Abstract

Huntington's disease is caused by an expanded polyglutamine repeat in the huntingtin protein (HTT), but the pathophysiological sequence of events that trigger synaptic failure and neuronal loss are not fully understood. Alterations in N-methyl-D-aspartate (NMDA)-type glutamate receptors (NMDARs) have been implicated. Yet, it remains unclear how the HTT mutation affects NMDAR function, and direct evidence for a causative role is missing. Here we show that mutant HTT redirects an intracellular store of juvenile NMDARs containing GluN3A subunits to the surface of striatal neurons by sequestering and disrupting the subcellular localization of the endocytic adaptor PACSIN1, which is specific for GluN3A. Overexpressing GluN3A in wild-type mouse striatum mimicked the synapse loss observed in Huntington's disease mouse models, whereas genetic deletion of GluN3A prevented synapse degeneration, ameliorated motor and cognitive decline and reduced striatal atrophy and neuronal loss in the YAC128 Huntington's disease mouse model. Furthermore, GluN3A deletion corrected the abnormally enhanced NMDAR currents, which have been linked to cell death in Huntington's disease and other neurodegenerative conditions. Our findings reveal an early pathogenic role of GluN3A dysregulation in Huntington's disease and suggest that therapies targeting GluN3A or pathogenic HTT-PACSIN1 interactions might prevent or delay disease progression.

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Acknowledgements

We thank S. Finkbeiner (Gladstone Institute, University of California San Francisco), H. Zoghbi (Baylor College of Medicine), T. Yamamoto (University of Tokyo) and M. Ehlers (Pfizer Neuroscience) for providing reagents, A. Zandueta, C. Rodríguez-Viña, F. Ballesteros, X. Remírez and M. Montañana for excellent technical help, M. Galarraga for advice with image analysis, and M. Ehlers, M. Arrasate, T. Aragón, J.A. Esteban and B.D. Philpot for critical readings of the manuscript. This work was funded by the Unión Temporal de Empresas (UTE) project at the Centro de Investigación Médica Aplicada, Gobierno de Navarra, and Spanish Ministry of Science grants (SAF2010-20636 and CSD2008-00005 to I.P.-O., BFU2009-12160 to J.F.W. and SAF2011-29507 to J.A.), grants from the Hereditary Disease Foundation (to I.P.-O. and D.C.L.), US National Institutes of Health grants P01 HD29587, P01 ES016738 and R01 EY05477 (to S.A.L.) and grants from the Cure Huntington's Disease Initiative and the Canadian Institutes of Health Research (to M.R.H.). M.R.H., a Killam University Professor, holds a Canada Research Chair in Human Genetics.

Author information

Author notes

    • Rona K Graham

    Present address: Research Center on Aging, Department of Physiology and Biophysics, University of Sherbrooke, Sherbrooke, Quebec, Canada.

    • Sonia Marco
    •  & Albert Giralt

    These authors contributed equally to this work.

Affiliations

  1. Cellular Neurobiology and Neurophysiology Laboratories, Centro de Investigación Médica Aplicada (CIMA) y Universidad de Navarra, Pamplona, Spain.

    • Sonia Marco
    • , Milos M Petrovic
    • , Rebeca Martínez-Turrillas
    • , John F Wesseling
    •  & Isabel Pérez-Otaño
  2. Department of Cellular Biology, Immunology and Neuroscience, Medical School, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), University of Barcelona, Barcelona and Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), Spain.

    • Albert Giralt
    • , Jesús Torres-Peraza
    •  & Jordi Alberch
  3. Institute of Medical Physiology, School of Medicine, University of Belgrade, Belgrade, Serbia.

    • Milos M Petrovic
  4. Centre For Molecular Medicine and Therapeutics, Child and Family Research Institute, University of British Columbia, Vancouver, Canada.

    • Mahmoud A Pouladi
    • , Rona K Graham
    •  & Michael R Hayden
  5. Translational Laboratory in Genetic Medicine (TLGM), Department of Medicine, National University of Singapore and Agency for Science, Technology and Research (A*STAR), Singapore.

    • Mahmoud A Pouladi
    •  & Michael R Hayden
  6. Departamento de Ciencias Médicas, Centro Regional de Investigaciones Biomédicas (CRIB), Facultad de Medicina, Universidad de Castilla-La Mancha, Albacete, Spain.

    • José Martínez-Hernández
    •  & Rafael Luján
  7. Center for Drug Discovery and Department of Neurobiology, Duke University Medical Center, Durham, North Carolina, USA.

    • Linda S Kaltenbach
    •  & Donald C Lo
  8. Department of Anatomy, Hokkaido University School of Medicine, Sapporo, Japan.

    • Masahiko Watanabe
  9. Del E. Webb Center for Neuroscience, Aging and Stem Cell Research, Sanford-Burnham Medical Research Institute, La Jolla, California, USA.

    • Nobuki Nakanishi
    •  & Stuart A Lipton
  10. Teva Pharmaceutical Industries Ltd, Petah Tikva, Israel.

    • Michael R Hayden

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Contributions

S.M. performed and analyzed the biochemical experiments in cultured neurons, recombinant cells and YAC128 mice, the imaging experiments in cultured neurons and the spine morphology measurements and collaborated with A.G. on neuropathological studies. A.G. performed and analyzed biochemical experiments in humans and R6/1 and knock-in mice and the behavioral experiments. M.M.P. performed electrophysiological recordings. M.A.P. performed and analyzed behavioral experiments. R.M.-T. contributed to mouse genotyping and conducted biochemical fractionation and real-time PCR assays. J.M.-H. and R.L. performed and analyzed electron microscopy studies. L.S.K. and D.C.L. performed and analyzed slice culture experiments. J.T.-P. performed the initial biochemical experiments in R6/1 mice. M.R.H. and R.K.G. provided the YAC128 mice. N.N. and S.A.L. provided the Grin3a−/− mice. M.W. made the GluN3A-specific antibody used in biochemical and immunocytochemical analyses. J.A. designed and supervised experiments. J.F.W. designed and analyzed electrophysiological and fluorescence colocalization experiments and contributed to the manuscript writing. I.P.-O. conceived the study, designed experiments, analyzed data and wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Isabel Pérez-Otaño.

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https://doi.org/10.1038/nm.3246

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