Pupil fluctuations track rapid changes in adrenergic and cholinergic activity in cortex methods

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    "description": "Evidence-backed execution summary for Pupil fluctuations track rapid changes in adrenergic and cholinergic activity in cortex methods from Pupil fluctuations track rapid changes in adrenergic and cholinergic activity in cortex.",
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        "position": 1,
        "name": "Methods",
        "text": "All procedures were carried out in accordance with the ethical guidelines of the National Institutes of Health and were approved by the Institutional Animal Care and Use Committee (IACUC) of Baylor College of Medicine, Yale University and Columbia University. In this study, we used a total of 21 mice, including male ( n =13) and female ( n =8) mice age 5 weeks to 5 months, and three rats. Twelve animals were injected with CNiFERs (see below), including one animal injected with α1a alone, six animals injected with M1 alone, and five animals injected with both α1a and M1. Several of these mice were SST-Cre/Ai9 ( n =6) or PV-Cre/Ai9 ( n =2) crosses on a C57Bl/6 background. For imaging of ChAT and DBH axons, we used six ChAT-Cre (Jackson labs strain B6;129S6-Chattm1(cre)Lowl/J ) and three DBH-Cre mice (MMRRC Stock#036778-UCD B6.FVB(Cg)-Tg(Dbh-cre) KH212Gsat/Mmucd ). Expression o..."
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      {
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        "position": 2,
        "name": "Methods",
        "text": "Viral injections were performed stereotactically through a burr hole under isoflurane anaesthesia. Injections were targeted to the basal forebrain ( ChAT-Cre mice, AAV1-Syn-FLEX-GCaMP6s ) by a vertical penetration 4 mm lateral and 0.5 mm posterior of Bregma, and ∼1 ul of virus was injected over a period of 10-15 min at a depth of 4-4.5 mm. Injections targeting the LC ( DBH-Cre mice, AAV5-CAG-DIO-GCaMP6s ) were performed similarly at coordinates 0.5 mm just above and below those used in a previous study to target the LC. Injections were performed 4-6 weeks before imaging to allow time for viral expression."
      },
      {
        "@type": "HowToStep",
        "position": 3,
        "name": "Locomotion and Pupillometry",
        "text": "After surgery, the mouse was allowed to recover on the heating pad and then transferred to the microscope. The animal's head was restrained under the microscope objective, and their body was supported by a treadmill that allowed free movement along a single axis of rotation. Treadmill motion was measured using a rotary optical encoder with a resolution of 8,000 counts/revolution. Running periods were defined as periods of two seconds or more with treadmill speeds greater than 0.5 cm s -1 after filtering the treadmill trace at 5 Hz (Hamming). Multiple running periods separated by less than two seconds were lumped together as a single running epoch."
      }
    ],
    "tool": [
      {
        "@type": "HowToTool",
        "name": "CNiFER imaging of ACh and NE release in V1"
      },
      {
        "@type": "HowToTool",
        "name": "CNiFERs"
      },
      {
        "@type": "HowToTool",
        "name": "Locomotion and Pupillometry"
      },
      {
        "@type": "HowToTool",
        "name": "Locomotion and Pupillometry"
      },
      {
        "@type": "HowToTool",
        "name": "Imaging"
      },
      {
        "@type": "HowToTool",
        "name": "Preprocessing of calcium imaging data"
      }
    ],
    "supply": [
      {
        "@type": "HowToSupply",
        "name": "CNiFERs"
      }
    ],
    "isBasedOn": {
      "@type": "ScholarlyArticle",
      "headline": "Pupil fluctuations track rapid changes in adrenergic and cholinergic activity in cortex",
      "datePublished": "2016",
      "author": [
        {
          "@type": "Person",
          "name": "Jacob Reimer"
        },
        {
          "@type": "Person",
          "name": "Matthew J McGinley"
        },
        {
          "@type": "Person",
          "name": "Yang Liu"
        },
        {
          "@type": "Person",
          "name": "Charles Rodenkirch"
        },
        {
          "@type": "Person",
          "name": "Qi Wang"
        },
        {
          "@type": "Person",
          "name": "David A McCormick"
        },
        {
          "@type": "Person",
          "name": "Andreas S Tolias"
        }
      ],
      "identifier": "10.1038/ncomms13289"
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