Bx50wi

Parasagittal cerebellar slices (250-µm thick) were prepared from Dox-treated and untreated mGluR1 cKO mice. Slices were perfused at 31 °C with standard saline containing (in mM): NaCl, 125; KCl, 2.5; CaCl₂, 2; MgSO₄, 1; NaH₂PO₄, 1.25; NaHCO₃, 26; glucose, 20; and bicuculline methochloride, 0.01 (Tocris Cookson, Bristol UK). This standard bathing solution was bubbled with 95% O₂ and 5% CO₂. Whole-cell recordings were made from visually identified PCs in lobules I-VIII using an upright microscope (Olympus BX50WI or Zeiss Axioskop). Detailed procedures are described in previous reports^{25 (link),55 (link)}.

To visualize S100a10 neurons, an adult male S100a10 bacTRAP transgenic mouse was perfused transcardially with saline followed by 4% paraformaldehyde (PFA)/0.1 M sodium phosphate buffer (PBS) under deep anesthesia. Brain was postfixed in 4% PFA overnight. Coronal sections (50 μM) were prepared by using the DSK vibratome. Free-floating sections were incubated with rabbit anti-GFP (1:500, Invitrogen), mouse anti-NeuN (1:500, Millipore) followed by goat anti-rabbit Alexa 488 (1:1000, Invitrogen), and goat anti-mouse Alexa 594 (1:1000, Invitrogen). For S100a10 staining, 40 µM sections through sensorimotor cortex were collected using a freezing microtome (Leica). Free-floating sections were incubated with chicken anti-EGFP (1:1000, Abcam) and goat anti-S100a10 (1:200, R&D Systems) followed by donkey anti-chicken Alexa 488 (1:2000, Invitrogen) and donkey anti-goat Alexa 568 (1:2000, Invitrogen). Images were obtained using either an epifluorescent microscope (Olympus BX50WI) or Zeiss LSM700.