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Proteostasis of polyglutamine varies among neurons and predicts neurodegeneration

Nature Chemical Biology volume 9, pages 586–592 (2013)Cite this article

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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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Figure 1: Optical pulse labeling to measure protein turnover in individual neurons.
Figure 2: PolyQ expansion destabilizes diffuse mHttex1 and full-length Htt.
Figure 3: Proteostasis of mhttex1 determines degeneration.
Figure 4: Nrf2, a stress-activated transcription factor, shortens the mean lifetime of mHttex1-Q46-Dendra2 and increases survival of striatal neurons.
Figure 5: Inhibiting the ubiquitin-proteasome system or autophagy differentially affects the mean lifetimes of Httex1-Q25-Dendra2 and mHttex1-Q46-Dendra2.
Figure 6: Neuron type–specific proteostasis of mHttex1 contributes to their susceptibility to degeneration.

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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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  1. Montserrat Arrasate

    Present address: Present address: Division of Neuroscience, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.,

Authors and Affiliations

  1. Gladstone Institute of Neurological Disease, San Francisco, California, USA

    Andrey S Tsvetkov, Montserrat Arrasate, Sami Barmada, Punita Sharma & Steven Finkbeiner

  2. Taube-Koret Center for Neurodegenerative Disease Research, San Francisco, California, USA

    Andrey S Tsvetkov, Montserrat Arrasate, Punita Sharma & Steven Finkbeiner

  3. Biomedical Sciences Graduate Program, University of California–San Francisco, San Francisco, California, USA

    D Michael Ando & Steven Finkbeiner

  4. Department of Statistical Science, Duke University, Durham, North Carolina, USA

    Benjamin A Shaby

  5. Graduate Programs in Neuroscience and Biomedical Sciences, University of California–San Francisco, San Francisco, California, USA

    Steven Finkbeiner

  6. Program in Biological Sciences and Medical Scientist Training Program, University of California–San Francisco, San Francisco, California, USA

    Steven Finkbeiner

  7. Department of Neurology and Physiology, University of California–San Francisco, San Francisco, California, USA

    Steven Finkbeiner

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Contributions

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.

Corresponding author

Correspondence to Steven Finkbeiner.

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The authors declare no competing financial interests.

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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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