In this study, seven-week-old male Sprague Dawley® rats were used. Animals were allocated in groups and were allowed to accommodate for one week in the animal house. Rats were provided with a standard chow diet and were given water ad libitum. During the experimental period, the animals were kept at a temperature of 22 ± 3 °C, a 12-h light/dark cycle, and a relative humidity varied between 40% and 60%. Animals were treated with either of six different conditions: (1) vehicle; (2) scopolamine (1 mg/kg) (Sigma Aldrich, Inc.); (3) scopolamine with 50 mg/kg/day of PS as a positive control; (4) scopolamine with 75 mg/kg/day of SC+RF (SR (75)); (5) scopolamine with 150 mg/kg/day of SC+RF [SR (150)]; and (6) scopolamine with 300 mg/kg/day of SC+RF (SR (300)). Rats were orally administrated SC+RF for 28 days. The animal research protocol was approved by the Animal Care and Use Committee of the Ajou University School of Medicine, and all experiments were conducted in accordance with the institutional guidelines established by the Committee.
Scopolamine
Manufactured by Merck Group
Sourced in United States, United Kingdom, Germany, Sao Tome and Principe, France, India, China
Scopolamine is a chemical compound used in various laboratory applications. It functions as a muscarinic antagonist, which means it blocks the effects of the neurotransmitter acetylcholine on muscarinic receptors. This property makes it useful in research and scientific experiments, but its specific applications and intended uses should not be extrapolated upon.
Automatically generated - may contain errors
Lab products found in correlation
Donepezil, by Merck Group (16 mentions)
Pilocarpine, by Merck Group (11 mentions)
Acetylthiocholine iodide, by Merck Group (6 mentions)
Lithium chloride (licl), by Merck Group (6 mentions)
Piracetam, by Merck Group (5 mentions)
Mecamylamine, by Merck Group (5 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (5 mentions)
Ascorbic acid, by Merck Group (4 mentions)
Curcumin, by Merck Group (4 mentions)
Tween 80, by Merck Group (4 mentions)
Methanol, by Merck Group (4 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (4 mentions)
Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (4 mentions)
Hyoscyamine, by Merck Group (3 mentions)
Diazepam, by Merck Group (3 mentions)
Mk 801, by Merck Group (3 mentions)
Oxotremorine, by Merck Group (3 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (3 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
130 protocols using scopolamine
1
Scopolamine-Induced Cognitive Decline: Neuroprotective Effects of SC+RF
2
Scopolamine-Induced Memory Impairment Model
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The rats were divided into five groups (10 animals per group): (1) control group received saline treatment (0.9% NaCl); (2) Scopolamine (Sco) alone-treated group received Scopolamine (Sigma, Chicago, IL) 30 min after saline pretreatment daily, for 12 continuous days and 30 min prior to performing behavioural testing on test day (Fernandes et al. 2008 (link); Hritcu et al. 2012 (link); Xiang et al. 2012 (link)) at a dose of 0.1 mg/kg intraperitoneally (Kay et al. 2010 (link)); (3), (4), and (5) L. officinalis extract-pretreated groups received L. officinalis extract with concentrations of 100, 200 and 400 mg/kg (Kashani et al. 2011 (link); Rahmati et al. 2012 , 2013 (link)), per day, 30 min before each Scopolamine injection and 1 h before each behavioural testing on test day. Materials were administered intraperitoneally (i.p.) in a volume of 0.3 mL. Scopolamine (0.1 mg/kg) was used for the induction of memory impairment, anxiety and depression-like behaviour for 12 continuous days (Hritcu et al. 2012 (link)).
3
Microfluidic Worm Attraction Assay
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Microfluidic chips (60 μm deep) were based on a design by McCormick et al. (2011) (link). Fluid flow was gravity driven and controlled with manual one-way stopcocks (Cole-Parmer). Animals were exposed to streams of NGM buffer and 100 μM IAA on either side of their heads. For scopolamine treatment, animals were immersed in NGM buffer containing 100 μM scopolamine (Sigma) for 5 min before and during the assay. Worms were loaded individually into the chip, and once in place, their movements were recorded for 2 min, with a switch between buffer and IAA at the 1-min mark, using a USB camera (Aven Mighty Scope, #2700-200) and VirtualDub software.
