Imaging And Quantification Of Amyloid Fibrillation In The Cell Nucleus
Xenobiotics, as well as intrinsic processes such as cellular aging, contribute to an environment that constantly challenges nuclear organization and function. While it becomes increasingly clear that proteasome-dependent proteolysis is a major player, the topology and molecular mechanisms of nuclear protein homeostasis remain largely unknown. We have shown previously that (1) proteasome-dependent protein degradation is organized in focal microenvironments throughout the nucleoplasm and (2) heavy metals as well as nanoparticles induce nuclear protein fibrillation with amyloid characteristics. Here, we describe methods to characterize the landscape of intranuclear amyloid on the global and local level in different systems such as cultures of mammalian cells and the soil nematode Caenorhabditis elegans. Application of discrete mathematics to imaging data is introduced as a tool to develop pattern recognition of intracellular protein fibrillation. Since stepwise fibrillation of otherwise soluble proteins to insoluble amyloid-like protein aggregates is a hallmark of neurodegenerative protein-misfolding disorders including Alzheimer's disease, CAG repeat diseases, and the prion encephalopathies, investigation of intracellular amyloid may likewise aid to a better understanding of the pathomechanisms involved. We consider aggregate profiling as an important experimental approach to determine if nuclear amyloid has toxic or protective roles in various disease processes.
F. Arnhold et al., "Imaging And Quantification Of Amyloid Fibrillation In The Cell Nucleus," Methods in Molecular Biology, vol. 1228, pp. 187 - 202, Springer, Jan 2015.
The definitive version is available at https://doi.org/10.1007/978-1-4939-1680-1_15
Keywords and Phrases
Aging; Amyloid; Congo red; Fibrillation; Huntington's disease; Neurodegeneration; Nucleus; Polyglutamine (polyQ) expansion diseases; Protein aggregation
International Standard Serial Number (ISSN)
Article - Journal
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01 Jan 2015