Journal: bioRxiv

Article Title: Astrocytic Foxo1 regulates hippocampal spinogenesis and synaptic plasticity to enhance fear memory

doi: 10.1101/2023.05.01.538923

Figure Lengend Snippet: (A) Representative scheme depicting the bilateral stereotaxic injections of AAV5-GFAP(0.7)-mCherry-2A-m-FOXO1 virus in the hippocampal CA1 area of C57BL/6J mice. (B) Quantitative RT-PCR for the Foxo1, Itp2 and Aldh1l1 genes of CA1 hippocampal astrocytes from FOXO1+ and control mice. Data are presented as mean ± SEM. * p < 0.05. (C-F) Confocal micrographs of immuno-histochemistry in brain slices illustrating co-expression of mCherry reporter with astrocytic (S100β and GFAP), but not neuronal (NeuN) or microglia (Iba1) markers. (C) mCherry (red) reporter gene is co-expressed with GFAP (green). (D) mCherry (red) reporter gene is co-expressed with S100β (green). (E) mCherry (red) expression is absent in NeuN + neurons (green). (F) mCherry (red) expression is absent in Iba1 + microglia (green). DAPI staining, blue. Scale bars depicted in representative images.

Article Snippet: These steps were followed by the incubation with primary antibodies overnight at 4°C in a solution of 2% NGS in PBS 1x: chicken anti-mCherry (1:1000, HenBiotech, Portugal) and rabbit anti-NeuN (1:100, Cell Signaling, USA), rabbit anti-Iba1 (1:750, FUJIFILM Wako Chemicals, Hong Kong), rabbit anti-GFAP (1:200, DakoCytomation, USA), or rabbit anti-S100β (1:200, DakoCytomation, USA).

Techniques: Quantitative RT-PCR, Immunohistochemistry, Expressing, Staining

Journal: bioRxiv

Article Title: Astrocytic Foxo1 regulates hippocampal spinogenesis and synaptic plasticity to enhance fear memory

doi: 10.1101/2023.05.01.538923

Figure Lengend Snippet: (A) Confocal micrographs of immuno-histochemistry in brain slices illustrating an EGFP+ CA1 pyramidal neural dendrites in the territory of a mCherry+ astrocyte that overexpresses FOXO1. (B) Percentage of each spine type at apical dendrites (n=10-14 neurons per group). (C) The scheme shows the experimental configuration for electrical stimulation in stratum radiatum and the whole-cell recording of CA1 pyramidal neurons. (D) Relative EPSCs before and 30 min after LFS in acute hippocampal slices. (E) Average relative changes versus time during an LTD experiment with control slices (black, n=5) and slices from FOXO1+ mice (green, n=10). Time zero corresponds to the onset of the LTD protocol. (F-H) Control and FOXO1+ mice were tested in a contextual fear conditioning paradigm (n= 7-12 per group).(F) Training session depicting baseline activity (left) and freezing acquired after light-shock exposure (right) at context A. (G) Re-exposure (24 h later) to context A (left) enhanced freezing response in FOXO1+ mice. After switching to a new context, FOXO1+ mice still displayed a higher freezing response than control mice (context B, right). (H) Presentation of a light stimulus in the new context led to similar freezing responses between groups. Control mice are represented in black, and FOXO1+ mice are in green bars. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: These steps were followed by the incubation with primary antibodies overnight at 4°C in a solution of 2% NGS in PBS 1x: chicken anti-mCherry (1:1000, HenBiotech, Portugal) and rabbit anti-NeuN (1:100, Cell Signaling, USA), rabbit anti-Iba1 (1:750, FUJIFILM Wako Chemicals, Hong Kong), rabbit anti-GFAP (1:200, DakoCytomation, USA), or rabbit anti-S100β (1:200, DakoCytomation, USA).

Techniques: Immunohistochemistry, Activity Assay