rmMFG-E8 promotes neural stem cell migration via netrin-1.

(A) rmMFG-E8 upregulates netrin-1 via integrin αV. Pre-treatment of mouse embryonic neural stem cells with 1 mg/ml anti-Itgav antibody prevented rmMFG-E8 mediated upregulation of netrin-1, but not 1 mg/ml control IgG. Data are presented as mean ± SEM. *p<0.05 vs PBS; #p<0.05 vs IgG; n = 3/group. (B) Quantification of the mouse embryonic neural stem cell migration through the transwell showed that rmMFG-E8 promotes embryonic neural stem cell migration through netrin-1. Both rmNtn-1 and rmMFG-E8 increased cell migration compared with PBS. Pre-treatment of the mouse embryonic neural stem cells with anti-Ntn-1 antibody blocked the effects of both rmNtn-1 and rmMFG-E8 on cell migration. Treatment of the mouse embryonic neural stem cells with a combination of rmNtn-1 and rmMFG-E8 had an additive effect in increasing cell migration. Data are presented as mean ± SEM. *p<0.05 vs PBS; #p<0.05 vs rmNtrn1/rmMFG-E8; n = 3. (C) The effect of rhMFG-E8 on neural precursor migration in rats. Doublecortin and BrdU double immunofluorescent staining of the striatum was performed to identify migrating neuroblast (Doublecortin+BrdU+) at 7 days after tMCAO. Representative images are shown at the top of the figure. tMCAO increased the number of Doublecortin+BrdU+ cells in the striatum compared to sham baseline. rhMFG-E8 treatment significantly increased the number of Doublecortin+BrdU+ cells compared with the vehicle group. Data are presented as mean ± SEM. *p<0.05 vs sham; #p<0.05 vs vehicle; n = 3/group.