Systemic Inflammation Induces Acute Behavioral and Cognitive Changes and Accelerates Neurodegenerative Disease methods

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        "name": "Methods and Materials",
        "text": "Female C57BL/6 mice, n = 233 (Harlan Olac, United Kingdom), were housed in cages of five at 21°C with a 12:12 hour light-dark cycle (lights on 0700 to 1900) with free access to food and water. They were anesthetized intraperitoneally (IP) with Avertin (Sigma, Poole, United Kingdom) and positioned in a stereotaxic frame. One microliter of a 10% wt/vol scrapie-infected (ME7 strain) C57BL/6 brain homogenate (or 10% wt/vol normal brain homogenate [NBH]) was injected into both dorsal hippocampi (from bregma: anterior-posterior -2.5 mm, lateral -1.7 mm, depth -1.6 mm) using a microsyringe (Hamilton, Reno, Nevada). Further animals ( n = 9) were injected intracerebrally (IC) with N -methyl-D-aspartate (NMDA) (10 mg/mL) to ablate the hippocampus. This surgery was performed under isoflurane anesthesia (approximately 2%) with perisurgical analgesia (carprophen, 5 mg/kg). C..."
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      {
        "@type": "HowToStep",
        "position": 2,
        "name": "Quantitative PCR",
        "text": "Animals were terminally anesthetized and transcardially perfused with heparinized saline 2 hours postadministration of LPS or saline. Brains were rapidly removed and the hippocampi were dissected out, snap frozen in liquid nitrogen, and stored at -80°C. Total RNA was extracted using Qiagen RNeasy mini columns (Qiagen, Crawley, United Kingdom) according to the manufacturer's instructions. Contaminating genomic DNA was degraded during extraction with Qiagen DNase1 (Qiagen). Two hundred nanograms of total RNA were reverse-transcribed in a 10 µL reaction volume and 1 µL of the reverse transcription (RT) reaction was used for polymerase chain reaction (PCR). Equipment and reagents for quantitative PCR were supplied by Applied Biosystems (Warrington, United Kingdom). Assays for IL-1β, tumor necrosis factor-alpha (TNF-α), and interferon-beta (IFN-β) were d..."
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      {
        "@type": "HowToStep",
        "position": 3,
        "name": "Immunohistochemistry",
        "text": "Immunohistochemistry for cyclooxygenase-2 (COX-2) and IL-1β was carried out on formalin-fixed, paraffin-embedded sections. Sections were dewaxed, rehydrated, quenched with 1% hydrogen peroxide (H 2 O 2 ) in absolute methanol, and washed briefly in phosphate buffered saline (PBS) before antigen retrieval by microwaving in citrate buffer (pH 6) for 2 × 5 minutes. Sections were blocked with 10% horse serum (for COX-2) or 20% normal goat serum (for IL-1β) before overnight incubation with the primary antibody at 1/1000 (goat polyclonal anti-COX-2, Santa Cruz Biotechnology Inc., Santa Cruz, California) or 1/50 (rabbit polyclonal anti-IL-1β, Peprotech, London, United Kingdom) prepared in 20% serum to block nonspecific interactions. The sections were washed and then incubated with the appropriate biotinylated secondary antibody. Labeling was visualized using the avidin-bio..."
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    "isBasedOn": {
      "@type": "ScholarlyArticle",
      "headline": "Systemic Inflammation Induces Acute Behavioral and Cognitive Changes and Accelerates Neurodegenerative Disease",
      "datePublished": "2009",
      "author": [
        {
          "@type": "Person",
          "name": "Colm Cunningham"
        },
        {
          "@type": "Person",
          "name": "Suzanne Campion"
        },
        {
          "@type": "Person",
          "name": "Katie Lunnon"
        },
        {
          "@type": "Person",
          "name": "Carol L. Murray"
        },
        {
          "@type": "Person",
          "name": "Jack F.C. Woods"
        },
        {
          "@type": "Person",
          "name": "Robert M.J. Deacon"
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        {
          "@type": "Person",
          "name": "J. Nicholas P. Rawlins"
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        {
          "@type": "Person",
          "name": "V. Hugh Perry"
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      "identifier": "10.1016/j.biopsych.2008.07.024"
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