Fig. 5. Reelin and APP cooperatively regulate dendritic spine density in vitro and in vivo. (A) Primary hippocampal neurons were transfected with GFP plasmid and PLL (control) or GFP plasmid and APP shRNA on DIV 19 and treated with 1.4 μM Reelin or vehicle (Mock medium) on DIV 20. Then, dendritic spine density was measured (GFP plasmid+PLL+vehicle: n=33 dendrites from 13 neurons, GFP plasmid+PLL+Reelin: n=31 dendrites from 14 neurons, GFP plasmid+APP shRNA+Vehicle: n=34 dendrites from 14 neurons, GFP plasmid+APP shRNA+Reelin: n=19 dendrites from 9 neurons). Scale bar=10 μm. (B, C) Three-month-old APP KO mice received bilateral intraventricular injections of 210 nM Reelin or saline. After 5 days, Golgi staining was performed, and dendritic spine density was measured in the AO and BS regions of hippocampal area CA1 (for AO, vehicle: n=18 dendrites from 18 neurons, Reelin: n=12 dendrites from 12 neurons; for BS, vehicle: n=18 dendrites from 18 neurons, Reelin: n=16 dendrites from 16 neurons, n=4 mice/group) and cortical layer II/III (for AO, vehicle: n=27 dendrites from 27 neurons, Reelin: n=29 dendrites from 29 neurons; for BS, vehicle: n=30 dendrites from 30 neurons, Reelin: n=33 dendrites from 33 neurons, n=4 mice/group). *p<0.05, **p<0.01. Scale bar=20 μm.
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