(A) Images of dendrites from EGFP-expressing hippocampal neurons at 7–10 DIV before and after the addition of myristoylated PKI 14–22 (20 μM), Rp-cAMPS (5 μM), or KN-93 (30 μM) at t = 0 (black arrow). Yellow arrows indicate new spines. (B) Inhibition of PKA with PKI 14–22 (dark gray bar; 58 spines, 7 cells; p = 0.9) or Rp-cAMPS (light gray bar; 66 spines, 7 cells; p = 0.4) did not alter rates of spine outgrowth relative to vehicle-treated controls (black bar; 54 spines, 7 cells). In contrast, inhibition of CaMKs with KN-93 decreased spine outgrowth (light green bar; 40 spines, 7 cells; p < 0.001) but did not further decrease spine outgrowth in cells transfected with Rpt6-S120A (dark green bar; 34 spines, 7 cells; p = 0.6). (C) Images of dendrites from EGFP-expressing WT or GluN2B L1298A/R1300Q knockin (GluN2B KI) mouse neurons at 8–11 DIV, before and after treatment with vehicle, bicuculline (30 μM), or lactacystin (10 μM) at t = 0 (black arrow). Yellow arrows indicate new spines. (D) In neurons from WT mice, treatment with bicuculline increased spine outgrowth (solid blue bar; 76 spines, 5 cells; p = 0.01) relative to vehicle-treated controls (solid black bar; 40 spines, 4 cells), while lactacystin decreased spine outgrowth (solid green bar; 16 spines, 5 cells) relative to controls (solid black bar; 54 spines, 5 cells). In contrast, in neurons from GluN2B KI mice, neither bicuculline (open blue bar; 41 spines, 5 cells; p = 0.6) nor lactacystin (open green bar; 42 spines, 5 cells; p = 0.8) altered spine outgrowth relative to vehicle-treated controls (open black bar; 45 spines, 5 cells). Error bars represent SEM.