Pharmacological targeting of CSF1R inhibits microglial proliferation and prevents the progression of Alzheimer's-like pathology methods

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    "name": "Pharmacological targeting of CSF1R inhibits microglial proliferation and prevents the progression of Alzheimer's-like pathology methods",
    "description": "Evidence-backed execution summary for Pharmacological targeting of CSF1R inhibits microglial proliferation and prevents the progression of Alzheimer's-like pathology methods from Pharmacological targeting of CSF1R inhibits microglial proliferation and prevents the progression of Alzheimer's-like pathology.",
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        "name": "Animals",
        "text": "For the evaluation of the effects of treatment with GW2580, mice were 6 months of age when treatment began ( n = 6-8). Mice were fed with a control diet (RM1) or a diet containing GW2580 [Modified LabDiet® PicoLab EURodent Diet 14%, 5L0W (5LF2) with 0.1% (1000 ppm) GW2580 (LC Laboratories); TestDiet] for 3 months, before behavioural tasks were conducted. Alternatively, to test the effects of increasing doses of GW2580 on microglial survival, GW2580 (LC Laboratories) was suspended in 0.5% hydroxypropylmethylcellulose and 0.1% Tween 80 and was dosed orally at 0.2 ml per mouse (75 mg/kg), daily for five consecutive days to wild-type mice. Mice weight was monitored during all experiments. Mice received one injection of intraperitoneal bromodeoxyuridine (BrdU) (Sigma-Aldrich; 7.5 mg/ml, 0.1 ml/10 g weight in sterile saline), 1 day before the end of the experiment. All procedures..."
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        "name": "Behavioural tests",
        "text": "The open-field tests were carried out using activity monitor software (Med Associated Inc.). The mice were placed in individual cages of 27 × 27 × 0.3 cm for a period of 5 min, to further analyse the total distance travelled (cm) and the number of rears (vertical counts), using the average speed as an internal control of the mouse motor abilities, during the test period (5 min). For measuring anxiety-related behaviour, differential exploratory activity (distance travelled or number of entries) was analysed in a residual and an open zone, using activity monitor software (Med Associated Inc.), as previously described ( )."
      },
      {
        "@type": "HowToStep",
        "position": 3,
        "name": "Behavioural tests",
        "text": "Discrete trial spontaneous alternation in the T-maze was performed as previously described ( ). The apparatus for this test consisted of a grey T-shaped maze with 30 × 10 × 29-cm arms, with a central partition extending 7 cm into the start arm from the back of the maze, and two guillotine doors each having the potential to block off the left or right goal arms. Mice were placed in the start arm of the maze (facing the wall) and allowed to make a choice to enter the left or right goal arm. Following their choice, they were then enclosed in that arm for 30 s to facilitate habituation by sliding down the appropriate guillotine door. Mice were then taken out of the maze, the central partition removed and guillotine doors reopened. Mice were once again placed in the start arm and allowed to make another free choice of either goal arm. Whether or not the mouse alternated was noted..."
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      {
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        "position": 4,
        "name": "Immunohistochemistry",
        "text": "The general immunohistochemistry protocol was modified for the detection of BrdU, adding a DNA denaturation step with 2 N HCl (30 min, 37 ° C), as previously described (, ). For the detection of amyloid-β plaques (6E10), sections were pretreated with 95% formic acid (Sigma-Aldrich) for 10 min at room temperature. All other histological stains (i.e. Congo red) were done according to standard laboratory procedures."
      },
      {
        "@type": "HowToStep",
        "position": 5,
        "name": "Immunohistochemistry",
        "text": "The protocol used for immunohistochemistry on human sections was a modification of the general protocol (DAB + alkaline phosphatase), with antigen unveiling in citrate buffer being performed for 25 min, as previously described ( )."
      },
      {
        "@type": "HowToStep",
        "position": 6,
        "name": "Golgi-Cox staining",
        "text": "A subgroup of APP/PS1 or wild-type mice treated with control (RM1) or GW2580 diet ( n = 4), as stated before, were deeply anaesthetized with sodium pentobarbital and then transcardially perfused with artificial CSF. Brains were then rapidly dissected and sliced with a vibrating microtome (200 µm; Leica). The hippocampal slices were incubated in rapid Golgi-Cox solutions, following manufacturer's instructions (FD Rapid Golgi Stain Kit, FD Neurotechnologies). The slices were infused with a solution containing potassium dichromate, potassium chromate and mercuric chloride for 2 weeks, to be further developed into the Golgi-Cox staining on free-floating plates. The slices were mounted onto gelatinized slides, dried, dehydrated, cleared with xylene and mounted with DPX. Golgi-treated slices were analysed with a Leica CTR 5000 microscope, coupled to a Leica DFC300FX microscope c..."
