drd3 polyclonal antibody  (Bioss)

Journal: Frontiers in Cardiovascular Medicine

Article Title: Loss of Function in Dopamine D3 Receptor Attenuates Left Ventricular Cardiac Fibroblast Migration and Proliferation in vitro

doi: 10.3389/fcvm.2021.732282

Figure Lengend Snippet: The expression and distribution of dopamine receptors, D1R and D3R in mouse hearts: (A) Representative photomicrographs showing immunofluorescence staining of D1R and D3R which co-localized with vimentin, a cardiac fibroblast-specific marker, in the heart tissue sections. No primary antibody control sections were incubated with the antibody diluent alone and no primary antibody, followed by incubation with secondary antibodies and DAPI (the nuclear stain), which confirms the antibody specificity of D1R and D3R. (B) Gene expression was determined using TaqMan primers by real-time qPCR. The normalized -dCT values indicate that there is relatively higher expression of D1R compared to D3R in cardiac tissue. (C) Receptor protein expression determined by western blot using receptor specific antibodies as described in methods section.

Article Snippet: Both cells and tissues were then incubated with primary antibodies for Vimentin [M0725, lot #027(102), Dako, 1:500 dilution], D1R (NB110-60017AF488, lot #B-3-101620, Novus Biologics, 1:500 dilution), and D3R (bs-1743R-Cy5, lot #AF12125641, Bioss, 1:500 dilution) overnight at 4°C.

Techniques: Expressing, Immunofluorescence, Staining, Marker, Incubation, Western Blot

Journal: Frontiers in Cardiovascular Medicine

Article Title: Loss of Function in Dopamine D3 Receptor Attenuates Left Ventricular Cardiac Fibroblast Migration and Proliferation in vitro

doi: 10.3389/fcvm.2021.732282

Figure Lengend Snippet: The expression and distribution of dopamine receptors, D1R and D3R in mouse primary cardiac fibroblasts: (A) Representative photomicrographs showing immunofluorescence staining of D1R and D3R which co-localized with cardiac fibroblasts specific marker vimentin staining in the isolated WT and D3KO primary cardiac fibroblasts in culture. Cardiac fibroblast expression of D1R and D3R is not lost when primary cells are maintained in cell culture. As expected, there is no observable staining for D3R in D3KO cardiac fibroblasts. No primary antibody control images show the specificity of the primary antibodies used. (B) Gene expression determined using TaqMan primers by real-time qPCR. The normalized -dCT values indicate that there is relatively higher expression D1R compared to D3R in cardiac fibroblasts. (C) Receptor protein expression determined by western blot using receptor specific antibodies as described in methods section.

Article Snippet: Both cells and tissues were then incubated with primary antibodies for Vimentin [M0725, lot #027(102), Dako, 1:500 dilution], D1R (NB110-60017AF488, lot #B-3-101620, Novus Biologics, 1:500 dilution), and D3R (bs-1743R-Cy5, lot #AF12125641, Bioss, 1:500 dilution) overnight at 4°C.

Techniques: Expressing, Immunofluorescence, Staining, Marker, Isolation, Cell Culture, Western Blot

Journal: Frontiers in Cardiovascular Medicine

Article Title: Loss of Function in Dopamine D3 Receptor Attenuates Left Ventricular Cardiac Fibroblast Migration and Proliferation in vitro

doi: 10.3389/fcvm.2021.732282

Figure Lengend Snippet: Cardiac fibroblast cell proliferation and migration studies: (A) Cardiac fibroblast cell proliferation between WT + vehicle (black circle), WT treated with SB (closed blue square), and D3KO + vehicle (red circle) over a time-period of 36 h. Cells were incubated in DMEM/F-12 growth media supplemented with 10% FBS; n = 3 for each treatment at each time-point, * p < 0.05 statistically significant. Bars represent standard error of mean. (B) Cardiac fibroblast cell migration distance in response to a scratch injury over a 24-h time-period. Migration distance in μm between WT + vehicle (black circle), D3KO + vehicle (red circle), and WT cells treated with D3R specific agonist, pramipexole (blue circle). Cells were incubated in DMEM/F-12 growth media supplemented with 0.5% FBS; n = 9 for each treatment at each time-point, * p < 0.05 statistically significant. Bars represent standard error of mean. Statistical significance between WT treatment and D3KO + vehicle. (C) Cardiac fibroblast cell migration distance in response to a scratch injury over a 24-h time-period. Migration distance in μm of D3KO + vehicle cells (red circle), D3KO cells treated with pramipexole (closed blue square), and D3KO cell treated with SB (green square), compared against WT + vehicle cells (black circle); n = 9 for each treatment at each time-point, * p < 0.05 statistically significant. Bars represent standard error of mean.

