Chemical LTP enhances proliferation/survival and neuronal differentiation in NSC-neuron co-cultures.

<p>(<i>A</i>) Schematic representation of the experimental design. GFP-labeled NSCs were co-cultured with dissociated hippocampal neurons as described in the methods. (<i>B</i>) The cLTP-inducing protocol reliably induces LTP in co-cultures. Western blotting of biotinylated plasma membrane proteins revealed that in comparison with control (PBS), the cLTP-inducing treatment (cLTP) resulted in a specific increase in the level of GluA1 and GluA2 subunits of AMPARs, but not β-LRP1 (as a plasma membrane protein control), expressed on the plasma membranes 10 min after the treatment, and the increase was prevented by 50 µM NMDAR blocker APV (cLTP+APV), confirming the successful induction of NMDAR-dependent LTP. ***p<0.001, n = 7. (<i>C</i>) Representative Immunostaining images showing enhanced BrdU (top panel, red) and MAP2-positive (bottom panel, red) GFP-labeled NSCs following cLTP induction in co-cultures. (<i>D</i>) Bar graph summarizing data from multiple experiments shown in C. n = 5 and *p<0.05 for left panel; and n = 10 and ***p<0.001 for right panel. Statistical analyses were performed with One Way ANOVA. <i>Post hoc</i> testing revealed a significant difference between the cLTP group and other two groups (<i>B</i>) or three groups (<i>D</i>). Data are mean ± SEM.