Skf81297 hydrobromide

Manufactured by Bio-Techne

Sourced in United Kingdom

SKF81297 hydrobromide is a chemical compound that acts as a selective dopamine D1 receptor agonist. It is commonly used in research applications to study the effects of D1 receptor activation on various biological processes. The compound has a molecular formula of C17H23BrN2O and a molecular weight of 383.30 g/mol.

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SKF81297 (44 citations)

18 protocols using skf81297 hydrobromide

Seizure Behavior Evaluation in Mice

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Mice were ad random divided over experimental groups. Mice received one of the following treatments 30 min prior to SKF81297 injection: saline, JMV-1843, JMV-2959 (provided by Prof. Fehrentz), or YIL781 hydrochloride (Tocris, Bristol, UK). All drugs were dissolved in 0.9 % (w/v) NaCl (5 mg/kg; body volume of 10 mL/kg) and administered i.p. The D1R agonist SKF81297 hydrobromide (5 mg/kg i.p., Tocris) was dissolved in 0.9 % (w/v) NaCl and administered according to the treatment regimen previously described (Figure 1A) [32 (link)].
After SKF81297 administration, animals were evaluated for behavioral seizures during 45 min and assigned a score based on the following observations: 1 = transient disruption of exploratory behavior by tonic immobility/rigidity; 2 = rearing with forepaw myoclonus; 3 = generalized clonus; 4 = tonic–clonic seizure or rapid jumping and wild running [32 (link)]. All conducting researchers were blinded for treatment.

Buckinx A., Van Den Herrewegen Y., Pierre A., Cottone E., Ben Haj Salah K., Fehrentz J.A., Kooijman R., De Bundel D, & Smolders I. (2019). Differential Effects of a Full and Biased Ghrelin Receptor Agonist in a Mouse Kindling Model. International Journal of Molecular Sciences, 20(10), 2480.

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Electrophysiology Experiments: Pharmacological Agents

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All chemicals used for electrophysiology experiments were obtained from Sigma-Aldrich except D-L-AP5 and NBQX (from Ascent Scientific). The D1 Receptor agonist SKF 81297 hydrobromide (6-chloro-2,3,4,5-tetrahydro-1-phenyl-1H-3-benzazepine hydrobromide) and the D1R antagonist SCH 23390 hydrochloride ((R)-(+)-7chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine hydrochloride) were from Tocris. SKF81297 and SCH23390 were dissolved in water at a 10 mM concentration for stock solutions. Drugs were bath applied for 15 minutes before a “drug recording”.

Meunier C.N., Callebert J., Cancela J.M, & Fossier P. (2015). Effect of Dopaminergic D1 Receptors on Plasticity Is Dependent of Serotoninergic 5-HT1A Receptors in L5-Pyramidal Neurons of the Prefrontal Cortex. PLoS ONE, 10(3), e0120286.

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Pharmacological Modulation of Neural Signaling

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All drugs were purchased from Sigma Aldrich (St. Louis, MO), or Tocris (Bristol, UK). SKF 81297 hydrobromide (Tocris, 1447) was applied either as puff (500 µM) or flow in (10 µM) depending on experiment type. All other drugs were bath applied: SCH-39166 hydrobromide (Tocris,2299), SCH-23390 hydrochloride (Tocris, 0925), SKF-83566 hydrobromide (Tocris, 1586), Strychnine (Sigma-Aldrich, 8753), Tetraethylammonium chloride (Sigma-Aldrich, 86614), TTX (Tocris Bioscience, 1069), (RS)-CPP (Tocris Bioscience, 0173), NBQX (Tocris Bioscience, 0373), SR 95531 hydrobromide (Tocris Bioscience, 1262), MNI-caged-L-glutamate (Tocris Biosciences 1490), L-741,626 (Tocris Bioscience, 1003), Pyr-3 (Tocris,3753), prazosin (Sigma-Aldrich, P7791), propranolol (Sigma-Aldrich, 40543), and quinpirole hydrochloride (Tocris Bioscience, 1061). For experiments requiring pharmacological agents dissolved in DMSO the concentration never exceeded 0.02% DMSO.

Canton-Josh J.E., Qin J., Salvo J, & Kozorovitskiy Y. (2022). Dopaminergic regulation of vestibulo-cerebellar circuits through unipolar brush cells. eLife, 11, e76912.

