All modulators were dissolved in dimethyl sulfoxide (DMSO), if not indicated otherwise. Dimethyl sulfoxide concentrations in the final solution never exceeded 0.14% at the highest test concentration of the respective modulator and were further reduced by serial dilution of the compounds. Pregnenolone sulfate, allyl isothiocyanate (AITC), CFA (containing 1 mg·mL−1 of heat-killed and dried mycobacterium), diclofenac, maprotiline, primidone, and clotrimazole were purchased from Sigma-Aldrich (Deisenhofen, Germany). For behavioural studies, primidone was suspended in 0.5% Tween 20 (Sigma-Aldrich) which was also used as vehicle control.
Primidone
Manufactured by Merck Group
Sourced in Germany
Primidone is a laboratory instrument manufactured by Merck Group. It is a pharmaceutical drug that is primarily used as an anticonvulsant medication for the treatment of epilepsy. The core function of Primidone is to prevent and control seizures in individuals diagnosed with epilepsy.
Automatically generated - may contain errors
Lab products found in correlation
Clotrimazole, by Merck Group (3 mentions)
Carbamazepine, by Merck Group (3 mentions)
Ethosuximide, by Merck Group (3 mentions)
Tween 20, by Merck Group (2 mentions)
Diclofenac, by Merck Group (2 mentions)
Pregnenolone sulfate, by Merck Group (2 mentions)
Lamotrigine, by Merck Group (2 mentions)
Levetiracetam, by Merck Group (2 mentions)
Oxcarbazepine, by Merck Group (2 mentions)
Isosakuranetin, by Carl Roth (2 mentions)
Valproate, by Merck Group (2 mentions)
Maprotiline, by Merck Group (1 mentions)
Allyl isothiocyanate, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Magnesium sulfate, by Merck Group (1 mentions)
Gabapentin, by Merck Group (1 mentions)
Vigabatrin, by Merck Group (1 mentions)
Tiagabine, by Merck Group (1 mentions)
Topiramate, by Merck Group (1 mentions)
Antibiotic antimycotic solution, by Thermo Fisher Scientific (1 mentions)
7 protocols using primidone
1
Dissolution and Dilution of Modulators
2
Reagent Sources for Cell Culture
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EMEM powder and antibiotic/antimycotic solution were obtained from Gibco, Grand Island, NY. FBS and phosphate buffered saline (PBS) were obtained from Hyclone through Thermo Scientific. Dimethyl sulfoxide (DMSO), ethanol (99 %) (EtOH), TMZ, DEX, VPA, carbamazepine (CBZ), ethosuximide (ESX), gabapentin (GBP), lamotrigine (LTG), levetiracetam (LEV), magnesium sulfate (MgSO4), oxcarbazepine (OXC), phenytoin (5, 5 diphenylhydantoin) (PHT), primidone (PRM), tiagabine (TGB), topiramate (TPM), and vigabatrin (VBT) were obtained from Sigma-Aldrich, St. Louis, MO. R-(−)-MHC and S-(+)-MHC were obtained from Santa Cruz, Dallas, TX.
3
Quantitative CBZ Immunoassay Development
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CBZ, valproate sodium, phenytoin sodium, primidone, Tween-20 and bovine serum albumin (BSA) were purchased from Sigma-Aldrich (Shanghai, China). CBZ antibody clone CA1 was obtained from Bio-Rad Laboratories (Shanghai, China). CBZ bovine serum albumin conjugate was purchased from Unibiotest Co., Ltd. (Wuhan, China). ZebaTM spin desalting columns (7 K MWCO, 0.5 mL), 3,3′- diaminobenzidine (DAB) enhanced liquid substrate system tetrahydrochloride, and SuperBlock™ (PBS) blocking buffer were obtained from Thermo Scientific (Shanghai, China). The biotinylating kit was obtained from Genemore (Suzhou, China). The horseradish peroxidase (HRP) Conjugation Kit-Lightning-Link® (ab102890) was from Abcam (Shanghai, China). Single donor human serum off the clot and human whole blood was obtained from Innovative Research (Novi, MI, USA). The 96-well polystyrene black microplates were obtained from Greiner Bio-One GmbH (Shanghai, China). The Octet® K2 2-channel system and streptavidin sensors were purchased from Sartorius Stedim Biotech GmbH (Gottingen, Germany). Phosphate-buffered saline (PBS, 10 mM, pH 7.4), PBS buffer containing 0.02% (v/v) Tween-20 and 0.1% BSA (referred to as sample diluent (SD) buffer), and SD buffer containing 274 mM NaCl (referred to as high-salt SD buffer) were used in this study.
