phospho thr 34 darpp 32  (Cell Signaling Technology Inc)

Journal: The European journal of neuroscience

Article Title: Dopamine depletion alters phosphorylation of striatal proteins in a model of Parkinsonism

doi: 10.1111/j.1460-9568.2005.04190.x

Figure Lengend Snippet: Effects of 6-OHDA lesion on striatal protein phosphatases. (A and B) Representative immunoblots and summary graphs quantifying total striatal levels of protein phosphatase catalytic subunits and PP1 targeting and regulatory proteins. Phosphorylation of DARPP-32 at Thr75 was significantly elevated in dopamine-depleted striatal samples (F2,45 = 3.5, P = 0.038). (C) The increase in Thr75 phosphorylation of DARPP-32 was reversed by L-DOPA administration (F1,27 = 22.13, P = 0.0005).

Article Snippet: The following primary antibodies were used for immunoblotting: goat anti-CaMKII α / β ( McNeill & Colbran, 1995 ; 1 : 4000), mouse anti-CaMKIIα (ABR, 1 : 4000), rabbit anti-phospho-Thr 286 -CaMKIIα (Promega, 1 : 2500), rabbit anti-DARPP-32 (Cell Signalling, 1 : 4000), rabbit anti-phospho-Thr 34 -DARPP-32 (Cell Signalling, 1 : 250), rabbit anti-phospho-Thr 75 -DARPP-32 (Cell Signalling, 1 : 500), rabbit anti-GluR1 (Upstate, 1 : 4000), rabbit anti-phospho-Ser 831 -GluR1 (Upstate, 1 : 500), rabbit anti-phospho-Ser 845 -GluR1 (Upstate, 1 : 2000), rabbit anti-neurabin (1 : 2500) ( MacMillan et al ., 1999 ), mouse anti-NR1 (Chemicon, 1 : 3000), rabbit or mouse anti-NR2B (Molecular Probes, 1 : 500), sheep anti-PP1γ1 (1 : 1000; Colbran et al ., 2003 ), mouse anti-PSD-95 (Upstate, 1 : 1000), rabbit anti-spinophilin (1 : 2000; MacMillan et al ., 1999 ) and mouse anti-tyrosine hydroxylase (ImmunoStar, 1 : 1000).

Techniques: Western Blot

Journal: The European journal of neuroscience

Article Title: Dopamine depletion alters phosphorylation of striatal proteins in a model of Parkinsonism

doi: 10.1111/j.1460-9568.2005.04190.x

Figure Lengend Snippet: Changes in PP1γ1 and PP1-regulatory proteins during normal ageing. Quantification of protein levels in dorsolateral striatal homogenates from normal rats at 4–6, 12–14 or 21–23 months of age. PP1γ1 was elevated only at 21–23 months (F2,47 = 12.02, P < 0.0001), DARPP-32 was decreased only at 12–14 months (F2,45 = 7.98, P = 0.001), while both spinophilin (F2,45 = 7.77, P = 0.001) and neurabin (F2,43 = 10.46, P = 0.0002) were decreased at 21–23 months.

Article Snippet: The following primary antibodies were used for immunoblotting: goat anti-CaMKII α / β ( McNeill & Colbran, 1995 ; 1 : 4000), mouse anti-CaMKIIα (ABR, 1 : 4000), rabbit anti-phospho-Thr 286 -CaMKIIα (Promega, 1 : 2500), rabbit anti-DARPP-32 (Cell Signalling, 1 : 4000), rabbit anti-phospho-Thr 34 -DARPP-32 (Cell Signalling, 1 : 250), rabbit anti-phospho-Thr 75 -DARPP-32 (Cell Signalling, 1 : 500), rabbit anti-GluR1 (Upstate, 1 : 4000), rabbit anti-phospho-Ser 831 -GluR1 (Upstate, 1 : 500), rabbit anti-phospho-Ser 845 -GluR1 (Upstate, 1 : 2000), rabbit anti-neurabin (1 : 2500) ( MacMillan et al ., 1999 ), mouse anti-NR1 (Chemicon, 1 : 3000), rabbit or mouse anti-NR2B (Molecular Probes, 1 : 500), sheep anti-PP1γ1 (1 : 1000; Colbran et al ., 2003 ), mouse anti-PSD-95 (Upstate, 1 : 1000), rabbit anti-spinophilin (1 : 2000; MacMillan et al ., 1999 ) and mouse anti-tyrosine hydroxylase (ImmunoStar, 1 : 1000).

