Fig. 11

CA140 promotes dendritic spine formation through DRD1/CaMKII/ERK signaling in primary hippocampal neurons (PHNs). A, B DRD1 levels in GFP-transfected PHNs treated with vehicle (1% DMSO) or CA140 (5 μM) for 24 h were measured by immunostaining with anti-DRD1 antibodies (Veh, n = 152; CA140, n = 155). C, D Dendritic spine number in GFP-transfected PHNs pretreated with DRD1 inhibitor (LE300, 10 μM) or vehicle (1% DMSO) for 1 h and treated with CA140 (5 μM) or vehicle (1% DMSO) for 23 h (Veh, n = 79; CA140, n = 54; LE300, n = 72; LE300 + CA140, n = 62). E–H Immunostaining of p-CaMKIIα or p-ERK in GFP-transfected PHNs treated with CA140 (5 μM) or vehicle (1% DMSO) for 24 h (p-CaMKIIα: Veh, n = 93; CA140, n = 98; p-ERK: Veh, n = 29; CA140, n = 28). I–L Immunostaining of p-CaMKIIα or p-ERK in GFP-transfected PHNs treated with LE300 (10 μM) or vehicle (1% DMSO) for 24 h (p-CaMKIIα: Veh, n = 99; LE300, n = 85; p-ERK: Veh, n = 78; LE300, n = 93). Scale bar = 10 μm. M–P GFP-transfected PHNs were pretreated with vehicle (1% DMSO), CaMKIIα inhibitor (10 μM), or ERK inhibitor (PD98059) for 1 h and treated with CA140 (5 μM) or vehicle (1% DMSO) for 23 h. Then, dendritic spine number was measured (KN93: Veh, n = 37; CA140, n = 41; KN93 + Veh, n = 36; KN93 + CA140, n = 33; PD98059: Veh, n = 49; CA140, n = 45; PD98059 + Veh, n = 24; PD98059 + CA140, n = 34). Scale bar = 20 μm. *p < 0.05, **p < 0.01, ***p < 0.001