Neuropeptide content of the D1, D2 and D1/D2 MSNs was assessed on singly labeled sections for ENK or DYN, taken from the 2 colchicine-injected mice. Briefly, sections were incubated with either a mouse anti-ENK (1/50, NOC1; Medicorp, Montreal, QC, Canada) or a rabbit anti-DYN (1/200, catalog no. SP1215; Acris, San Diego, CA, USA) antibody overnight at 4 oC. Sections were then incubated with either a horse anti-mouse (1/200, catalog no. BA-2000; Vector Laboratories) biotinylated secondary antibodies for 2 h at RT, followed by streptavidin-Cy5 (1/200, catalog no. SA1011; Invitrogen, Carlsbad, CA, USA) for 1 h at RT in the case of ENK, or directly with a goat anti-rabbit-Cy5 (1/200, catalog no. 111-175-003; Jackson ImmunoResearch) for 2 h at RT in the case of DYN. Images were acquired using a confocal microscope (Zeiss, LSM 700; Oberkochen, Germany).
