Vector sg grey

For detailed methods, see Additional file 1. In brief, paraffin-embedded tissue blocks of the LC were cut into 4 × 20 µm-thick consecutive sections, and stained with monoclonal rabbit anti-dopamine-beta hydroxylase (DBH, dilution 1:400, Abcam, Cambridge, UK), rabbit anti-phosphorylated-Ser129 α-syn (clone EP1536Y, dilution 1:4000, Abcam), mouse anti-p-tau (clone AT8, dilution 1:800, ThermoFisher, Pittsburgh, PA) or mouse anti-Aβ (clone 4G8, dilution 1:5000, BioLegend, San Diego, CA) antibody. The paraffin-embedded cortical tissue blocks, the ACC, DLPFC, M1 and hippocampus, were cut at 6 µm and processed for immunohistochemistry with rabbit anti-DBH (dilution 1:400, Abcam). The DLPFC sections were stained with an alternative DBH antibody for validation (dilution 1:100, Novus, Cambridge, UK, Additional file 1). The primary antibodies were incubated at 4°C overnight and visualized with Vector SG grey (Vector, Newark, CA), 3,3′-diaminovenzidine (DAB, Sigma-Aldrich, Darmstadt, Germany), or DAB + Nickel, followed by counter-staining with nuclear fast red (Vector). The sections were dehydrated in a series of ethanol, xylene, and then mounted with Entellan.

In brief, paraffin-embedded tissue blocks of the SN were cut into 6 μm-thick consecutive sections with a microtome and incubated with mouse-anti tyrosine hydroxylase (TH, dilution 1:800 in TBS-Trition 0.1%, Immunostar, Hudson, USA) at 4°C overnight. TH immunoreactivity was visualized with Vector SG grey (Vector, California, United State) followed by counter-staining with fast red (Vector, California, United State), and dehydrated in a series of ethanol, xylene, then mounted with Entellan.