Adult C57 mice were injected with AAV2/9-hSyn-GCaMP6s into the DG followed by optic fiber implantation 6 weeks postnatally. A 200-μm-diameter optical fiber was glued into a short cannula with the fiber tip extended approximately 2.5 mm out of the cannula. The preprocessed fiber was inserted through a small craniotomy made at the DG region (bregma coordinates: AP, –2.1 mm; ML, ±1.7 mm; DV, –2.05 mm). Last, the cannula was secured to the skull using dental cement. Mice were individually housed for at least 1 week to recover from fiber photometry. The rescue experiments were conducted 2 weeks after the virus was completely expressed.

For the running test, mice were first habituated to the fiber photometry apparatus for 30 min and then tested on a subsequent day. The fiber photometry was conducted by a fiber photometry system purchased from Thinker Tech Nanjing BioScience Inc. The analog voltage signals were digitized at 50 Hz. All the Ca2+ signals and behavior videos were synchronized offline with event marks. The Ca2+ signal data were segmented on the basis of behavior events with individual trials, which were marked at the time of experimental mice contact. We derived the values of fluorescence change (ΔF/F) by calculating (FF0)/F0, where F0 is the baseline fluorescence signal averaged over a 2-s-long control time window. The ΔF/F data were presented with average plots with a shaded area indicating SEM.

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