antibodies against d1r  (Proteintech)

Journal: Molecular Pain

Article Title: Dopamine receptor D2, but not D1, mediates the reward circuit from the ventral tegmental area to the central amygdala, which is involved in pain relief

doi: 10.1177/17448069221145096

Figure Lengend Snippet: Pain relief increased dopaminergic D2 receptor expression in injured mice. D2 receptor expression in the CeA was significantly decreased in incised mice treated with PNB compared with incised mice treated with saline (n = 6; * p < 0.05 vs the incision+saline group; one-way ANOVA,Interaction::F (3, 20) = 6.274). The expression of D1 receptor was not significantly changed. (b) In the CeA, D1 receptor was mainly expressed in glutamatergic excitatory neurons (positive for glutaminase). In the CeA, D2 receptor was predominantly expressed in GABAergic inhibitory neurons (positive for GABA). Scale bar, 50 μm.

Article Snippet: After being blocked with 3% bovine serum albumin (BSA) (Solarbio, Beijing, China) and incubated overnight at 4°C with primary antibodies against D1R (1:1000; catalog No. MAB5290, Millipore), D2R (1:1000 catalog No. AB5084P, Millipore), and GAPDH (catalog No. RM2002 L, Beijing Ray), the membranes were washed six times for 10 min each in Tris-buffered saline and 1% Tween.

Techniques: Expressing

Journal: Molecular Pain

Article Title: Dopamine receptor D2, but not D1, mediates the reward circuit from the ventral tegmental area to the central amygdala, which is involved in pain relief

doi: 10.1177/17448069221145096

Figure Lengend Snippet:

Article Snippet: After being blocked with 3% bovine serum albumin (BSA) (Solarbio, Beijing, China) and incubated overnight at 4°C with primary antibodies against D1R (1:1000; catalog No. MAB5290, Millipore), D2R (1:1000 catalog No. AB5084P, Millipore), and GAPDH (catalog No. RM2002 L, Beijing Ray), the membranes were washed six times for 10 min each in Tris-buffered saline and 1% Tween.

Techniques:

Journal: Drug Design, Development and Therapy

Article Title: Targeting the D1-N-methyl- d -aspartate receptor complex reduces l -dopa-induced dyskinesia in 6-hydroxydopamine-lesioned Parkinson’s rats

doi: 10.2147/DDDT.S93487

Figure Lengend Snippet: Effects of intrastriatal administration of Tat-fusion interfering peptide (Tat-D1Ri) on D1R–GluN1 interactions. Notes: Tat-D1Ri, Tat-D1Rc, or saline were locally injected into the striatum of the normal adult rat at a rate of 0.2 µL/min. Rat striatal tissues were then used for coimmunoprecipitation experiments to validate the efficacy and selectivity of interfering peptide. The intrastriatal injection of Tat-D1Ri rather than Tat-D1Rc caused reduction of D1R–GluN1 interactions, which demonstrated the effectiveness of Tat-D1Ri. Abbreviations: Tat-D1Rc, Tat-fusion control peptide; Ig, Immunoglobulin; IP, immunoprecipitation; IB, immunoblot.

Article Snippet: Samples were incubated with a rabbit antibody against D1R (EMD Millipore, Billerica, MA, USA) or a mouse antibody against GluN1 (Millipore) overnight at 4°C.

Techniques: Injection, Immunoprecipitation, Western Blot

Journal: Drug Design, Development and Therapy

Article Title: Targeting the D1-N-methyl- d -aspartate receptor complex reduces l -dopa-induced dyskinesia in 6-hydroxydopamine-lesioned Parkinson’s rats

