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Chromogranin-mediated secretion of mutant superoxide dismutase proteins linked to amyotrophic lateral sclerosis

Nature Neuroscience volume 9, pages 108118 (2006) | Download Citation

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Abstract

Here we report that chromogranins, components of neurosecretory vesicles, interact with mutant forms of superoxide dismutase (SOD1) that are linked to amyotrophic lateral sclerosis (ALS), but not with wild-type SOD1. This interaction was confirmed by yeast two-hybrid screen and by co-immunoprecipitation assays using either lysates from Neuro2a cells coexpressing chromogranins and SOD1 mutants or lysates from spinal cord of ALS mice. Confocal and immunoelectron microscopy revealed a partial colocalization of mutant SOD1 with chromogranins in spinal cord of ALS mice. Mutant SOD1 was also found in immuno-isolated trans-Golgi network and in microsome preparations, suggesting that it can be secreted. Indeed we report evidence that chromogranins may act as chaperone-like proteins to promote secretion of SOD1 mutants. From these results, and our finding that extracellular mutant SOD1 can trigger microgliosis and neuronal death, we propose a new ALS pathogenic model based on the toxicity of secreted SOD1 mutants.

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Acknowledgements

We thank R. Janvier for sample preparation for immunoelectron microscopy and B. Gentil for advice on experimental procedures. The technical help from G. Soucy, S.A. Ezzi (Laval University) and J. Kurisu (RIKEN Brain Science Institute) is appreciated. We thank D. Cleveland (University of California San Diego) for the G37R SOD1 transgenic mice and Y. Imai for the DsRed-Golgi plasmid. This work was supported by the Canadian Institutes of Health Research (CIHR), the Robert Packard Centre for ALS Research at Johns Hopkins, the ALS Association (USA), the ALS Society of Canada, the Japan Society for the Promotion of Science (JSPS) and the Japan Foundation for Neuroscience and Mental Health. J.-P.J. holds a Canada Research Chair in Neurodegeneration. M.U. is a recipient of a Uehara Memorial Foundation research fellowship and a postdoctoral fellowship from CIHR.

Author information

Affiliations

  1. Department of Anatomy and Physiology, Laval University, Centre de Recherche du Centre Hospitalier de l' Université Laval, 2705 boulevard Laurier, Sainte-Foy, Quebec G1V 4G2, Canada.

    • Makoto Urushitani
    •  & Jean-Pierre Julien
  2. Department of Psychiatry, Centre de Recherche Université Laval Robert-Giffard, 2601 de la Canardiere, Quebec, Quebec G1J 2G3, Canada.

    • Attila Sik
  3. Laboratory for Structural Neuropathology, RIKEN Brain Science Institute, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.

    • Takashi Sakurai
    •  & Nobuyuki Nukina
  4. Motor System Neurodegeneration, RIKEN Brain Science Institute, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.

    • Ryosuke Takahashi

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Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Jean-Pierre Julien.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Amino acid sequence alignment data.

  2. 2.

    Supplementary Fig. 2

    Expression of chromogranin mRNA and proteins the mouse spinal cord.

  3. 3.

    Supplementary Fig. 3

    Immunoelectron microscope showing different distribution pattern of G37R and wild-type SOD1 in the spinal motor neuron of human SOD1 transgenic mice

  4. 4.

    Supplementary Fig. 4

    Generation of antibody recognizing amino-terminal peptides of mouse mature CgA.

  5. 5.

    Supplementary Fig. 5

    CgA is expressed in reactive astrocytes in spinal anterior horn of the G93A SOD1 mutant transgenic mice.

  6. 6.

    Supplementary Fig. 6

    A model of pathogenesis consistent with the notion of non-cell autonomous toxicity of mutant SOD1.

  7. 7.

    Supplementary Table 1

    Cloning primer pairs for mouse chromogranin A (CgA), B (CgB), or deletion mutants of mouse chromogranin B (amino terminus deletion; ΔN, carboxyl terminus deletion; ΔC, inter-conserved domain deletion; ΔIBC) and for recombinant mouse chromogranin A (Rec CgA).

  8. 8.

    Supplementary Table 2

    Primer pairs used for semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR).

  9. 9.

    Supplementary Methods

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DOI

https://doi.org/10.1038/nn1603

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