Journal: Psychopharmacology
Article Title: Energizing effects of bupropion on effortful behaviors in mice under positive and negative test conditions: modulation of DARPP-32 phosphorylation patterns
doi: 10.1007/s00213-021-05950-4
Figure Lengend Snippet: Effect of treatment (Veh-Veh, TBZ-Veh, Veh-BUP, or TBZ-BUP) on number of cell counts positive for pDARPP-32-(Thr34) and pDARPP-32(Thr75) immunoreactivity on Nacb core. Upper left: Diagram showing the effect of TBZ on DARPP-32 phosphorylation in D2-receptor containing cells, and coronal section with bregma coordinates showing location of the brain area for immunohistochemistry taken from Paxino and Franklin’s mouse atlas. TBZ, which depletes DA, increases D2-receptor stimulation by increasing cyclic adenosine monophosphate (c-AMP) production and protein kinase A (PKA) activity yielding to the increase of the dephosphorylation of pDARPP-32(Thr34) and no change or a decrease in the phosphorylation of pDARPP-32(Thr75) by blocking the protein phosphatase (PP1) (based on results from Bateup et al. ; Nunes et al. ; López-Cruz et al. ). Bottom left: photomicrographs of individual representative brain sections. Right: mean ± S.E.M of number of pDARPP-32-(Thr34) and pDARPP-32-(Thr75) counts in a 100um 2 ROI. * p < 0.05, ** p < 0.01 significantly different from Veh-Veh. ## p < 0.01 significant differences from TBZ-Veh
Article Snippet: Equal amounts (30ug) of striatum protein samples were separated by 12.5% SDS-PAGE and transferred to nitrocellulose membrane (Bio-rad) for 90 min at 30 V. Filtering membranes were incubated in 5% of bovine serum albumin (BSA, Sigma-Aldrich) dissolved in TBS-T (Tris Base 200 Mm and 5 M NaCl) containing 0.1% Tween 20, and then incubated overnight at 4 °C with polyclonal rabbit anti-DARPP32-Thr75 antibody (1:500, cell signaling).
Techniques: Phospho-proteomics, Immunohistochemistry, Activity Assay, De-Phosphorylation Assay, Blocking Assay
