Journal: Frontiers in Cellular Neuroscience
Article Title: Dopamine signaling negatively regulates striatal phosphorylation of Cdk5 at tyrosine 15 in mice
doi: 10.3389/fncel.2013.00012
Figure Lengend Snippet: Effects of L-dopa on striatal phosphorylation of Cdk5 and DARPP-32 in MPTP mice. Saline- or MPTP-treated mice received vehicle or L-dopa 30 min before sacrifice. Striatal tissue extracts and sections were prepared and subjected to western blotting and immunohistochemical staining (see “Materials and Methods”). (A) Western blot analysis of striatal levels of Cdk5-pTyr15 and Cdk5 in mice treated with saline + vehicle, saline + L-dopa, MPTP + vehicle, or MPTP + L-dopa. Values are means ± S.E.M. ( n = 5–6). * P < 0.001 versus mice treated with saline + vehicle, saline + L-dopa, or MPTP + L-dopa; One-Way ANOVA [ F (3.18) = 31.06] followed by Bonferroni–Dunn test. (B) Immunohistochemical study on striatal labeling for Cdk5-pTyr15 in mice treated with saline + vehicle, saline + L-dopa, MPTP + vehicle, or MPTP + L-dopa. Representative images of the striatal sections (0.0–1.0 mm anterior to bregma) stained for Cdk5-pTyr15 from each group are shown. Densitometric analysis was made on a striatal section from each mouse in a group of saline + vehicle, saline + L-dopa, MPTP + vehicle or MPTP + L-dopa. Values are means ± S.E.M. ( n = 5). * P < 0.005 versus mice treated with saline + vehicle, saline + L-dopa, or MPTP + L-dopa; One-Way ANOVA [ F (3.16) = 26.84] followed by Fisher's PLSD test. (C) Western blot analysis of striatal levels of DARPP-32-pThr75, DARPP-32-pThr34, and DARPP-32 in mice treated with saline + vehicle, saline + L-dopa, MPTP + vehicle, or MPTP + L-dopa. Values are means ± S.E.M. ( n = 4–7). * P < 0.01 versus mice treated with saline + vehicle, saline + L-dopa, or MPTP + L-dopa; One-Way ANOVA [ F (3.18) = 6.96] followed by Bonferroni–Dunn test. (D) Immunohistochemical study on striatal labeling for DARPP-32-pThr75 in mice treated with saline + vehicle ( n = 5), saline + L-dopa ( n = 5), MPTP + vehicle ( n = 5), or MPTP + L-dopa ( n = 5). Representative images of the striatal sections (0.0–1.0 mm anterior to bregma) stained for DARPP-32-pThr75 from each group are shown. Densitometric analyses were made on a striatal section from each mouse in a group of saline + vehicle, saline + L-dopa, MPTP + vehicle or MPTP + L-dopa. Values are means ± S.E.M ( n = 5). * P < 0.005 versus mice treated with saline + vehicle, saline + L-dopa, or MPTP + L-dopa; One-Way ANOVA [ F (3.16) = 19.62] followed by Fisher's PLSD test.
Article Snippet: Primary antibodies against Cdk5-pTyr15 (1:20,000; Santa Cruz), tyrosine hydroxylase (TH, 1:100,000) (Sato et al., ), and DARPP-32-Thr75 (1:20,000; Cell Signaling) (Sako et al., ) were used.
Techniques: Phospho-proteomics, Saline, Western Blot, Immunohistochemical staining, Staining, Labeling