4
Scopolamine-Induced Memory Impairment in Rats
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A total of 30 male albino Wistar rats, aged 6 weeks and weighing 150–200 g (BrlHan: WIST@Jcl (GALAS), Nomura Siam International, Bangkok, Thailand) were caged at room temperature (25 °C) with a 12/12-h light/dark cycle and were provided with food and water ad libitum. The experimental protocol was approved by the Animal Ethic Committee of the Faculty of Medicine, Chiang Mai University (approved protocol no., 27/2562). The rats were randomly divided into five groups (six rats per group). The control group received intraperitoneal (i.p.) injections of 1 mL of saline for 15 days. The scopolamine (or AD) group and the other three AM groups received 1 mg/kg of scopolamine (Sigma, Lot no. LRAB7821, St. Louis, MO, USA) for 5 days. Next, the AM200 [20 ,24 (link)], AM400 [20 ,24 (link)], and AM600 groups were treated with the AM leaf extract for 10 days at doses of 200, 400, and 600 mg/kg of body weight, respectively. All animals were tested for their motor and memory behaviors and their hippocampi were investigated for synaptophysin, dendritic spine, and histological features. All the protocols and experiments in this study are shown in Figure 1 .
5
Pilocarpine-Induced Seizure Model in Rodents
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To induce seizures in the experimental animals, the following procedure was conducted. 19 h prior to the administration of pilocarpine, lithium chloride (127 mg/kg, i.p, Sigma-Aldrich Co., St. Louis, MO, USA) was injected intraperitoneally. 30 min before the administration of pilocarpine, scopolamine (2 mg/kg, i.p, Sigma-Aldrich Co., St. Louis, MO, USA) was injected intraperitoneally. After the 30-min interval following scopolamine administration, pilocarpine (25 mg/kg, i.p, Sigma-Aldrich Co., St. Louis, MO, USA) was injected intraperitoneally. The stages of seizure were assessed using the Racine scale, which consists of five stages: Stage 1: Mouth and facial movement. Stage 2: Head nodding. Stage 3: Forelimb clonus. Stage 4: Rearing with forelimb clonus. Stage 5: Rearing and falling with forelimb clonus. Diazepam (10 mg/kg, French, France Neuilly Sur Valium, Hoffman, Seine La Roche) was administered 1 h after the onset of seizures. If recurrent seizures persisted, additional doses of diazepam were injected. This protocol was designed to induce seizures in the animals and evaluate their seizure behavior according to the Racine scale. Diazepam was used as an anticonvulsant to terminate prolonged or recurrent seizures.
6
Pilocarpine-Induced Seizure Model: Cerebrolysin Effects
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To confirm the effect of cerebrolysin on neuronal death after pilocarpine-induced seizure, the rats were administered lithium chloride (127 mg/kg, i.p, Sigma-Aldrich Co., St. Louis, MO, United States) 19 h before the administration of pilocarpine. Scopolamine (2 mg/kg, i.p., Sigma-Aldrich Co., St. Louis, MO, United States) was administered 30 min before the administration of pilocarpine (Biagini et al., 2009 (link)). Thirty minutes after Scopolamine administration, status epilepticus (SE) was induced by the intraperitoneal administration of pilocarpine (25 mg/kg, i.p., Sigma-Aldrich Co., St. Louis, MO, United States). SE is observed according to the presence of five symptoms (1. mouth and facial movement, 2. head nodding, 3. forelimb clonus, 4. rearing with forelimb clonus, and 5. rearing and falling with forelimb clonus) that occur progressively in Racine’s method. The animals were placed in individual cages for ease of observation. SE usually occurred within 20–30 min after pilocarpine injection (Persinger et al., 1988 (link)). Diazepam (10 mg/kg, i.p., Valium, Hoffman la Roche, Neuilly sur-Seine, France) was injected intraperitoneally 2 h after the last Racine’s stage occurred (Biagini et al., 2001 (link)). If the animals presented consistent recurrent seizures, additional diazepam was injected (2 mg/kg, i.p.) (Kim et al., 2013 (link)).
