(±)-SKF-38393 hydrochloride (Sigma), a specific D1 receptor agonist (Guo et al., 2015 (link)), was injected (18 nl, 10−7 mol l−1) into the CX (n=12). Spontaneous behavior was evaluated 2 h after injection.
Skf 38393 hydrochloride
Manufactured by Merck Group
Sourced in United States, Germany
SKF-38393 hydrochloride is a chemical compound used in laboratory research. It is a selective agonist of the D1 dopamine receptor. The product is intended for use in scientific investigations and experiments.
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14 protocols using skf 38393 hydrochloride
1
Selective D1 Receptor Agonist Regulates Spontaneous Behavior
2
Pharmacological Agents for Neuroscience Research
3
Chronic Effects of D1 Receptor Modulation and Estrogen on Ovariectomized Rats
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D1 receptor agonist, SKF-38393 hydrochloride, purchased from Sigma (D-047, Sigma Chemical Co., USA) and D1 receptor antagonist, SCH-23390, purchased from Sigma (D-054, Sigma Chemical Co., USA) were dissolved in sterile saline (0.9%). The estrogen 17β-E2, purchased from Sigma (E-8875, Sigma Chemical Co., USA), was dissolved in sterile sesame oil. All solutions were freshly prepared before each experimental series. Rats received SKF-38393 in dose of 0.1 mg/kg, i.p., SCH-23390 in dose of 0.1 mg/kg, i.p., and 17β-E2 in dose of 5.0 μkg/rat, s.c. All preparations were chronically injected for 14 days once daily. Drugs were administered in 2 weeks after postoperative period following ovariectomy. Saline was injected i.p. to control 1 (intact rats) with the same procedure. Control 2 (OVX rats) received oil solvent injection in the same volume. All animals were gently handled by experienced keepers from the facility each day for a week prior to experimental procedures. Any environmental or physical stress was avoided in order to habituate the rats to manipulation. Rats subjected to drug or saline administration received an injection volume of 0.1 mL. All behavioral experiments were carried out in 45 min after the last injection of drug.
4
Evaluating cAMP and CREB Signaling
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For cAMP and CREB experiments, primary samples or AML cell lines were exposed to small molecules including (±)-SKF-38393 hydrochloride, R(+)-SCH-23390 hydrochloride, Thioridazine (all sourced from Sigma), Forskolin (Abcam), or antibody for 30 minutes in serum-free conditions, followed by cell lysis in HCL 1N. The supernatant containing cAMP was collected and applied to cAMP direct immunoassay kit (Millipore/Calbiochem) as per the manufacturer’s instructions.
5
In Vitro Immune Cell Activation
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Bovine serum albumin (BSA) and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) were purchased from Sigma, Italy. RPMI 1640, heat-inactivated fetal bovine serum (FBS), glutamine, and penicillin/streptomycin were obtained from Euroclone, Italy. Ficoll-Paque Plus was from Pharmacia Biotech (Uppsala, Sweden). Purified mouse ab anti-human CD3 (code 555330, clone UCHT1, Mouse IgG1, κ) and purified mouse ab anti-human CD28 (code 555726, clone CD28.2, Mouse IgG1, κ) were obtained from Becton Dickinson, Italy. (±)SKF-38,393 hydrochloride (cod. D047), R(+)7-OH-DPAT hydrobromide (code H168), PD-168,077 maleate (code P233), pramipexol dihydrochloride (code A1237), and dopamine hydrochloride (code H8502) were all from Sigma, Italy. Human recombinant α-synuclein and its fibrillar form were a kind gift from Dr. Lars Kjær and Dr. Daniel Otzen (iNANO - Interdisciplinary Nanoscience Center, Aarhus University, Aarhus, Denmark), and were prepared as published before55 (link).
6
Electrophysiological Recordings in Brain Slices
7
Intracerebral Infusions for Behavioral Studies
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Infusions were delivered through an injector cannula extending 1 mm beyond the tip of the guide cannula. The volume infused was 1 μl per side and the infusion rate was 0.5 μl/min. The injector was left in place for an additional minute after infusion to allow diffusion and to prevent reflux. Doses were as follow: emetine 50 μg/μl, muscimol 0.1 μg/μl, SCH23390 hydrochloride 1.5 μg/μl or SKF38393 hydrochloride 12.5 μg/μl (Sigma Aldrich, St Louis, MO). Drugs were dissolved in sterile 0.9% saline, except for SKF38393 hydrochloride which was dissolved in 10% DMSO. The doses utilized were determined based on previous studies showing the effect of each compound on learning or behavioral performance (Majchrzak and Di Scala, 2000 (link); Lima et al., 2009 (link); Kramar et al., 2014 (link)).
8
Pharmacological Manipulation of Dopamine Signaling
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Dopamine dihydrochloride (Sigma-Aldrich, St Louis, MO), SCH23390 (Tocris-Bioscience, Bristol, UK), (S)-(–)-sulpiride (Sigma-Aldrich), and SKF38393 hydrochloride (Sigma-Aldrich) were dissolved at 10 mM in water and stocked at 4°C. Immediately before use, they were diluted to the final concentration with aCSF containing (in mM): 127 NaCl (Nacalai Tesque, Kyoto, Japan), 1.6 KCl (Wako, Tokyo, Japan), 1.24 KH2PO4 (Nacalai Tesque), 1.3 MgSO4 (Nacalai Tesque), 2.4 CaCl2 (Wako), 26 NaHCO3 (Wako), and 10 d -glucose (Wako).
9
Pharmacological Manipulation of Neuronal Signaling
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The D1/D5 receptor agonist SKF‐38393 hydrochloride [(±)‐1‐Phenyl‐2,3,4,5‐ tetrahydro‐(1H)‐3‐benzazepine‐7,8‐diol hydrochloride] (SKF; #D047; Sigma‐Aldrich, Singapore) was stored at −20 °C as a 50‐mm stock in deionised water. The stocks were used within a week. The Zn2+ chelator TPEN [N,N,N′,N′‐Tetrakis (2‐pyridylmethyl) ethylenediamine] (Tocris Bioscience, Bristol, UK) was stored as a 25‐mm stock in dimethyl sulfoxide (DMSO) at −20 °C. Emetine dihydrochloride hydrate (Sigma‐Aldrich, Singapore) and d ‐2‐amino‐5‐phosphonopentanoic acid (AP‐5) (Tocris Bioscience, Bristol, UK) were prepared as concentrated stock solutions in DMSO and were diluted in ACSF to obtain a final concentration of 20 and 50 μm , respectively. Light‐sensitive drugs were protected from light during storage and bath application. Prior to application, the drug stocks were diluted to the final concentration in ACSF, equilibrated with carbogen, and bath‐applied for specified durations. Whenever the stocks are prepared in DMSO, the final DMSO concentration was kept below 0.1%, a concentration which has been shown to not affect basal synaptic responses (Navakkode et al., 2005 ).
10
Pharmacological Reagents for Neuroscience Research
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The dr ugs (±)-isoprenaline tar trate, dopamine hydrochloride, (±)-SKF 38393 hydrochloride, and R(+)-SCH23390 hydrochloride were purchased from Sigma-Aldrich (Germany). All other chemicals were of the highest purity grade commercially available. Deionized water was used throughout the experiments. Stock solutions were prepared fresh daily.
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