Techniques:

Journal: The European journal of neuroscience

Article Title: Dopamine depletion alters phosphorylation of striatal proteins in a model of Parkinsonism

doi: 10.1111/j.1460-9568.2005.04190.x

Figure Lengend Snippet: Effects of chronic dopamine depletion on striatal proteins. (A–C) Long-term dopamine depletion caused an enduring decrease in TH at 9–11 months (t12 = 6.82, P = 0.0001) and at 18–20 months (t5 = 7.99, P = 0.0005). This was paralleled by an enduring increase in phosphorylation of both CaMKIIα at Thr286 at 9–11 months (t11 = 2.28, P = 0.043) and 18–20 months (t5 = 3.50, P = 0.017) and of DARPP-32 at Thr75 at 9–11 months (t6 = 5.03, P = 0.0024) and at 18–20 months (t5 = 3.46, P = 0.018).

Article Snippet: The following primary antibodies were used for immunoblotting: goat anti-CaMKII α / β ( McNeill & Colbran, 1995 ; 1 : 4000), mouse anti-CaMKIIα (ABR, 1 : 4000), rabbit anti-phospho-Thr 286 -CaMKIIα (Promega, 1 : 2500), rabbit anti-DARPP-32 (Cell Signalling, 1 : 4000), rabbit anti-phospho-Thr 34 -DARPP-32 (Cell Signalling, 1 : 250), rabbit anti-phospho-Thr 75 -DARPP-32 (Cell Signalling, 1 : 500), rabbit anti-GluR1 (Upstate, 1 : 4000), rabbit anti-phospho-Ser 831 -GluR1 (Upstate, 1 : 500), rabbit anti-phospho-Ser 845 -GluR1 (Upstate, 1 : 2000), rabbit anti-neurabin (1 : 2500) ( MacMillan et al ., 1999 ), mouse anti-NR1 (Chemicon, 1 : 3000), rabbit or mouse anti-NR2B (Molecular Probes, 1 : 500), sheep anti-PP1γ1 (1 : 1000; Colbran et al ., 2003 ), mouse anti-PSD-95 (Upstate, 1 : 1000), rabbit anti-spinophilin (1 : 2000; MacMillan et al ., 1999 ) and mouse anti-tyrosine hydroxylase (ImmunoStar, 1 : 1000).

Techniques:

Journal: The European journal of neuroscience

Article Title: Changes in Nucleus Accumbens and Neostriatal c-Fos and DARPP-32 Immunoreactivity During Different Stages of Food-Reinforced Instrumental Training

doi: 10.1111/j.1460-9568.2012.08036.x

Figure Lengend Snippet: Table 2

Article Snippet: Free-floating sections were permeabilized with 0.1% Triton X-100 in PBS for 10 min, blocked in 5% donkey serum in PBS/0.1% Triton X-100 for 45 min, and incubated with the following primary antibodies: rabbit anti-cFos polyclonal antisera (1:2000; Calbiochem, Germany); mouse monoclonal antiserum Leu5-enkephalin (1:400; Clone NOC1, Chemicon, Millipore Corp, Billerica, MA, USA); rabbit anti-DARPP-32 (Thr 34 ) (1:500; Santa Cruz Biotechnology, CA, USA).

Techniques:

Journal: The European journal of neuroscience

Article Title: Changes in Nucleus Accumbens and Neostriatal c-Fos and DARPP-32 Immunoreactivity During Different Stages of Food-Reinforced Instrumental Training

doi: 10.1111/j.1460-9568.2012.08036.x

Figure Lengend Snippet: Table 2

Article Snippet: The sections were next incubated for 24 hr in a cocktail of an affinity-purified rabbit anti-cFos polyclonal antisera (1:2000; Calbiochem, Germany), goat anti-substance P polyclonal antisera (1:400; Santa Cruz Biotechnology, Inc, Santa Cruz, CA, USA), Inc) (used to label striatonigral neurons), rabbit anti-DARPP-32 (Thr 34 ) polyclonal antisera (1:500; Santa Cruz Biotechnology, Inc), goat anti-substance P polyclonal antisera (1:400; Santa Cruz Biotechnology, Inc), rabbit anti-cFos polyclonal antisera (1:2000; Calbiochem, Germany), and an affinity-purified goat anti-DARPP-32 (Thr 34 ) polyclonal antisera (1:500; Santa Cruz Biotechnology, CA, USA) containing 5% NDS and 0.1 % Triton X-100 in 0.1 M phosphate buffer, pH 7.4 over night at 4° C with gentle agitation on a rotating shaker.

Techniques: