d1r agonist skf-38393  (Tocris)

Journal: The Journal of Clinical Investigation

Article Title: A midbrain–cortical circuit mediated by a claustrum neuronal ensemble orchestrates drug-paired context memory processing

doi: 10.1172/JCI196944

Figure Lengend Snippet: ( A ) Experimental design. ( B ) Schematic of viral transfection. ( C ) Representative images of virus injection and fiber implant in CL. ( D ) Heatmap of DA3h fluorescence (left), quantification (middle), and AUC (right) of ΔF/F in CL. n = 5 mice/group. ( E ) The protein levels of D1R in CL. The protein levels of D1R following the METH CPP-Test. n = 3 mice/group. ( F ) Immunofluorescence of c-Fos + CaMKII + D1R + neurons in CL. Mean gray value of D1R CaMKII following METH CPP-Test. n = 6 mice/group. * P < 0.05. Two-tailed unpaired t test ( D – F ). Scale bars: 100 μm ( C and F ).

Article Snippet: The DA hydrochloride (ASL279, MedChemExpress) or D1R agonist (±)-SKF-38393 hydrochloride (SKF-38393, MedChemExpress) was diluted to 10 mM in 0.9% saline by shaking.

Techniques: Transfection, Virus, Injection, Fluorescence, Immunofluorescence, Two Tailed Test

Journal: The Journal of Clinical Investigation

Article Title: A midbrain–cortical circuit mediated by a claustrum neuronal ensemble orchestrates drug-paired context memory processing

doi: 10.1172/JCI196944

Figure Lengend Snippet: ( A ) Experimental design and timeline. ( B ) Schematic of viral transfection. ( C ) Representative images of virus injection and cannula implantation in VTA or CL. ( D ) Analysis of CPP behavior of time spent in CPP apparatus. S-CNO+Veh/S-CNO+DA group, n = 6 mice/group; M-CNO+Veh/M-CNO+DA group, n = 8 mice/group. ( E ) Representative heatmap of time spent in CPP apparatus during CPP-Test. ( F ) Experimental design and timeline. ( G ) Schematic of viral transfection. ( H ) Representative images of virus injection and cannula implantation in VTA or CL. ( I ) Analysis of CPP behavior of time spent in CPP apparatus. S-CNO+Veh/S-CNO+SKF group, n = 5 mice/group; M-CNO+Veh/M-CNO+SKF group, n = 6 mice/group. ( J ) Representative heatmap of time spent in CPP apparatus during CPP-Test. ( K ) Experimental design and timeline. ( L ) Schematic of viral transfection. ( M ) Representative images of virus injection in CL. ( N ) Analysis of CPP behavior and heatmap of time spent in CPP apparatus. S-CTRL/S-KD group, n = 6 mice/group; M-CTRL/M-KD group, n = 8 mice/group. ( O ) Representative heatmap of time spent in CPP apparatus during CPP-Test. ( P ) Immunofluorescence of c-Fos + D1R + EGFP + neurons in CL following CPP-Test. The percentage of c-Fos + EGFP + neurons in EGFP + neurons of CL. n = 6 mice/group. NS, P > 0.05, * P < 0.05, ** P < 0.01. Two-way ANOVA with Šidák’s multiple-comparison test ( D , I , N , and P ). Scale bars: 100 μm ( C , H , M , and P ).

Article Snippet: The DA hydrochloride (ASL279, MedChemExpress) or D1R agonist (±)-SKF-38393 hydrochloride (SKF-38393, MedChemExpress) was diluted to 10 mM in 0.9% saline by shaking.

Techniques: Transfection, Virus, Injection, Immunofluorescence, Comparison

Journal: bioRxiv

Article Title: Dopamine D1 and D4 receptors contribute to light adaptation in ON-sustained retinal ganglion cells

doi: 10.1101/2020.10.29.361147

Figure Lengend Snippet: D1R activation decreases the magnitude and times to peak of L-EPSCs in ON-s ganglion cells. A . Example L-EPSC traces from the same cell at increasing light intensities before (black) and after (blue) application of the D1R agonist SKF-38393 (20 µM). The gold bars represent 30 ms light stimuli. Stimulus intensities in total rod photoisomerizations are given above each set of traces. B-E . Comparison of average peak amplitude (B), Q (C), time to peak (D) and D37 (E) between dark-adapted (DA) and D1R-agonized (DA+D1) conditions. All values are normalized on a cell-by-cell basis to the maximum values of each parameter recorded for each cell. Average normalized values of light-adapted ganglion cells from are included for reference. Main effects of agonist treatment p-values for two-way ANOVAs are shown above each graph. * = significantly different main effect between dark-adapted and D1R agonist-treated state. # = significantly different pairwise comparison between dark-adapted and D1R agonized states. † = significantly different pairwise comparison between D1R agonized and light-adapted states. Shaded regions represent 95% confidence intervals. For all of our D1R L-EPSC experiments, n = 9 cells from 8 animals.

Article Snippet: The partial D1R agonist SKF-38393 (SKF, 20 μM; Tocris, Bristol, United Kingdom) was used to selectively activate D1Rs, and the D4R agonist PD-168077-maleate (PD, 500 nM; Sigma) was used to selectively activate D4Rs.

