Arecoline hydrobromide (Sigma, 31593-250MG), 1 mM for killing assays and 5 mM for other experiments; Scopolamine (Sigma PHR1470-500MG), 1 mM for killing assays and 5 mM for other assays; Oxotremorine (sigma, O100-100MG) 1 mM, LiCl (Sigma 203637-10G) 100 mM). For killing assays and qRT-PCR, all drug treatments were performed on solid NGM plates supplemented with drug. Animals were treated at the young adult stage and incubated at 25 °C for 24 h. After 16 h, animals were harvested for killing assays or qRT-PCR. For reporter gene quantification, all drug treatments were performed in NGM liquid culture for 16 h. Subsequently, animals were washed with M9 buffer and prepared for imaging.
Oxotremorine
Manufactured by Merck Group
Sourced in United Kingdom
Oxotremorine is a laboratory chemical used as a muscarinic acetylcholine receptor agonist. It is commonly used in scientific research for its pharmacological effects on the nervous system.
Automatically generated - may contain errors
Lab products found in correlation
Scopolamine, by Merck Group (3 mentions)
Arecoline hydrobromide, by Merck Group (1 mentions)
Clozapine n oxide cno, by Cayman Chemical (1 mentions)
Nicotine, by Merck Group (1 mentions)
Muscarine, by Merck Group (1 mentions)
Mecamylamine, by Merck Group (1 mentions)
Acetylcholine chloride, by Merck Group (1 mentions)
Methacholine, by Merck Group (1 mentions)
Carbachol, by Merck Group (1 mentions)
Arecoline, by Merck Group (1 mentions)
Xanomeline, by Bio-Techne (1 mentions)
Pilocarpine, by Merck Group (1 mentions)
N desmethylclozapine, by Merck Group (1 mentions)
Mcn a 343, by Merck Group (1 mentions)
Serotonin, by Merck Group (1 mentions)
Borax solution, by Merck Group (1 mentions)
Artificial cerebrospinal fluid, by Merck Group (1 mentions)
8 protocols using oxotremorine
1
Cholinergic Modulation of C. elegans
2
Neuropharmacological Compound Infusion
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Cocaine (Takeda Pharmaceutical Companies), nicotine (Sigma-Aldrich), oxotremorine (Sigma-Aldrich), dihydro-β-erythroidin (DHβE; Sigma-Aldrich), atropine (Sigma-Aldrich), and clozapine N-oxide (CNO; Cayman Chemical) were dissolved in Ringer’s solution for local infusion.
3
Pharmacological Evaluation of Cholinergic Agents
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Acetylcholine chloride (ACh), arecoline, muscarine, methacholine, oxotremorine, carbachol, mecamylamine, and atropine were purchased from Sigma-Aldrich Chemical Company (St. Louis, MO). Fresh ACh stock solutions were made in Ringer's solution each day of experimentation. Stock solutions of the test drugs were made in Ringer's solution and kept at 4°C and used within two days. Working solutions were prepared freshly at the desired concentration from the stored stock.
4
Muscarinic Agonist Receptor Activation Study
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Muscarinic agonists arecoline, carbachol furmethide, iperoxo, McN-A343, N-desmethylclozapine, oxotremorine, pilocarpine (Sigma, St.Louis, MO, USA), xanomeline (Tocris Bioscience, Bristol, UK), JR-6, and JR-7 (synthesized at Barry University, Miami Shores, FL, USA [19 (link)]) were used in this study. Structures of all used agonists are in the Supplementary Material (Figure S2) .
5
Superfusion System for Pharmacological Studies
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A custom-built superfusion system incorporating a U120 peristaltic pump (Watson-Marlow Pumps, Falmouth, UK) was used to superfuse physiological saline over preparations at a rate of 0.1 mL/s. Drugs were bath applied and washed out by switching between different gravity-fed inflows. For most experiments, control periods typically lasted 300–600 s, drug applications typically lasted 400 s, and wash periods lasted 400–800 s. In some experiments involving ablation of CNS regions, drug application periods were sometimes shortened to 200 s. It typically took ∼45–60 s for full bath exchange after switching feeds. Analysis windows were adjusted to account for time of bath exchange. Minimum time of analysis window in any experiment was 200 s. In ablation experiments, we typically waited at least 8–10 min after each cut before conducting experiments to reduce the chances of artifacts due to injury firing. The muscarinic agonist oxotremorine (Sigma-Aldrich, Irvine, UK) was applied at concentrations ranging from 10−4 to 10−6 M. The muscarinic antagonist scopolamine (Sigma-Aldrich, Irvine, UK) was applied at concentrations ranging from 3 × 10−4 to 3 × 10−7 M.
