Abstract
In polyglutamine (polyQ) diseases, only certain neurons die, despite widespread expression of the offending protein. PolyQ expansion may induce neurodegeneration by impairing proteostasis, but protein aggregation and toxicity tend to confound conventional measurements of protein stability. Here, we used optical pulse labeling to measure effects of polyQ expansions on the mean lifetime of a fragment of huntingtin, the protein that causes Huntington's disease, in living neurons. We show that polyQ expansion reduced the mean lifetime of mutant huntingtin within a given neuron and that the mean lifetime varied among neurons, indicating differences in their capacity to clear the polypeptide. We found that neuronal longevity is predicted by the mean lifetime of huntingtin, as cortical neurons cleared mutant huntingtin faster and lived longer than striatal neurons. Thus, cell type–specific differences in turnover capacity may contribute to cellular susceptibility to toxic proteins, and efforts to bolster proteostasis in Huntington's disease, such as protein clearance, could be neuroprotective.
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Acknowledgements
This work was supported by grants R01 3NS039746 and 2R01 NS045191 from the US National Institute of Neurological Disease and Stroke; grant P01 2AG022074 from the National Institute on Aging; by the Huntington's Disease Society of America (made possible with a gift from the James E. Bashaw Family); the Taube-Koret Center for Neurodegenerative disease and the Gladstone Institutes (S.F.); the Milton Wexler Award and a fellowship from the Hereditary Disease Foundation (A.S.T.); a fellowship from the Hillblom Foundation (M.A.); a fellowship from California Institute for Regenerative Medicine (P.S.), and in part by DMS-0914906 from the US National Science Foundation (B.A.S.). Gladstone Institutes received support from a US National Center for Research Resources Grant RR18928-01. We thank Y. Dabaghian, I. Kelmanson, A. Gelfand and members of the Finkbeiner laboratory for helpful discussions. The animal care facility was partly supported by a US National Institutes of Health Extramural Research Facilities Improvement Project (C06 RR018928). K. Nelson provided administrative assistance, and G.C. Howard, A.L. Lucido and S. Ordway edited the manuscript.
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A.S.T., M.A. and S.F. designed the study. A.S.T. and S.F. wrote the manuscript. B.A.S. performed statistical analysis and wrote the statistical analysis section of the manuscript. A.S.T., M.A., P.S., S.B. and D.M.A. wrote scripts for automated photoswitching and imaging. A.S.T. cloned all of the constructs used in the study. A.S.T. and M.A. cultured primary neurons and performed transfections, automated microscopy, fluorescence intensity measurements and data analysis. A.S.T. performed detergent extraction, metabolic labeling and photobleaching experiments. A.S.T. and P.S. performed survival analyses.
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Tsvetkov, A., Arrasate, M., Barmada, S. et al. Proteostasis of polyglutamine varies among neurons and predicts neurodegeneration. Nat Chem Biol 9, 586–592 (2013). https://doi.org/10.1038/nchembio.1308
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DOI: https://doi.org/10.1038/nchembio.1308
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