 0.0001). These kinetics result in (F) increased global linear flow rate (F_3,201 = 109.5, p < 0.0001) and (G) increased anterograde net flux (F_3,201 = 103.0, p < 0.0001) of axonal vesicles that stabilize after DIV 14 (DIV 14 vs DIV 21, n.s). All bar graphs recapitulate data obtained at early (DIV 4) and late (DIV 21) stages of neuronal culture. ***p < 0.0001.