Salts used in ND96, HEPES, GABA, propofol, and 5β-pregnan-3α-ol-20-one (3α5βP) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Etomidate was purchased from Toronto Research Chemicals (Toronto, ON, Canada). NS-1738 was obtained from Cayman Chemical (Ann Arbor, MI, USA) and Adooq Bioscience (Irvine, CA, USA). PAM-2 was synthesized as described previously [33 (link)]. A stock solution of 500 mM GABA in ND96 was stored at 4 °C. All other stock solutions were made in dimethyl sulfoxide (Sigma-Aldrich) with 200 mM propofol and 20 mM 3α5βP stocks stored at room temperature and 200 mM Etomidate, 100 mM NS-1738, and 100 mM PAM-2 stocks stored at −20 °C. Final dilutions were made on the day of the experiment.
Etomidate
Manufactured by Merck Group
Sourced in Denmark, United States
Etomidate is a short-acting intravenous anesthetic agent used for the induction of general anesthesia. It is a positive allosteric modulator of the GABA(A) receptor and exerts its effects by enhancing the activity of this receptor. Etomidate is known for its rapid onset of action and relatively short duration of anesthetic effects.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (3 mentions)
Mz125, by Leica (2 mentions)
Propofol, by Merck Group (1 mentions)
Ns 1738, by Cayman Chemical (1 mentions)
Hepes, by Merck Group (1 mentions)
Rnalater, by Merck Group (1 mentions)
Menthol, by Merck Group (1 mentions)
4α pdd, by Merck Group (1 mentions)
Lipofundin, by B. Braun (1 mentions)
Capsaicin, by Merck Group (1 mentions)
Progesterone, by Steraloids (1 mentions)
Androstenedione, by Steraloids (1 mentions)
Testosterone, by Steraloids (1 mentions)
Unlabeled pregnenolone, by Steraloids (1 mentions)
17α hydroxyprogesterone, by Steraloids (1 mentions)
3h pregnenolone, by DuPont (1 mentions)
3h progesterone, by DuPont (1 mentions)
3h androstenedione, by DuPont (1 mentions)
3h testosterone, by DuPont (1 mentions)
11 protocols using etomidate
1
Preparation of Solutions for Neuroscience Experiments
2
Zebrafish CBD Exposure Study
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A total of 30 zebrafish was divided into three experimental groups (each containing 2x5 fish): 1) pure water control (facility water), 2) solvent control (0.4% ethanol) and 3) CBD (CBD in 0.4% ethanol in water). Before experimental start each duplicate group was acclimatized for 1h in a 1.8L tank containing water from the facility whereafter individual fish were placed separately in a well (6-well plate, Greiner bio-one, CELLSTAR) containing 10 ml of the test solution. After 30 min exposure (water, ethanol or CBD) fish were euthanized by immersion into an overdose (30 mg/L) etomidate (Sigma-Aldrich, Denmark) solution, subsequently dissected under a stereomicroscope (Leica MZ12.5, Leica Denmark) and internal organs (intestine, liver, spleen) transferred to RNAlater (Sigma-Aldrich, Denmark). Preserved material was kept at 4°C for 24h and then frozen at -20°C until processing for gene expression studies.
3
Etomidate Modulates AGE-Induced Chondrocyte Responses
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SW1353 chondrocytes were obtained from ATCC (ATCC, Manassas, USA) and cultured in DMEM supplemented with 5% FBS at 37°C and 5% CO2 [19 (link)]. A specific siRNA was designed and transfected into human SW1353 chondrocytes together with the transfection agent lipofectamine 3000 (Thermo, MA, USA) according to the instructions of the manufacturer, followed by verifying the efficacy of silencing SOX-9 using Western blotting assay. Cells were stimulated with 1.25, 2.5, and 5 μM Etomidate (#E6530, Sigma-Aldrich, USA) for 24 hours [20 (link)]. Etomidate was dissolved in DMSO. Cells were then stimulated with 100 μg/mL AGEs (Glycosylated BSA, # bs-1158P, Beijing Biosynthesis Biotechnology, China).
4
Zebrafish Immune Response to Lipopolysaccharide and Exopolysaccharide
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We used a total number of 80 adult AB wild-type zebrafish, age 11 months [mean body weight of 387.8 (SD: 104.4) mg and mean body length of 29.9 (SD: 2.28) mm] which were provided by the Panum Institute, University of Copenhagen and reared in a thermostat-controlled room at 28°C with a 12 h light: 12 h dark cycle. The fish were divided into eight experimental groups each containing 10 fish in duplicate (5 fish/tank); PBS-injected group (control), LPS-injected group using phenol-purified LPS from E. coli 0111:B4 (L2630, Sigma) (1.5 mg/mL), three ES-injected groups treated with different concentrations (low: 50 μg/mL, medium: 500 μg/mL, high: 1000 μg/mL), and three LPS+ES-injected groups treated with different concentrations of ES as explained earlier.
Fish were anesthetized by immersion into a solution of 2 mg/L etomidate (Sigma-Aldrich, Denmark). In each group, individual fish were injected intraperitoneally (i.p.) with a total volume of 20 μL solution. Intraperitoneal injection was performed using a Biohit automatic pipette (Dandiag, Denmark) mounted with a sterile BD MicrolanceTM 3 needle (BD, Denmark) on a modified 200 μL pipette tip (Almeco A/S, Denmark) under a stereomicroscope (Leica MZ12.5, Leica Denmark). After injection, fish were returned to their tanks and observed every 2 hours in order to remove and euthanize any moribund fish from the tanks.
