Computational protocol: Interaction Effect of Social Isolation and High Dose Corticosteroid on Neurogenesis and Emotional Behavior

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Protocol publication

[…] Sholl analysis was applied to investigate the dendritic complexity (Wang et al., ). Ten DCX-ir cells with tertiary dendrites in DG were selected randomly in each animal and they were observed under 400× magnification. Microphotographs were taken and imported into ImageJ software (National Institutes of Health, Bethesda, MD, USA) with Sholl analysis plug-in (The Ghosh Lab, UCSD, La Jolla, CA, USA). Concentric circles were drawn on the neurons in every 10 μm radii increment and up to 200 μm away from soma when the neurons were traced in ImageJ. Dendritic complexity is assessed based on the numbers of intersections formed by the dendrites and concentric circles. A higher number of intersections indicate a more complex dendritic structure. [...] All statistical analyses were conducted with IBM SPSS Statistics version 23 (IBM Corporation, CA, USA). The dataset was firstly analyzed by non-parametric Kruskal-Wallis followed by Dunn-Bonferroni post hoc test for pairwise comparison. The physiological, behavioral and cell quantifications were presented in box-plot graphs which the median is used for the central tendency and with the interquartile range of upper 75% and lower 25% percentiles. Repeated measures ANOVA was used to evaluate the changes in dendritic complexity from 10 to 200 μm away from soma of immature neurons in each group. Based on the prior knowledge that the branches of dendrites grow more extensively starting from 50 ± 10 μm away from soma (Po et al., ), a two-stage analysis (Tabachnick and Fidell, ) was employed to locate the area where between-group difference in dendritic complexity occurred. The level of significance was set at p < 0.05. The sampling distribution of each variable was expressed as mean ± standard error of mean (SEM). […]

Pipeline specifications

Software tools Sholl analysis, ImageJ, SPSS
Applications Miscellaneous, Microscopic phenotype analysis
Organisms Homo sapiens, Rattus norvegicus
Chemicals Bromodeoxyuridine, Corticosterone