dopamine receptor d1 drd1  (Proteintech)

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: Dopamine Receptor 1 Treatment Promotes Epithelial Repair of Corneal Injury by Inhibiting NOD-Like Receptor Protein 3-Associated Inflammation.

doi: 10.1167/iovs.65.1.49

Figure Lengend Snippet: FIGURE 1. Cell viability after exposure to IL-1β. Control iMCECs had no additional experimental interventions. (A) The effect of different concentrations of IL-1β (0, 50, 100, 200 ng/mL) at different times (12, 24, 48, 72 hours) of intervention on the cell viability of iMCECs. (B) DRD1 was successfully overexpressed in iMCECs by pcDNA_DRD1. (C) Quantification analysis of iMCEC proliferation during homeostatic conditions via CCK-8 assay. (D) Quantification analysis of iMCEC proliferation in an inflammatory environment via CCK-8 assay. Data are represented as mean ± SEM. Experiments were repeated three or four times, and data in each group are from triplicate or quadruplicate wells. *P < 0.05, **P < 0.01, ***P < 0.005, ****P < 0.001; ns, not significant.

Article Snippet: Five microliters of 0.1% DRD1 agonist (A68930; MedChemExpress, Monmouth Junction, NJ, USA) or PBS was applied topically to the injured eyes using a micropipette three times a day for up to 5 days post-injury.

Techniques: Control, CCK-8 Assay

Journal: Investigative ophthalmology & visual science

Article Title: Dopamine Receptor 1 Treatment Promotes Epithelial Repair of Corneal Injury by Inhibiting NOD-Like Receptor Protein 3-Associated Inflammation.

doi: 10.1167/iovs.65.1.49

Figure Lengend Snippet: FIGURE 2. Overexpressed DRD1 inhibited NLRP3-related inflammatory factors in iMCECs. (A) Representative confocal microscopic images of the NLRP3+ iMCECs. Scale bars: 100 μm. (B) Quantification analysis of NLRP3 protein expression via immunofluorescence staining. (C) Quantification analysis of IL-1β mRNA expression via real-time qPCR. (D) Quantification analysis of IL-6 mRNA expression via real-time qPCR. Data are represented as mean ± SEM. Experiments were repeated four times, and data in each group are from quadruplicate wells. *P < 0.05, **P < 0.01, ***P < 0.005, ****P < 0.001; ns, not significant.

Article Snippet: Five microliters of 0.1% DRD1 agonist (A68930; MedChemExpress, Monmouth Junction, NJ, USA) or PBS was applied topically to the injured eyes using a micropipette three times a day for up to 5 days post-injury.

Techniques: Expressing, Immunofluorescence, Staining

Journal: Investigative ophthalmology & visual science

Article Title: Dopamine Receptor 1 Treatment Promotes Epithelial Repair of Corneal Injury by Inhibiting NOD-Like Receptor Protein 3-Associated Inflammation.

doi: 10.1167/iovs.65.1.49

Figure Lengend Snippet: FIGURE 3. DRD1 agonist promoted pMCEC proliferation and inhibited expression of NLRP3. The DRD1 agonist was A68930, and the DRD1 antagonist was SCH23390. (A) The effect of different concentrations of A68930 (0, 10, 20, 30 μM) on the cell viability of pMCECs. (B) Representative immunoblot images of the expression of NLRP3 and GAPDH protein. (C) Quantification analysis of NLRP3 protein expression via western blotting. (D) Representative confocal microscopic images of the Ki-67+ and p63+ pMCECs. Scale bars: 100 μm. (E) Quantification analysis of Ki-67+ pMCECs via immunofluorescent staining. (F) Quantification analysis of p63+ pMCECs via immunoflu- orescent staining. Data are represented as mean ± SEM. Experiments were repeated four or five times, and data in each group are from quadruplicate or quintuplicate wells. *P < 0.05, **P < 0.01, ***P < 0.005, ****P < 0.001; ns, not significant.

Article Snippet: Five microliters of 0.1% DRD1 agonist (A68930; MedChemExpress, Monmouth Junction, NJ, USA) or PBS was applied topically to the injured eyes using a micropipette three times a day for up to 5 days post-injury.

Techniques: Expressing, Western Blot, Staining

Journal: Investigative ophthalmology & visual science

Article Title: Dopamine Receptor 1 Treatment Promotes Epithelial Repair of Corneal Injury by Inhibiting NOD-Like Receptor Protein 3-Associated Inflammation.

doi: 10.1167/iovs.65.1.49

Figure Lengend Snippet: FIGURE 4. Administration of 0.1% A68930 promoted MCEC proliferation and expression of DRD1 in vivo. (A) Representative confocal micro- scopic images of the Ki-67+ and p63+ cells. Scale bars: 100 μm. (B) Quantification analysis of Ki-67+ cells in corneas via immunofluorescent staining. (C) Quantification analysis of p63+ cells via immunofluorescent staining. (D) Quantification analysis of DRD1 mRNA expression via real-time qPCR. (E) Representative immunoblot images of the expression of DRD1 and GAPDH protein. (F) Quantification analysis of DRD1 protein expression via western blotting. Data are represented as mean ± SEM. Experiments were repeated three to eight times. *P < 0.05, **P < 0.01, ***P < 0.005, ****P < 0.001; ns, not significant.

