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Sonic Hedgehog is Cytoprotective against Oxidative Challenge in a Cellular Model of Amyotrophic Lateral Sclerosis

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Abstract

We have previously demonstrated that primary cilia on spinal motor neurons are reduced in G93A SOD1 (mSOD) mice, a mouse model of amyotrophic lateral sclerosis (ALS). Sonic hedgehog (Shh) signaling involves the primary cilium and Shh has been shown to be cytoprotective in models of other neurodegenerative diseases. Thus, the Shh signaling pathway may bear further study in ALS. Accordingly, we established that interference with the Shh pathway (with the Shh antagonist cyclopamine or with miRNA 3245p) sensitized HT22 cells, while augmentation of the Shh pathway (with Shh or the Shh agonist purmorphamine) protected cells against hydrogen peroxide (H2O2) challenge. We ectopically expressed mSOD, human wild-type SOD1 (wtSOD), or an empty vector in HT22 cells. Compared to empty vector, wtSOD decreased cell death and mSOD increased cell death in response to H2O2 challenge. Treatment with cyclopamine or miRNA 3245p sensitized all three transfections to H2O2 challenge. Treatment with recombinant human Shh or purmorphamine decreased cell death after H2O2 challenge, an effect more pronounced in mSOD cells. Compared with empty vector, overexpression of wtSOD increased Shh and Gli transcript levels and increased activity in a Gli-responsive reporter assay. Overexpression of mSOD did not change Shh transcript levels, but decreased Gli transcript levels, especially Gli3, and reduced activity in a Gli reporter assay. These results suggest that overexpression of mSOD but not wtSOD reduces signaling in the Shh pathway and renders mSOD cells more susceptible to H2O2 challenge, and that treatment with Shh or Shh agonists is cytoprotective to mSOD cells. Shh or Shh agonists merit further consideration as potential therapy in ALS.

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Acknowledgment

The authors wish to thank Mr. David Hunt, whose generosity has helped fund this research.

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Authors and Affiliations

  1. Department of Medicine, McMaster University Medical Centre, Rm 4U7, 1200 Main St West, Hamilton, Ontario, L8N 3Z5, Canada

    Randy Peterson & John Turnbull

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  1. Randy Peterson
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Correspondence to John Turnbull.

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Fig. S1

Relative transfection efficiency in HT22 cells. 24 h after transfection, three wells per transfection group were used to quantitate GFP florescence. Five fields per well were counted, and a minimum of 1,000 cells scored. Data represent the mean ± SD (DOC 1049 kb)

Fig. S2

The relative percent viability of treated or transfected HT22 cells in the absence of H2O2 challenge. 24 h post-treatment or transfection, cellular viability was measured via the MTT assay. Data represents the mean ± SD (DOC 71 kb)

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Peterson, R., Turnbull, J. Sonic Hedgehog is Cytoprotective against Oxidative Challenge in a Cellular Model of Amyotrophic Lateral Sclerosis. J Mol Neurosci 47, 31–41 (2012). https://doi.org/10.1007/s12031-011-9660-x

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  • DOI: https://doi.org/10.1007/s12031-011-9660-x

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