Mouse anti parv

Brain sections were pre-treated with 0.3% H₂O₂ in 10 mM PBS (pH 7.4, 4°C) for 30 min. Separate series of sections were respectively incubated at 4°C for 48 h with one of the following primary antibodies: mouse anti-NeuN (1∶500, Millipore), rabbit anti-Darpp32 (1∶200, Cell Signaling, Danvers, MA), mouse anti-Parv (1∶1,000, Sigma), rabbit anti-Cr (1∶2,000, Millipore), rabbit anti-NPY (1∶5,000, ABCAM) and rabbit anti-ChAT (1∶1,000, Millipore). After rinsed in 10 mM PBS for three times (5 min/time), the sections were applied with secondary antibodies anti-mouse IgG or anti-rabbit IgG (both 1∶200, Sigma) at room temperature for 4 h, followed by three rinses (5 min/time) in 10 mM PBS and incubation with homologous peroxidase-antiperoxidase (PAP) complex (1∶200, Sigma) at room temperature for 2 h. The peroxidase reaction was performed using 3, 3′-diaminobenzidine (DAB, 0.05% in 10 mM PBS, pH 7.4, Sigma) for 2–8 min, and then the sections were mounted onto gelatin-coated slides, routinely dehydrated, cleared and covered with neutral balsam for microscopic detection.