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: The Journal of Neuroscience

Article Title: Fear Memory Recall Potentiates Opiate Reward Sensitivity through Dissociable Dopamine D1 versus D4 Receptor-Dependent Memory Mechanisms in the Prefrontal Cortex

doi: 10.1523/JNEUROSCI.3113-17.2018

Figure Lengend Snippet: Schematic summary of experimental timeline and behavioral conditioning procedures combining olfactory associative fear conditioning and morphine CPP assays. Intra-PFC D1R or D4R pharmacological activation was administered immediately before fear memory recall testing or acquisition phases, respectively.

Article Snippet: The selective D4R agonist, PD 168077 (5–50 ng/0.5 μl), the D1R agonist SKF81297 hydrobromide (10–100 ng/0.5 ml; Tocris Bioscience) and ERK 1/2 inhibitor, U0126 (1 μg/0.5 μl) were first dissolved in dimethyl-sulfoxide (DMSO) and then diluted with physiological saline to a final concentration of 5% DMSO in physiological saline.

Techniques: Activation Assay

Journal: The Journal of Neuroscience

Article Title: Fear Memory Recall Potentiates Opiate Reward Sensitivity through Dissociable Dopamine D1 versus D4 Receptor-Dependent Memory Mechanisms in the Prefrontal Cortex

doi: 10.1523/JNEUROSCI.3113-17.2018

Figure Lengend Snippet: Histological analysis of PFC injector placements. A, Representative microphotograph showing a typical intra-PFC injector placement. B, Schematic representation of bilateral intra-PFC injector placements. Figure symbols represent the following experimental groups: ● = PFC VEH controls receiving CS+ presentations during the fear recall test; □ = rats receiving SKF81297 100 ng/0.5 μl; ■ = rats receiving intra-PFC PD168077 50 ng/0.5 μl.

Article Snippet: The selective D4R agonist, PD 168077 (5–50 ng/0.5 μl), the D1R agonist SKF81297 hydrobromide (10–100 ng/0.5 ml; Tocris Bioscience) and ERK 1/2 inhibitor, U0126 (1 μg/0.5 μl) were first dissolved in dimethyl-sulfoxide (DMSO) and then diluted with physiological saline to a final concentration of 5% DMSO in physiological saline.

Techniques:

Journal: The Journal of Neuroscience

Article Title: Fear Memory Recall Potentiates Opiate Reward Sensitivity through Dissociable Dopamine D1 versus D4 Receptor-Dependent Memory Mechanisms in the Prefrontal Cortex

doi: 10.1523/JNEUROSCI.3113-17.2018

Figure Lengend Snippet: Effects of PFC D1R activation on the recall of suprathreshold fear memory and morphine reward CPP behaviors. A, Relative to VEH (n = 6) controls, intra-PFC D1R activation with SKF81297 immediately before the recall test, dose-dependently blocks the recall of suprathreshold fear memories demonstrated by significantly lower levels of associative freezing in rats receiving 10 ng (n = 8) or 100 ng (n = 8) of intra-PFC SKF81297. B, Blockade of suprathreshold fear memory recall with SKF81297 similarly blocks the potentiation of subthreshold morphine reward CPP, demonstrated by attenuated morphine environment preferences in rats receiving 10 or 100 ng intra-PFC SKF81297. **p < 0.01.

Article Snippet: The selective D4R agonist, PD 168077 (5–50 ng/0.5 μl), the D1R agonist SKF81297 hydrobromide (10–100 ng/0.5 ml; Tocris Bioscience) and ERK 1/2 inhibitor, U0126 (1 μg/0.5 μl) were first dissolved in dimethyl-sulfoxide (DMSO) and then diluted with physiological saline to a final concentration of 5% DMSO in physiological saline.

Techniques: Activation Assay

Journal: The Journal of Neuroscience

Article Title: Fear Memory Recall Potentiates Opiate Reward Sensitivity through Dissociable Dopamine D1 versus D4 Receptor-Dependent Memory Mechanisms in the Prefrontal Cortex

doi: 10.1523/JNEUROSCI.3113-17.2018

Figure Lengend Snippet: Effects of PFC D4R activation on the acquisition of subthreshold fear memory and morphine reward CPP behaviors. A, Relative to VEH controls (n = 9), intra-PFC D4R activation with PD 168077 during fear memory acquisition, dose-dependently potentiates the formation of subthreshold fear memories demonstrated by significantly higher levels of associative freezing in rats receiving 50 ng intra-PFC PD 168077 (n = 7) but not a lower dose of 5 ng (n = 6). This effect is blocked by intra-PFC D1R activation with SKF 81297 (100 ng) immediately before recall testing (n = 6). B, Potentiation of normally subthreshold fear memory with PD 168077 similarly potentiated the reward salience of normally sub-reward threshold conditioning doses of morphine (0.05 mg/kg, i.p.) in these same groups. Again, this effect is also blocked when fear memory recall is prevented with intra-PFC SKF 81297 immediately before recall testing. *p < 0.05, **p < 0.01.

