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. Author manuscript; available in PMC: 2010 Apr 1.
Published in final edited form as: Nat Neurosci. 2009 Sep 6;12(10):1275–1284. doi: 10.1038/nn.2386

Figure 5.

Figure 5

Dopamine D1/D5-like receptors modulate Rap activity, spine morphology, and GluR2 surface expression. (a) Rap activation by SKF-38393 (20 µM, 30 min) in cortical neurons. Fold Rap activation compared to control: 1.62±08 fold increase, *P<0.05, n = 4 experiments. (b) Effect of SKF-38393 (20 µM, 30 min) on B-Raf phosphorylation in situ in dendrites. B-Raf immunofluorescence (a.u.): control, 68.52±4.6; SKF-38393, 141.55±15.2, *P<0.001, n = 6–9 cells per condition, 2–3 experiments. (c) Effect of SKF-38393 (20 µM, 30 min) on spine morphology and surface GluR2 in spines in cortical neurons (div 28). Epac2 knockdown prevents SKF-38393-dependent spine remodeling and AMPAR removal. GluR2 was detected using an antibody to its extracellular N-terminus (GluR2-n) in non-permeabilized cells. (arrowhead, clusters in spines; open arrowhead, shafts) (d) Quantification of spine areas in e; area (µm2): control, 0.74±0.02; SKF-38393, 0.59±0.01, Epac2 RNAi, 0.86±0.03; Epac RNAi+SKF-38393, 0.90±0.04, *P<0.001, n = 308–426 spines from 9–13 cells per condition, 4 experiments. (e) Quantification of surface GluR2 (GluR2-n) clusters in e; GluR2-n immunofluorescence (a.u.): control, 60.6±3.53; SKF-38393, 36.9±1.65; Epac2 RNAi, 67.7±4.48; Epac2 RNAi+SKF-38393, 59.6±3.94, *P < 0.001, n = 9–13 cells per condition, 4 experiments. Error bars: s.e.m. Scale bars: 5µm.