Acetylcholine chloride, bradykinin, carbamylcholine chloride, cyclopiazonic acid, DL-dithiothreitol, L-NAME (NG nitro L-arginine methyl ester), indomethacin, sodium nitroprusside, DMSO (dimethyl sulfoxide), and all required chemicals to prepare physiological solutions were purchased from Sigma-Aldrich (Oakville, ON, Canada). Fura-2 acetoxymethylester was obtained from Molecular Probes/Invitrogen. Euthanyl (sodium pentobarbital, 250 mg/mL) was purchased from Bimeda-MTC Animal Health Inc, Cambridge, ON, Canada. SKA-31 (naphtho [1, 2-d] thiazole-2-ylamine) was synthesized as previously described [27 (link)]. SKA-31 and indomethacin were prepared as 10 mM stock solutions in DMSO and then diluted directly into the external bath solution. The final concentration of DMSO reaching the tissue was typically 0.05% (vol/vol) or less. In control experiments, bath application of 0.2% (v/v) DMSO had no effect on either developed myogenic tone or arterial responsiveness to acetylcholine and SKA-31 in Goto-Kakizaki cremaster arteries (n = 2) (data not shown).
Carbamylcholine chloride
Manufactured by Merck Group
Sourced in United States, Brazil
Carbamylcholine chloride is a chemical compound used as a laboratory reagent. It is a cholinergic agonist, meaning it activates receptors for the neurotransmitter acetylcholine. Carbamylcholine chloride is commonly used in research and experimental settings, but a detailed description of its core function without interpretation or extrapolation cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Cyclopiazonic acid, by Merck Group (2 mentions)
Acetylcholine chloride, by Merck Group (2 mentions)
Magnesium chloride hexahydrate, by Vetec (2 mentions)
Calcium chloride dihydrate, by Vetec (2 mentions)
Sodium bicarbonate, by Vetec (2 mentions)
Bradykinin, by Merck Group (1 mentions)
Fura 2 acetoxymethyl ester, by Thermo Fisher Scientific (1 mentions)
Ng nitro l arginine methyl ester l name, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Indomethacin, by Merck Group (1 mentions)
Dl dithiothreitol, by Merck Group (1 mentions)
Sodium nitroprusside, by Merck Group (1 mentions)
Isoproterenol hydrochloride, by Merck Group (1 mentions)
Fluo 4 direct calcium assay kit, by Thermo Fisher Scientific (1 mentions)
Hyd hybrid detector, by Leica camera (1 mentions)
Hexamethonium, by Merck Group (1 mentions)
Yohimbine, by Merck Group (1 mentions)
Prazosin, by Merck Group (1 mentions)
L name, by Merck Group (1 mentions)
Verapamil, by Bio-Techne (1 mentions)
11 protocols using carbamylcholine chloride
1
Studying Vasodilatory Mechanisms in Arteries
2
Calcium Imaging of Cell Aggregates
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The aggregates were transferred to uncoated 8-well µ-slides (Ibidi). Calcium transients were measured using the Fluo-4 Direct Calcium Assay kit (Thermo Fisher Scientific). Fluo-4 was pre-warmed at 37 °C and loaded into the cells by adding an equal volume to the culture medium present in the well. The cells were incubated at 37 °C for 30 min. In experiments using chemical receptor agonists, isoproterenol hydrochloride (Sigma-Aldrich, Burlington, MA, USA) or carbamylcholine chloride (Sigma-Aldrich, Burlington, MA, USA) were diluted in pre-warmed Fluo-4 solution and loaded into cells at a final concentration of 1 µM. The cells were incubated at 37 °C for 15 to 30 min.
The samples were measured using the confocal microscope previously described with a dry objective 10.0× g magnification (numerical aperture of 0.40), with images obtained at a resolution of 128 × 128 pixels, with a scan speed of 700 Hz, and with frame intervals of 97 msec during 30 to 60 s. Fluo-4 excitation was performed using a 488 nm line with an argon ion laser and the fluorescence emission was collected (500–650 nm) using a Leica HyD hybrid detector. The samples were measured within 4–5 min after leaving the incubator using a heating stage set at 37 °C to maintain the temperature before and during measurements.
The samples were measured using the confocal microscope previously described with a dry objective 10.0× g magnification (numerical aperture of 0.40), with images obtained at a resolution of 128 × 128 pixels, with a scan speed of 700 Hz, and with frame intervals of 97 msec during 30 to 60 s. Fluo-4 excitation was performed using a 488 nm line with an argon ion laser and the fluorescence emission was collected (500–650 nm) using a Leica HyD hybrid detector. The samples were measured within 4–5 min after leaving the incubator using a heating stage set at 37 °C to maintain the temperature before and during measurements.
