On test days, rats were injected i.p. with either saline, or the adenosine receptor antagonist caffeine (Sigma-Aldrich, St. Louis, MO; 50 mg/kg) or the partial inverse agonist at the benzodiazepine allosteric site on the GABAA receptor, FG-7142 (Sigma-Aldrich, St. Louis, MO; 7.5 mg/kg). Both caffeine and FG-7142 have been shown to be anxiogenic in rodents: caffeine, 50 mg/kg [19 (link)–22 (link)] and FG-7142, 7.5 mg/kg [22 (link),23 (link)]. FG-7142 was dissolved in (40%w/v) 2-hydroxypropyl cyclodextrin (Sigma-Aldrich, St. Louis, MO) in 0.05N HCl to increase its solubility as done in previous studies [24 (link),25 (link)]. Each of these drugs (caffeine and FG-7142) have been reported to cause increased vigilance and arousal behaviors in home cage environment of rats from when compared to vehicle-injected controls [25 (link),26 (link)].
Fg 7142
Manufactured by Merck Group
Sourced in United States
FG-7142 is a laboratory instrument used for analytical purposes. It is designed to perform specific measurements and analyses as part of research and testing procedures. The core function of FG-7142 is to provide accurate and reliable data to support scientific investigations and product development.
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7 protocols using fg 7142
1
Modulation of Rodent Anxiety Behaviors
2
Pharmacological Modulation of SphK1 and S1P Receptors
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All drugs were administered by intraperitoneal injection (I.P.). FG-7142 (Sigma-Aldrich, #E006-100MG) was used as an anxiolytic drug, and administered as described previously (Evans and Lowry, 2007 (link)) at a final concentration of 5 mg/kg body weight 30 min before behavioral testing. The SphK1 antagonist PF-543 (Cayman Chemicals, #17034) was dissolved in 2% DMSO/PBS 0.01M and administered at a final dosage of 10mg/kg. Fingolimod (FTY-720, Sigma-Aldrich, #SML0700-5MG) was used as an S1P receptor agonist, dissolved in saline solution and administered at a final dosage of 0.1 mg/kg, as described (Deogracias et al., 2012 (link)).
3
Preparation and Storage of FG-7142 Solution
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FG-7142 was purchased from Sigma-Aldrich Canada Co. (Oakville, ON, Canada) and stored in a dry and dark drug storage cabinet at an ambient room temperature of ~ 23 °C within the key-locked behavioural neuroscience lab at MacEwan University. FG-7142’s carboxamide structure makes it naturally hydrophobic and insoluble in water without an appropriate vehicle1 ,8 (link), thus a stock treatment solution of 14.0 mM FG-7142 dissolved in 10% DMSO as a vehicle was prepared based on a factor of four36 (link) or five and a half41 (link) of the concentrations used in previous studies with zebrafish. The solution was prepared in a MacEwan university organic chemistry laboratory with appropriate skin and respiratory protection underneath a fume hood. The stock solution was refrigerated at ~ 3.0 °C until experimentation. Because the freezing point of DMSO is 19 °C, each vial was only thawed once to retain compound integrity and minimize the risk of solution separation via oiling-out. Any unused solution was kept in a room temperature (~ 23 °C) dry and dark drug cabinet and to be used within the next testing day. Thawed stock solution that was left unused for more than 5 days was marked and discarded.
4
Pharmacological Induction of Anxiety
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To achieve an anxiogenic state, the animals received an intraperitoneal injection of FG-7142 (beta-Carboline-3 carboxylic acid-N-methyl amide; Sigma Aldrich, St. Louis, MO, USA), an inverse partial agonist of benzodiazepines, in a single dose of 7.5 mg/kg, dissolved in one-Cyclodextrin solution (Sigma Aldrich, St. Louis, MO, USA) dissolved in a sterile saline solution acidified to 1 M hydrochloric acid (HCl) pH5 to a final volume of 2 ml/kg following the procedures of Lawther et al. (2015) (link).
5
Anxiolytic Agents in Mice Behavior
6
Dose-dependent Effects of FG7142 on Cued Behavior
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Seven male and 11 female animals reached criterion on both the cued-approach and cued-avoidance tasks and were put into the full task. The effect of FG7142 on performance in the full task was tested in these animals. FG7142 (Sigma-Aldrich, St. Louis, MO, USA) was dissolved in sterile 0.09% saline with 15% v/v Tween 80 (Sigma-Aldrich) for intraperitoneal injection of 1 mg/kg (FG1), 3 mg/kg (FG3), and 10 mg/kg (FG10). The vehicle for the control treatment was sterile 0.09% saline with 15% v/v Tween 80. Each animal received each of the four doses in a Latin square design over a period of 7 days, with wash days (no injection) between each injection day. FG1, FG3, FG10, or vehicle was delivered 10 min prior to the beginning of the behavioral session. Animals were monitored for evidence of immobility or seizure.
7
FG-7142 Induces Acetylcholine Release for Anxiety
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As a control, before the administration of the anxiogenic drug, the animals were injected with 2 mL/kg saline solution ip FG-7142 (Sigma-Aldrich, St. Louis, MO, USA), which was administered ip at a single dose of 7.5 mg/kg diluted in 12.5 mL of β-cyclodextrin and 1 M chlorohydric acid (HCl), pH 5.0, to a final volume of 2 mL/kg [74 (link)].
Under urethane, FG-7142 induces the release of acetylcholine in the vHPC [100 (link)], an effect that was recently found under chronic restraint stress, which is a model of anxiety [101 (link)].
Under urethane, FG-7142 induces the release of acetylcholine in the vHPC [100 (link)], an effect that was recently found under chronic restraint stress, which is a model of anxiety [101 (link)].
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