Journal: Neuropsychiatric Disease and Treatment

Article Title: The abnormal activation of D1R/Shp-2 complex involved in levodopa-induced dyskinesia in 6-hydroxydopamine-lesioned Parkinson’s rats

doi: 10.2147/NDT.S162562

Figure Lengend Snippet: PD rats show different AIMs scores after the administration of l -DOPA, D1R antagonist or agonist. Notes: Axial ( A ), limb ( B ), orolingual ( C ) and locomotor ( D ). AIMs scores were observed at 2, 7, 12, 17 and 22 days. Increased AIMs scores in PD rats were observed after the administration of l -DOPA. Meanwhile, the group treated with l -DOPA and D1R agonist showed increasing AIMs scores; however, rats treated with D1R antagonist, SCH23390, plus l -DOPA showed no increasing AIMs scores compared to rats peritoneally injected with l -DOPA alone. Data are presented as mean ± SD. # p < 0.05, versus day 2; * p < 0.05, versus l -DOPA group. Data are statistically analyzed by one-way ANOVA test. Abbreviations: AIMs, abnormal involuntary movements; ANOVA, analysis of variance; d, days; D1R, D1 dopamine receptor; l -DOPA, L-3,4-dihydroxyphenylalanine; PD, Parkinson’s disease.

Article Snippet: Samples were incubated with a mouse antibody against D1R (EMD Millipore, Billerica, MA, USA) or a rabbit antibody against Shp-2 (Proteintech, Rosemont, IL, USA) overnight at 4°C.

Techniques: Injection

Journal: Neuropsychiatric Disease and Treatment

Article Title: The abnormal activation of D1R/Shp-2 complex involved in levodopa-induced dyskinesia in 6-hydroxydopamine-lesioned Parkinson’s rats

doi: 10.2147/NDT.S162562

Figure Lengend Snippet: D1R interacts with Shp-2 in the striatal neurons. Notes: Striatal proteins were coimmunoprecipitated with anti-D1R and anti-Shp-2 antibodies ( A and B ). Representative immunoblots showing D1R and Shp-2 interactions in striatal neurons as detected by coimmunoprecipitation. No precipitating antibody, an irrelevant IgG was used in L2, L3, respectively. The D1R antibody (L4A) or Shp-2 antibody (L4B) was used in L4 for normal rats and L5 for LID rats, respectively. No striatal proteins and antibodies were used in L1. Abbreviations: IP, immunoprecipitation; D1R, D1 dopamine receptor; L1, lane 1; L2, lane 2; L3, lane 3; L4, lane 4; L5, lane 5; LID, levodopa-induced dyskinesia.

Article Snippet: Samples were incubated with a mouse antibody against D1R (EMD Millipore, Billerica, MA, USA) or a rabbit antibody against Shp-2 (Proteintech, Rosemont, IL, USA) overnight at 4°C.

Techniques: Western Blot, Immunoprecipitation

Journal: Neuropsychiatric Disease and Treatment

Article Title: The abnormal activation of D1R/Shp-2 complex involved in levodopa-induced dyskinesia in 6-hydroxydopamine-lesioned Parkinson’s rats

doi: 10.2147/NDT.S162562

Figure Lengend Snippet: Molecular events underlying LID involving D1R/Shp-2 complex and its downstream signaling factors, such as ERK1/2 and mTOR. Notes: The bands represent immunoblot images detected by antibodies against p-Shp-2 ( A ), p-ERK ( B ) and p-mTOR ( C ). Proteins were analyzed from the sham group (1), l -DOPA group (2), SCH23390 + l -DOPA group (3), SKF38393 group (4). Repeated administration of l -DOPA increased the level of p-Shp-2, p-ERK1/2 and p-mTOR. SKF38393 increased the levels similarly. Conversely, SCH23390 plus L-DOPA prevented the increase. Data are presented as mean ± SD. * p < 0.05, versus sham group; # p < 0.05, versus l -DOPA group. Data are statistically analyzed by one-way ANOVA test. Abbreviations: ANOVA, analysis of variance; D1R, D1 dopamine receptor; ERK1/2, extracellular signal-regulated kinases 1 and 2; l -DOPA, l -3,4-dihydroxyphenylalanine; LID, levodopa-induced dyskinesia.

Article Snippet: Samples were incubated with a mouse antibody against D1R (EMD Millipore, Billerica, MA, USA) or a rabbit antibody against Shp-2 (Proteintech, Rosemont, IL, USA) overnight at 4°C.

Techniques: Western Blot

Journal: Neuropsychiatric Disease and Treatment

Article Title: The abnormal activation of D1R/Shp-2 complex involved in levodopa-induced dyskinesia in 6-hydroxydopamine-lesioned Parkinson’s rats

doi: 10.2147/NDT.S162562

Figure Lengend Snippet: PD rats show different AIMs scores after the administration of l -DOPA, D1R antagonist or agonist. Notes: Axial ( A ), limb ( B ), orolingual ( C ) and locomotor ( D ). AIMs scores were observed at 2, 7, 12, 17 and 22 days. Increased AIMs scores in PD rats were observed after the administration of l -DOPA. Meanwhile, the group treated with l -DOPA and D1R agonist showed increasing AIMs scores; however, rats treated with D1R antagonist, SCH23390, plus l -DOPA showed no increasing AIMs scores compared to rats peritoneally injected with l -DOPA alone. Data are presented as mean ± SD. # p < 0.05, versus day 2; * p < 0.05, versus l -DOPA group. Data are statistically analyzed by one-way ANOVA test. Abbreviations: AIMs, abnormal involuntary movements; ANOVA, analysis of variance; d, days; D1R, D1 dopamine receptor; l -DOPA, L-3,4-dihydroxyphenylalanine; PD, Parkinson’s disease.

