We used several pharmacological agents including Hm1a (Alomone Labs STH-601), linopirdine (Sigma L134), Retigabine (Alomone D-23129), the cholinomimetic carbamoylcholine chloride (carbachol; Sigma C4382), muscarine (Sigma M6532), and the nicotinic agonist 4-Acetyl-1,1-dimethylpiperazinium iodide (Tocris 0352). Hm1a, muscarine, and 4-Acetyl-1,1-dimethylpiperazinium iodide were dissolved in deionized water, aliquoted, and frozen at −20° C. carbachol was stored as a powder at RT, and both linopirdine and Retigabine were stored as 100 mM stock solutions in DMSO (Sigma) aliquoted in −20°C. For pharmacological experiments in which drug was dissolved in DMSO, the same concentration of DMSO was present in control external solution. After a baseline recording, drugs were perfused in at 3 mL/min while continuously recording membrane potential; repeat measurements were performed 10 min after wash-in. A subset of cells were recorded for up to 1 hr following washout of each drug, as needed in some cases to observe complete or near-complete washout. For experiments using carbachol, repeat measurements were made 5 min after wash-in.
Muscarine
Manufactured by Merck Group
Sourced in United States, Germany, Japan
Muscarine is a laboratory-grade product used for various research and analytical purposes. It serves as a chemical standard and reference material for analytical techniques. The core function of Muscarine is to provide a reliable and well-characterized compound for research, testing, and calibration applications.
Automatically generated - may contain errors
Lab products found in correlation
Carbachol, by Merck Group (4 mentions)
Mecamylamine, by Merck Group (2 mentions)
Glutamate, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
4 damp, by Merck Group (1 mentions)
Methoctramine, by Merck Group (1 mentions)
Tetrodotoxin ttx, by Bio-Techne (1 mentions)
Idazoxan, by Merck Group (1 mentions)
Idazoxan, by Bio-Techne (1 mentions)
Scopolamine, by Merck Group (1 mentions)
Naloxone, by Abcam (1 mentions)
Mk 801, by Hello Bio (1 mentions)
Uk14304 tartrate, by Bio-Techne (1 mentions)
Met5 enkephalin, by Merck Group (1 mentions)
Endomorphin 1, by Merck Group (1 mentions)
Hexamethonium, by Merck Group (1 mentions)
Methyllycaconitine, by Merck Group (1 mentions)
Aspartate, by Merck Group (1 mentions)
Taurine, by Merck Group (1 mentions)
Nicotine, by Merck Group (1 mentions)
13 protocols using muscarine
1
Pharmacological Agents for Membrane Potential
2
Electrophysiological Protocols for Neuronal Recordings
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The dissecting aCSF contained (in mM): 25 NaCl, 188 sucrose, 1.9 KCl, 1.2 NaH2PO4, 10 MgSO4, 1 CaCl2, 26 NaHCO3, 25 glucose and 1.5 kynurenic acid. The recovery solution contained (in mM): 119 NaCl, 1.9 KCl, 1.2 NaH2PO4, 10 MgSO4, 1 CaCl2, 26 NaHCO3, 20 glucose and 1.5 kynurenic acid. The recording aCSF contained (in mM): 127 NaCl, 3 KCl, 1.25 NaH2PO4, 1 MgCl2, 2 CaCl2, 26 NaHCO3, 10 glucose. The intracellular solution for patch-clamp recordings was made of (in mM): 140 KMeSO4, 10 NaCl, 1 CaCl2, 10 HEPES, 1 EGTA, 3 Mg-ATP and 0.4 GTP-Na2 (pH 7.2–7.3, adjusted with KOH). Muscarine, 4-DAMP, methoctramine, NMDA, DA and 5-HT were supplied by Sigma-Aldrich®; tetrodotoxin (TTX) by Bio-Techne®. All drugs were dissolved in H2O except for 4-DAMP which was dissolved in DMSO to a concentration that did not exceed 0.1% (vol/vol) in working solutions.
