Figure 2.

Hesperidin increases synapse formation in vivo and in vitro. Adult mice treated i.p with vehicle (control group) or 10 mg/kg of hesperidin were submitted to immunohistochemistry analysis for the pre- and post-synaptic markers, synaptophysin and drebrin, respectively (A–C) in the hippocampal CA1 area, and western blotting analysis of the levels of the synaptic proteins, synaptophysin, postsynaptic density protein 95 (PSD-95), drebrin and α-amino-3-hydroxy-5- methyl-4-isoxazolepropionic acid receptors (AMPARs) (D,E). Increased synapse formation (A–C) and levels of synaptic proteins (D,E) were observed in the hippocampus of mice treated with hesperidin. Cortical neurons treated with hesperidin were analyzed by immunocytochemistry for Cleaved Caspase 3 and β-Tubulin-III (F–H), synaptophysin and PSD-95 (I–K) and were submitted to the FM1-43 assay (L–N). Neurons treated with the flavonoid showed reduced cell death (F–H), and increased synaptogenesis (I–K,I′,J′), as well as increased pre-synaptic activity (L–N). Scale bars, 20 μm, 20 μm and 10 μm, respectively. *p < 0.050, **p < 0.010 and ***p < 0.001, n = 3–5 animals per experimental group for the in vivo experiments and n = 3–6 independent neuronal cultures for the in vitro experiments. Student’s t test.