7
Optical Stimulation and Locomotor Activity
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Optical Stimulation: Mice were placed in a square box (47 cm x 47 cm x 21 cm) for 20 min with a floor of filter paper that was changed between mice. During the 3 rd 5 min bin of the session, mice received optical stimulation (20 sec on/off, 20 Hz, 25 ms pulses). Locomotor activity was recorded via overhead camera and analyzed in 5 min bins with EthoVision.
Antagonists: Locomotor data was collected using an Accuscan Instruments (Columbus, Ohio) behavior monitoring system and software. Mice were individually tested in empty cages, with bedding and nesting material removed to prevent obstruction of infrared beams. Mice were injected (i.p.) with saline, mecamylamine (1 mg/kg, Sigma), scopolamine (0.5 mg/kg, Sigma), or mecamylamine+scopolamine (1 mg/kg and 0.5 mg/kg, respectively) 30 min before locomotor testing. Locomotion was monitored for 20 min using 13 photocells placed 4 cm apart to obtain an ambulatory activity count, consisting of the number of beam breaks recorded during a period of ambulatory activity (linear motion rather than quick, repetitive beam breaks associated with behaviors such as scratching and grooming).
Antagonists: Locomotor data was collected using an Accuscan Instruments (Columbus, Ohio) behavior monitoring system and software. Mice were individually tested in empty cages, with bedding and nesting material removed to prevent obstruction of infrared beams. Mice were injected (i.p.) with saline, mecamylamine (1 mg/kg, Sigma), scopolamine (0.5 mg/kg, Sigma), or mecamylamine+scopolamine (1 mg/kg and 0.5 mg/kg, respectively) 30 min before locomotor testing. Locomotion was monitored for 20 min using 13 photocells placed 4 cm apart to obtain an ambulatory activity count, consisting of the number of beam breaks recorded during a period of ambulatory activity (linear motion rather than quick, repetitive beam breaks associated with behaviors such as scratching and grooming).
8
Molecular Signaling Pathways in Neurological Research
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Primary antibodies were purchased as follows: P-PRAS40 (Thr246; cat. no. 2997; Cell Signaling Technology, Inc.), PRAS40 (cat. no. 2691; Cell Signaling Technology, Inc.), p-mTOR (Ser2448; cat. no. 5536; Cell Signaling Technology, Inc.), mTOR (cat. no. 2983; Cell Signaling Technology, Inc.), p-AKT (Ser473; cat. no. 4060; Cell Signaling Technology, Inc.), AKT (cat. no. 4685; Cell Signaling Technology, Inc.), p-P70S6K (Thr389; cat. no. 9234; Cell Signaling Technology, Inc.), P70S6K (cat. no. 14130; Cell Signaling Technology, Inc.), Light chain 3-I/II (LC3-I/II; cat. no. 12741; Cell Signaling Technology, Inc.), 14-3-3 (cat. no. 8312; Cell Signaling Technology, Inc.), P62 (cat. no. 18420-1-AP; ProteinTech Group, Inc.) and GAPDH (cat. no. sc-365062; 1:2,000; Santa Cruz Biotechnology, Inc.). Goat anti-rabbit horseradish peroxidase (HRP)-conjugated secondary antibody (cat. no. sc-2054; Santa Cruz Biotechnology, Inc.). The following drugs were obtained from commercial sources as follows: Pilocarpine (cat. no. S4231; Selleck Chemicals), lithium chloride (cat. no. L9650; Sigma-Aldrich; Merck KGaA), Scopolamine (cat. no. S2508; Selleck Chemicals) and LY3023414 (cat. no. S8322; Selleck Chemicals).
9
Donepezil and Scopolamine Cognitive Protocol
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Donepezil (acetylcholinesterase inhibitor) and scopolamine (cognitive defects inducer) were purchased from Merck (Merck KGaA, Darmstadt, Germany). All other chemicals procured for the experiment were of analytical grade.
10
Pharmacological Effects of Neuroactive Agents
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Gabapentin, pregabalin, baclofen and scopolamine were purchased from Merck Co. (Germany). These three drugs were freshly dissolved in distilled water and scopolamine solution was prepared freshly in saline and administered by intraperitoneal (i.p) injection. All drug solutions were freshly prepared before each experiment. Drug injection was made between 8-10 A.M every day.
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