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. 2017 Oct 27;8:1158. doi: 10.1038/s41467-017-01046-w

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© The Author(s) 2017

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. 5 — Delayed callosal projection neurons display reduced synaptic input and reduced neuronal activity. a Activity-reporter (SAREdGFP) shows decreased GFP expression in late-arrived neurons (arrowheads mark activated cells, magnified at right). Graph shows the proportion of GFP-positive activated cells during axonal arborization (P10) and at P21; n = 6 and 7 brains (Control vs. dnTCF4, respectively) at P10 and n = 5 brains at P21, Mann–Whitney, P > 0.9999 for P21. b Excitatory connections labeled by PSD-95-specific FingR-GFP showing a decreased number of synaptic puncta in neurons with delayed migration (dnTCF4); n = 18 and 21 cells from 3 and 4 brains (Control vs. dnTCF4, respectively), Mann–Whitney. c mEPSCs recording samples of layer 2/3 pyramidal neurons for control (top trace) and dnTCF4 (bottom trace) P14-P15 animals. No significant difference in amplitude and a significant increase in mEPSCs inter-event interval for dnTCF4 animals; n = 7 animals per group, 1 cell per animal, Student’s t test (bar graphs, P > 0.9999 for P21), Kolmogorov–Smirnov test (cumulative distribution). d Rabies-mediated monosynaptic retrograde tracing (schematized at left) shows reduced afferent inputs on late arrived (dnTCF4) starter cells, magnified inserts display equal density of starter cells. e Quantification of input/starter cells shows a major reduction of inputs from the granular layer; n = 8 and 14 brains (Control and dnTCF4, respectively), Mann–Whitney, P = 0.1876 for supragranular layer. f The timeline showing the experimental design used for rescuing aberrant callosal projections via hM3Dq DREADD receptor activation. Daily clozapine-n-oxide (CNO) injections were performed during axonal arborization. Chemogenetic stimulation of late-arrived CPNs rescues contralateral arbors compared to non-activated neurons (PBS injection); n = 11, 24, and 11 brains (Control, PBS, CNO, respectively), Kruskal–Wallis, Control vs. CNO: P = 0.5455 for L5A P > 0.9999 for L2/3. Graphs display mean ± s.e.m. ns, non-significant, *P < 0.05, **P < 0.01. Bars = 5 μm (b), 20 μm (a and d, insert), 50 μm (a, overview) and 100 μm (d and f, overview)