Figure 4:

Molecular characterizations of PRV- labeled RVM neurons in adult naïve rats. Representative images of 2-color immunohistochemistry (GFP-green and TpH2-magenta) analyses of bladder specific RVM neurons are shown in (a1-a3). White arrows represent GFP+ neurons expressing TpH2, and the scale bar is 100µm. Representative images of 3-color in situ hybridization (GFP-green, Gad1-red, and PENK-white) analyses of bladder specific RVM neurons are shown in (b1-b4). The white arrows represent GFP+ neurons having both GAD1 and PENK expression while the yellow arrow represents GFP+ cell with either GAD1 or PENK expression. Similarly, representative images of 3-color in situ hybridization (GFP-green, RFP-red, and VGAT-white) analyses are shown in (c1-c4). The white and yellows arrows represent converging neurons with and without VGAT expression respectively. Scale bar is 25µm.