Skip to main content
. 2022 Jan 24;11(3):390. doi: 10.3390/cells11030390

Figure 1.

Figure 1

In vivo longitudinal analysis of astrocytic markers in the dorsal hippocampal dentate gyrus (dDG) of an animal model of depression and in in vitro primary cultures. (a) Illustrative experimental timeline. (b) Hippocampal DG coronal section immunostained for bromodeoxyuridine (BrdU) (in red), glial fibrillary acidic protein (in green), and DAPI (in blue). Additionally, a hippocampal DG coronal section immunostained for BrdU (in red), GFAP (in green), S100β (in grey), and DAPI (in blue). Quantitative analysis of GFAP+ cells in the dorsal dentate gyrus (dDG) at tp1 (c) and at tp2 (g), after a six-week uCMS protocol including antidepressants (ADs) treatment, fluoxetine, and imipramine. (d,h) Quantitative analysis of the number of S100β+ cells in the dDG, both at tp1 (d) and tp2 (h). (e,i) Quantitative analysis of GFAP+BrdU+ cells, both immediately after stress exposure, tp1, (e) and 4 weeks after stress exposure, tp2 (i). (f,j) Analysis of the number of GFAP+BrdU+ cells per total number of BrdU+ cells, both at tp1 (f) and at tp2 (j). (k,m) Representative image of immunocytochemistry of hippocampal primary cells in a control plate (k), and incubated with BrdU (m), with neurons labelled with β3-tubulin, astrocytes with GFAP proliferating cells with BrdU and cell nucleus with DAPI. (l,n) In vitro analysis of hippocampal DG primary cultures of p3–5 animals, regarding the number of GFAP+ astrocytic cells (l) and astrocytes differentiation—GFAP+BrdU+ (n) after incubation of the primary cell cultures with dexamethasone (DEX), norfluoxetine (NORFLX), or desipramine (DESIP) and BrdU. * Represents uCMS effect analyzed by Student’s t-test. # Represents ADs effect, by comparison of treatment and SAL animals; δ represents differences between ADs, analyzed by one-way analysis of variance (ANOVA). Data are represented as mean ± s.e.m. Scale bars represent 50 μm.*, # p < 0.05, ## p < 0.01, δδδδ p < 0.0001; Sample size: TP1: CTRL: 5–7; CMS: 5–7; FLX: 6–8; IMIP: 4–7; TP2: CTRL: 6–8; CMS: 6–9; FLX:6–8; IMIP: 6–8. Abbreviations: GFAP, Glial Fibrillary Acidic Protein; CTRL, non-stressed animals; SAL, animals exposed to uCMS and injected with saline; IMIP, animals exposed to uCMS and treated with imipramine; FLX, animals exposed to uCMS and treated with fluoxetine; dDG, dorsal dentate gyrus; DEX, dexamethasone; NORFLX, Norfluoxetine; DESIP, Desipramine; β3tub, β3-tubulin; DAPI, 4′,6′-diamino-2-fenil-indol; BrdU, Bromodeoxyuridine; tp1, time point 1 (6 weeks; immediately after the stress protocol cessation); and tp2, time point 2 (10 weeks; 4 weeks after the stress protocol cessation).