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. 2017 Jul 14;11:202. doi: 10.3389/fncel.2017.00202

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Copyright © 2017 Maddaloni, Bertero, Pratelli, Barsotti, Boonstra, Giorgi, Migliarini and Pasqualetti.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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3D-reconstruction and analysis of serotonergic fibers in PND 28 brain. (A) Coronal tables adapted from adult mouse brain atlas showing the precise anatomical localization of the region analyzed (Franklin and Paxinos, 2008), as indicated by asterisks corresponding to high magnification images in (B–I). (B–I) Representative high power confocal images of GFP-immunostained serotonergic fibers of a single analyzed block, and their 3D-reconstructions. (J) Histogram comparing the occupancy index across the regions, and heatmap showing color-coded p-values of the differences in occupancy index among the analyzed regions. Box plots show the log₂ fold changes increase/decrease in the occupancy index at PND 28, as compared to PND 14. (K) Histogram showing the total length of 5-HT-fibers in the analyzed areas, and heatmap representing color-coded p-values relative to the differences in fiber length among the regions. Box plots show log₂ fold change increase/decrease in fiber length in each region at PND 28, as compared to PND 14. (L) Histogram showing the mean diameter of serotonergic fibers, and heatmap representing color-coded p-value relative to the differences in fiber diameter. Box plots show the log₂ fold changes increase/decrease in the mean diameter calculated at PND 28, as compared to PND 14. (M) ED distribution graph, composed by curves depicting the frequency of EDs (from 0.1 μm to 1.4 μm, 0.05 μm step; x-axis) in each region. Data are expressed as mean ± SEM. Statistical significance was calculated with one way ANOVA, with multiple comparisons and Tukey’s p-values correction. Two tailed Student’s t test for unpaired data statistical significance was reported in box-whisker graphs. ns: p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. Scale bar: 10 μm. mPFC, medial Prefrontal Cortex; GP, Globus Pallidus; CPu, Caudate/Putamen; S1BF, Primary Somatosensory Cortex, Barrel Field; BLA, Basolateral Amygdala; Hp, Lacunosum moleculare layer of the dorsal CA1 of hippocampus; DLG, Dorsal Lateral Geniculate Nucleus; SN, Substantia Nigra.