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Tau Fibril Formation in Cultured Cells Compatible with a Mouse Model of Tauopathy

Neurofibrillary tangles composed of hyperphosphorylated tau protein are primarily neuropathological features of a number of neurodegenerative diseases collectively termed tauopathy. To understand the mechanisms underlying the cause of tauopathy, precise cellular and animal models are required. Recen...

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Autores principales: Matsumoto, Gen, Matsumoto, Kazuki, Kimura, Taeko, Suhara, Tetsuya, Higuchi, Makoto, Sahara, Naruhiko, Mori, Nozomu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5983680/
https://www.ncbi.nlm.nih.gov/pubmed/29772786
http://dx.doi.org/10.3390/ijms19051497
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author Matsumoto, Gen
Matsumoto, Kazuki
Kimura, Taeko
Suhara, Tetsuya
Higuchi, Makoto
Sahara, Naruhiko
Mori, Nozomu
author_facet Matsumoto, Gen
Matsumoto, Kazuki
Kimura, Taeko
Suhara, Tetsuya
Higuchi, Makoto
Sahara, Naruhiko
Mori, Nozomu
author_sort Matsumoto, Gen
collection PubMed
description Neurofibrillary tangles composed of hyperphosphorylated tau protein are primarily neuropathological features of a number of neurodegenerative diseases collectively termed tauopathy. To understand the mechanisms underlying the cause of tauopathy, precise cellular and animal models are required. Recent data suggest that the transient introduction of exogenous tau can accelerate the development of tauopathy in the brains of non-transgenic and transgenic mice expressing wild-type human tau. However, the transmission mechanism leading to tauopathy is not fully understood. In this study, we developed cultured-cell models of tauopathy representing a human tauopathy. Neuro2a (N2a) cells containing propagative tau filaments were generated by introducing purified tau fibrils. These cell lines expressed full-length (2N4R) human tau and the green fluorescent protein (GFP)-fused repeat domain of tau with P301L mutation. Immunocytochemistry and super-resolution microscopic imaging revealed that tau inclusions exhibited filamentous morphology and were composed of both full-length and repeat domain fragment tau. Live-cell imaging analysis revealed that filamentous tau inclusions are transmitted to daughter cells, resulting in yeast-prion-like propagation. By a standard method of tau preparation, both full-length tau and repeat domain fragments were recovered in sarkosyl insoluble fraction. Hyperphosphorylation of full-length tau was confirmed by the immunoreactivity of phospho-Tau antibodies and mobility shifts by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). These properties were similar to the biochemical features of P301L mutated human tau in a mouse model of tauopathy. In addition, filamentous tau aggregates in cells barely co-localized with ubiquitins, suggesting that most tau aggregates were excluded from protein degradation systems, and thus propagated to daughter cells. The present cellular model of tauopathy will provide an advantage for dissecting the mechanisms of tau aggregation and degradation and be a powerful tool for drug screening to prevent tauopathy.
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spelling pubmed-59836802018-06-05 Tau Fibril Formation in Cultured Cells Compatible with a Mouse Model of Tauopathy Matsumoto, Gen Matsumoto, Kazuki Kimura, Taeko Suhara, Tetsuya Higuchi, Makoto Sahara, Naruhiko Mori, Nozomu Int J Mol Sci Article Neurofibrillary tangles composed of hyperphosphorylated tau protein are primarily neuropathological features of a number of neurodegenerative diseases collectively termed tauopathy. To understand the mechanisms underlying the cause of tauopathy, precise cellular and animal models are required. Recent data suggest that the transient introduction of exogenous tau can accelerate the development of tauopathy in the brains of non-transgenic and transgenic mice expressing wild-type human tau. However, the transmission mechanism leading to tauopathy is not fully understood. In this study, we developed cultured-cell models of tauopathy representing a human tauopathy. Neuro2a (N2a) cells containing propagative tau filaments were generated by introducing purified tau fibrils. These cell lines expressed full-length (2N4R) human tau and the green fluorescent protein (GFP)-fused repeat domain of tau with P301L mutation. Immunocytochemistry and super-resolution microscopic imaging revealed that tau inclusions exhibited filamentous morphology and were composed of both full-length and repeat domain fragment tau. Live-cell imaging analysis revealed that filamentous tau inclusions are transmitted to daughter cells, resulting in yeast-prion-like propagation. By a standard method of tau preparation, both full-length tau and repeat domain fragments were recovered in sarkosyl insoluble fraction. Hyperphosphorylation of full-length tau was confirmed by the immunoreactivity of phospho-Tau antibodies and mobility shifts by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). These properties were similar to the biochemical features of P301L mutated human tau in a mouse model of tauopathy. In addition, filamentous tau aggregates in cells barely co-localized with ubiquitins, suggesting that most tau aggregates were excluded from protein degradation systems, and thus propagated to daughter cells. The present cellular model of tauopathy will provide an advantage for dissecting the mechanisms of tau aggregation and degradation and be a powerful tool for drug screening to prevent tauopathy. MDPI 2018-05-17 /pmc/articles/PMC5983680/ /pubmed/29772786 http://dx.doi.org/10.3390/ijms19051497 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Matsumoto, Gen
Matsumoto, Kazuki
Kimura, Taeko
Suhara, Tetsuya
Higuchi, Makoto
Sahara, Naruhiko
Mori, Nozomu
Tau Fibril Formation in Cultured Cells Compatible with a Mouse Model of Tauopathy
title Tau Fibril Formation in Cultured Cells Compatible with a Mouse Model of Tauopathy
title_full Tau Fibril Formation in Cultured Cells Compatible with a Mouse Model of Tauopathy
title_fullStr Tau Fibril Formation in Cultured Cells Compatible with a Mouse Model of Tauopathy
title_full_unstemmed Tau Fibril Formation in Cultured Cells Compatible with a Mouse Model of Tauopathy
title_short Tau Fibril Formation in Cultured Cells Compatible with a Mouse Model of Tauopathy
title_sort tau fibril formation in cultured cells compatible with a mouse model of tauopathy
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5983680/
https://www.ncbi.nlm.nih.gov/pubmed/29772786
http://dx.doi.org/10.3390/ijms19051497
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