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11 protocols using atomoxetine

1

Cocaine Seeking Modulated by Progesterone and Atomoxetine

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Cocaine HCl, supplied by the National Institute of Drug Abuse (Research Triangle Institute, Research Triangle Park, NC), was diluted in 0.9% NaCl (saline; SAL) to a concentration of 1.6 mg/ml and then refrigerated. Heparin (5 USP/ml) was added to the cocaine solution to aid catheter patency. The syringe pump in the experimental chamber delivered 0.025 ml/s, and the duration of the pump was set to provide 0.3 mg/kg/infusion. Progesterone (Sigma-Aldrich, St. Louis, MO) was dissolved to 0.625 mg/ml in peanut oil (USP; Sigma-Aldrich) as a vehicle (VEH) and administered subcutaneously (SC) at a 0.5 mg/kg dose. Atomoxetine (Sigma-Aldrich) was dissolved in saline (SAL; 30 mg/ml) and delivered intraperitoneally (IP) at a 1.5 mg/kg dose. PRO, ATO, and control treatments (VEH or SAL) were administered approximately 30 min prior to the start of experimental sessions. The PRO dose was based on previous work that found it was effective in reducing cocaine and cue-primed reinstatement as well as reducing breakpoints for cocaine on a progressive ratio schedule (e.g., Larson et al. 2007 (link); Zlebnik et al. 2014 (link)). The ATO dose was selected based on previous research showing it reduced cocaine seeking (Swavle et al. 2016 ; Zlebnik and Carroll 2014 (link)) and impulsive choice at doses of 1–2 mg/kg (e.g., Bizot et al. 2011 (link); Robinson et al. 2008 ).
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2

T-Maze Spontaneous Alternation Assay

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T-maze spontaneous alternation is an established assay to assess cognitive performance in rodents [40 (link)]. The alternation performance is drastically reduced with administration of drugs such as Scopolamine and this effect is reversible with cognitive-enhancing [41 (link), 42 (link)] drugs. The T-maze assay and apparatus has been described previously [43 (link)]. Seven groups of mice (n = 10) were used: (1) Vehicle + Saline; (2) Vehicle + Scopolamine; (3) Scopolamine + Donepezil (0.3 mg/kg); (4) Scopolamine + Atomoxetine (3 mg/kg); (5) Scopolamine + Moxonidine (0.1 mg/kg); (6) Scopolamine + Moxonidine (0.3 mg/kg) and (7) Scopolamine + Moxonidine (1 mg/kg). Scopolamine, Atomoxetine, and Donepezil were used at 1, 3, and 0.3 mg/kg, respectively. All compounds were dissolved in saline and were administered with Scopolamine using i.p. injection 30 min prior to the assay except for donepezil which was administrated p.o. 60 min prior to assay. (Scopolamine and Atomoxetine: Sigma-Aldrich, Saint-Quentin-Fallavier, France; Donepezil: Tocris Biotechene, Noyal Chatillon sur Seiche, France).
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3

Operant Conditioning and Social Interaction Protocol

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Testing occurred in operant conditioning chambers from Med Associates (St. Albans, VT). The relevant components of these chambers were a house light, one response lever, and an attached social compartment. The social compartment was located directly opposite the lever and separated from the main chamber by a guillotine door that could be raised to allow social contact between the two rats. A metal screen was affixed in the opening between the two chambers that permitted visual, auditory, olfactory, and limited tactile contact between the two rats, but prevented each rat from traversing from one compartment to the other (see description in Venniro and Shaham, 2020 (link)). A white noise machine was also used throughout training and testing.
Cocaine, d-amphetamine, MDMA, and WIN-35,428 were generously supplied by the National Institute on Drug Abuse (Research Triangle Institute, Research Triangle Park, NC, United States). Fluoxetine and atomoxetine were purchased from Sigma-Aldrich (St. Louis, MO, United States) and Tocris Bioscience (Minneapolis, MN, United States), respectively. All drugs were dissolved in sterile saline for injection.
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4

Esmolol Modulates Audiogenic Seizures in DBA/1 Mice

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The susceptibility of DBA/1 mice to S-IRA was confirmed 24 h before treatment with atomoxetine or vehicle. For intraperitoneal (IP) administration, atomoxetine (Ca #Y0001586, Sigma-Aldrich) and esmolol (H19991058; Qilu Pharmaceutical Co., Ltd.) were dissolved in saline. DBA/1 mice received either atomoxetine (15 mg/kg, treatment group) or saline (vehicle control group) by IP injection 120 min prior to acoustic stimulation. The mice in both groups were then treated (115 min later) with different dosages of esmolol (25 or 50 mg/kg) or vehicle 5 min before acoustic stimulation (n = 5–8 mice per group). The mice were video recorded for offline analysis of the incidence of S-IRA, latency to AGSZs, duration of wild running, clonic seizures, tonic-clonic seizures, and seizure scores, as described previously.35 (link),36 (link),37 (link) To observe the incidence of death among DBA/1 mice as well as heart rate changes due to esmolol, the group of mice (n = 6) used to confirm S-IRA 24 h before the start of the experiment was pre-treated with esmolol (50 mg/kg, IP) without administration of atomoxetine and subjected to acoustic stimulation in the same manner. For DBA/1 mice in all of the different pre-treatment groups, ECG was performed before acoustic stimulation and after S-IRA.
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5