Article Snippet: The selective D4R agonist, PD 168077 (5–50 ng/0.5 μl), the D1R agonist SKF81297 hydrobromide (10–100 ng/0.5 ml; Tocris Bioscience) and ERK 1/2 inhibitor, U0126 (1 μg/0.5 μl) were first dissolved in dimethyl-sulfoxide (DMSO) and then diluted with physiological saline to a final concentration of 5% DMSO in physiological saline.

Techniques: Activation Assay

Journal: The Journal of Neuroscience

Article Title: Fear Memory Recall Potentiates Opiate Reward Sensitivity through Dissociable Dopamine D1 versus D4 Receptor-Dependent Memory Mechanisms in the Prefrontal Cortex

doi: 10.1523/JNEUROSCI.3113-17.2018

Figure Lengend Snippet: VTA histological analysis and effects of intra-PFC D1R or D4R activation on fear memory processing and intra-VTA morphine reward CPP behaviors. A, Microphotograph showing typical intra-VTA injector location. B, Schematic summary of bilateral intra-VTA cannulae injector locations; ○ = intra-PFC VEH; ● = intra-PFC PD168077 (50 ng/0.5 μl); ■ = intra-PFC SKF81297 (100 ng/0.5 μl). C, Intra-PFC SKF81297 administration immediately before fear memory recall significantly attenuates freezing behaviors. D, Relative to VEH controls (n = 7), rats receiving intra-PFC SKF81297 (n = 8) do not display morphine CPP. E, Intra-PFC PD168077 (n = 7) administration during fear memory acquisition significantly potentiates subthreshold associative fear memory and F, potentiates intra-VTA morphine reward CPP relative to VEH controls (n = 8). **p < 0.01.

Article Snippet: The selective D4R agonist, PD 168077 (5–50 ng/0.5 μl), the D1R agonist SKF81297 hydrobromide (10–100 ng/0.5 ml; Tocris Bioscience) and ERK 1/2 inhibitor, U0126 (1 μg/0.5 μl) were first dissolved in dimethyl-sulfoxide (DMSO) and then diluted with physiological saline to a final concentration of 5% DMSO in physiological saline.

Techniques: Activation Assay

Journal: The Journal of Neuroscience

Article Title: Fear Memory Recall Potentiates Opiate Reward Sensitivity through Dissociable Dopamine D1 versus D4 Receptor-Dependent Memory Mechanisms in the Prefrontal Cortex

doi: 10.1523/JNEUROSCI.3113-17.2018

Figure Lengend Snippet: Effects of PFC D1R activation on total and phosphorylated ERK 1/2 levels in the PFC and effects of intra-PFC ERK inhibition on PFC D1R modulation of fear and morphine reward behaviors. A, Sample Western blots showing PFC pERK 1/2 bands relative to loading controls in intra-PFC VEH (n = 6), SKF 81297 (100 ng; n = 6), or PD 168077 (50 ng; n = 5) -treated rats. B, C, Normalized densitometry values comparing average levels of pERK 1 or 2 following intra-PFC VEH, SKF 81297, or PD 168077 administration. D, Sample Western blots showing PFC tERK 1/2 bands relative to loading controls in intra-PFC VEH, SKF 81297 (100 ng), or PD 168077 (50 ng) -treated rats. E, F, Normalized densitometry values comparing average levels of tERK 1 or 2 following intra-PFC VEH, SKF 81297, or PD 168077 administration. G, Coadministration of SKF 81297 (100 ng) with the MEK 1/2 inhibitor U0126 (1 μg; n = 6) reverses the effects of intra-PFC D1R activation on blockade of fear memory recall, with U0126 having no effect on its own (n = 8), relative to SKF 81297 (100 ng) on its own (n = 7). H, Similarly, reversing the block of D1R-mediated memory recall with U0126 restores the potentiation of sub-reward threshold morphine reward CPP in these same groups. Rats receiving U0126 alone showed a normal potentiated morphine CPP. *p < 0.05, **p < 0.01.

Article Snippet: The selective D4R agonist, PD 168077 (5–50 ng/0.5 μl), the D1R agonist SKF81297 hydrobromide (10–100 ng/0.5 ml; Tocris Bioscience) and ERK 1/2 inhibitor, U0126 (1 μg/0.5 μl) were first dissolved in dimethyl-sulfoxide (DMSO) and then diluted with physiological saline to a final concentration of 5% DMSO in physiological saline.

Techniques: Activation Assay, Inhibition, Western Blot, Blocking Assay

Journal: Frontiers in Behavioral Neuroscience

Article Title: Activation of D1/5 Dopamine Receptors in the Dorsal Medial Prefrontal Cortex Promotes Incubated-Like Aversive Responses

doi: 10.3389/fnbeh.2017.00209

Figure Lengend Snippet: Activation of mPFC D1/5R after exposure of a neutral, context-independent paradigm did not generate an aversive response. (A) Schema of the modified conditioned taste aversion (CTA) protocol used. (B) Animals were infused with D1R agonist SKF 38393 in the mPFC immediately after being trained on saccharin consumption. Test was performed 7 days later. No significant differences were found between groups ( p > 0.05, n = 6).

