Research Paper Volume 11, Issue 15 pp 5628—5645

Figure 2. Alk5i+OT treatment improves hippocampal neurogenesis and attenuates inflammation in the brains of old mice. (A) Immunofluorescence was performed on serial 25-micron brain sections with anti-Ki67 (proliferation marker) and anti-Sox-2 (neural stem cell marker), using Hoechst to stain all nuclei. A representative image of ki67 (red)/Sox2 (green)/Hoechst (blue) triple positive cells in the hippocampal Dentate Gyrus of a young control animal treated with HBSS for 7 consecutive days is shown. Scale bar=50 µm. Isotype-matched IgG negative controls exhibited minimal background fluorescence. (B) Immunofluorescence was performed on serial 25-micron brain sections with anti-Ki67 (proliferation marker), using Hoechst to stain all nuclei, imaging the cells in the SGZ of the hippocampal Dentate Gyrus. Representative images of Ki67 (red)//Hoechst (blue) positive cells in the hippocampal Dentate Gyrus. Arrows point to these double-positive cells. Scale bar=50 µm. (C) Immunofluorescence was performed on serial 25-micron brain sections with anti-CD68 (monocyte/microglia marker), using Hoechst to stain all nuclei. Representative images of CD68 (red)/Hoechst (blue) double positive cells. Scale bar=50 µm. (D) The numbers of Ki67+/Hoechst+ cells in the SGZ of DG were quantified through entire hippocampi of each cohort and were found to decline with age as expected, and to increase in the Alk5i+OT old cohort, as compared to the control vehicle-treated old cohort. Young control (Yc n=5), old control (Oc n=5), Alk5i+OT (OA5iOT n=6) *p Oc & OA5iOT = 0.043, **p Yc & OA5iOT = 0.00159, mean and SE are shown. (E) The number of CD68+ brain cells were quantified in all cohorts and were found to increase with age and to decline in Alk5i+OT-treated old muscle, as compared to the vehicle-treated old control. N young control (Yc n=5), old control (Oc n=5), Alk5i+OT (OA5iOT n=6). ***p<0.001, **p=0.0226.