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Neurotransmitter Stock Solution Preparation

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Dopamine (DA) hydrochloride (1 M stock, H8602, Sigma-Aldrich), serotonin hydrochloride (50 mM stock, 14332, CAY), and L-adrenaline (epinephrine) (5 mM stock, A0173, TCI) were dissolved in 10 mM HCl. L-noradrenaline bitartrate monohydrate (1 M stock, A0906, TCI), sodium L-glutamate monohydrate (10 mM stock, G0188, TCI), 4-aminobutyric acid (100 mM stock, A0282, TCI), histamine (100 mM stock, 18111–71, Nacalai Tesque), acetylcholine chloride (10 mM stock, A6625, Sigma-Aldrich), R( +)-SCH 23390 hydrochloride (10 mM stock, D054, Sigma-Aldrich), and octopamine hydrochloride (10 mM stock, O0413, TCI) were each dissolved separately in distilled water. SKF 81297 hydrobromide (10 mM stock, 1447, TOCRIS), haloperidol hydrochloride (20 mM stock, 0931, TOCRIS), yohimbine hydrochloride (20 mM stock, 1127, TOCRIS), and tyramine (10 mM stock, A0302, TCI) were dissolved in DMSO. Compound solutions were then subdivided into aliquots and stored at − 20 °C until use. A working solution of 1 M DA was stored at 4 °C for 3 weeks prior to use.

Nakamoto C., Goto Y., Tomizawa Y., Fukata Y., Fukata M., Harpsøe K., Gloriam D.E., Aoki K, & Takeuchi T. (2021). A novel red fluorescence dopamine biosensor selectively detects dopamine in the presence of norepinephrine in vitro. Molecular Brain, 14, 173.

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Dopamine Receptor Agonists Reinstate Cocaine Seeking

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Testing conditions were identical to training except the syringe containing sucrose solution was absent. Rats were randomly assigned to drug dose conditions within each experiment. As rats had already been randomly assigned to treatment conditions, this resulted in cohorts of rats (typically 8–12 subjects) each assigned to one of a variety of testing conditions. Drugs were stored at 4 °C in the dark and used within 48 h of preparation. SKF 81297 hydrobromide (Tocris Bioscience, Minneapolis, MN, USA) and (−) quinpirole hydrochloride (R & D Systems, Minneapolis, MN, USA) were dissolved in sterile 0.9 % saline and administered according to body weight (1 ml/ kg) to provide the doses indicated in Figs. 1–4. Doses were selected from the lowest range of each drug that has been shown to reinstate extinguished cocaine seeking in rats (Self et al. 1996 (link); Alleweireldt et al. 2002 (link)). Rats received handling injections of saline (1 ml/kg) in the vivarium in the afternoon during the 2 days prior to testing. Testing day injections occurred immediately prior to testing in the room containing the operant conditioning chambers. SKF 81297 was administered SC; quinpirole, IP.

Glueck E., Ginder D., Hyde J., North K, & Grimm J.W. (2016). Effects of dopamine D1 and D2 receptor agonists on environmental enrichment attenuated sucrose cue reactivity in rats. Psychopharmacology, 234(5), 815-825.

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Live-Cell Imaging of GPCR Endocytosis

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Live-cell imaging was carried out using Nikon spinning disk confocal microscope with a ×60, 1.4 numerical aperture, oil objective and a CO₂ and 37°C temperature-controlled incubator. A 488, 568 nm and 640 Voltran was used as light sources for imaging GFP, mRFP/mApple, and Snap-647 signals, respectively. Cells expressing both Snap-tagged receptor (2 μg) and the indicated nanobody–GFP (200 ng) were plated onto glass coverslips. Receptors were surface labeled by addition of Snap-Cell 647 SiR (1:1000, New England Biolabs) to the media for 20 min, as described previously. Live-cell images where endocytosis was inhibited were carried out by incubating the cells in 30 μM Dyngo 4a (ab120689) at 37°C for 30 min before indicated agonist was added. HEK293 PKA-Cat-GFP knock-in cells were a generous gift from the Huang Lab. Indicated agonists (dopamine hydrochloride [Sigma], SKF81297 hydrobromide [Tocris]) were added and cells were imaged every 20 s for 20 min in DMEM without phenol red supplemented with 30 mM HEPES, pH 7.4. NPEC-caged-dopamine (Tocris) was incubated for 10 min before cells were stimulated with 3.2 μW/cm² blue light. Time-lapse images were acquired with a CMOS camera (Photometrics) driven by Nikon Imaging Software (NS Elements).

Puri N.M., Romano G.R., Lin T.Y., Mai Q.N, & Irannejad R. (2022). The organic cation transporter 2 regulates dopamine D1 receptor signaling at the Golgi apparatus. eLife, 11, e75468.