Article Snippet: Five microliters of 0.1% DRD1 agonist (A68930; MedChemExpress, Monmouth Junction, NJ, USA) or PBS was applied topically to the injured eyes using a micropipette three times a day for up to 5 days post-injury.

Techniques: Expressing, In Vivo, Staining, Western Blot

Journal: Investigative ophthalmology & visual science

Article Title: Dopamine Receptor 1 Treatment Promotes Epithelial Repair of Corneal Injury by Inhibiting NOD-Like Receptor Protein 3-Associated Inflammation.

doi: 10.1167/iovs.65.1.49

Figure Lengend Snippet: FIGURE 7. Administration of 0.1% A68930 promoted the corneal wound healing process. (A) The PBS treatment group and the 0.1% A68930 treatment group were treated with the same corneal injury. (B) After injury, expression of DRD1 was downregulated and NLRP3, IL-1β, and IL-6 levels were upregulated. (C) The 0.1% A68930 promoted corneal wound healing by upregulation of DRD1 and downregulation of NLRP3-associated inflammatory factors.

Article Snippet: Five microliters of 0.1% DRD1 agonist (A68930; MedChemExpress, Monmouth Junction, NJ, USA) or PBS was applied topically to the injured eyes using a micropipette three times a day for up to 5 days post-injury.

Techniques: Expressing

Journal: Journal of Psychiatry & Neuroscience : JPN

Article Title: Reviving: restoring depression-like behaviour through glial cell–derived neurotrophic factor treatment in the medial prefrontal cortex

doi: 10.1503/jpn.230079

Figure Lengend Snippet: Effect of glial cell–derived neurotrophic factor (GDNF) in the medial prefrontal cortex (mPFC) and regional dopamine receptor D1 (DRD1) pathway activity in relieving depression-like behaviour in Parkinson disease mouse models. (A) Experimental timeline. (B) Western blot analysis of DRD1 expression in the mPFC of control mice, mice treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and mice treated with MPTP plus GDNF (MPTP v. control: p = 0.0048; MPTP v. MPTP + GDNF: p = 0.0053). (C) Immunofluorescent staining showed immunoreactive puncta (red) of DRD1 in the mPFC of control, MPTP-treated and MPTP plus GDNF–treated mice (4′,6-diamidino-2-phenylindole [DAPI] in blue; scale 50 μm) (normalized positive area: MPTP v. control: p < 0.0001; MPTP v. MPTP + GDNF: p < 0.001). (D–F) Western blot analysis of protein kinase A (PKA), and phospho-PKA (pPKA) expression in the mPFC of control, MPTP-treated or MPTP plus GDNF–treated mice (PKA: MPTP v. control: p = 0.5826; MPTP v. MPTP + GDNF: p = 0.7182,) (pPKA: MPTP v. control: p = 0.0005; MPTP v. MPTP + GDNF: p = 0.0029), (pPKA/PKA, MPTP v. control: p = 0.0003, MPTP v. MPTP + GDNF: p = 0.0011). (G, H) Western blot analysis of pPKA and PKA expression in the mPFC of mice treated with MPTP plus GDNF or with MPTP, GDNF and DRD1 antagonist (pPKA: t = 4.273, p = 0.0238) (pPKA/PKA: t = 7.328, p = 0.0429). (I) Performance of mice treated with MPTP plus GDNF or with MPTP, GDNF and DRD1 antagonist in the forced swimming test ( p = 0.0028) and (J) the sucrose preference test. ( p = 0.0051). (K–P) Western blot analysis of dopamine receptor D2 (DRD2), arrestin, glycogen synthase kinase-3β (GSK-3β) and protein phosphatase 2A (PP2A) expression in the mPFC of control, MPTP-treated and MPTP plus GDNF–treated mice (DRD2: MPTP v. control: p = 0.3152; MPTP v. MPTP + GDNF: p = 0.4269) (arrestin: MPTP v. control: p = 0.0065; MPTP v. MPTP + GDNF: p = 0.0267) (PP2A: MPTP v. control: p = 0.4127; MPTP v. MPTP + GDNF: p = 0.0381) (GSK-3: MPTP v. control: p = 0.0039; MPTP v. MPTP + GDNF: p = 0.0058). ACC = anterior cingulate cortex; IL = infralimbic cortex; PL = prelimbic cortex; SEM = standard error of the mean.

Article Snippet: After intraperitoneal injection of the DRD1 antagonist (SCH-23390 MedChemExpress) at a dose of 0.1 mg/kg in the MPTP plus GDNF injection group, the mice were sacrificed within 30 minutes.

Techniques: Derivative Assay, Activity Assay, Western Blot, Expressing, Control, Staining