Figure 5.

Functional NMDA receptors are localized on the surface of HPASMCs. Immunofluorescence staining was performed on non-permeabilized HPASMCs. Both GluN1 and GluN2 (2B and 2D) subunits were detected on the surface of HPASMCs (A,B). GluN1 co-localized with GluN2B (A) and GluN2D (B) on the surface of HPASMCs, respectively. Results are representative images taken from at least three separate experiments. Scale bar = 100 µM. (C,D) are representative scatter plots of each fluorescent pixel from confocal images with GluN1 green fluorescent intensity along the x-axis and GluN2 red fluorescent intensity along the y-axis. The shaded area in the upper right quadrant represents colocalized pixels above background with the associated Pearson correlation coefficient indicated for all colocalized pixels. (E) Representative traces for cultured HPASMCs in response to 100 μM NMDA and 10 μM glycine treatment. Cells were clamped at − 50 mV and whole cell recordings were performed. (F) NMDA and glycine treatment evoked an inward whole-cell current of 10.9 ± 1.7 pA (Mean ± SE, **p < 0.01, n = 16). (G) NMDA and glycine treatment evoked increased AUC compared with baseline (*p < 0.05, n = 16).