Figure 4.

Incubation of ONPs with the triple administration schedule of melatonin/ketamine combination elicited cluster formation. Olfactory neuronal precursors were cultured with the vehicle added at time 0, 17 h and 23.5 h (V+V+V); or with VEH, VEH and KET; VEH, VEH, and MEL; MEL, MEL, and MEL; VEH, VEH, and KET/MEL or MEL, MEL, and KET/MEL combination (M+M+K/M). Panel A shows representative images of cells administered three times with the VEH (a) or with MEL+ MEL + KET/MEL combination (b). After fixation, cells were stained with DAPI (blue), and with an anti-nestin antibody (green). Single cells are noted using an arrow. Bar= 50 µm. Panel B: The graph shows the number of clusters counted in 10 fields in cultures treated as described and normalized by total cells in the field counted by DAPI staining. Each point in the graph represents the number of clusters counted in one field in the cultures incubated with the indicated treatments. Data are expressed as mean ± standard deviation obtained in ten fields. The experiment was repeated twice— and data were analyzed using a Kruskal–Wallis test followed by the Dunn´s post-test. * p = 0.007, when compared with the vehicle control group (V+V+V).