C57BL/6 mice were purchased from Charles River Laboratories (Kingston, NY) and housed in the Florida State University Laboratory Animal Resources facility. The facility is a temperature-and humidity-controlled environment maintained on a 12-h light-dark cycle (lights off at 7:00 a.m. and on at 7:00 p.m.). The mice had food and water available ad libitum. Breeding age (8-12 weeks old) female mice were randomly assigned to one of the 3 experimental groups based on the type of drinking water supplied: PNE group was provided with water containing nicotine (100 μg/mL (-)-nicotine; Sigma Chemical Co., St. Louis, MO; Cat# N3876) and 2% saccharin (Alfa Aesar, Heysham, UK; Cat# A15530); the saccharin group was provided with water containing 2% saccharin (Fig. 1a). To control for the potential effects of saccharin, a third group of mice received plain water [27] [28] [29] 37] . After 3 weeks, female mice in each group were bred with drug-naïve male mice. Based on the presence of a vaginal plug, the day of successful mating was designated embryonic day 0 (E0) and the day of birth postnatal day 0 (P0). The litter size was standardized to contain 6-8 offspring on the day of birth. The 3 types of water were continued until the pups were weaned on P21. Throughout pregnancy, each female mouse was singly housed. Upon weaning, same sex offspring were housed 2-4 per cage.
Saccharin
Manufactured by Thermo Fisher Scientific
Sourced in United States, United Kingdom
Saccharin is a laboratory reagent used as a non-nutritive sweetener. It is a white crystalline powder with a sweet taste. Saccharin is commonly used in various research and analytical applications, including food science, biochemistry, and analytical chemistry.
Automatically generated - may contain errors
Lab products found in correlation
C57bl 6 mice, by Charles River Laboratories (1 mentions)
Nicotine, by Merck Group (1 mentions)
Polysorbate 80, by Thermo Fisher Scientific (1 mentions)
Sucrose, by Thermo Fisher Scientific (1 mentions)
Ethanol, by Decon Labs (1 mentions)
Quinine, by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Thermo Fisher Scientific (1 mentions)
Methanol, by Thermo Fisher Scientific (1 mentions)
Acetonitrile, by Thermo Fisher Scientific (1 mentions)
Isopropanol, by Thermo Fisher Scientific (1 mentions)
Sodium lauryl sulfate, by Merck Group (1 mentions)
Salicylic acid, by Merck Group (1 mentions)
Sodium hydroxide pellets, by Avantor (1 mentions)
Anhydrous carbamazepine, by Merck Group (1 mentions)
Milli q plus water system, by Merck Group (1 mentions)
Indomethacin, by Merck Group (1 mentions)
Furosemide, by Merck Group (1 mentions)
P aminobenzoic acid, by Merck Group (1 mentions)
Carprofen, by Zoetis (1 mentions)
Grk2i, by Bio-Techne (1 mentions)
11 protocols using saccharin
1
Maternal Nicotine Exposure in Mice
2
Saccharin Regulates Laser-Induced CNV
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The animal experiments complied with the guidelines of the ARVO statement for the use of animals in ophthalmic and vision research. They were also approved by the local government authorities (Landesamt für Gesundheit und Soziales, LaGeSo, Berlin). Twelve male C57BL/6J mice purchased from Charles River (Germany) have been used for the described experiments. All of them were 10 to 14 weeks old and weighed more than 22 g (mean 25.5 g) at the time of the laser-induced CNV. The animals were housed in a 12-hour day/night cycle and got food and water ad libitum. One day before the laser treatment, we added 0.03% saccharin (Thermo Fisher Scientific, Waltham, MA, USA) to the drinking water of our saccharin intervention group (Sa; n = 6). The Sa group got the saccharin-containing water for 15 days (until the end of the experiments) while the control group (Co; n = 6) got pure water.
3
THC Suspension Preparation and Administration
4
Binge-Like Ethanol Consumption in Mice: The DID Paradigm
5
Olfactory Mixtures and Gustatory Responses
6
Odor Stimuli and Intraoral Delivery
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Unisensory odor stimuli were exemplars of monomolecular odorants (obtained from Sigma-Aldrich; >98% purity) that have been used in similar behavioral (Blankenship et al., 2019 (link)) and neural recording (Maier, 2017 (link)) paradigms in previous studies: methyl valerate and 2-hexanone in aqueous solution (0.025% volume/volume in distilled water). Saccharin (0.2%; Fisher Scientific) was used as an unconditioned stimulus during conditioning sessions. All stimuli were presented intraorally to allow, as much as possible, natural sensory stimulation dynamics associated with intraoral evaluation of flavor. There is no empirical evidence that the odorants used here completely lack gustatory and/or trigeminal qualities. However, similar concentrations of odor solutions have been shown to be below nonolfactory behavioral detection thresholds (Slotnick et al., 1997 (link); Gautam and Verhagen, 2012 (link)) and are therefore unlikely to have contributed to the observed responses.
7
Synthesis and Characterization of Carbamazepine Polymorphs
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Anhydrous carbamazepine (CBZ), salicylic
acid (SLC), and sodium lauryl sulfate (SLS) were purchased from Sigma
Chemical Company (St. Louis, MO) and used as received. Carbamazepine
dihydrate (CBZD) was prepared by slurrying anhydrous CBZ in deionized
water for 24 h, and solid was obtained through vacuum filtration.
Saccharin (SAC) was purchased from Acros Organics (Pittsburgh, PA)
and used as received. Isopropanol, acetonitrile, methanol, and hydrochloric
acid were purchased from Fisher Scientific (Pittsburgh, PA). Sodium
hydroxide pellets were purchased from J.T. Baker (Philipsburg, NJ).
Water used in this study was filtered through a double deionized purification
system (Milli Q Plus Water System) from Millipore Co. (Bedford, MA).
acid (SLC), and sodium lauryl sulfate (SLS) were purchased from Sigma
Chemical Company (St. Louis, MO) and used as received. Carbamazepine
dihydrate (CBZD) was prepared by slurrying anhydrous CBZ in deionized
water for 24 h, and solid was obtained through vacuum filtration.
Saccharin (SAC) was purchased from Acros Organics (Pittsburgh, PA)
and used as received. Isopropanol, acetonitrile, methanol, and hydrochloric
acid were purchased from Fisher Scientific (Pittsburgh, PA). Sodium
hydroxide pellets were purchased from J.T. Baker (Philipsburg, NJ).
Water used in this study was filtered through a double deionized purification
system (Milli Q Plus Water System) from Millipore Co. (Bedford, MA).
8
Synthesis of Pharmaceutical Compounds
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Indomethacin, furosemide and p-aminobenzoic acid were purchased from Sigma Aldrich (St. Louis, MO, USA). Saccharin was acquired from Acros Organics (Morris, NJ, USA). The solvents were obtained from POCH (Polish Chemical Reagents, Gliwice, Poland). The purity of substances was ≥98%, the purity of solvent ≥99.8%.
9
Opioid Pharmacology: Antagonist and Analgesic
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Saccharin (Acros Organics #149005000) was diluted to several concentrations (the highest being 0.1%) in tap water. Oxycodone HCl was supplied by the NIDA drug sharing program. Oxycodone was diluted at various concentrations in either tap water for self-administration experiments, or sterile saline for injections and osmotic minipump loading. Naloxone Hydrochloride was purchased from Spectrum Chemical (#N1231), dissolved in sterile saline, and injected at 1 mg/kg intraperitoneal (i.p.) immediately prior to assessing jumping behavior. Lidocaine (Covetrus, North America VINB-0024-6800) was injected locally at 4 mg/kg subcutaneously (s.c.) prior to incision for minipump implantation. Carprofen (Zoetis) was injected i.p. at 5 mg/kg prior to incision for minipump implantation, and again at 24 and 48 h after surgery.
10
Corticosterone Hemisuccinate Solution Preparation
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Corticosterone hemisuccinate (CORT; 4-pregnen-11β,21-diol-3,20-dione21-hemisuccinate, Steraloids, Newport, RI, USA) was prepared fresh every three days. CORT was dissolved in tap water and stirred overnight at a pH of 10–11 and neutralized to a pH of 7.0–7.4 prior to use. Ethanol (EtOH; Decon Labs, King of Prussia, PA, USA) and saccharin (Acros Organics, Pittsburgh, PA, USA) were diluted in tap water to concentrations of 10% (v/v) and 0.1% (w/v), respectively, to make the sweetened EtOH solution. Yohimbine (Sigma, St. Louis, MO, USA) was dissolved in double distilled water to a concentration of 1.25 mg/mL. GRK2i (GRK2 inhibitory polypeptide; Tocris, Minneapolis, MN, USA) was dissolved in saline to a concentration of 2 mM.
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