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4 protocols using rs ampa hydrobromide

1

Pharmacological Agents in Neurophysiology

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Dyes and drugs were purchased from the following suppliers: (RS)-AMPA hydrobromide (AMPA), Kainate (KA), GYKI 52466 hydrochloride (GYKI), DL-threo-β-Benzyloxyaspartic acid (TBOA), NBQX disodium salt (NBQX), DL-AP5 sodium salt (APV), dynamin inhibitory peptide (QVPSRPNRAP), dynamin inhibitory peptide myristoylated (scrambled), dynamin inhibitory peptide myristoylated, cyclothiazide, mecamylamine, dhβe, and Dynasore (Tocris, Ellisville, MO, USA); N-Methyl-D-Aspartate (NMDA), tetrodotoxyn (TTX), Strychnine, Bicuculline and L-glutamate, atropine (Sigma, St Louis, MO); Fluoro-Gold (Fluorochrome, Inc., Denver, CO, USA); Lucifer Yellow (LY), Acridine Orange 10-Nonyl (AO), 6-methoxy-N-ethylquinolinium iodide (MEQ), and Florescien-5-isothiocyan (FITC) were from Invitrogen. Dynasore was dissolved in DMSO. When bath applied to the cord, the concentration of DMSO was less than 0.1%. All other drugs were dissolved in artificial cerebrospinal fluid (aCSF).
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2

Preparation of Compounds for Electrophysiology

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BAPTA-AM, NMDA and (R,S)-AMPA hydrobromide, zolpidem, cyclothiazide, pregnenolone sulfate sodium salt, ifenprodil hemitartrate and (+)-MK801 maleate were obtained from Tocris (Bristol, U.K.); γ-aminobutyric acid (GABA) and (-)bicuculline methiodide were from Sigma-Aldrich (Poole, U.K.). Compound LY3130481 was synthesised internally at Eli Lilly and Company (Gardinier et al., 2016 ). Concentrated stock solutions of BAPTA-AM, zolpidem, cyclothiazide, pregnenolone sulfate, ifenprodil and MK801 were prepared in DMSO and stocks of NMDA, AMPA hydrobromide, GABA and bicuculline were prepared in distilled water. The stock solutions were stored in aliquots at −20 °C. Final dilutions in oocyte recording solution were made on the day of the experiments.
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3

AMPA and Dexmedetomidine Larval Assay

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(RS)-AMPA hydrobromide (Tocris; Bristol, United Kingdom) was dissolved in filtered sea water to a stock concentration of 7.5 mM and then diluted to a final concentration of 500 μM. Dexmedetomidine (Tocris) was dissolved in filtered sea water to a stock concentration of 6.75 mM and then diluted to a final concentration of 20 μM. Larvae were incubated with the drug for about 10 min before beginning assays and remained in the drug solution through the entirety of the assay.
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4

AMPA Receptor Activation in Larvae

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( RS )-AMPA hydrobromide (Tocris) was dissolved in filtered sea water to a stock concentration of 7.5 mM and then diluted to a final concentration of 500 µM. Larvae were incubated with the drug for about 10 min before beginning assays.
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