Carbamoylcholine chloride carbachol

DAMGO, U50488H, SNC80, CGS21680, AVP (arginine vasopressin), IBMX, Forskolin, stromal cell-derived factor 1 (SDF-1 or CXCL12), carbamoylcholine chloride (carbachol), quinpirole hydrochloride, SKF 81297, ionomycin calcium salt, were purchased from Tocris Bioscience (Bristol, UK) and diluted in DMSO (diméthylsulfoxyde) at 10⁻² M (except SDF1 at 125 µM and IBMX at 200 mM) for frozen stock aliquots and coelenterazine H substrate from Interchim (Montluçon, France) was diluted in 100% ethanol and kept at −20°C. Protease/Phosphatase Inhibitor Cocktail were purchased from Cell Signaling Technology (Leiden, Netherlands). Pertussis toxin (PTX), purchased from Tocris, were diluted in water at 0.1 µg/µl and stored at 4°C. Morphine HCl was purchased from Francopia (Paris, France). Phenylmethanesulfonyl fluoride (PMSF) was diluted in isopropyl alcohol at 200 mM and stored at −20°C. Compound 19 was generously synthetized by Domain Therapeutics (Illkirch, France). For in vitro studies, it was diluted in DMSO (diméthylsulfoxyde) at 10⁻² M and frozen at −20°C. For in vivo studies, Compound 19 was kept as a powder at 4°C and diluted in a saline solution before ICV administration (NaCl 9%).

Soluble Aβ42 oligomers or soluble scrambled Aβ42 oligomers were prepared as described (32 (link), 104 (link)). One milligram of lyophilized human Aβ42 (Anaspec) or scrambled Aβ42 (Anaspec) was dissolved in 1 ml of 1,1,1,3,3,3-hexafluoro-2-propanol (Fisher Scientific) to prevent aggregation, portioned into 10-μg aliquots, air dried, and stored at −80 °C. For use in experiments, an aliquot was thawed at room temperature and then dissolved in 100% dimethyl sulfoxide, then diluted into PBS to make a 100 μM solution. The solution was incubated for 16 h at 4 °C and then diluted to a final concentration for use in experiments. The following agonists were used in this study: 1 μM PNU-120596 (Alomone Labs), 2 μM RJR-2403 Oxalate (Alomone labs), 1 μM NS-3861 (Tocris Bioscience), and 1 μM Carbamoylcholine chloride (carbachol) (Tocris Bioscience). Aβcore (YEVHHQ) and inactive Aβcore (SEVAAQ) peptides were prepared as described (43 (link)).