Fig. 7: Intranasal administration of EOF2 induce neuroblast migration from the neurogenic regions to the damage area.

a Scheme of BrdU administration. Experimental procedure followed to label proliferating neural precursors with BrdU exclusively in neurogenic niches and not in the injured area: mice received BrdU injections on days 6, 5 and 4 before performing the cortical injury; then we waited three more days to allow for complete withdrawal of BrdU from the animal organism. Lesion was performed on day 0. Intranasal EOF2 (5 µM) or only vehicle was administered for 14 days. b Representative confocal microscopy images of the injured cortex of adult mice after bearing cortical lesions and the intranasal administration of EOF2 or only vehicle processed for the immunohistochemical detection of the proliferation marker BrdU and the neuroblast marker doublecortin (DCX). The dotted line indicates the limit of the lesion (L) and the scale bar represent 100 µm in the low magnification pictures and 50 µm in the high magnification picture. c Graph shows the number of proliferating cells marked with BrdU per mm³ in the peri-lesional area of the indicated animal groups. Data shown are the mean ± S.E.M.; n = 6 animals per group. Statistical analysis: *p = 0.0377 in two tailed unpaired Student’s t test comparing EOF2 with the control. d Quantification of DCX⁺ cells/mm³ in the peri-lesional area of the indicated animal groups. Data shown are the mean ±S.E.M.; n = 6 animals per group. Statistical analysis: *p = 0.0012 in two tailed unpaired Student’s t test comparing EOF2 with the control. e Percentage of BrdU⁺ cells that co-expressed DCX in the peri-lesional area. Data shown are the mean ± S.E.M.; n = 6 animals per group. Statistical analysis: *p = 0.0100 in two tailed unpaired Student’s t test comparing EOF2 with the control. f Representative confocal microscopy images of the injured cortex of adult mice in the previously indicated groups processed for the immunohistochemical detection of the proliferation marker BrdU and the glial marker GFAP. The dotted line indicates the limit of the lesion (L) and the scale bar represent 100 µm in the low magnification pictures and 50 µm in the high magnification picture. g Percentage of BrdU⁺ cells that co-expressed GFAP in the peri-lesional area. Data shown are the mean ± S.E.M.; n = 6 animals per group. Statistical analysis: *p = 0.0214 in two tailed unpaired Student’s t test comparing EOF2 with the control. h Graph shows the GFAP burden within the peri-lesional area, expressed as a percentage of the total lesion area. No statistically significant differences were found. Data shown are the mean ± S.E.M.; n = 6 animals per group. Statistical analysis: p = 0.4464 in two tailed unpaired Student’s t test comparing EOF2 with the control.