Giuseppe Battaglia1,2, Paola Coan3,4, Giacomo E Barbone5,6, Alberto Bravin7,8, Alberto Mittone7,9, Alexandra Pacureanu7, Giada Mascio10, Paola Di Pietro10,11, Markus J Kraiger12, Marina Eckermann5,13, Mariele Romano5, Martin Hrabě de Angelis12,14,15, Peter Cloetens7, Valeria Bruno10,16. 1. Department of Molecular Pathology, I.R.C.C.S. Neuromed, Pozzilli, Italy. giuseppe.battaglia@neuromed.it. 2. Department of Physiology and Pharmacology, University Sapienza, Piazzale Aldo Moro 5, 00185, Rome, Italy. giuseppe.battaglia@neuromed.it. 3. Department of Medical Physics, Faculty of Physics, Ludwig-Maximilians-Universität München, Am Coulombwall 1, 85748, Garching, Germany. paola.coan@physik.uni-muenchen.de. 4. Department of Clinical Radiology, Faculty of Medicine, Ludwig-Maximilians-Universität München, Munich, Germany. paola.coan@physik.uni-muenchen.de. 5. Department of Medical Physics, Faculty of Physics, Ludwig-Maximilians-Universität München, Am Coulombwall 1, 85748, Garching, Germany. 6. Department of Clinical Radiology, Faculty of Medicine, Ludwig-Maximilians-Universität München, Munich, Germany. 7. ESRF, The European Synchrotron, Grenoble, France. 8. Department of Physics, University Milano Bicocca, Milan, Italy. 9. CELLS-ALBA Synchrotron, Cerdanyola del Vallès, Spain. 10. Department of Molecular Pathology, I.R.C.C.S. Neuromed, Pozzilli, Italy. 11. Department of Medicine, Surgery and Dentistry, "Scuola Medica Salernitana", University of Salerno, Baronissi, Italy. 12. Institute of Experimental Genetics, German Mouse Clinic, Helmholtz Zentrum München, German Research Center for Environmental Health, Neuherberg, Germany. 13. Institut Für Röntgenphysik, Georg-August-Universität Göttingen, Göttingen, Germany. 14. Chair of Experimental Genetics, TUM School of Life Sciences, Technische Universität München, Freising, Germany. 15. German Center for Diabetes Research, Helmholtz Zentrum München, German Research Center for Environmental Health, Neuherberg, Germany. 16. Department of Physiology and Pharmacology, University Sapienza, Piazzale Aldo Moro 5, 00185, Rome, Italy.
Abstract
PURPOSE: Modern neuroimaging lacks the tools necessary for whole-brain, anatomically dense neuronal damage screening. An ideal approach would include unbiased histopathologic identification of aging and neurodegenerative disease. METHODS: We report the postmortem application of multiscale X-ray phase-contrast computed tomography (X-PCI-CT) for the label-free and dissection-free organ-level to intracellular-level 3D visualization of distinct single neurons and glia. In deep neuronal populations in the brain of aged wild-type and of 3xTgAD mice (a triply-transgenic model of Alzheimer's disease), we quantified intracellular hyperdensity, a manifestation of aging or neurodegeneration. RESULTS: In 3xTgAD mice, the observed hyperdensity was identified as amyloid-β and hyper-phosphorylated tau protein deposits with calcium and iron involvement, by correlating the X-PCI-CT data to immunohistochemistry, X-ray fluorescence microscopy, high-field MRI, and TEM. As a proof-of-concept, X-PCI-CT was used to analyze hippocampal and cortical brain regions of 3xTgAD mice treated with LY379268, selective agonist of group II metabotropic glutamate receptors (mGlu2/3 receptors). Chronic pharmacologic activation of mGlu2/3 receptors significantly reduced the hyperdensity particle load in the ventral cortical regions of 3xTgAD mice, suggesting a neuroprotective effect with locoregional efficacy. CONCLUSIONS: This multiscale micro-to-nano 3D imaging method based on X-PCI-CT enabled identification and quantification of cellular and sub-cellular aging and neurodegeneration in deep neuronal and glial cell populations in a transgenic model of Alzheimer's disease. This approach quantified the localized and intracellular neuroprotective effects of pharmacological activation of mGlu2/3 receptors.
PURPOSE: Modern neuroimaging lacks the tools necessary for whole-brain, anatomically dense neuronal damage screening. An ideal approach would include unbiased histopathologic identification of aging and neurodegenerative disease. METHODS: We report the postmortem application of multiscale X-ray phase-contrast computed tomography (X-PCI-CT) for the label-free and dissection-free organ-level to intracellular-level 3D visualization of distinct single neurons and glia. In deep neuronal populations in the brain of aged wild-type and of 3xTgAD mice (a triply-transgenic model of Alzheimer's disease), we quantified intracellular hyperdensity, a manifestation of aging or neurodegeneration. RESULTS: In 3xTgAD mice, the observed hyperdensity was identified as amyloid-β and hyper-phosphorylated tau protein deposits with calcium and iron involvement, by correlating the X-PCI-CT data to immunohistochemistry, X-ray fluorescence microscopy, high-field MRI, and TEM. As a proof-of-concept, X-PCI-CT was used to analyze hippocampal and cortical brain regions of 3xTgAD mice treated with LY379268, selective agonist of group II metabotropic glutamate receptors (mGlu2/3 receptors). Chronic pharmacologic activation of mGlu2/3 receptors significantly reduced the hyperdensity particle load in the ventral cortical regions of 3xTgAD mice, suggesting a neuroprotective effect with locoregional efficacy. CONCLUSIONS: This multiscale micro-to-nano 3D imaging method based on X-PCI-CT enabled identification and quantification of cellular and sub-cellular aging and neurodegeneration in deep neuronal and glial cell populations in a transgenic model of Alzheimer's disease. This approach quantified the localized and intracellular neuroprotective effects of pharmacological activation of mGlu2/3 receptors.
Authors: M A Beltran; D M Paganin; K K W Siu; A Fouras; S B Hooper; D H Reser; M J Kitchen Journal: Phys Med Biol Date: 2011-11-02 Impact factor: 3.609
Authors: Giacomo E Barbone; Alberto Bravin; Pantaleo Romanelli; Alberto Mittone; Domenico Bucci; Thomas Gaaβ; Géraldine Le Duc; Sigrid Auweter; Maximilian F Reiser; Markus J Kraiger; Martin Hrabě de Angelis; Giuseppe Battaglia; Paola Coan Journal: Int J Radiat Oncol Biol Phys Date: 2018-04-07 Impact factor: 7.038