Dorsal pancreatic buds were dissected on E13.5 under a stereoscope and cultured on 0.4 μm pore‐sized filters (Millicell‐CM; Millipore) in DMEM (Gibco) supplemented with N2 (Gibco) and streptomycin–penicillin–glutamine (Gibco) (Serafimidis et al, 2017 ). Equivalent to E14.5, E15.5, and E16.5 (i.e., 1–3 days in vitro (DIV)), explants were transferred to fresh medium that had optionally been supplemented with amphetamine (10 μM), cocaine (10 μM), methamphetamine (10 μM), or 5‐HT (0.5 μM; Tocris). On DIV4, all explants were transferred to fresh medium and cultured for 2 more days. Culture media were then collected with explants either fixed for immunohistochemical analysis or snap frozen for gene expression profiling. Quantitative analysis was performed using ImageJ1.45 with appropriate plug‐ins on n = 4–6 explants/group.
Amphetamine
Manufactured by Bio-Techne
Sourced in Canada, United States
Description not available
Automatically generated - may contain errors
Lab products found in correlation
Tyramine, by Merck Group (2 mentions)
Millicell cm, by Merck Group (1 mentions)
Glutamine penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Alprenolol, by Merck Group (1 mentions)
Ephedrine, by Merck Group (1 mentions)
Fluoxetine, by Bio-Techne (1 mentions)
Tryptamine, by Merck Group (1 mentions)
Isoproterenol, by Merck Group (1 mentions)
Indole, by Merck Group (1 mentions)
β pea, by Merck Group (1 mentions)
Bupropion hydrochloride, by LGC (1 mentions)
Evodiamine, by Merck Group (1 mentions)
Dopamine, by Merck Group (1 mentions)
Water purification system, by Merck Group (1 mentions)
Potassium chloride (kcl), by Merck Group (1 mentions)
Caffeine, by Merck Group (1 mentions)
Sodium phosphate dibasic, by Merck Group (1 mentions)
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
Potassium phosphate monobasic, by Merck Group (1 mentions)
4 protocols using amphetamine
1
Pancreatic Bud Cultures Exposed to Psychoactive Drugs
2
Synthesis and Characterization of Psychoactive Compounds
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Bupropion hydrochloride and 6-hydroxybupropion were obtained from Toronto Research Chemicals (Toronto, Canada), ibogaine from Ibogaworld, cocaine from Medisca (Montreal, Canada), β-PEA, pinoline, indole, tryptamine, ephedrine, frovatriptan, alprenolol, isoproterenol, evodiamine, and tyramine from Sigma (Oakville, Canada), fluoxetine and amphetamine were from Tocris Bioscience (Bristol, United Kingdom). PAL analogs, RTI analogs, and bicifadine were synthesized at RTI in the Blough laboratory as described previously (Carroll et al., 2009 (link), 2011 (link); Blough et al., 2011 , 2014 (link)). Noribogaine (Batch No. 606950002) was from Deborah Mash, Ph.D. (Obach et al., 1998 ). Ibogamine was obtained from Specs (Zoetermeer, The Netherlands). Sodium phenylbutyrate was purchased from Enzo Life Sciences Inc. (Farmingdale, NY). Ibogaminalog, ibogainealog, noribogainalog, fluorogainalog, and tabernanthalog were synthesized in the lab of David E. Olson (Cameron et al., 2020 (link)).
3
Detailed Procedure for Electrochemical Analysis
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All commercial reagents were of analytical grade and were used without further purification. All aqueous solutions were prepared using ultrapure water with resistivity not less than 18.2 MΩ cm at 298 K (Millipore water purification system, Burlington, MA, USA). Potassium hexacyanoferrate (III), potassium hexacyanoferrate (II) trihydrate, o-PD, dopamine, caffeine, tyramine, and potassium chloride (KCl) were obtained from Sigma-Aldrich, St. Louis, MO, USA and amphetamine from Tocris Bioscience, Bristol, UK. Phosphate buffered saline (PBS), 0.17 mol L–1, pH 7.4, was prepared using sodium phosphate dibasic and potassium phosphate monobasic, all obtained from Sigma-Aldrich. MDPV hydrochloride was purchased online (www.sensearomatics.eu ) and was fully characterized by MS and elemental analysis (shown in the supplementary data of a recent publication [63 (link)]). The analytical data were consistent with the expected structure, with the salt having a purity of 99.5%, the salt consisted in a racemic (R and S enantiomers) mixture.
4
Amphetamine-Induced Hyperlocomotion in Mice
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Following a 180-min habituation period in the activity cage, mice received an i.p. injection of vehicle (saline, 10 mL/Kg) and were monitored for another 60 min. Afterwards, the procedure was repeated but, instead of saline, animals were challenged with amphetamine (3 mg/kg; Tocris Bioscience). The hyperlocomotion response to the drug was recorded for 180 min. After a three week wash-out period, the experiment was repeated with a lower dose of amphetamine (0.75 mg/kg).
In another set of experiments, ORX + placebo-treated mice were subjected to the former procedure but, prior to the challenge with 3 mg/kg amphetamine, they received either intranasal T (2 mg/kg, 12 µL per mouse) or vehicle (sesame oil). A subgroup of animals was euthanized 25 min after T or vehicle administration and the levels of T in serum and brain extracts were determined by liquid chromatography tandem mass spectrometry (LC-MS/MS).
In another set of experiments, ORX + placebo-treated mice were subjected to the former procedure but, prior to the challenge with 3 mg/kg amphetamine, they received either intranasal T (2 mg/kg, 12 µL per mouse) or vehicle (sesame oil). A subgroup of animals was euthanized 25 min after T or vehicle administration and the levels of T in serum and brain extracts were determined by liquid chromatography tandem mass spectrometry (LC-MS/MS).
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