6
Pharmacological Modulation of Neurophysiology
7
Superfusion System for Pharmacological Studies
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A custom built superfusion system incorporating a U120 peristaltic pump (Watson-Marlow Pumps, Falmouth, United Kingdom) was used to superfuse physiological saline over preparations at a rate of 0.1 ml/s. Drugs were bath applied and washed out by switching between different gravity fed inflows. Control periods typically lasted 5 -7 minutes, drug applications lasted 6 minutes, and wash periods lasted 6 -10 minutes. It typically took ~45 -60 seconds for full bath exchange after switching feeds. In ablation experiments, we typically waited at least 8 -10 minutes after each cut before conducting experiments to reduce the (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprint this version posted March 9, 2021. ; https://doi.org/10.1101/2021.03.08.432150 doi: bioRxiv preprint chances of artifacts due to injury firing. The muscarinic agonist oxotremorine (Sigma-Aldrich, Irvine, United Kingdom) was applied at concentrations ranging from 10 -4 to 10 -6 M. The muscarinic antagonist scopolamine (Sigma-Aldrich, Irvine, United Kingdom) was applied at concentrations ranging from 3 x 10 -4 to 3 x 10 -7 M.
The copyright holder for this preprint this version posted March 9, 2021. ; https://doi.org/10.1101/2021.03.08.432150 doi: bioRxiv preprint chances of artifacts due to injury firing. The muscarinic agonist oxotremorine (Sigma-Aldrich, Irvine, United Kingdom) was applied at concentrations ranging from 10 -4 to 10 -6 M. The muscarinic antagonist scopolamine (Sigma-Aldrich, Irvine, United Kingdom) was applied at concentrations ranging from 3 x 10 -4 to 3 x 10 -7 M.
8
Anesthetized Rat Brain Microinjection
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Each rat was anesthetized with an i.p. injection of achloralose (100 mg/kg) and urethane (500 mg/kg) dissolved in a borax solution (2%) (Sigma-Aldrich, Steinheim, Germany). Left femoral artery and vein were catheterized for the measurement of arterial pressure and for the administration of anesthetics, respectively. The catheter of the artery was connected to a pressure transducer and the signals were acquired using the MP-36 Biopac system (Biopac Systems, Santa Barbara, CA). Mean arterial pressure (MAP) was calculated as follows: MAP 5 diastolic pressure 1 pulse pressure/3. Body temperature was maintained at 368C using a feedbackcontrolled heating pad (Diagnostic & Research Instruments, Taipei, Taiwan).
After been fixed in a stereotaxic device, the rat's skull was exposed and a 2-mm-diameter hole was drilled on the right site for injection. The injection coordinates for the hypothalamic area were 3.0 mm caudal and 1.4 mm lateral to bregma and 8.5 mm deep from the surface of the cortex. The pretreatment chemical was unilaterally injected into the right hypothalamus 15 min before the injection of oxotremorine (1 nmol in 100 nL artificial cerebrospinal fluid, Sigma-Aldrich Chemicals). The injection site was verified with histological examination for the location of a dye injected at the end of the experiment. Each rat was used for only one experiment.
After been fixed in a stereotaxic device, the rat's skull was exposed and a 2-mm-diameter hole was drilled on the right site for injection. The injection coordinates for the hypothalamic area were 3.0 mm caudal and 1.4 mm lateral to bregma and 8.5 mm deep from the surface of the cortex. The pretreatment chemical was unilaterally injected into the right hypothalamus 15 min before the injection of oxotremorine (1 nmol in 100 nL artificial cerebrospinal fluid, Sigma-Aldrich Chemicals). The injection site was verified with histological examination for the location of a dye injected at the end of the experiment. Each rat was used for only one experiment.
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