3
Pharmacological Manipulation of Opioid Signaling
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Morphine sulfate and morphine alkaloid were obtained from the National Institute on Drug Abuse (NIDA), Neuroscience Center (Bethesda, MD). Naloxone was purchased from Abcam (Cambridge, MA), MK-801, from Hello Bio (Princeton, NJ), UK14304 tartrate and idazoxan (Ida) from Tocris (Bio-Techne Corp. Minneapolis, MN). Potassium methanesulfonate was from Alfa Aesar (Ward Hill, MA). [Met5] enkephalin (ME), endomorphin-1, muscarine, scopolamine, idazoxan and other reagents were from Sigma (St. Louis, MO). Caged-enkephalin (CYLE) and Caged-Naloxone (CNV-Nal) were gifts from Mathew Banghart.
Morphine alkaloid was converted to salt form with 0.1 M HCl and made up a stock solution in water. The working solution was diluted in artificial cerebrospinal fluid (ACSF) and applied during incubation or superfusion. Naloxone, endomorphin-1, muscarine, scopolamine, UK14304 tartrate and idazoxan were dissolved in water, diluted in ACSF and applied by bath superfusion. Bath perfusion of ME was with bestatin (10 mM) and thiorphan (1 mM) to limit breakdown of ME.
Morphine alkaloid was converted to salt form with 0.1 M HCl and made up a stock solution in water. The working solution was diluted in artificial cerebrospinal fluid (ACSF) and applied during incubation or superfusion. Naloxone, endomorphin-1, muscarine, scopolamine, UK14304 tartrate and idazoxan were dissolved in water, diluted in ACSF and applied by bath superfusion. Bath perfusion of ME was with bestatin (10 mM) and thiorphan (1 mM) to limit breakdown of ME.
4
Neurotransmitter Assay Protocol with Diffusion Marker
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All candidate neurotransmitters, agonists, and antagonists, except sNPF, were prepared in adult hemolymph-like saline, AHLS (Wang et al., 2003 (link); 2 mM CaCl2, 5 mM KCl, 8.2 mM MgCl2, 108 mM NaCl, 4 mM NaHCO3, 1 mM NaH2PO4, 10 mM sucrose, and 5 mM trehalose) to a concentration of 100 mM. Aspartate, glutamate, taurine, GABA, ACh, nicotine, muscarine, mecamylamine, methyllycaconitine, and hexamethonium were all from Sigma Aldrich. Short neuropeptide F (sNPF; Ala-Gln-Arg-Ser-Pro-Ser-Leu-Arg-Leu-Arg-Phe-NH2) was synthesized by PolyPeptide (Strasbourg, France) to a purity of >95% and was dissolved in DMSO. Each solution delivered in the micropipette also contained 5 μM Texas red dextran (TXR; Life Technologies) to monitor diffusion.
5
Muscarinic Receptor Modulation of T Cell Apoptosis and Proliferation
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PBT cells (1×106) were cultured in 300 µl of complete medium in 96-well plates. The cells were cultured in RPMI 1640 supplemented with 10% fetal calf serum in a 5% CO2 humid atmosphere at 37°C. PBT cells were cultured in the presence of medium alone, CSE or mAChR agonist/antagonist. The mAChRs were activated with muscarine (50 µM) and inhibited with atropine (100 µM) (Sigma-Aldrich or Tocris). In the proliferation assays, 1 µg/ml PHA and 50 ng/ml PMA (both from Sigma-Aldrich, St. Louis, MO) were added to the medium to stimulate T cell proliferation. After 5 days, the cells were harvested and then stained with APC-conjugated Annexin V and propidium iodide (Annexin V Apoptosis Detection Kit APC; eBioscience) at room temperature in the dark for 10 min. Finally, the proportion of apoptotic T cells was determined by flow cytometry. For T cell proliferation assays, intracellular staining with Alexa Fluor 647 conjugated anti-human Ki-67 (eBioscience) was performed.
