Abstract
Parkinson’s disease (PD) is a neurodegenerative disorder characterized with the progressive loss of dopaminergic (DA) neurons within the substantia nigra pars compacta (SNc). Quantitative analysis of neuronal loss including neuronal processes, axons and dendrites, would advance the understanding of the pathogenesis of PD. ScaleS, an aqueous tissue clearing method, provides stable tissue preservation while maintaining potent clearing capability, allowing quantitative three-dimensional (3D) imaging of biological tissues. In this chapter, we describe detailed procedures for 3D imaging of brain slice tissues with ScaleS technique. These include brain slice preparation, tissue clarification, chemical and immunohistochemical labeling (ChemScale and AbScale), and observation of labeled tissues using a confocal laser scanning microscope (CLSM).
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References
Kalia LV, Lang AE (2015) Parkinson’s disease. Lancet 386(9996):896–912. https://doi.org/10.1016/S0140-6736(14)61393-3
Richardson DS, Lichtman JW (2015) Clarifying tissue clearing. Cell 162(2):246–257. https://doi.org/10.1016/j.cell.2015.06.067
Ueda HR, Dodt HU, Osten P, Economo MN, Chandrashekar J, Keller PJ (2020) Whole-brain profiling of cells and circuits in mammals by tissue clearing and light-sheet microscopy. Neuron 106(3):369–387. https://doi.org/10.1016/j.neuron.2020.03.004
Ueda HR, Erturk A, Chung K, Gradinaru V, Chedotal A, Tomancak P, Keller PJ (2020) Tissue clearing and its applications in neuroscience. Nat Rev Neurosci 21(2):61–79. https://doi.org/10.1038/s41583-019-0250-1
Hama H, Hioki H, Namiki K, Hoshida T, Kurokawa H, Ishidate F, Kaneko T, Akagi T, Saito T, Saido T, Miyawaki A (2015) ScaleS: an optical clearing palette for biological imaging. Nat Neurosci 18(10):1518–1529. https://doi.org/10.1038/nn.4107
Miyawaki A, Hama H, Hioki H, Namiki K, Hoshida T, Kurokawa H (2016) Deep imaging of cleared brain by confocal laser-scanning microscopy. Protocol Exchange. https://doi.org/10.1038/protex.2016.019
Acknowledgments
This study was supported by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) and the Japan Society for the Promotion of Science (JSPS) for Scientific Research (JP20K07231 to K.Y; JP16H04663 to H.H.) and Scientific Research on Innovative Area “Resonance Bio” (JP18H04743 to H.H.). This study was also supported by the Japan Agency for Medical Research and Development (AMED) (JP20dm0207064 to H.H.), Grants-in-Aid from the Research Institute for Diseases of Old Age at the Juntendo University School of Medicine (X2016 to K.Y.; X2001 to H.H.), and MEXT Private University Research Branding Project (Juntendo University).
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Yamauchi, K., Takahashi, M., Hioki, H. (2021). Application of a Tissue Clearing Method for the Analysis of Dopaminergic Axonal Projections. In: Imai, Y. (eds) Experimental Models of Parkinson’s Disease. Methods in Molecular Biology, vol 2322. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1495-2_14
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DOI: https://doi.org/10.1007/978-1-0716-1495-2_14
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