Fig. 4: CRHR1 antagonists rescue autophagy-related hippocampal synaptic loss and memory impairment after thoracic trauma.

a Comparison of the number of excitatory synapses in the hippocampal CA1 and CA3 regions of young and old (empty columns) mice (Shank2 and Vglut1 were used as post- and presynaptic markers). b CRH expression (green, scale bar = 20 μm, magnification scale bar = 5 μm) in the hippocampus of aged mice. c IHC for Shank2 (green) and Vglut1 (red) and colocalization analysis in the CA1 and CA3 regions of the treated groups (see internal legend) (scale bar = 15 µm, magnification scale bar = 1 µm). Arrowheads indicate synaptic colocalization. d Immunoblot of BDNF in the P2 hippocampal fraction and the relative quantification of the treatment groups. e Schematic of (left to right) the behavioral experiment, analysis of memory and learning before trauma, and analysis of auditory and contextual memory 5 and 18 days after TxT. f IHC for p62 in young (left) and old (right) mice in the hippocampal CA1 and CA3 regions (scale bar = 5 μm). g Immunoblot of Beclin-1 and LC3 in hippocampal homogenates from young and old mice 5 days after trauma with or without CRHR1 blockage. N = 3–6; the error bars represent the SEMs; one-way ANOVA and Bonferroni’s post hoc comparison test were always performed (*P ≤ 0.05, **P ≤ 0.005, ***P ≤ 0.0005, ****P ≤ 0.0001).