Oil hydraulic micromanipulator

All stereotaxic surgeries were conducted as described in our animal study protocol. Mice were anesthetized with a Ketamine/Xylazine solution (100mg/10mg in PBS) and a stereotaxic device (Stoelting, USA) was used for viral injections at the following stereotaxic coordinates: cLVM, -2.50 mm from Lambda, ±1.40 mm lateral from midline, and -5.30 mm vertical from cortical surface. LC, -5.50 mm from Bregma, 0.95 mm lateral from midline, and -3.50 mm vertical from cortical surface. PVT, -1.6 mm from Bregma, -0.06 mm lateral from midline with a 6-dgree angle, and -3.0 mm from cortical surface. AAVs were injected with an oil hydraulic micromanipulator (Narishige). AAVs were injected at a total volume of 0.1 μl in the cVLM. All other AAVs were injected at approximately 0.2-0.3 μl. Following stereotaxic injections, AAVs were allowed 2-3 weeks for maximal expression. Optical fibers with diameters of 200 μm (0.48 NA) and 400 μm (0.66 NA), were used for optogenetics and fiber photometry experiments, respectively (Doric Lenses). These fibers were implanted over the cLVM or LC immediately after viral injection and cemented using C&B Metabond Quick Adhesive Cement System (Parkell, Inc.). Mice received subcutaneous injections with Ketoprofen (5 mg/kg) for analgesia and anti-inflammatory purposes pre- and post-operatively and were allowed to recover on a heating pad.