3
Pharmacological Modulation of Cellular Signaling
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The following drugs were used: Carbamylcholine chloride (Carbachol, CCh), propranolol,
yohimbine, prazosin, L-NAME, calcium chloride (CaCl2), and hexamethonium were
purchased from Sigma Chemical Co. (Sigma-Aldrich, USA). Atropine was supplied by Research
Biochemical Incorporated, USA and Verapamil by Tocris, USA. All chemicals used were of the
analytical grade available and solubilized in distilled water.
yohimbine, prazosin, L-NAME, calcium chloride (CaCl2), and hexamethonium were
purchased from Sigma Chemical Co. (Sigma-Aldrich, USA). Atropine was supplied by Research
Biochemical Incorporated, USA and Verapamil by Tocris, USA. All chemicals used were of the
analytical grade available and solubilized in distilled water.
4
Preparation of Physiological Solutions
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Calcium chloride dihydrate (CaCl2.2H2O), magnesium chloride
hexahydrate (MgCl2.6H2O), potassium chloride (KCl) and sodium
bicarbonate (NaHCO3) were purchased from VETEC (Brazil). Monosodium phosphate
1-hydrate (NaH2PO4.H2O), glucose
(C6H12O6), magnesium sulfate monohydrate
(MgSO4.H2O) and hydrochloric acid (HCl) were purchased from
Nuclear (Brazil). Sodium chloride (NaCl) was purchased from Dinâmica (Brazil).
Carbamylcholine chloride (CCh), aminophylline, trichloroacetic acid, arachidonic acid
(AA), and thiobarbituric acid were purchased from Sigma-Aldrich (Brazil). Carbogenic
mixture (95% O2 and 5% CO2) was purchased from White Martins
(Brazil).
hexahydrate (MgCl2.6H2O), potassium chloride (KCl) and sodium
bicarbonate (NaHCO3) were purchased from VETEC (Brazil). Monosodium phosphate
1-hydrate (NaH2PO4.H2O), glucose
(C6H12O6), magnesium sulfate monohydrate
(MgSO4.H2O) and hydrochloric acid (HCl) were purchased from
Nuclear (Brazil). Sodium chloride (NaCl) was purchased from Dinâmica (Brazil).
Carbamylcholine chloride (CCh), aminophylline, trichloroacetic acid, arachidonic acid
(AA), and thiobarbituric acid were purchased from Sigma-Aldrich (Brazil). Carbogenic
mixture (95% O2 and 5% CO2) was purchased from White Martins
(Brazil).
5
Rat Aortic Vascular Reactivity
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Calcium chloride dihydrate (CaCl2.2H2O), magnesium chloride
hexahydrate (MgCl2.6H2O), potassium chloride (KCl), and sodium
bicarbonate (NaHCO3) were purchased from VETEC (Brazil). Monosodium
phosphate 1-hydrate (NaH2PO4.H2O), glucose
(C6H12O6), magnesium sulfate monohydrate
(MgSO4.H2O), and hydrochloric acid (HCl PA) were purchased
from Nuclear (Brazil). Sodium chloride (NaCl) was purchased from Dinâmica (Brazil).
Carbamylcholine chloride, acetylcholine (ACh), and phenylephrine (Phe) were purchased
from Sigma-Aldrich (USA). The carbogenic mixture (95% O2 and 5%
CO2) was purchased from White Martins (Brazil).
hexahydrate (MgCl2.6H2O), potassium chloride (KCl), and sodium
bicarbonate (NaHCO3) were purchased from VETEC (Brazil). Monosodium
phosphate 1-hydrate (NaH2PO4.H2O), glucose
(C6H12O6), magnesium sulfate monohydrate
(MgSO4.H2O), and hydrochloric acid (HCl PA) were purchased
from Nuclear (Brazil). Sodium chloride (NaCl) was purchased from Dinâmica (Brazil).
Carbamylcholine chloride, acetylcholine (ACh), and phenylephrine (Phe) were purchased
from Sigma-Aldrich (USA). The carbogenic mixture (95% O2 and 5%
CO2) was purchased from White Martins (Brazil).
6
Resveratrol Vasorelaxant Mechanism Evaluation
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Resveratrol (5-[(1E)-2-(4-Hydroxyphenyl)ethenyl]-1,3,benzenediol) was purchased from abcr GmbH (Karlsruhe, Germany). Carbamylcholine chloride ((2-Hydroxyethyl)trimethylammonium chloride carbamate; carbachol), NG-Methyl-L-Arg (Nω-Nitro-L-arginine methyl ester hydrochloride, L-NAME), N5-(Nitroamidino)-L-2,5-diaminopentanoic acid, NG-NO2-L-Arg (Nω-Nitro-L-arginine, L-NNA), 1H-[1,2,4]Oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), iberiotoxin (IbTX), charybdotoxin (ChTX), apamin, glibenclamide, N,N,N,N-Tetraethylammonium chloride (TEA), 4-Aminopiridine (4AP) and tamoxifen were purchased from the Sigma Chemical Company, were purchased from Sigma (St. Louis, MO).