doi: 10.2147/DDDT.S93487

Figure Lengend Snippet: Effects of intrastriatal administration of Tat-D1Ri on membrane D1R subunit expression. Notes: Protein levels were evaluated by Western blotting of proteins extracted from the lesioned striatum of the rat brains. They were assessed in extracts from 6-OHDA-lesioned rats treated with pulsatile l -dopa plus intrastriatal administration of Tat-D1Rc or Tat-D1Ri. ( A ) Representative Western blot analysis of D1R in the membrane fraction; ( B ) densitometric analysis of two blots with specific protein signals normalized to the corresponding GAPDH levels. Data are expressed as means ± SEM. Statistical analysis was conducted by independent-samples t -test. * P =0.05 versus LID + Tat-D1Rc. Abbreviations: 6-OHDA, 6-hydroxydopamine; l -dopa, L-3,4-dihydroxyphenylalanine; LID, l -dopa-induced dyskinesia; SEM, standard error of mean; Tat-D1Ri, Tat-fusion interfering peptide; Tat-D1Rc, Tat-fusion control peptide; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Samples were incubated with a rabbit antibody against D1R (EMD Millipore, Billerica, MA, USA) or a mouse antibody against GluN1 (Millipore) overnight at 4°C.

Techniques: Expressing, Western Blot

Journal: Drug Design, Development and Therapy

Article Title: Targeting the D1-N-methyl- d -aspartate receptor complex reduces l -dopa-induced dyskinesia in 6-hydroxydopamine-lesioned Parkinson’s rats

doi: 10.2147/DDDT.S93487

Figure Lengend Snippet: Effects of intrastriatal administration of Tat-fusion interfering peptide (Tat-D1Ri) on D1R–GluN1 interactions. Notes: Tat-D1Ri, Tat-D1Rc, or saline were locally injected into the striatum of the normal adult rat at a rate of 0.2 µL/min. Rat striatal tissues were then used for coimmunoprecipitation experiments to validate the efficacy and selectivity of interfering peptide. The intrastriatal injection of Tat-D1Ri rather than Tat-D1Rc caused reduction of D1R–GluN1 interactions, which demonstrated the effectiveness of Tat-D1Ri. Abbreviations: Tat-D1Rc, Tat-fusion control peptide; Ig, Immunoglobulin; IP, immunoprecipitation; IB, immunoblot.

Article Snippet: Membranes were blocked in 5% nonfat milk for 1 hour at room temperature and incubated with a rabbit primary antibody against D1R (Millipore) or a mouse primary antibody against GluN1 (Millipore) overnight at 4°C.

Techniques: Injection, Immunoprecipitation, Western Blot

Journal: Drug Design, Development and Therapy

Article Title: Targeting the D1-N-methyl- d -aspartate receptor complex reduces l -dopa-induced dyskinesia in 6-hydroxydopamine-lesioned Parkinson’s rats

doi: 10.2147/DDDT.S93487

Figure Lengend Snippet: Effects of intrastriatal administration of Tat-D1Ri on membrane D1R subunit expression. Notes: Protein levels were evaluated by Western blotting of proteins extracted from the lesioned striatum of the rat brains. They were assessed in extracts from 6-OHDA-lesioned rats treated with pulsatile l -dopa plus intrastriatal administration of Tat-D1Rc or Tat-D1Ri. ( A ) Representative Western blot analysis of D1R in the membrane fraction; ( B ) densitometric analysis of two blots with specific protein signals normalized to the corresponding GAPDH levels. Data are expressed as means ± SEM. Statistical analysis was conducted by independent-samples t -test. * P =0.05 versus LID + Tat-D1Rc. Abbreviations: 6-OHDA, 6-hydroxydopamine; l -dopa, L-3,4-dihydroxyphenylalanine; LID, l -dopa-induced dyskinesia; SEM, standard error of mean; Tat-D1Ri, Tat-fusion interfering peptide; Tat-D1Rc, Tat-fusion control peptide; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Membranes were blocked in 5% nonfat milk for 1 hour at room temperature and incubated with a rabbit primary antibody against D1R (Millipore) or a mouse primary antibody against GluN1 (Millipore) overnight at 4°C.

Techniques: Expressing, 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: 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