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: Hypertension research : official journal of the Japanese Society of Hypertension

Article Title: Antihypertensive effect of etamicastat in dopamine D2 receptor-deficient mice.

doi: 10.1038/s41440-018-0041-5

Figure Lengend Snippet: Fig. 8 Filtered dopamine, as well as dopamine produced in the renal proximal tubule, stimulates dopamine receptors to increase sodium excretion under conditions of normal or moderate sodium load. Deletion of any dopamine receptor gene increases blood pressure. Etamicastat inhibits dopamine β hydroxylase (DBH), decreasing the conversion of norepinephrine to dopamine, allowing the levels of dopamine to increase. The increase in filtered dopamine and dopamine produced by the kidney (presumably from the renal nerves; DBH is not expressed in renal tubules) stimulate the other dopamine receptor subtypes, D1R, D3R, D4R, and D5R, which overcomes the absence or decreased expression of D2R. The increased production of reactive oxygen species (ROS) is normalized, sodium excretion (UNaV) is increased, and blood pressure is normalized in mice without D2R in the body or mice with decreased renal D2R expression

Article Snippet: We have reported the specificity of our D1R antibody [18, 20], D3R antibody [19], D4R antibody [17], D5R antibody [21, 22], D1R antibody from Origene (Rockville, MD) [22], and D2R from EMD Millipore [13].

Techniques: Produced, Expressing

Journal: American Journal of Physiology - Renal Physiology

Article Title: G protein-coupled receptor 37L1 regulates renal sodium transport and blood pressure

doi: 10.1152/ajprenal.00289.2018

Figure Lengend Snippet: In human renal proximal tubule cells (hRPTCs), overexpression of GPR37L1 increases while silencing of GPR37L1 decreases intracellular sodium. hRPTCs were cultured in 12-well Transwell plates and transfected with the indicated plasmids. Two days later, the cells were serum starved overnight and treated with the indicated drugs at the luminal side for 30 min before loading with the sodium fluorescent dye. A: hRPTCs were transfected with control plasmid (empty vector, pcDNA), plasmid carrying cDNA encoding GPR37L1 (pCMV-GPR37L1), luciferase (LUC), β-galactosidase (GAL), or human DRD1 (pCMV-DRD1). n = 3/group; *P < 0.05 GPR37L1 vs. the rest, **P < 0.05 DRD1 vs. pcDNA. B: hRPTCs transfected with plasmids pcDNA or pCMV-GPR37L1 were treated at the luminal membrane side with vehicle (Veh), nonselective NHE isoform inhibitor 5-(N-ethyl-N-isopropyl) amiloride (EIPA, 10 μM), or NHE3-selective inhibitor 3-[2-(3-guanidino-2-methyl-3-oxopropenyl)-5-methyl-phenyl]-N-isopropylidene-2-methyl-acrylamide dihydrochloride (S3226, 10 µM). n = 3/group; +P < 0.05 vs. pcDNA-Veh; n = 3/group. C: hRPTCs transfected with nonsilencing shRNA (NS-shRNA) or GPR37L1-shRNA (37L1-shRNA) and treated with Veh or S3226 at the luminal side similar to B. n = 3/group; ++P < 0.05 vs. NS-shRNA-Veh. D: hRPTCs transfected with plasmids pcDNA or pCMV-GPR37L1 were treated at the luminal membrane side with Veh, INM, PMA, FSK, or S3226. n = 3/group; $P < 0.05 vs. pcDNA-Veh, #P < 0.05 vs. GPR37L1-Veh. DRD1, dopamine receptor D1; FSK, forskolin; GPR37L1, G protein-coupled receptor 37L1; INM, ionophore ionomycin; NHE, Na+/H+ exchanger; CMV, cytomegalovirus.

Article Snippet: A plasmid encoding human GPR37L1 (pCMV- GPR37L1 ]) or dopamine receptor D1 (pCMV- DRD1 ), under the control of cytomegalovirus promoter, was obtained from Origene Technology (Rockville, MD; cat. no. GPR37L1 : SC117166; DRD1 : RC210389).

Techniques: Over Expression, Cell Culture, Transfection, Control, Plasmid Preparation, Luciferase, Membrane, shRNA