The standard Ringer’s solution used for wide-field Ca2+ imaging studies contained the following: 155 mM NaCl, 4.5 mM KCl, 10 mM d-glucose, 5 mM Hepes, 1 mM MgCl2, and 2 mM CaCl2. The Ca2+-free Ringer’s solution contained 3 mM MgCl2, 1 mM EGTA (Sigma-Aldrich), and no added CaCl2. pH was adjusted to 7.4 with 1 N NaOH. Stock solutions of TG, 2-APB, and YM-58483 (BTP2; Calbiochem) were dissolved in dimethyl sulfoxide and used at the indicated concentrations. ATP (Sigma-Aldrich), UTP (Sigma-Aldrich), and l-glutamic acid (Sigma-Aldrich) were all dissolved in water and used at the indicated concentrations. Thrombin was dissolved in 0.1% albumin and used at the indicated concentrations. EGTA-AM and BAPTA-AM were obtained from Invitrogen and loaded into astrocytes by incubating the cells for 35 to 45 min with 5 μM AM buffer at 37°C.

For GCaMP6f imaging and electrophysiological recordings of slices, the external artificial cerebrospinal fluid (aCSF) solution contained the following: 125 mM NaCl, 2.4 mM KCl, 1.2 mM Na2PO4, 25 mM NaHCO3, 25 mM glucose, 2 mM CaCl2, and 1 mM MgCl2, equilibrated with 95% O2 and 5% CO2. Ca2+-free aCSF solution contained the following: 125 mM NaCl, 2.4 mM KCl, 1.2 mM Na2PO4, 25 mM NaHCO3, 25 mM glucose, and 3 mM MgCl2, with 1 mM EGTA, equilibrated with 95% O2 and 5% CO2. The internal solution for electrophysiological recordings contained the following: 95 mM CsF, 25 mM CsCl, 10 mM Hepes, 10 mM EGTA, 2 mM Mg-ATP, 0.3 mM Na3–guanosine triphosphate, 10 mM QX-314, 5 mM tetraethylammonium chloride (TEA-Cl), and 5 mM 4-aminopyridine (4-AP) (pH 7.3 with CsOH).

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