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Dose-Dependent Modulation of Reversal Learning

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All drugs were dissolved in saline and injected via the intraperitoneal route. Raclopride (Tocris Bioscience, Bristol, UK) was administered at 0, 0.015, 0.03, and 0.06 mg/kg, 20 min before testing. SCH39166 hydrobromide (Tocris Bioscience) was administered at 0, 0.025, 0.05, and 0.1 mg/kg, 20 min before testing. Note that SCH39166 was chosen for D1R antagonism to avoid off-target effects at the serotonin 5-HT_2C receptor, since activity at this receptor affects visual reversal learning (Alsiö et al. 2015 (link)). SKF81297 hydrobromide (Tocris Bioscience) was administered at 0, 0.1, and 0.25 mg/kg, 30 min before testing. (−)-Quinpirole hydrochloride (Sigma-Aldrich, St. Louis, MO, USA) was administered at 0, 0.01, 0.025, 0.1, 0.25, and 0.5 mg/kg, 60 min before testing. A broad dose range of quinpirole was chosen to allow detection of potential differential effects of canonical presynaptic and postsynaptic doses (Eilam and Szechtman 1989 (link)). No adverse reactions to repeated injections were observed in any experiments.

Alsiö J., Phillips B.U., Sala-Bayo J., Nilsson S.R., Calafat-Pla T.C., Rizwand A., Plumbridge J.M., López-Cruz L., Dalley J.W., Cardinal R.N., Mar A.C, & Robbins T.W. (2019). Dopamine D2-like receptor stimulation blocks negative feedback in visual and spatial reversal learning in the rat: behavioural and computational evidence. Psychopharmacology, 236(8), 2307-2323.

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CAMP Signaling Pathway Modulation

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8-(4-chlorophenylthio) adenosine-3’,5’-cyclic monophosphate (8-CPT-cAMP) was purchased from BIOLOG Life Science Institute. The inhibitors 9-(tetrahydro-2-furanyl)-9H-purin-6-amine (SQ22536), 1,4-diamino-2,3-dicyano-1,4-bis[2-aminophenylthio]butadiene (U0126), N-[2-[[3-(4-bromophenyl)-2-propenyl-]amino]ethyl]-5-isoquinolinesulfonamide dihydrochloride (H-89), and α-[amino[(4-aminophenyl)thio]methylene]-2-(trifluoromethyl)benzeneacetonitrile (SL327), and D1 receptor ligands SKF81297 hydrobromide and SCH-23390 hydrochloride were purchased from Tocris. Reserpine and phorbol 12-myristate 13-acetate (PMA) were obtained from Sigma. Drug stocks were prepared at 50 or 100 mM in DMSO, followed by dilution in culture media to final concentrations of 100 μM 8-CPT-cAMP, 10 μM Reserpine, 10 μM U0126, 30 µM H89, 1 mM SQ22536, 10 µM SCH-23390, 10 µM SKF 81297, and 100 nM PMA.

Jiang S.Z., Xu W., Emery A.C., Gerfen C.R., Eiden M.V, & Eiden L.E. (2017). NCS-Rapgef2, the Protein Product of the Neuronal Rapgef2 Gene, Is a Specific Activator of D1 Dopamine Receptor-Dependent ERK Phosphorylation in Mouse Brain. eNeuro, 4(5), ENEURO.0248-17.2017.

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Dopaminergic Signaling Modulation Assay

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CNO (Tocris) was first diluted in DMSO (Sigma), then in Dulbecco’s Phosphate Buffered Saline (PBS, Gibco) to a 0.5% DMSO final concentration and injected intraperitoneally at 3 mg/kg 15 min before behavioral testing. Dopamine hydrochloride (Tocris, #3548), SKF81297 hydrobromide (Tocris, #1447) and (−)-Quinpirole hydrochloride (Tocris, #1061) were diluted in ACSF (see below) to respectively 10, 50 and 10 µM, extemporaneously for each day of recording, and kept protected from light for the duration of the bath perfusion.

Godino A., Salery M., Minier-Toribio A.M., Patel V., Fullard J.F., Parise E.M., Martinez-Rivera F.J., Morel C., Roussos P., Blitzer R.D, & Nestler E.J. (2023). Dopaminoceptive D1 and D2 neurons in ventral hippocampus arbitrate approach and avoidance in anxiety. bioRxiv.

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Pharmacological Modulation of Acute Slicing

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For acute slicing experiments, drugs were applied via bath perfusion as previously described^{73 (link)}. Final concentrations are indicated in parentheses: (+)-muscarine-iodide (10 μM) and SKF 81297 hydrobromide (1 μM) were obtained from Tocris. FSK (50 µM) was obtained from either Tocris or Cayman Chemicals.

Tilden E.I., Maduskar A., Oldenborg A., Sabatini B.L, & Chen Y. (2024). A Cre-dependent reporter mouse for quantitative real-time imaging of protein kinase A activity dynamics. Scientific Reports, 14, 3054.

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