Journal:

Article Title:

doi:

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: 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