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        "position": 7,
        "name": "Pharmacological targeting of CSF1R activation with an orally-available inhibitor blocks microglial proliferation in A...",
        "text": "As CSF1R is likely to drive microglial proliferation in APP/PS1 mice, we targeted the activation of CSF1R with a selective inhibitor (GW2580) from 6 to 9 months of age, to evaluate target engagement and efficacy of a CSF1R-inhibitory approach ( ). To determine if microglia would be the only target of CSF1R inhibitors, we used APP/PS1/Macgreen mice as reporters of CSF1R expression. Using immunofluorescence and confocal microscopy we found no evidence of expression of CSF1R in neurons (NeuN + ) or astrocytes (GFAP + ) ( ), supporting that CSF1R expression is exclusive to microglia, as previously described (;; ). While a dose of GW2580 of 75 mg/kg has been used in the past without causing significant changes in the survival of microglia ( ), recent evidence supports that CSF1R inhibitors could cause the depletion of the microglial population ( ). Therefore, we assessed the effects of r..."
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        "position": 8,
        "name": "Pharmacological targeting of CSF1R activation with an orally-available inhibitor blocks microglial proliferation in A...",
        "text": "CSF1R inhibition prevents behavioural deficits in APP/PS1 mice. ( A ) Spontaneous alternation in the T-maze of APP/PS1 and wild-type mice at 9 months of age, after treatment for 3 months with a control diet (RM1) or a diet containing GW2580. Alternation was measured as % election of the alternative arm in the second test (short-term memory). ( B and C ) Analysis of the behaviour in the open field, measured as total distanced travelled ( B ) or number of entries in the open zone ( C ) of APP/PS1 and wild-type (WT) mice at 9 months of age, after treatment for 3 months with a control diet (RM1) or a diet containing GW2580. Exploratory activity was measured as distance travelled (cm) in the open field test, analysing the locomotor activity on an open zone versus residual zone as a correlate of anxiety. ( D ) Burrowing behaviour, a measure of sickness behaviour, was measured as weight disp..."
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        "@type": "HowToTool",
        "name": "Behavioural tests"
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        "@type": "HowToTool",
        "name": "Immunohistochemistry"
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        "@type": "HowToTool",
        "name": "Golgi-Cox staining"
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        "name": "Quantification and image analysis"
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        "@type": "HowToTool",
        "name": "Analysis of gene expression by reverse transcriptase PCR"
      },
      {
        "@type": "HowToTool",
        "name": "Analysis of gene expression by reverse transcriptase PCR"
      },
      {
        "@type": "HowToTool",
        "name": "Multiplex analysis of soluble amyloid-β 38, amyloid-β 40 and amyloid-β 42"
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    ],
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        "@type": "HowToSupply",
        "name": "Immunohistochemistry"
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        "@type": "HowToSupply",
        "name": "Immunohistochemistry"
      },
      {
        "@type": "HowToSupply",
        "name": "Golgi-Cox staining"
      },
      {
        "@type": "HowToSupply",
        "name": "Analysis of gene expression by reverse transcriptase PCR"
      },
      {
        "@type": "HowToSupply",
        "name": "Analysis of gene expression by reverse transcriptase PCR"
      },
      {
        "@type": "HowToSupply",
        "name": "Multiplex analysis of soluble amyloid-β 38, amyloid-β 40 and amyloid-β 42"
      },
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        "@type": "HowToSupply",
        "name": "Pharmacological targeting of CSF1R activation with an orally-available inhibitor blocks microglial proliferation in A..."
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        "name": "Pharmacological targeting of CSF1R activation with an orally-available inhibitor blocks microglial proliferation in A..."
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      "@type": "ScholarlyArticle",
      "headline": "Pharmacological targeting of CSF1R inhibits microglial proliferation and prevents the progression of Alzheimer's-like pathology",
      "datePublished": "2016",
      "author": [
        {
          "@type": "Person",
          "name": "Adrian Olmos-Alonso"
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        {
          "@type": "Person",
          "name": "Sjoerd T. T. Schetters"
        },
        {
          "@type": "Person",
          "name": "Sarmi Sri"
        },
        {
          "@type": "Person",
          "name": "Katharine Askew"
        },
        {
          "@type": "Person",
          "name": "Renzo Mancuso"
        },
        {
          "@type": "Person",
          "name": "Mariana Vargas-Caballero"
        },
        {
          "@type": "Person",
          "name": "Christian Holscher"
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        {
          "@type": "Person",
          "name": "V. Hugh Perry"
        },
        {
          "@type": "Person",
          "name": "Diego Gomez-Nicola"
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      "identifier": "10.1093/brain/awv379"
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