N methylsulfonyl 2 2 propynyloxy benzenehexanamide msppoh

FITC-dextran (2000KDa), rhodamine B isothiocyanate-dextran (Rhodamine-B-dextran, 70KDa), dexamethasone 21-phopshate disodium, Chremophore® EL, DMSO were obtained from Sigma Aldrich. D-(-)-2-Amino-5-phosphonopentanoic acid (AP5) and Compound 21 dihydrochloride was purchased from Hello Bio Inc. (Princeton, NJ). U46619, N-(methylsulfonyl)-2-(2-propynyloxy)-benzenehexanamide (MSPPOH), medotomidine hydrochloride, and atipamezole hydrochloride were ordered from Cayman Chemicals. BAPTA-tetrapotassium and Alexa 488 hydrazide were obtained from Invitrogen, isoflurane from Fresenius Kabi Canada Ltd., Pluronic F127 and Rhod-2/AM from Biotium, buprenorphine from Champion Alstoe Animal Health, and enrofloxacin from Bayer, meloxicam from Boehringer Ingelheim, ketamine from Vétoquinol N.-A. Inc. AAV2/5.GfaABC₁D.GCamP6f and AAV9.hSyn.GCaMP6f were ordered from Penn Vector Core, PA, USA, pZac2.1-GfaABC₁D-mCherry-hPMCA2w/b plasmid was ordered from Addgene.

The following chemicals were used in Krebs–Henseleit buffer preparation (in mmol/L): NaCl, 118.3; NaHCO₃, 25; glucose, 11; KCl, 4.7; CaCl₂, 2.5; MgSO₄, 1.2 and KH₂PO₄, 1.2 (all from Carl Roth GmbH, Karlsruhe, Germany). Drugs used in the vasoreactivity study are as follows: N^ω-nitro L-arginine methyl ester (L-NAME), indomethacin, acetylcholine hydrochloride (ACh), phenylephrine and baicalein (Sigma-Aldrich Chemie GmbH, Steinheim, Germany), quinacrine, sulphaphenazole, N-(methylsulfonyl)-2-(2-propynyloxy) benzenehexanamide (MS-PPOH), N-(4-butyl-2-methylphenyl)-N'-hydroxy-methanimidamide (HET0016), 12-[[(tricyclo[3.3.1.13,7]dec-1-ylamino)carbonyl]amino]-dodecanoic acid (AUDA), N-[1-(1-oxopropyl)-4-piperidinyl]-N’-[4-(trifluoromethoxy)phenyl)-urea (TPPU), 14,15-epoxyeicosa-5(Z)-enoic acid (14,15-EEZE), and all four EET regioisomers i.e. ( ±)5,6-, ( ±)8,9-, ( ±)11,12-, and ( ±)14,15-EET were purchased from Cayman Chemical (Ann Arbor, MI, USA). Ethanol in 14, 15-EEZE and EETs was evaporated and dissolved in dimethyl sulfoxide (DMSO). L-NAME was dissolved in phosphate buffer saline (PBS), whereas indomethacin, baicalein, quinacrine, sulphaphenazole, MS-PPOH, TPPU and AUDA were dissolved in DMSO, according to the manufacturer’s instructions. DMSO at ≤ 0.2% (v/v) did not influence vasoreactivity, as reported previously^{3 (link),71 (link),72 (link)}.