Hippocampal slices were cut from the brains of 3- to 5-week-old mice for electrophysiology and 14 to 22 days after viral injections for GCamP6f imaging. Animals were deeply anesthetized with isoflurane, and the brains were removed quickly. Horizontal slices, either 250 μm (for GCamP6f imaging) or 300 μm (for slice electrophysiology), were cut using a tissue slicer (Compresstome model VF-200-0Z, Precisionary Instruments) in ice-cold sucrose aCSF. For the electrophysiological slice experiments, the composition of this solution was as follows: 85 mM NaCl, 2.5 mM KCl, 1.25 mM NaH2PO4, 25 mM NaHCO3, 25 mM glucose, 75 mM sucrose, 0.5 mM CaCl2, and 4 mM MgCl2, saturated with 95% O2 and 5% CO2. For GCamP6f imaging, slices were cut in a solution containing the following: 110 mM choline chloride, 2.5 mM KCl, 25 mM NaHCO3, 1.25 mM NaH2PO4, 25 mM d-glucose, 11.6 mM ascorbic acid, 3.1 mM pyruvic acid, 7 mM MgCl2, and 0.5 mM CaCl2, saturated with 95% O2 and 5% CO2. Slices were then quickly transferred into a recovery chamber with aCSF solution maintained at 30°C and containing the following: 125 mM NaCl, 2.4 mM KCl, 1.2 mM Na2PO4, 25 mM NaHCO3, 25 glucose, 1 mM CaCl2, and 2 mM MgCl2, saturated with 95% O2 and 5% CO2. After 30 min in the recovery chamber, slices were subsequently transferred into a storage chamber with aCSF containing 2 mM CaCl2, where they were stored for 0.5 to 6 hours until they were used for calcium imaging using 2PLSM or electrophysiology.

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