A New Tool for the Analysis of the Effect of Intracerebrally Injected Anti-Amyloid-β Compounds

Abstract

Background: A wide range of techniques has been developed over the past decades to characterize amyloid-β (Aβ) pathology in mice. Until now, no method has been established to quantify spatial changes in Aβ plaque deposition due to targeted delivery of substances using ALZET ® pumps. Objective: Development of a methodology to quantify the local distribution of Aβ plaques after intracerebral infusion of compounds. Methods: We have developed a toolbox to quantify Aβ plaques in relation to intracerebral injection channels using Zeiss AxioVision ® and Microsoft Excel ® software. For the proof of concept, intracerebral stereotactic surgery was performed in 50-day-old APP-transgenic mice injected with PBS. At the age of 100 days, brains were collected for immunhistological analysis. Results: The toolbox can be used to analyze and evaluate Aβ plaques (number, size, and coverage) in specific brain areas based on their location relative to the point of the injection or the injection channel. The tool provides classification of Aβ plaques in pre-defined distance groups using two different approaches. Conclusion: This new analytic toolbox facilitates the analysis of long-term continuous intracerebral experimental compound infusions using ALZET ® pumps. This method generates reliable data for Aβ deposition characterization in relation to the distribution of experimental compounds.

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“…Formalin-fixed hemispheres were embedded in paraffin and cut into 4-μm-thick coronal sections using a rotation microtome (HM355S, Leica Biosystems GmbH, Nußloch, Germany) as described previously [ 75 , 76 , 77 , 204 , 205 , 206 , 207 , 208 , 209 , 210 ]. Sections (bregma +0.8 mm and −1.8 mm) were stained for microglia (IBA1, 1:1000, FUJIFILM Wako Chemicals Europe GmbH, 019–19741), astrocytes (GFAP, 1:500, Agilent, Santa Clara, CA, USA, Z033401-2) using a BOND-III ® automated immunostaining system (Leica Biosystems GmbH, Nußloch, Germany) with a hematoxylin counterstain (provided with the staining kit, Bond Polymer Refine Detection, DS9800).…”

Time- and Sex-Dependent Effects of Fingolimod Treatment in a Mouse Model of Alzheimer’s Disease

“…Formalin-fixed hemispheres were embedded in paraffin and sliced into four-micrometerthick coronal sections using a rotation microtome (HM355S, Leica Biosystems GmbH, Nussloch, Germany), as described previously [38,[51][52][53][55][56][57][58][59][60][61]. The sections (bregma−2.0 mm) were stained for microglia (anti-IBA1, 1:1000, FUJIFILM Wako Chemicals Europe GmbH, 019-19741) or astrocytes (anti-GFAP, 1:500, Agilent, USA, Z033401-2) using a BOND-III ® automated immunostaining system (Leica Biosystems GmbH, Nussloch, Germany) with a haematoxylin counterstain (provided with the staining kit, Bond Polymer Refine Detection, DS9800).…”