For pharmacological inactivation, ≈500-μm-diameter craniotomies were drilled over cool (32–22 °C)-responsive areas in S1 and pIC in mice anaesthetized with isoflurane (1.5–2% in O2). Both regions were identified functionally using the widefield calcium imaging response to thermal stimuli. Following the craniotomy, the dura was covered with transparent silicone gel (3-4680, Dow Corning). A 300 nl volume of muscimol or Ringer’s solution was injected at a rate of 100 nl min−1 300–500 μm below the pial surface using a pulled glass pipette and a hydraulic injection system (MO-10, Narashige). muscimol (Abcam, ab120094) was dissolved in Ringer’s solution to a concentration of 5 mM. Imaging sessions were carried out >10 min following the end of Ringer’s solution or muscimol injection (Fig. 1f,g).
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