4
Standardized Compound Procurement Protocol
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Clotrimazole, [D‐Ala2, N‐Me‐Phe4, Gly5‐ol]‐Enkephalin acetate salt (DAMGO), Diclofenac, Naloxone, Nifedipine, N‐methyl‐D‐glucamine, Pregnenolone sulphate sodium salt and Primidone were purchased from Sigma Aldrich. CIM0216 was purchased from Tocris Bioscience. Isosakuranetin was purchased from Carl Roth and Ionomycin was purchased from Fisher Scientific. N‐methyl‐D‐glucamine was directly diluted in external recording solutions. Stock solutions were prepared from all other drugs by diluting them in DMSO according to distributor instructions.
5
Synthesis and Characterization of Anticonvulsant Compounds
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Ethyl ketopentenoate (EKP) was synthetized according to a procedure described in supplementary materials. Pentylenetetrazol (PTZ), lamotrigine (LMT), carbamazepine (CBZ), primidone (PMD), ethosuximide (ETS), levetiracetam (LVT), valproate (VPA), oxcarbazepine (OXC), zonisamide (ZSM), topiramate (TPM) and aspirin (ASP) were purchased from Sigma. Other compounds used were: phenytoin (PHT) (Acros), tiagabine (TGB) (Chemos), perampanel (PER) (Eisai), retigabine (RGB) (Valeant Pharmaceuticals/GlaxoSmithKline) and coelenterazine-h (CLZN-h) (NanoLight® Technologies). All compounds were dissolved in dimethyl sulfoxide (DMSO) and kept as a stock at −20 °C or −80 °C. Stock solutions were further diluted in embryo medium for locomotor activity assay and local field potential recordings, or E3 medium (5 mM NaCl, 0.17 mM KCl, 0.33 mM CaCl2 and 0.33 mM MgSO4) for neuroluminescence recording to achieve a final DMSO concentration of 1% w/v. As vehicle control (VHC) 1% w/v DMSO in embryo or E3 medium was used.
6
Anticonvulsant Drug Characterization
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Purified CA I and CA II, HEPES, p-nitrophenol, carbamazepine, phenytoin, valproate, primidone, clonazepam, and ethosuximide were purchased from SIGMA (Deisenhofen, Germany). The antiepileptic drugs used were: Tegretol (Ciba, Basel, Switzerland), Phenhydan (Desitin, Hamburg, Germany), Depakine (Sanofi, Paris, France), Mysoline (ICI-Zeneca, London, UK), Rivotril (Hoffmann-La Roche, Basel, Switzerland), and Pentinimid (Gerot, Vienna, Austria).
7
Ratiometric Fura-2 Fluorimetry for Intracellular Calcium
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Changes in intracellular calcium concentration were monitored using ratiometric Fura-2-based fluorimetry. Cells were loaded with 2 µM Fura-2-acetoxymethyl ester (Alexis Biochemicals) for 30 min at 37°C. Fluorescence was measured during alternating illumination at 340 and 380 nm using Eclipse Ti (Nikon) fluorescence microscopy system, and absolute calcium concentration was calculated from the ratio of the fluorescence signals at these two wavelengths (R = F340/F380) as [Ca2+]=Km × (R-Rmin)/(Rmax-R), where Km, Rmin and Rmax were estimated from in vitro calibration experiments with known calcium concentrations. The bath solution contained (in mM) 138 NaCl, 5.4 KCl, 2 CaCl2, 2 MgCl2, 10 glucose, and 10 HEPES, pH 7.4. Pregnenolone sulfate, clotrimazole and primidone were obtained from Sigma-Aldrich, isosakuranetin was obtained from Carl Roth. The chemical ligands were dissolved in bath solution from a stock diluted in DMSO.
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