1：Experimental Design and Method Selection:

The study used viral transfection to express melanopsin in astrocytes, combined with calcium imaging and electrophysiology to monitor Ca2+ signals and synaptic responses. Two-photon imaging and patch-clamp techniques were employed.

2：Sample Selection and Data Sources:

Hippocampal slices from wild-type and Ip3r2?/? mice (P20–P30) were used. Mice were transfected with AAV vectors containing melanopsin, ChR2, or control constructs.

3：List of Experimental Equipment and Materials:

Equipment included a vibratome (Leica Vibratome VT1200S), microscopes (Olympus BX50WI, Nikon Eclipse FN1, Zeiss LSM510 two-photon microscope), patch-clamp setup (PC-ONE amplifier, DigiData 1440 interface), lasers (473 nm blue laser, Spectra Physics Mai-Tai laser), and viral vectors (AAV2/5 serotype). Materials included drugs like TTX, D-AP5, and various antagonists.

4：Experimental Procedures and Operational Workflow:

Viral injections were performed in neonatal mice, followed by slice preparation after 2 weeks. Calcium imaging was done using GCaMP6f, and electrophysiological recordings were made from CA1 neurons and astrocytes. Light stimulation at various durations and frequencies was applied, and data were analyzed for Ca2+ events and synaptic changes. In vivo behavioral tests (Novel Object Location) were conducted with light stimulation.

5：Data Analysis Methods:

Data were analyzed using ImageJ, MATLAB, and pCLAMP software. Statistical analyses included ANOVA, t-tests, and post-hoc tests. Calcium signals were quantified as ΔF/F, and synaptic changes were assessed using coefficient of variation and holding current indices.