Methylmercury

The following reagents were purchased from Thermo Fisher Scientific (Waltham, MA, USA): B-27^® supplement 50x, Fluo-4 AM cell permeant, formaldehyde 37%, GlutaMAX™ supplement, human recombinant epidermal growth factor (EGF), MEM non-essential amino acids solution 100x, N-2 supplement 100x, and phosphate buffered saline 1x (PBS). The following reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA): 2,5-hexanedione, γ-Aminobutyric acid (GABA), bovine serum albumin (BSA), dimethyl sulfoxide (DMSO), Dulbecco’s Modified Eagle Medium (DMEM), endosulfan, methylmercury, penicillin-streptomycin solution 100x, Triton™ X-100, and TWEEN^® 20. Matrigel^® was purchased from Corning (Corning, NY, USA). Fetal calf serum (FCS) was purchased from ATCC (Manassas, VA, USA). Human recombinant fibroblast growth factor basic (bFGF) was purchased from Peprotech (Rocky Hill, NJ, USA). Tetrodotoxin (TTX) was purchased from Tocris Bioscience (Bristol, UK).

Valproic acid (VPA), deferoxamine (DFX), acetaminophen (APAP), nicotine (NCT), dexamethasone (DEX), chlorpyrifos (CPF), and methyl mercury (MeHg) were purchased from Sigma (St. Louis, MO, USA). Cyclopamine (CPM) and retinoic acid (RA) were purchased from Wako (Osaka, Japan). Trichostatin A (TSA), carbamazepine (CBZ), bisphenol A (BPA), and saccharin (SAC) were purchased from Tokyo Kasei (Tokyo, Japan). With the exception of CPM, DFX, and APAP, stock solutions were prepared in dimethyl sulfoxide (DMSO; Nacalai, Kyoto, Japan). CPM was dissolved in ethanol (Wako), and DFX and APA were dissolved in in 0.3× Danieau’s solution (19.3 mM NaCl, 0.23 mM KCl, 0.13 mM MgSO₄, 0.2 mM Ca(NO₃)₂, 1.7 mM HEPES, pH 7.2). For experiments, chemicals were serially diluted in 0.3× Danieau’s solution. Controls contained the same final concentrations of vehicle.