Alcohol (95% w/v) was diluted in distilled water to a concentration of 20% (v/v) and administered IG, with volumes varied by weight to obtain the desired dose. Fluoro-Gold (FG; Fluorochrome, LLC, Denver, Colorado) was dissolved in 0.9% saline (w/v)/2% (v/v) FG per manufacturer instructions (Schmued & Fallon, 1986 (link)). Muscimol and baclofen (R&D systems, Minneapolis, Minnesota) were dissolved in sterile 0.9% saline to produce a cocktail of 0.1mM Muscimol + 1mM baclofen, and the doses were chosen based on previous work and our own pilot studies (Lasseter et al., 2011 (link); Chaudhri et al., 2013 (link)).
Muscimol
Manufactured by R&D Systems
Muscimol is a potent GABAA receptor agonist. It is a naturally occurring compound found in certain species of mushrooms. Muscimol acts as a selective agonist for the GABAA receptor, which is an important inhibitory neurotransmitter receptor in the central nervous system.
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6 protocols using muscimol
1
Alcohol, FG, Muscimol, and Baclofen Administration
2
Muscimol Preparation and Dosing
3
Alcohol, Sucrose, and Neuropharmacological Cocktail Protocol
4
Muscimol Dosage Protocol for Experiments
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5
Pharmacological Reagents Protocol
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Reagents were purchased from Sigma except for isradipine, L–NNA, DTT, ryanodine, orexin, muscimol, and CGP37157 were purchased from R&D Systems; TMRM, Alexa568, Alexa594, and Fluo4 obtained from Life Technologies; genipin was obtained from Wako Reagents, and Ru360 obtained from EMD Biosciences. L–NAME was purchased from Sigma–Aldrich. Drugs stock solutions were prepared in deionized water, DMSO, ethanol, or methanol as instructed by manufacturer on the day of experiment. Stocks were diluted to final concentrations in ACSF to achieve a final solvent concentration of <0.01% v/v.
6
Evaluating TLR4 Inhibitor and GABAA Agonist in Epilepsy
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We used TAKC-242 as TLR4 inhibitor, which can selectively bind TLR4 and block the interaction between TLR4 and joint molecules, thus inhibiting TLR4 signal transduction and its downstream signal pathways [23 (link)]. muscimol is a potent GABAA receptor agonist. Intramuscular injection of muscimol inhibits acetylcholine, histamine, or the vagus nerve by activating GABAA channels [24 (link)].
From the second day, before administration of pilocarpine or IS, TAK-242 (3 mg/Kg, dissolved in DMSO, APExBIO) and muscimol (1 mg/Kg, dissolved in sterile distilled water, R&D Systems) were injected intraperitoneally before 1.5 h and 0.5 h, respectively (Table 2 ). The epileptic latency and seizure levels of rats were recorded during the epilepsy period, and the number of scratches and the pain thresholds of rats was observed during the comorbid period.
From the second day, before administration of pilocarpine or IS, TAK-242 (3 mg/Kg, dissolved in DMSO, APExBIO) and muscimol (1 mg/Kg, dissolved in sterile distilled water, R&D Systems) were injected intraperitoneally before 1.5 h and 0.5 h, respectively (
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