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A cell culture model for investigation of Hirano bodies

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Abstract

Hirano bodies are paracrystalline F-actin-rich aggregations associated with a variety of conditions including aging, and neurodegenerative diseases. The composition and structure of these inclusions have been described by immunohistochemistry and ultrastructure, respectively. However, studies of the physiological function and dynamics of Hirano bodies have been hindered due to lack of a facile in vitro experimental system. We have developed a model for formation of Hirano bodies in mammalian cell cultures by expression of the carboxy-terminal fragment (CT) of a 34-kDa actin-bundling protein. Expression of the CT protein induces F-actin rearrangement in HEK 293, HeLa, Cos7 cells, neuroblastoma and astrocytic cells, and in primary neurons. We have termed these structures model Hirano bodies, since their composition and ultrastructure is quite similar to that reported in vivo. Model Hirano bodies in cell cultures sometimes appeared to be formed of a number of smaller domains, suggesting that small aggregates are intermediates in the formation of Hirano bodies. Stable lines expressing CT and bearing model Hirano bodies exhibit normal growth, morphology, and motility. This model provides a valuable system for the study of the dynamics of Hirano bodies, and their role in disease processes.

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Acknowledgments

Most of the microscopy was performed using the facilities of the University of Georgia Center for Advanced Ultrastructural Research. H4 astrocytoma and N2A cells with model Hirano bodies were kindly provided by Sangdeuk Ha, and Nisha Gupta, respectively. We thank Dr. Richard McCann for antibody to talin 1 produced in his laboratory, and for a panel of commercial antibodies to focal adhesion components. This work was supported by awards to RF and MF from NSF (MCB 98-08748), the Alzheimer’s Association (IIRG-00-2436), and NIH (1R01-NS04645101).

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  1. Department of Cellular Biology, University of Georgia, 724 Biological Sciences, Athens, GA, 30602, USA

    Richard C. Davis, Ruth Furukawa & Marcus Fechheimer

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  1. Richard C. Davis
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  2. Ruth Furukawa
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Correspondence to Marcus Fechheimer.

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Davis, R.C., Furukawa, R. & Fechheimer, M. A cell culture model for investigation of Hirano bodies. Acta Neuropathol 115, 205–217 (2008). https://doi.org/10.1007/s00401-007-0275-9

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  • DOI: https://doi.org/10.1007/s00401-007-0275-9

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