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4 protocols using s bayk8644

1

Striatal Slice Preparation and Neuromodulation

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Mice were anesthetized using isofluorane and then decapitated. Brains were removed, cut into left and right hemispheres, and then 300 μm coronal slices were made at 1-4°C in oxygenated (95% v/v O2, 5% v/v CO2) dissecting solution (208 mM sucrose, 2.5 mM KCl, 1 mM CaCl2, 4 mM MgCl2, 4 mM MgSO4, 1.6 mM NaH2PO4, 26 mM NaHCO3, 10 mM glucose, and 3 mM Na-pyruvate) using a Vibratome 3000 (The Vibratome Company). Typically, a total of 5-7 striatal hemi-slices (left/right hemisphere combined) were obtained from each mouse brain (1.1 to 0.14 mm from Bregma). Slices were allowed to recover on a nylon mesh for 1 h at 30°C in oxygenated ACSF (113 mM NaCl, 2.5-5 mM KCl, 2.5 mM CaCl2, 1.2 mM MgSO4, 1 mM NaH2PO4, 26 mM NaHCO3, 20 mM glucose, and 3 mM Na-pyruvate) followed by addition of picrotoxin (50 μM) for 30 min. Slices were then transferred to oxygenated 30°C ACSF solutions supplemented with vehicle or drug for 1-30 min, as defined in the figure legends. The following drugs were used and dissolved in water or DMSO based on manufacturer's instructions: BAPTA (Sigma), (S)-BayK8644 (Tocris), FPL64176 (Tocris), isradipine (National Institute of Mental Health Chemical Synthesis and Drug Supply Program), KCl (Sigma), mibefradil (Tocris), Nickel Chloride (Sigma), nimodipine (MP Biomedicals), SNX-482 (Peptides International), and TTX (Tocris).
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2

Comprehensive Compound Acquisition for Research

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ML297 and AM095 were purchased from
Sigma-Aldrich (St. Louis, MO). (S)-(−)-BAY-K-8644,
(R)-(−)-BAY-K-8644, and SAK3 were obtained
from Tocris Bioscience (Bristol, UK). lysoPA18:1 was purchased from
Avanti Polar Lipids Inc. (Alabaster, AL). hC3a was purchased from
Complement Technology Inc. (Tyler, TX). Fluo-4AM was purchased from
Thermo Fisher Scientific Inc. (Waltham, MA). MK-8998, ST101, TTA-I1,
and TTA-A2 were synthesized in house. All other compounds were obtained
from Alomone Labs (Jerusalem, Israel).
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3

Pharmacological Inhibition of Ion Channels

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(S)-Bay K8644, nifedipine, D-AP5, CNQX, CGP52432, and QX-314 were purchased from Tocris Bioscience. PTX was purchased from Sigma-Aldrich. (S)-Bay K8644, nifedipine, CNQX were dissolved in DMSO and stored at −20°C before use. Other drugs were dissolved in nanopure H2O. All drugs were applied by perfusion in the bath solution.
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4

Pharmacological characterization of vascular responses

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Equilin, 17β-estradiol, phenylephrine, acetylcholine, indomethacin, L-NAME (Nω-nitro-L-arginine methyl ester), and KCl were purchased from Sigma-Aldrich (St. Louis, MO, USA). Tetraethylammonium, glibenclamide, 4-amynopiridine, ouabain, (S)-(−)-Bay K 8644, U46619 (9,11-dideoxy-11α,9α-epoxymethanoprostaglandin F2α), ICI 182,780, KT 5720, KT 5823, SQ 22,536 (9-(tetrahydro-2-furanyl)-9H-purin-6-amine), and ODQ (1H-[1 (link)–2 (link),4 (link)]oxadiazolo [4,3-alpha]-quinoxalin-1-one) were obtained from Tocris Bioscience (Bristol, UK). Caffeine was purchased from Roth (Karlsruhe, Germany).
Equilin and 17β-estradiol were dissolved in 70% v/v ethanol with further dilution in distilled water before use. Working concentrations of ethanol in the bath were < 0.01% (v/v).
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