Article Snippet: Both cells and tissues were then incubated with primary antibodies for Vimentin [M0725, lot #027(102), Dako, 1:500 dilution], D1R (NB110-60017AF488, lot #B-3-101620, Novus Biologics, 1:500 dilution), and D3R (bs-1743R-Cy5, lot #AF12125641, Bioss, 1:500 dilution) overnight at 4°C.

Techniques: Migration, Incubation

Journal: International Journal of Molecular Sciences

Article Title: Alteration of the Dopamine Receptors’ Expression in the Cerebellum of the Lysosomal Acid Phosphatase 2 Mutant (Naked–Ataxia ( NAX )) Mouse

doi: 10.3390/ijms21082914

Figure Lengend Snippet: RNA sequencing data analysis in nax ( n = 5) and wild–type (wt) ( n = 6) mice. Data analysis shows dopamine receptor D1 ( Drd1, A ), Drd3 ( C ), and Drd4 ( D ) are significantly increased in the nax cerebellum. Although an increase in Drd2 ( B ) and Drd5 ( E ) is apparent, statistical significance was not reached. The data in the bar graph are presented as the mean ± SEM, and statistical analysis was performed using an unpaired t –test (* p < 0.05 and ** p < 0.01). RPKM: Reads Per Kilobase of transcript per Million mapped reads.

Article Snippet: D1: rabbit polyclonal anti–D1 dopamine receptor (TA328798, anti–Drd1, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA); D2: rabbit polyclonal anti–D2 dopamine receptor (TA328800, anti–Drd2, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 2 (DR2); D3: rabbit polyclonal anti–D3 dopamine receptor (TA328800, anti–Drd3, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 3 (DR3); D4: rabbit polyclonal anti–D4 dopamine receptor (TA321202, anti–DRD4, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor D4 (DRD4); and D5: rabbit polyclonal anti–D5 dopamine receptor (TA328802, anti–Drd5, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 5 (DR5).

Techniques: RNA Sequencing Assay

Journal: International Journal of Molecular Sciences

Article Title: Alteration of the Dopamine Receptors’ Expression in the Cerebellum of the Lysosomal Acid Phosphatase 2 Mutant (Naked–Ataxia ( NAX )) Mouse

doi: 10.3390/ijms21082914

Figure Lengend Snippet: Frontal sections of wt and nax mouse cerebella: DRD2 expression at P5 and P17. Immunoperoxidase staining for DRD2 in wt ( A ) and nax ( B ) cerebella at P5 shows weak immunoreactivity in the Pcl of wt but relatively strong immunoreactivity in PC somata of the nax cerebellum. ( C , D ) Immunoperoxidase staining for DRD2 in wt ( C ) and nax ( D ) cerebella at P17 shows weak immunoreactivity in PCs, with no differences between the two groups. ( E ) Western blot analysis of whole cerebellar lysates revealed no significant differences in DRD2 protein expression between wt and nax cerebella at P5 and P17 (wt: n = 3 and nax : n = 3). The data in the bar graphs are presented as the means of three independent experiments ± SEM; statistical analysis was performed using two–way ANOVA. P; postnatal. Scale bars: 100 μm in A, C, and D and 50 μm in B.

Article Snippet: D1: rabbit polyclonal anti–D1 dopamine receptor (TA328798, anti–Drd1, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA); D2: rabbit polyclonal anti–D2 dopamine receptor (TA328800, anti–Drd2, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 2 (DR2); D3: rabbit polyclonal anti–D3 dopamine receptor (TA328800, anti–Drd3, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 3 (DR3); D4: rabbit polyclonal anti–D4 dopamine receptor (TA321202, anti–DRD4, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor D4 (DRD4); and D5: rabbit polyclonal anti–D5 dopamine receptor (TA328802, anti–Drd5, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 5 (DR5).