6
Rearing and Dissection of Fall Armyworm
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Mythimna separata larvae were initially obtained from the Biorational Pesticides Research and Development Center, Northwest A&F University, Shaanxi, China, and reared on corn leaves at 25 ± 2°C, 50 ± 5% relative humidity, and a photoperiod of 16 h light: 8 h darkness. The moths were supplied with a 5% honey solution as nutrient. Different developmental stages of M. separata were selected and immediately placed into RNAlater (Ambion, Austin, TX, United States) and stored at −70°C until use. For tissue collection, male and female adults at 3 days after eclosion were selected and chilled at 4°C for 30 min to sedate the moths. Then the moths were dissected in the 1 × phosphate buffered saline (PBS), the head, midgut, fat body, Malpighian tubules, ovary, and testis were harvested by micro scalpel and tweezers. Tissues were immersed immediately into RNAlater and stored at −70°C until used for RNA extraction. Carbachol, muscarine, and atropine were purchased from Sigma (St. Louis, MO, United States). All other chemicals of research grade were obtained from commercial sources.
7
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.
8
Muscarinic Receptor Agonist Testing in hSCs
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Human SCs were treated with M2 selective agonist arecaidine propargyl ester hydrobromide (APE, A140; Sigma-Aldrich) at the final concentration ranging from 25 to 100 µM. Albeit APE was reported as preferential M1–M4 agonist [25 (link)], several modifications of the molecule suggested a preferential agonistic effect on M2 receptor subtype [26 (link),27 (link)]. Our works have been focused on the characterization of APE hydrobromide by pharmacological competition studies with M2 antagonists methoctramine or gallamine [8 (link),17 (link),32 (link)]. Moreover, the silencing of M2 receptors by siRNAs have largely demonstrated the selectivity of APE for M2 receptor in several cell lines (i.e., human glioblastoma, neuroblastoma, and urothelial bladder cell lines [16 (link),29 (link),30 (link),31 (link),33 (link)].
To study all muscarinic receptors activation, hSCs were treated with the non-selective muscarinic agonist, muscarine (Sigma-Aldrich), at the final concentration ranging from 25 to 100 µM.
All experiments were performed in technical and experimental triplicate.
To study all muscarinic receptors activation, hSCs were treated with the non-selective muscarinic agonist, muscarine (Sigma-Aldrich), at the final concentration ranging from 25 to 100 µM.
All experiments were performed in technical and experimental triplicate.
9
Detailed Neuromuscular Receptor Pharmacology
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The following reagents were used at the following concentrations: P2Y1R agonists ATP and ADP (Sigma; 10 or 20 μM); P2Y1R antagonist MRS2500 (Tocris; 1 μm); P1R agonist adenosine (Sigma; 100 μM); pan-P2 antagonist suramin (Sigma; 100 μM); pan-muscarinic agonist muscarine (Sigma; 10 μM); pan-muscarinic blocker atropine (Sigma; 10 μM); pan-nicotinic agonist nicotine (Sigma; 50 μM); pan-nicotinic antagonist curare (Sigma; 200 μM); pan-cholinesterase inhibitor neostigmine (Sigma; 1 μM); sarco-/endoplasmic reticulum Ca2+-ATPase (SERCA) inhibitor, cyclopiazonic acid (Sigma; CPA; 10 μM); potassium chloride (Sigma; 2–10 mM); GIIIb μ-conotoxin (Peptides International; 2.3 μM); skeletal muscle myosin-blocker 3-(N-butylethanimidoyl)−4-hydroxy-2H-chromen-2-one (BHC; Hit2lead; 100 μM); 488-, 594-, 633-conjugated-α-bungarotoxin (α-BTX; Biotium; 1 μg/mL).
10
Pharmacological Regulation of Synaptic Signaling
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Muscarine, Ro25-6981, NMDA, glycine, carbachol, PACAP38, sodium orthovanadate, SKF81297 and DMSO were bought from Sigma (St Louis, MO). TCN201, pirenzepine and xanomeline oxalate were bought from Tocris Bioscience (Bristol, UK). Xestospongin C, PP2 and PP3 were bought from Cayman Chemicals (Ann Arbor, MI). Src(40–58) were provided by Dr. MW Salter (Hospital for Sick Children, Toronto, Canada). Fyn(39–57) was synthesized by the Advanced Protein Technology Centre (Toronto, Canada). PACAP and cell permeable EGTA/AM were from Calbiochem (San Diego, CA). The STEP inhibitor TC-2153 was synthesized as described previously33 (link).
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