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. 2019 Mar 25;9:5037. doi: 10.1038/s41598-019-41661-9

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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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Firing properties in both cortical layers develop largely in parallel. (a) Example voltage traces in response to suprathreshold depolarising current step that elicits 10 or more action potentials. Inset: first two action potentials of the same voltage trace. Black lines indicate the amplitude of the current step that elicited the voltage response. (b) Action potential halfwidth decreases rapidly during the second postnatal week, decreasing even further afterwards (L3, Welch’s F(2,18) = 25.61, p < 0.001; post-hoc w1-w2, p = 0.001; w2 vs w4, p = 0.011; L5, F(2,42) = 124.67, p < 0.001; post-hoc w1-w2, p < 0.001; w2 vs w4, p = 0.003). (c) Action potential amplitude increases during the second postnatal week in neurons of both layers (L3, F(2,37) = 46.85, p < 0.001; post-hoc w1-w2, p < 0.001; L5, F(2,43) = 43.78, p < 0.001; post-hoc w1-w2, p < 0.001). (d) Action potential threshold becomes more hyperpolarised between weeks 1 and 2 (L3, F(2,38) = 31.36, p < 0.001; post-hoc w1-w2, p < 0.001; L5, F(2,43) = 36.87, p < 0.001; post-hoc w1-w2, p < 0.001). (e) Rheobase increases during development of neurons in both layers (L3, F(2,35) = 23.13, p < 0.001; post-hoc w1-w2, p = 0.036; w2 vs w4, p < 0.001; L5, F(2,40) = 29.74, p < 0.001; post-hoc w1-w2, p < 0.001; w2 vs w4, p = 0.003). (f) Spike frequency adaptation represented through ISI ratios. Heatmap colours represent the ratio between the 9^th ISI and each of the 8 previous ISIs (numbered below). (g) ISI1/9 ratio decreases during development in L3 but not L5 neurons (L3, F(2,31) = 5.21, p = 0.011; post-hoc w1-w4, p = 0.008). (h) ISI4/9 ratio increases during development in L5 neurons (F(2,38) = 23.86, p < 0.001, post-hoc w1-w2, p < 0.001).