Article Snippet: Membranes were blocked in 5% nonfat milk for 1 hour at room temperature and incubated with a mouse primary antibody against D1R (EMD Millipore) or a rabbit primary antibody against Shp-2 (Proteintech) overnight at 4°C.

Techniques: Injection

Journal: Neuropsychiatric Disease and Treatment

Article Title: The abnormal activation of D1R/Shp-2 complex involved in levodopa-induced dyskinesia in 6-hydroxydopamine-lesioned Parkinson’s rats

doi: 10.2147/NDT.S162562

Figure Lengend Snippet: D1R interacts with Shp-2 in the striatal neurons. Notes: Striatal proteins were coimmunoprecipitated with anti-D1R and anti-Shp-2 antibodies ( A and B ). Representative immunoblots showing D1R and Shp-2 interactions in striatal neurons as detected by coimmunoprecipitation. No precipitating antibody, an irrelevant IgG was used in L2, L3, respectively. The D1R antibody (L4A) or Shp-2 antibody (L4B) was used in L4 for normal rats and L5 for LID rats, respectively. No striatal proteins and antibodies were used in L1. Abbreviations: IP, immunoprecipitation; D1R, D1 dopamine receptor; L1, lane 1; L2, lane 2; L3, lane 3; L4, lane 4; L5, lane 5; LID, levodopa-induced dyskinesia.

Article Snippet: Membranes were blocked in 5% nonfat milk for 1 hour at room temperature and incubated with a mouse primary antibody against D1R (EMD Millipore) or a rabbit primary antibody against Shp-2 (Proteintech) overnight at 4°C.

Techniques: Western Blot, Immunoprecipitation

Journal: Neuropsychiatric Disease and Treatment

Article Title: The abnormal activation of D1R/Shp-2 complex involved in levodopa-induced dyskinesia in 6-hydroxydopamine-lesioned Parkinson’s rats

doi: 10.2147/NDT.S162562

Figure Lengend Snippet: Molecular events underlying LID involving D1R/Shp-2 complex and its downstream signaling factors, such as ERK1/2 and mTOR. Notes: The bands represent immunoblot images detected by antibodies against p-Shp-2 ( A ), p-ERK ( B ) and p-mTOR ( C ). Proteins were analyzed from the sham group (1), l -DOPA group (2), SCH23390 + l -DOPA group (3), SKF38393 group (4). Repeated administration of l -DOPA increased the level of p-Shp-2, p-ERK1/2 and p-mTOR. SKF38393 increased the levels similarly. Conversely, SCH23390 plus L-DOPA prevented the increase. Data are presented as mean ± SD. * p < 0.05, versus sham group; # p < 0.05, versus l -DOPA group. Data are statistically analyzed by one-way ANOVA test. Abbreviations: ANOVA, analysis of variance; D1R, D1 dopamine receptor; ERK1/2, extracellular signal-regulated kinases 1 and 2; l -DOPA, l -3,4-dihydroxyphenylalanine; LID, levodopa-induced dyskinesia.

Article Snippet: Membranes were blocked in 5% nonfat milk for 1 hour at room temperature and incubated with a mouse primary antibody against D1R (EMD Millipore) or a rabbit primary antibody against Shp-2 (Proteintech) overnight at 4°C.

Techniques: Western Blot

Journal: PLoS Genetics

Article Title: Mutation of SLC35D3 Causes Metabolic Syndrome by Impairing Dopamine Signaling in Striatal D1 Neurons

doi: 10.1371/journal.pgen.1004124

Figure Lengend Snippet: Mice at 12 weeks of age were used in this study. ( A ) Immunohistochemical detection of D1R on the coronal brain sections of the striatum. In ros mice, visible immunoreactive cell bodies are present (arrows). Scale bar: 100 µm. ( B, C ) Representative immuno-EM pictures of D1R-labeled particles on plasma membrane (PM, upper panels) and endomembrane structures (EnM, lower panels) in wild-type (WT) and ros striatal neurons are shown without much difference (B). However, the quantification test revealed that the particles on PM in ros mice (34.0%, n = 103) is significantly lower than that in WT (51.7%, n = 87), ** P< 0.01. Scale bar: 200 nm. ( D ) SKF82958–induced cAMP accumulation in striatal membranes prepared from WT and ros mice. As compared with the WT (11.09±0.61 pmol/ml, n = 6), cAMP activity in ros membranes (7.11±0.45 pmol/ml, n = 6) was reduced about 36%. * P <0.001. ( E ) Immunoblot analysis of D1 receptor. left panel , Striatum tissues (20 µg) from WT and ros mice were probed with the monoclonal D1R antibody. β-actin was used as a loading control. The immunoblots shown are representative of three independent experiments. right panel , Normalized percentages of the band intensities shown in left panel are means ± SEM (n = 3). There is no significant difference of total D1R levels between WT and ros mice ( P >0.05).

Article Snippet: Mouse monoclonal antibody against D1 receptor (Chemicon, Temecula, CA, USA) was used for WB (1∶600) and ICC (1∶300).

Techniques: Immunohistochemical staining, Labeling, Activity Assay, Western Blot