Dulbecco s modified eagle medium

The rat glioma C6 cell line was obtained from the American Type Culture Collection (Manassas, VA, USA) and maintained at 37°C and 5% CO₂ in Dulbecco's Modified Eagle Medium (CULTILAB, São Paulo, Brazil) supplemented with 10% (v/v) fetal bovine serum (CULTILAB), penicillin (50 U/mL), and streptomycin (50 µg/mL). Approximately 1.5×10⁵ cells were seeded and maintained at 37°C for 24 h before exposure to MeHg, morphine, and/or naloxone.

The following reference chemicals were obtained from the sources specified: histamine dihydrochloride, carbachol (CCh) (Merck, EUA), Cremophor EL®, S-(-)-Bay K8644, cesium chloride (CsCl), EDTA, HEPES, methylthiotetrazole (MTT), dimethyl sulfoxide (DMSO), penicillin-streptomycin (Sigma-Aldrich, Brazil), ethanol PA (Reagen, Brazil), dulbecco’s modified eagle medium (DMEM), bovine fetal serum, L-glutamine and trypsin/EDTA solution (1:250) (Cultilab, Brazil) and Flou-4 NW dye mix (Invitrogen, EUA).
The following chemicals were used to make the physiological salt solutions: glucose, magnesium sulphate, calcium chloride (Vetec, Brazil), sodium bicarbonate (Fmaia, Brazil), sodium chloride, potassium chloride (Química Moderna, Brazil), monosodium phosphate-1-hydrate, sodium hydroxide and hydrochloric acid (Nuclear, Brazil)
All chemicals used were of the highest purity grade available. Stock solutions of all the chemicals were made in distilled water and the dilutions were made fresh on the day of the experiment.
Carbogen mixture (95 % O₂ and 5 % CO₂) was obtained from White Martins (Brazil).