L glutamatic acid

Diazepam, LY341495, LY404039, SAR218645, SSR180711 and volinanserin (Sanofi Medicinal Chemistry), levetiracetam (Advanced Technology & Industrial Co. Ltd, Hong Kong), pentylenetetrazol, MK-801 (Sigma RBI, St Quentin Fallavier, France), haloperidol (Bell Medical Services, Inc., Marlborough, NJ), clozapine (Tocris Bioscience, Bristol, UK), olanzapine (Interchim, Clichy, France), amphetamine, 2,5-dimethoxy-4-iodoamphetamine [(±)-DOI] hydrochloride, L-glutamatic acid (Sigma-Aldrich, US or France), were dissolved or suspended in distilled water with 0.6% methylcellulose and the addition of 5% Tween 80 (Sigma RBI) (unless otherwise indicated) in in vivo studies and suspended in DMSO at 10 mM in in vitro experiments. Doses refer to the weight of the free base. Volume of administration was 10 or 20 ml/kg in mice, 2 or 5 ml/kg in rats. All drug solutions were prepared fresh daily.

For murine chondrocyte experiments, chondrocytes were isolated from sterna of newborn pups (C57BL/6 J) age P1-P3 without consideration for sex. Cells were isolated by sequential digestion with pronase (2 mg/mL, PRON-RO, Roche) at 37°, followed by collagenase D (3 mg/mL, COLLD-RO, Roche) two times at 37°, and cultured in DMEM (Life Technologies, Carlsbad, CA, USA) containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (15140122, ThermoFisher, Waltham, MA, USA) and plated in tissue culture plates. For glutamine deprivation conditions, media was changed to high glucose DMEM containing glutamine or devoid of glutamine (Life Technologies, Carlsbad, CA, USA). For experiments, cells are treated with recombinant mouse IL-1β (211-11B, Peprotech, Cranbury, NJ, USA) at 10 ng/mL, CB-839 (S7655, Selleck, Chemicals, Houston, TX, USA), rapamycin (HY-10219, MedChem Express, Monmouth Junction, NJ, USA), ammonium chloride (A9434, Sigma-Aldrich, USA), L-asparagine (A0884, Sigma-Aldrich, St. Louis, MO, USA), or L-glutamatic acid (G1626, Sigma-Aldrich, St. Louis, MO, USA).