Journal: Psychopharmacology

Article Title: Nicotine withdrawal-induced inattention is absent in alpha7 nAChR knockout mice

doi: 10.1007/s00213-017-4572-2

Figure Lengend Snippet: Synaptosomal receptor expression (% of saline control with pump). Bolded indicates significant interaction between Saline/Nicotine vs. Pump On/Pump Off. Effect of chronic nicotine (40 mg/kg/day) treatment on mGluR1, mGluR5, and dopamine D 4 receptor expression. After 33 days of chronic nicotine treatment (“Pump On”) and 4 h after pump removal (“Pump Off”), a subset of WT mice were removed and levels of synaptosomal mGluR1, mGluR5 and, D 4 receptor expression were analyzed. There was a significant effect of nicotine on mGluR1 expression during nicotine administration and during withdrawal. Significant interactions between synaptosomal expression during nicotine (Pump On) and withdrawal (Pump Off) for mGluR5 (trend) and dopamine D 4 receptors were observed, though no significant post hoc differences between levels were observed.

Article Snippet: Blots were probed with primary antibodies overnight in a cold room using anti-mGluR1 (#ab27199 from Abcam, Cambridge, MA), mGluR5 (#ab76316 from Abcam), and dopamine D 4 receptor (#ADR-004, from Alomone Labs, Jerusalem, Israel) antibodies.

Techniques: Expressing

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 and sagittal sections of wt and nax mouse cerebella: DRD4 expression at P5 and P17. ( A , B ) Immunoperoxidase staining for DRD4 in wt ( A ) and nax ( B ) cerebella at P5 shows immunoreactivity in the external germinal zone (egz) and no or only weak immunoreactivity in the Pcl. ( C – G ) Sagittal section of P17 wt cerebellum immunostained for DRD4 reveals strong immunoreactivity within the cerebellar cortex. In the anterior zone, the expression of DRD4 is present in PC dendrites in the molecular layer (ml), while PC somata and dendrites exhibit immunoreactivity in the nodular zone ( C ). Frontal section of P17 wt cerebellum immunostained with DRD4 shows strong immunoreactivity in a subset of PCs (indicated by black arrowheads), while another subset of PCs displays no (white arrowhead) or only weak expression (arrow) in the cerebellar cortex ( D ). ( E – G ) Immunoperoxidase staining for DRD4 in the wt cerebellum at P17 shows a striped expression pattern in lobule IXd ( E , F ) and more uniform expression in PCs in lobule X ( G ). ( H – J ) DRD4 immunostaining of a frontal section of the nax cerebellum at P17 shows immunoreactivity in a subset of PCs (asterisks) that follows a similar stripes pattern in lobule III ( H ) and IX ( I ) but is uniform in lobule X ( J ). ( K ) Western blot analysis of DRD4 expression in wt and nax cerebellum at P5 and P17 shows an apparent increase in expression in nax cerebella at both time–points (wt: n = 3 and nax : n = 3); however, these differences did not reach statistical significance. The data in the bar graphs are presented as the means of three independent experiments ± SEM, and statistical analysis was performed using two–way ANOVA. P; postnatal. Scale bars: 100 μm in A; 50 μm in B; 500 μm in C; 20 μm in D; 200 μm in E, F, G, H, and I; and 500 μm in J.

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

Journal: Investigative Ophthalmology & Visual Science

Article Title: Distinct Transcriptomic Profiles of Cultured Anterior and Posterior Populations of Human Infant Scleral Fibroblasts: Including Dopamine Receptors

doi: 10.1167/iovs.66.5.29

Figure Lengend Snippet: Silencing DRD4 significantly enhances DA-mediated contraction inhibition, particularly in posterior scleral fibroblasts. ( a , b ) Seven-day gel contraction curves of DRD2 -knockdown anterior and posterior infant scleral fibroblasts, with or without 50-µM DA. Each condition was tested in triplicate for cells derived from two donors ( n = 6). The control group was transfected with non-targeting siRNA. ( c , e ) Western blot validation of DRD2 knockdown in anterior and posterior fibroblasts from two donors. ( d , f ) Comparison of contraction rates at day 7 for DRD2 -knockdown anterior and posterior cells, respectively. Statistical analysis: *** P < 0.001, **** P < 0.0001 (two-way ANOVA). ( g , h ) Seven-day gel contraction curves of DRD4 -knockdown anterior and posterior infant scleral fibroblasts, with or without 50-µM DA ( n = 6). ( i , k ) Western blot validation of DRD4 knockdown in anterior and posterior fibroblasts from two donors. ( j , l ) Comparison of contraction rates at day 7 for DRD4 -knockdown anterior and posterior cells, respectively. Statistical analysis: * P < 0.05, ** P < 0.01, **** P < 0.0001 (two-way ANOVA). ( m , o ) Percentage reduction in gel contraction on day 7 resulting from siRNA-knockdown of DRD2 or DRD4 , compared to non-targeting siRNA controls, in anterior and posterior scleral fibroblasts, respectively. Statistical analysis: * P < 0.05, **** P < 0.0001 ( t -test). ( n , p ) Percentage reduction in gel contraction on day 7 caused by DA treatment alone (compared to control), and by DRD2 or DRD4 knockdown combined with DA (compared to DRD2 or DRD4 knockdown without DA), in anterior and posterior fibroblasts, respectively ( n = 6). Whiskers represent minimum to maximum values. * P < 0.05, ** P < 0.01, **** P < 0.0001 (one-way ANOVA).

Article Snippet: The antibodies used were as follows: Dopamine Receptor D1 Rabbit mAb (381747; Zen-Bioscience), DRD2 polyclonal antibody (55084-1-AP; Proteintech, Rosemont, IL, USA), Dopamine Receptor D3 Rabbit mAb (382983; Zen-Bioscience), DRD4 Rabbit polyclonal antibody (28094-1-AP; Proteintech), Dopamine Receptor D5 Rabbit pAb (820522; Zen-Bioscience), GAPDH (7E4) Mouse mAb (200306-7E4; Zen-Bioscience), Beta Tubulin Polyclonal antibody (10094-1-AP; Proteintech), Goat anti-Rabbit IgG(H&L) (HRP conjugate) (511203; Zen-Bioscience), and Goat anti-Mouse IgG (H&L) (HRP conjugate) (511103; Zen-Bioscience).

Techniques: Inhibition, Knockdown, Derivative Assay, Control, Transfection, Western Blot, Biomarker Discovery, Comparison