Fish were anesthetized by immersion into a solution of 2 mg/L etomidate (Sigma-Aldrich, Denmark). In each group, individual fish were injected intraperitoneally (i.p.) with a total volume of 20 μL solution. Intraperitoneal injection was performed using a Biohit automatic pipette (Dandiag, Denmark) mounted with a sterile BD MicrolanceTM 3 needle (BD, Denmark) on a modified 200 μL pipette tip (Almeco A/S, Denmark) under a stereomicroscope (Leica MZ12.5, Leica Denmark). After injection, fish were returned to their tanks and observed every 2 hours in order to remove and euthanize any moribund fish from the tanks.
5
Preparation and Dilution of Pharmacological Agents
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Etomidat-® Lipuro, Lipofundin® (both B. Braun Melsungen AG, Germany) and
Hypnomidate® (JANSSEN-CILAG GmbH, Germany) were purchased from the
hospital pharmacy of the University Clinics of Erlangen. Etomidate
(Sigma-Aldrich, Germany), BCTC, HC-030031 (both Biotrend, Germany),
AP-18, 2-amino-phenyl borane (2-APB) (both Tocris Bioscience, UK) were
dissolved in dimethyl sulfoxide to give stock solutions of 1–100 mM.
AITC, capsaicin, menthol, and 4αPDD (all Sigma-Aldrich, Germany) were
dissolved in ethanol to give stock solutions of 1 to 100 mM. PG and
N-acetyl-L-cysteine (NAC) were purchased from Sigma-Aldrich, Germany.
All formulations, substances, and stock solutions were diluted with
standard physiological buffers (pH 7.4) immediately before
experimental use.
Hypnomidate® (JANSSEN-CILAG GmbH, Germany) were purchased from the
hospital pharmacy of the University Clinics of Erlangen. Etomidate
(Sigma-Aldrich, Germany), BCTC, HC-030031 (both Biotrend, Germany),
AP-18, 2-amino-phenyl borane (2-APB) (both Tocris Bioscience, UK) were
dissolved in dimethyl sulfoxide to give stock solutions of 1–100 mM.
AITC, capsaicin, menthol, and 4αPDD (all Sigma-Aldrich, Germany) were
dissolved in ethanol to give stock solutions of 1 to 100 mM. PG and
N-acetyl-L-cysteine (NAC) were purchased from Sigma-Aldrich, Germany.
All formulations, substances, and stock solutions were diluted with
standard physiological buffers (pH 7.4) immediately before
experimental use.
6
Steroid Metabolism in Rats
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[3H]Pregnenolone, [3H]progesterone, [3H]androstenedione, [3H]testosterone, [3H]dihydrotestosterone were purchased from DuPont-New England Nuclear (Boston, MA). Unlabeled pregnenolone, progesterone, 17α-hydroxyprogesterone, androstenedione and testosterone were obtained from Steraloids (Newport, RI). Etomidate was purchased from Sigma (St. Louis, MO). Male Sprague-Dawley rats (30-day-old) were purchased from Shanghai Animal Center (Shanghai, China). All animal procedures were approved by the Institutional Animal Care and Use Committee of Wenzhou Medical University and were performed in accordance with the Guide for the Care and Use of Laboratory Animals.
7
Pharmacological Tools for GABA and Glutamate Studies
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The following drugs were purchased from Tocris Bioscience (UK) and were used during the experiments: 6,7-dinitroquinoxaline-2,3-dione (DNQX; selective blocker of non-NMDA glutamate receptors), tetrodotoxin (TTX; sodium channel selective blocker), gaboxadol (THIP; superagonist for δ-containing extrasynaptic GABAARs), etomidate (enhancer of β2/3-containing GABAARs), L-655,708 (selective inverse agonist for α5-containing GABAARs), picrotoxin (PTX; non-specific GABAARs antagonist), and N-methyl-D-aspartic acid (NMDA; NMDARs selective agonist). The stock solutions of etomidate, L-655,708, and picrotoxin were dissolved in dimethyl sulfoxide (DMSO, Sigma) and then added into the ACSF during experiments [not exceeding the concentrations of DMSO > 0.1% v/v (see Lebida and Mozrzymas, 2017 (link))].
8
Differentiation of Mouse ESCs
9
Purification and Characterization of Human GABAARs
10
Semiconducting SWCNT Materials Synthesis
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Semiconducting 99% single-walled carbon nanotubes (swCNTs) were purchased from NanoIntegris, Inc. (Canada). γ-aminobutyric acid (GABA), bicuculine, etomidate, cytochalasin B, Dulbecco's phosphate buffered saline (DPBS), protease inhibitor cocktail, 1,2-dichlorobenzene, octadecyltrichlorosilane, and poly-D-lysine were purchased from Sigma Aldrich (USA).
Dulbecco's Modified Eagle Medium (DMEM) and fetal bovine serum were purchased from Biowest (France). Penicillin-streptomycin serum was purchased from Gibco (USA).
Lipofectamine 3000 was purchased from Invitrogen (USA).
Dulbecco's Modified Eagle Medium (DMEM) and fetal bovine serum were purchased from Biowest (France). Penicillin-streptomycin serum was purchased from Gibco (USA).
Lipofectamine 3000 was purchased from Invitrogen (USA).
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