Article Snippet: The DA D1/5R agonist SKF 38393 hydrochloride (SKF, 12.5 μg/μl, Sigma-Aldrich), the selective DA D1R agonist SKF 83822 hydrobromide (SKF 83822, 1 μg/μl, Sigma-Aldrich) and the selective DA D5R agonist SKF 83959 hydrobromide (SKF 83959, 10 μg/μl, Sigma-Aldrich) were all dissolved in sterile 0.9% physiological saline supplemented with DMSO (10% final concentration, Ernesto van Rossum y Cia., Buenos Aires, Argentina).

Techniques: Activation Assay, Modification

Journal: Frontiers in Behavioral Neuroscience

Article Title: Activation of D1/5 Dopamine Receptors in the Dorsal Medial Prefrontal Cortex Promotes Incubated-Like Aversive Responses

doi: 10.3389/fnbeh.2017.00209

Figure Lengend Snippet: The aversive effect observed is mediated through the activation of dopamine (DA) receptor D5 in the mPFC. Animals were infused with the selective D1R dopaminergic agonist SKF 83822, with selective dopaminergic agonist SKF 83959 or Vehicle in the mPFC region immediately after the exposure to the white compartment of the apparatus. Test was performed 7 days later. Animals infused with SKF 83959 showed an aversive behavior ( p = 0.047, n = 8–19) while the animals infused with SKF 83822 showed no changes in preference or aversive behavior ( p > 0.05, n = 8–19). * p < 0.05.

Article Snippet: The DA D1/5R agonist SKF 38393 hydrochloride (SKF, 12.5 μg/μl, Sigma-Aldrich), the selective DA D1R agonist SKF 83822 hydrobromide (SKF 83822, 1 μg/μl, Sigma-Aldrich) and the selective DA D5R agonist SKF 83959 hydrobromide (SKF 83959, 10 μg/μl, Sigma-Aldrich) were all dissolved in sterile 0.9% physiological saline supplemented with DMSO (10% final concentration, Ernesto van Rossum y Cia., Buenos Aires, Argentina).

Techniques: Activation Assay

Journal: Acta Neuropathologica Communications

Article Title: Regulation of dopamine neurotransmission from serotonergic neurons by ectopic expression of the dopamine D2 autoreceptor blocks levodopa-induced dyskinesia

doi: 10.1186/s40478-018-0653-7

Figure Lengend Snippet: D2R s -injected animals do not develop severe AIMs with DA agonist treatment. Animals were treated three times each with apomorphine ( a - c ) quinpirole ( d - f ) or SKF-81297 ( g - i ). a - c rAAV-D2R s -injected animals remained AIM resistant with 0.1 m/kg pan-DA agonist apomorphine treatment, while rAAV-GFP animals continued to exhibit dyskinetic behaviors. d - f 0.2 mg/kg quinpirole (D2 agonist) did not elicit AIMs in rAAV-D2R s animals, where rAAV-GFP animals continued to exhibit moderate to severe AIMs. g - i rAAV-D2R s began to show mild-to-moderate AIMs with 0.8 mg/kg of the D1 agonist SKF-81297 treatments, but remained significantly less severe than their rAAV-GFP counterparts. (* = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001)

Article Snippet: The same injection and rating paradigm was used for AIM evaluations with the non-selective DA agonist apomorphine (0.1 mg/kg, R&D Systems, Minneapolis, MN), the D2/D3 receptor agonist quinpirole (0.2 mg/kg, Sigma-Aldrich, St. Louis, MO) and the D1 receptor agonist SKF-81297 (0.8 mg/kg, Sigma-Aldrich, St. Louis, MO).

Techniques: Injection

Journal: PLoS ONE

Article Title: Sodium bicarbonate cotransporter NBCe2 gene variants increase sodium and bicarbonate transport in human renal proximal tubule cells

doi: 10.1371/journal.pone.0189464

Figure Lengend Snippet: NBCe1 protein expression is decreased following an increase in intracellular sodium (↑Na + ) (10 μmol monensin/L, 24 h) (N = 4, *P<0.001 vs VEH, one-way ANOVA, Tukey’s test) in WT SLC4A5 hRPTC. The combination of monensin (↑Na + ) and D 1 R/D 5 R agonist SKF38393 (SKF, 10 μmol/L, 24 h) further decreases NBCe1 protein (N = 4, **P<0.05 vs monensin (↑Na + ), # P<0.001 vs SKF, one-way ANOVA, Tukey’s test) that is blocked by the D 1 -like receptor antagonist LE300 (10 μmol/l, 24 h), which by itself has no effect. NBCe1 protein is not affected by SKF or LE300 in the absence of monensin.

Article Snippet: The response to dopaminergic stimulation (24 h) of hRPTCs was studied using the D 1 -like (D 1 R/D 5 R) dopamine receptor agonists fenoldopam (FEN, fenoldopam mesylate, Corlopam, Hospira, Inc, 1 μmol/L) and SKF38393 (Sigma, 10 μmol/L), and the D 1 R/D 5 R antagonist LE300 (Tocris, 10 μmol/L)[ , ].

Techniques: Expressing

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