Atomoxetine Modulation of Seizure Response

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A vehicle control group and treatment groups with different dosages of Atomoxetine were included in testing of S-IRA in acoustic stimulation model and PTZ model, respectively. S-IRA was confirmed 24 hr before Atomoxetine or vehicle administration in the acoustic stimulation model. Atomoxetine (1-50 mg/kg for acoustic stimulation model or 5-15 mg/kg for PTZ model) or vehicle was administered IP in DBA/1 mice 2 hr prior to acoustic stimulation or IP injection of PTZ. The occurrence of S-IRA was videotaped for offline analysis. Atomoxetine (Cat # Y0001586; Sigma-Aldrich) was dissolved in saline for IP administration.
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6

Osteogenic Differentiation of Cells With ADHD Drugs

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The ADHD medication modafinil (11.2 µg/mL), atomoxetine (0.9 µg/mL), and guanfacine (17.7 ng/mL) was ordered from Sigma-Aldrich (Steinheim, Germany) and used in max. therapeutic plasma concentration [15 (link),16 (link),17 (link)]. The medication of modafinil solution (1 mg/mL), atomoxetine hydrochloride solution (1 mg/mL) and guanfacine hydrochloride (500 mg) was purchased from Sigma-Aldrich (Steinheim, Germany) and dissolved in sterile DMSO. The osteogenic differentiation medium consisting of ascorbic acid-2-phosphate (200 µM, Cayman Chemical, Ann Arbor, MI, USA), dexamethasone (0.1 µM) and ß-glycerol phosphate (10 mM, Merck, Germany) was used. 10% DMSO was purchased from Merck, Germany.
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7

Neurochemical Modulation of Gut Microbiome

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A group of mice was injected with the NE reuptake inhibitor atomoxetine (Sigma, 0.1 mg kg−1) and the α2-adrenergic receptor antagonist yohimbine (Sigma, 1 mg kg−1) administered intraperitoneally (0.2 ml/mouse in total), once daily for three subsequent days. Another group of mice received a single intraperitoneal injection of 6-hydroxydopamine (6-OHDA) (0.2 ml, Sigma, 100 mg kg−1) followed by 0.2 ml sterile saline for two subsequent days. Control mice were administered 0.2 ml saline daily for three days. Mice were sacrificed 72 h after the first injection; the caecum was quickly removed and was kept at −80 °C until use.
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8

Cell Culture Protocol for Neural Cells

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Dulbecco's modified Eagle medium, penicillin/streptomycin, and glutamine were purchased from Gibco. Trypsin was obtained from Biochrom AG, Berlin, Germany. DNAse 1 was obtained from Invitrogen, Carlsbad, Germany; fetal calf serum was obtained from HyClone, Perbio Science, Bonn, Germany. Poly-L-ornithine was purchased from Sigma-Aldrich, Schnelldorf, Germany. Atomoxetine and all other chemicals were obtained from Sigma-Aldrich Chemie GmbH, Steinheim, Germany.
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9

Investigating Attention in Mice

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Methylphenidate (MPH, Cat # M325880, Toronto Research Chemicals) and Atomoxetine (ATX, Cat # Y0001586; Sigma-Aldrich) were dissolved in 0.9% (w/v) sodium chloride, administered intraperitoneally, 30 minutes prior to testing in an injection volume of 10 ml/kg and dose of 3 mg/kg. The doses of MPH and ATX were selected based on previous touchscreen studies that assessed attention in C57BL/6J mice [31] .
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10

Atomoxetine Effects on Rat Cognition

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During the test phase, the rats were tested twice a week (Tuesday and Friday). On the day prior to testing, rats were trained using the stimulus duration of the final training level to guarantee optimal performance of the animals. In the test phase, the rats were treated with atomoxetine. On testing days, the rats were injected with saline (control) or atomoxetine at a dose of 0.1, 0.5 or 1 mg/kg body weight (Sigma-Aldrich, Schnelldorf, Germany) 20 min prior to testing. atomoxetine doses were used on the basis of previous findings (Bari et al. 2009; Jentsch et al. 2009; Koda et al. 2010; Liu et al. 2015; Navarra et al. 2008; Robinson et al. 2008) . atomoxetine was dissolved in saline, and the volume injected was determined individually (1 ml/kg body weight). Both atomoxetine and saline were administered intraperitoneally. Each rat received one dose of atomoxetine twice a week and was exposed to all doses and the control condition. The testing period lasted 4 weeks. Saline and the three atomoxetine doses were administered in a balanced manner in order to avoid systematic carryover effects.
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