Fluoroacetate

All photothrombic strokes were induced using this protocol as previously described (Zheng et al., 2010 (link), 2013 (link)), except for the data presented in Figures 5A,C. Briefly, mice were anesthetized with 4% isoflurane and maintained at 2% isoflurane throughout the surgery. Hair was removed, and incision made on the dorsal scalp, and head mounted in a custom frame. Either a cranial window or a thin skull prep was performed. Rose Bengal (Sigma, Cat no 330000) dye was then injected intravenously, and a blood clot induced with a 568 nm laser on a Nikon (TE 200) microscope, with blood vessels between 30 and 40μM targeted for clotting. Mice were injected with drugs [MRS2365: Tocris Cat no 2157; MRS1523: Sigma Cat no M1809; Fluoroacetate: Sigma Cat no 62-74-8; MRS5698: Tocris Cat no 5428; Cl-IB-MECA: Tocris Cat no 1104; AST-004, synthesized at the National Institute of Diabetes, Digestive and Kidney Diseases, Bethesda, MD (Ravi et al., 2002 (link))] either before surgery or 30 min post-stroke, as described in the paper.

Pharmacological agents were purchased from: Sigma-Aldrich: Fluoroacetate, thapsigargin, BAPTA, bicuculline methbromide, Tricine, Zinc chloride, d-serine, and GDPβS; Tocris Bioscience: nimodipine, (+)-MK-801 maleate, d-AP5, NBQX, TTX citrate, PPDA, Ro 25-6981 maleate, MCPG, MPEP, LY341495, AM251, 2-AG, and FK506; and Abcam: UBP-141. Salts used for internal and external solutions were purchased from Sigma-Aldrich. Compounds were dissolved in H₂O or Ringer solution with the exception of thapsigargin, nimodipine, PPDA, FK506, THL, AM251, and 2-AG, which were dissolved in DMSO. Vehicle (DMSO) at the concentrations used did not affect baseline EPSP amplitudes and had no other detectable effects on the neurons. When investigating the effect of pharmacological agents on plasticity, all drugs were included in the superfusion fluid or patch pipette from the start of the experiment until completion (from 0 to 50 min in a standard plasticity experiment), except for Fluoroacetate, which was applied from 60 min before the start of recording. When determining the effect of a pharmacological agent on baseline condition, a stable baseline of at least 10 min was first recorded and then the drug was bath applied by switching to a different perfusion line.