Resveratrol was dissolved in 70% ethanol so that the final concentration of ethanol was never >0.1%, which did not affect basal contraction. The working solutions were prepared fresh on the day of the experiment by diluting the stock solution.
Stock solutions of carbachol, L-NNA, L-NAME, apamin, IbTX, ChTX, TEA, 4AP, and tamoxifen were prepared with bidistilled water, and glibenclamide and ODQ were dissolved in dimethyl sulphoxide (DMSO). The given concentrations were the calculated final concentrations in the organ bath solution. All reagents were added directly to the bath fluid containing a Tyrode’s solution composed of (mmol/L): NaCl 139.6; KCl 2.68; MgCl2 1.05; NaH2PO4 1.33; CaCl2 1.80; NaHCO3 25.0; and glucose 5.55.
Resveratrol was dissolved in 70% ethanol so that the final concentration of ethanol was never >0.1%, which did not affect basal contraction. The working solutions were prepared fresh on the day of the experiment by diluting the stock solution.
Stock solutions of carbachol, L-NNA, L-NAME, apamin, IbTX, ChTX, TEA, 4AP, and tamoxifen were prepared with bidistilled water, and glibenclamide and ODQ were dissolved in dimethyl sulphoxide (DMSO). The given concentrations were the calculated final concentrations in the organ bath solution. All reagents were added directly to the bath fluid containing a Tyrode’s solution composed of (mmol/L): NaCl 139.6; KCl 2.68; MgCl2 1.05; NaH2PO4 1.33; CaCl2 1.80; NaHCO3 25.0; and glucose 5.55.
7
Pharmacological Modulation of Calcium Signaling
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Prostaglandin E2, Serotonin hydrochloride(5-HT), SKF-96365, TPEN (N, N, N′, N′- tetrakis (2-pyridylmethyl) ethylenediamine), GSK-7975A, ouabain, HC067047, and GSK1016790A were purchased from MedChemExpress (MCE; Monmouth Junction, NJ, United States). Sigma (Saint Louis, MO, United States) supplied carbamylcholine chloride (CCh), nifedipine, gadolinium chloride, and cyclopiazonic acid (CPA). Tocris Bioscience (Ellisville, MO, United States) supplied 2-Aminoethoxydiphenyl borate (2-APB), while APExBIO Technology LLC (Houston, TX, United States) provided dantrolene. Fura-2 was purchased from Invitrogen (UT, United States), meantime DMEM and FBS were obtained from Hyclone (Logan, UT, United States). Trypsin and penicillin/streptomycin were purchased from Gibco (CA, United States). The other chemicals were obtained from BBI Life Science (Shanghai, China).
8
Acute Hemorrhage Resuscitation Strategies
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Experimental groups differed according to the combination of nitroglycerine (Nitrolingual infusion, G. Pohl-Boskamp & co KG, Hohenlockstedt, Germany), iloprost (5-cis Iloprost, Cayman Chemicals, Ann Arbor, MI, USA), carbachol (Carbamylcholine chloride, Sigma-Aldrich Corporation, St. Louis, USA) and saline application. All investigators were blinded to the pharmacological treatment. The experimental protocol of acute hemorrhage and retransfusion did not differ between the groups. All experimental groups - including those with a sole nitroglycerine or iloprost treatment - have been performed exclusively for the here addressed research questions and have not been published before.
9
Nicotinic Receptor Agonist-Induced Current Assay
10
Microinjection of Cholinergic and Dopaminergic Drugs
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Carbachol as a non-selective cholinergic receptor agonist (Carbamylcholine chloride; Sigma-Aldrich, St. Louis, Missouri, USA) was dissolved in sterile normal saline. D1-like dopamine receptor antagonist, SCH-23390, (R)-(+)-7-Chloro-8-hydroxy-3-methyl-1-phenyl-2, 3, 4, 5-tetrahydro-1H-3-benzazepine hydrochloride (Tocris Bioscience, Bristol, UK) was dissolved in sterile normal saline. D2-like dopamine receptor antagonist, Sulpiride, (S)-5-aminosulfonyl-N- [(1- ethyl-2 pyrrolidinyl) methyl]-2-methoxybenzamide (Tocris Bioscience, Bristol, UK) was dissolved in dimethyl sulfoxide (DMSO 12%), Sigma-Aldrich, Germany. Formalin 2.5% was prepared by diluting 37% formaldehyde (Merck, Germany) with sterile physiological saline (0.9%). All drugs or vehicle solutions were freshly prepared on the day of the experiment and infused slowly over 60 s. All microinjections were conducted in freely moving animals via a stainless-steel injector (30-gauge needle; 1 mm longer than guide cannula) which was connected to a 1-µL Hamilton syringe via a polyethylene tube (PE-20).
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