Techniques: Expressing, Immunoperoxidase Staining, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Alteration of the Dopamine Receptors’ Expression in the Cerebellum of the Lysosomal Acid Phosphatase 2 Mutant (Naked–Ataxia ( NAX )) Mouse

doi: 10.3390/ijms21082914

Figure Lengend Snippet: RNA sequencing data analysis in nax ( n = 5) and wild–type (wt) ( n = 6) mice. Data analysis shows dopamine receptor D1 ( Drd1, A ), Drd3 ( C ), and Drd4 ( D ) are significantly increased in the nax cerebellum. Although an increase in Drd2 ( B ) and Drd5 ( E ) is apparent, statistical significance was not reached. The data in the bar graph are presented as the mean ± SEM, and statistical analysis was performed using an unpaired t –test (* p < 0.05 and ** p < 0.01). RPKM: Reads Per Kilobase of transcript per Million mapped reads.

Article Snippet: D1: rabbit polyclonal anti–D1 dopamine receptor (TA328798, anti–Drd1, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA); D2: rabbit polyclonal anti–D2 dopamine receptor (TA328800, anti–Drd2, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 2 (DR2); D3: rabbit polyclonal anti–D3 dopamine receptor (TA328800, anti–Drd3, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 3 (DR3); D4: rabbit polyclonal anti–D4 dopamine receptor (TA321202, anti–DRD4, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor D4 (DRD4); and D5: rabbit polyclonal anti–D5 dopamine receptor (TA328802, anti–Drd5, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 5 (DR5).

Techniques: RNA Sequencing Assay

Journal: International Journal of Molecular Sciences

Article Title: Alteration of the Dopamine Receptors’ Expression in the Cerebellum of the Lysosomal Acid Phosphatase 2 Mutant (Naked–Ataxia ( NAX )) Mouse

doi: 10.3390/ijms21082914

Figure Lengend Snippet: Frontal sections of wt and nax mouse cerebella: DRD3 expression at P5 and P17. ( A , B ) Immunoperoxidase staining for DRD3 in wt ( A ) and nax ( B ) cerebella at P5 shows weak immunoreactivity in the Pcl and external germinal zone (egz). ( C – E ) Immunoperoxidase staining for DRD3 in the wt cerebellum at P17 demonstrates immunoreactivity in the whole cortex of the wt cerebellum, mostly in PC somata ( C , D ) within scattered cell bodies in the gl (arrows indicated in D ( d )) and in cerebellar nuclei neurons (CNs) ( C , E ). ( F – H ) DRD3 immunostaining of a frontal section of the nax cerebellum at P17 shows strong immunoreactivity in PC somata in the Pcl/ml ( F – H ), CNs ( F ), and a few cells in the gl (arrows in G , H ). ( E ) Western blot analysis of whole cerebellar lysates revealed no significant differences in DRD3 protein expression between wt and nax cerebella at P5 and P17 (wt: n = 3 and nax : n = 3). The data in the bar graph are presented as the mean of three independent experiments ± SEM, and statistical analysis was performed using two–way ANOVA. P; postnatal. Scale bars: 50 μm in A and B, 500 μm in C and F, 100 μm in D, 200 μm in E, 100 μm in G.

Article Snippet: D1: rabbit polyclonal anti–D1 dopamine receptor (TA328798, anti–Drd1, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA); D2: rabbit polyclonal anti–D2 dopamine receptor (TA328800, anti–Drd2, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 2 (DR2); D3: rabbit polyclonal anti–D3 dopamine receptor (TA328800, anti–Drd3, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 3 (DR3); D4: rabbit polyclonal anti–D4 dopamine receptor (TA321202, anti–DRD4, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor D4 (DRD4); and D5: rabbit polyclonal anti–D5 dopamine receptor (TA328802, anti–Drd5, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 5 (DR5).

Techniques: Expressing, Immunoperoxidase Staining, Immunostaining, Western Blot