Techniques: Activation Assay, Comparison

Journal: bioRxiv

Article Title: Dopamine D1 and D4 receptors contribute to light adaptation in ON-sustained retinal ganglion cells

doi: 10.1101/2020.10.29.361147

Figure Lengend Snippet: D1R activation decreases the frequency of sEPSCs in some but not all ON-s ganglion cells. A . Example sEPSC traces from the same cell before (black) and after (blue) application of the D1R agonist SKF 38393 (20 µM). B-C . Average cumulative histogram of sEPSC amplitudes (B) and inter-event intervals (C) before and after D1R activation. Shaded regions represent 95% confidence intervals. Bin frequencies were normalized to total number of events recorded under each condition, and amplitude/IEI values were normalized to maximum values recorded on a cell-by-cell basis. 2-way ANOVA main effects p-values for DA vs D1R are reported above each graph. D-E . Average changes in sEPSC amplitude (D) and frequency (E) after D1R agonist application or light adaptation (included from for reference). Average values for each treatment were normalized on a cell-by-cell basis to that recorded under dark-adapted conditions. Individual data points are marked by empty white circles. Error bars = +/-standard error. # = significant difference between dark-adapted and D1R agonist conditions. * = significant difference between D1R agonist and light-adapted conditions. For all D1R sEPSC experiments, n = 5 cells from 5 animals.

Article Snippet: The partial D1R agonist SKF-38393 (SKF, 20 μM; Tocris, Bristol, United Kingdom) was used to selectively activate D1Rs, and the D4R agonist PD-168077-maleate (PD, 500 nM; Sigma) was used to selectively activate D4Rs.

Techniques: Activation Assay

Journal: bioRxiv

Article Title: Dopamine D1 and D4 receptors contribute to light adaptation in ON-sustained retinal ganglion cells

doi: 10.1101/2020.10.29.361147

Figure Lengend Snippet: Additive effects of dopaminergic signaling do not fully explain light adaptation. A-B . Average percent of response remaining (as compared to dark-adapted conditions) after light adaptation (LA, gray), application of a D1R agonist (D1, blue), or D4R agonist (D4, red), as well as the theoretical reduction one would witness from the additive effects of D1R-D4R agonist co-application (D1+D4, dashed line). The additive effects of D1R/D4R agonist co-application approximate the effects of light adaptation in diminishing peak amplitude (A), but do not match the decline in charge transfer witnessed upon light adaptation (B). C-D . Average % of sEPSC amplitude (C) and frequency (D) compared to dark-adapted conditions after light adaptation (LA, grey), D1R activation (D1, blue), and D4R activation (D4, red). The hypothetical impact of combined D1R/D4R activation is also shown (D1+D4, gold). The combined effects of dopamine agonists approximate the effect of light adaptation on average sEPSC amplitude (C), but not average sEPSC frequency (D).

Article Snippet: The partial D1R agonist SKF-38393 (SKF, 20 μM; Tocris, Bristol, United Kingdom) was used to selectively activate D1Rs, and the D4R agonist PD-168077-maleate (PD, 500 nM; Sigma) was used to selectively activate D4Rs.

Techniques: Activation Assay

Journal: NPJ Regenerative Medicine

Article Title: An in vivo brain–bacteria interface: the developing brain as a key regulator of innate immunity

doi: 10.1038/s41536-020-0087-2

Figure Lengend Snippet: a Dopamine concentration (in pg/μL) in infected Control (Ctrl, white) and Brainless (BR – , gray) embryos at early stage 35, measured by liquid chromatography–mass spectrometry (LC–MS/MS). Unpaired t -test * P < 0.05. b Survival rates (in percentage) of Ctrl (white) and BR – (gray) infected embryos after pharmacological treatment with drugs targeting the type-1 or type-2 family of dopamine (DA) receptors (D1R, D2R). Two-way ANOVA P < 0.01; P values after Bonferroni post hoc test are indicated as ** P < 0.01, ns P > 0.05. a , b Data represent the mean and S.D. of three independent replicates of 30 embryos. Each replicate is shown by one dot. See also Supplementary Fig. . c , d Drawings represent the morphological and molecular events occurs in response to infection, in presence ( c ) or absence ( d ) of brain. Unique neural- and immune-related pathways are indicated in blue. Upregulation and downregulation of transcripts are indicated in black. E. coli is represented by red circles; apoptotic cells are represented by cyan circles; mmp7 + cells (early macrophages) are represented by pink stars; dopamine-activated macrophages are represented with a D on the pink star. Complete list of genes is presented in Supplementary Data . Green = down gene, Red = up gene. All measured genes found in a pathway are located in Supplementary Data .

Article Snippet: We used specific agonists and antagonists of each dopamine receptor: 10 μM SKF-38393, D1R agonist (SKF, Tocris 0922); 10 μM SCH-23390, D1R antagonist (SCH, Tocris 0925); 10 μM Quinpirole, D2R agonist (Quin, Tocris 1061); 1 μM L-741,626, D2R antagonist (L741, Tocris 1003).

Techniques: Concentration Assay, Infection, Control, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy