Rats were tested in 2 cohorts. With the exception of WAY 100635, which was administered subcutaneously, all drugs targeting the serotonergic system were injected i.p.. Cohort 1 (n = 16) received Citalopram hydrobromide (selective serotonin reuptake inhibitor), M100907 (5-HT2A receptor antagonist), and SB242,084 (5-HT2C receptor antagonist). Citalopram (Tocris Bioscience) was prepared in 0.9% saline. M100907 (Sigma Aldrich) was prepared in 0.9% saline with the pH adjusted to 6.5 with acetic acid. The vehicle for SB 242,084 (Tocris Bioscience) consisted of 240 mg of anhydrous citric acid and 4 g of cyclodextrine dissolved in 50 mL of 0.9% saline to yield an 8% cyclodextrin in 25 mM citric acid solution. The pH of this solution was then adjusted to 6.4 by adding 5 mL of 0.1 M NaOH. Cohort 2 (n = 18) received WAY-100635 (5-HT1A receptor antagonist) followed by 8-OH-DPAT (5-HT1A receptor agonist). WAY-100635 (Sigma Aldrich) was prepared in phosphate-buffered saline (PBS). 8-OH-DPAT (Sigma Aldrich) was prepared in 0.9% saline. The drug doses and timing of injections prior to testing were based on previous studies that used these drugs in comparable behavioral tests (Winstanley et al. 2004 (link)); (Bari et al. 2009 (link)); (Boulougouris, Glennon and Robbins 2008 (link), Zeeb et al. 2009 (link)).
Citalopram
Manufactured by Bio-Techne
Sourced in Spain
Citalopram is a laboratory reagent and analytical standard produced by Bio-Techne. It is a selective serotonin reuptake inhibitor (SSRI) used for analytical and research purposes. The product provides a consistent and reliable source of this compound for various applications in the life sciences industry.
Automatically generated - may contain errors
Lab products found in correlation
Sulpiride, by Bio-Techne (2 mentions)
Quinpirole, by Bio-Techne (2 mentions)
Sumatriptan, by Bio-Techne (2 mentions)
Tamoxetine, by Bio-Techne (2 mentions)
Gbr12783, by Bio-Techne (2 mentions)
Way 100635, by Abcam (2 mentions)
L 745870, by Bio-Techne (2 mentions)
Gabazine, by Abcam (2 mentions)
Kynurenic acid, by Abcam (2 mentions)
8 oh dpat, by Merck Group (1 mentions)
Sb242084, by Bio-Techne (1 mentions)
M100907, by Merck Group (1 mentions)
Way100635, by Merck Group (1 mentions)
Nisoxetine, by Bio-Techne (1 mentions)
3h dopamine, by PerkinElmer (1 mentions)
3h serotonin, by PerkinElmer (1 mentions)
125i rti 55, by PerkinElmer (1 mentions)
Fluvoxamine, by Bio-Techne (1 mentions)
3h norepinephrine, by PerkinElmer (1 mentions)
C57bl 6j mice, by Jackson ImmunoResearch (1 mentions)
6 protocols using citalopram
1
Serotonergic Modulation in Rat Behavior
2
Pharmacological Reagents Inventory
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Cocaine HCl was obtained from the National Institute on Drug Abuse. NBQX, CPP, sulpiride, quinpirole, sumatriptan, GBR12783, tamoxetine, citalopram, 5-HT, and L-745870 were purchased from Tocris. Gabazine, kynurenic acid (sodium salt), and WAY 100635 were obtained from Abcam. All other chemicals were from Sigma.
3
Molecular Modeling of Neurotransmitter Binding
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Molecular modeling was performed using MOE
v2010 and v2011.10 software from Chemical Computing Group (Montreal,
Quebec, CA). The radioligands [3H]serotonin (∼28
Ci/mmol), [3H]dopamine (∼26 Ci/mmol), [3H]norepinephrine (∼26 Ci/mmol), and [125I]RTI-55
(∼2200 Ci/mmol) were obtained from PerkinElmer Life and Analytical
Sciences (Foster City, CA). Nonradioactive citalopram, mazindol, nisoxetine,
and fluvoxamine were obtained from Tocris Bioscience (Ellisville,
MO). Virtual screening hit compounds were purchased from Ambinter
(Orleans, FR). C57BL/6J mice were obtained from The Jackson Laboratory
(Bar Harbor, ME). Data analysis was performed using GraphPad Prism
5.0 (GraphPad Software, San Diego, CA).
v2010 and v2011.10 software from Chemical Computing Group (Montreal,
Quebec, CA). The radioligands [3H]serotonin (∼28
Ci/mmol), [3H]dopamine (∼26 Ci/mmol), [3H]norepinephrine (∼26 Ci/mmol), and [125I]RTI-55
(∼2200 Ci/mmol) were obtained from PerkinElmer Life and Analytical
Sciences (Foster City, CA). Nonradioactive citalopram, mazindol, nisoxetine,
and fluvoxamine were obtained from Tocris Bioscience (Ellisville,
MO). Virtual screening hit compounds were purchased from Ambinter
(Orleans, FR). C57BL/6J mice were obtained from The Jackson Laboratory
(Bar Harbor, ME). Data analysis was performed using GraphPad Prism
5.0 (GraphPad Software, San Diego, CA).
4
Pharmacological Reagents Inventory
5
Serum 5-HT Measurement Post-Behavioral Test
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Serum 5-HT: Measurement following FST and MBT. 20 minutes after the FST (Behavioral Experiment 1; Fig. 1B ) or rotarod test (Behavioral Experiment 2; Fig. 1C ), animals were terminated and trunk blood collected. The blood samples were allowed to clot overnight at 4°C to release serotonin (5-HT) from platelets. Serum was separated by centrifugation at 12,000 rpm for 15 minutes at 4°C, and stored at -80°C until assayed. 5-HT levels were determined in duplicate using a commercial RIA kit according to the manufacturer’s instructions. The sensitivity of the RIA kit for serum 5-HT was 6.7 ng/mL and the interassay coefficient of variation was less than 5.0%. 5-HT RIA Fast Track kits were purchased from Rocky Mountain Diagnostics (Colorado Springs, CO). Citalopram was purchased from Tocris Biosciences (Ellisville, MO). Fluoxetine and all other chemicals, unless otherwise noted, were purchased from Sigma-Aldrich (St. Louis, MO).
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6
Microdialysis and Behavioral Assays
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All reagents used were of analytical grade and were obtained from Merck (Germany).
Veratridine and bicuculline were from Sigma-Aldrich-RBI (Madrid, Spain), citalopram from Tocris (Madrid, Spain) and ketamine from Hospital Clinic Pharmacy Service (Barcelona, Spain). Concentrated solutions (1 mM; pH adjusted to 6.5-7 with NaHCO3 when necessary) were stored at -80ºC and working solutions were prepared daily by dilution in aCSF. For microdialysis experiments, drugs were dissolved in aCSF (except for veratridine dissolved in aCSF + 1% of dimethyl sulfoxide -DMSO), and administered by reverse dialysis at the stated concentrations. For behavioral studies, drugs were dissolved in saline solution.
Veratridine and bicuculline were from Sigma-Aldrich-RBI (Madrid, Spain), citalopram from Tocris (Madrid, Spain) and ketamine from Hospital Clinic Pharmacy Service (Barcelona, Spain). Concentrated solutions (1 mM; pH adjusted to 6.5-7 with NaHCO3 when necessary) were stored at -80ºC and working solutions were prepared daily by dilution in aCSF. For microdialysis experiments, drugs were dissolved in aCSF (except for veratridine dissolved in aCSF + 1% of dimethyl sulfoxide -DMSO), and administered by reverse dialysis at the stated concentrations. For behavioral studies, drugs were dissolved in saline solution.
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