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. 2020 Nov 25;25(23):5508. doi: 10.3390/molecules25235508

Figure 4.

Figure 4

Effect of okadaic acid and melatonin on the number of neurons in hilar hippocampus. Rat hippocampal organotypic cultures (400 µm) were treated at the 8th day for 6 or 8 h; afterwards, they were fixed as described in methods section and 50 µm cryosections were processed for MAP2 staining by immunohistochemistry. Representative images of only three treatments are shown in the figure: (a) vehicle (VEH), (b) okadaic acid (OKA), and (c) simultaneous incubation with okadaic acid and melatonin (VEH = MEL). As MAP2 is distributed in the somatodendritic domain, the staining of this protein allowed us to detect neuronal somas shown with red circles and arrows. Fields were acquired randomly by treatment (4 fields/slice and 5 slices/treatment) and the total number of MAP2(+) cells at the hilus was counted and plotted in panel (d). Data represent the mean ± SEM and statistical analysis was performed with ANOVA on Ranks before Student-Newman-Keuls test for multiple comparisons. * represents p < 0.05. Scale bar in (b): 100 µm.