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: D1-like receptor activity promotes the inhibitory effect of DA on cell contraction, whereas D2-like receptor activity, particularly that of DRD2 in the posterior sclera, impedes it. ( a – p ) The cell contraction rates at 24 hours for anterior ( a – h ) and posterior ( i – p ) scleral fibroblasts from HSF12 and HSF13 were measured following treatment with a range of DA receptor antagonists and agonists, administered 1 hour prior to the addition of DA. One experiment was conducted with three replicate wells for each compound tested, using two biological samples (HSF12 and HSF13; total n = 6). Results from the two cell lines were averaged and presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (two-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: Activity Assay

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

Journal: Advanced materials (Deerfield Beach, Fla.)

Article Title: Intranasal Delivery of Near-infrared and Magnetic Dual-response Nanospheres to Rapidly Produce Antidepressant-like and Cognitive Enhancement Effects.

doi: 10.1002/adma.202405547

Figure Lengend Snippet: Figure 7. Investigation of the mechanism of therapeutic effect and biosafety assessment in vivo. a) Fluorescence staining of different brain regions. Blue: 4′,6-diamidino-2-phenylindole (DAPI), Red: TH or DRD2 or DRD1. The circle statistical charts show the fluorescence intensity ratio. b) Fluorescence intensity ratio of DRD2 or DRD1 in the PFC. c) Representative immunoblots of proteins in dopamine-related signaling pathways. d–l) Quantitative analyses of proteins in (c). Data are means ± s.d. (n ≥3), *p < 0.05, **p < 0.01.

Article Snippet: TH, DRD1, and DRD2 polyclonal antibodies were purchased from Proteintech Group, Inc. All other reagents were purchased from commercial resources and used as received unless otherwise noted.

Techniques: In Vivo, Fluorescence, Staining, Western Blot, Protein-Protein interactions