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Engrailed protects mouse midbrain dopaminergic neurons against mitochondrial complex I insults

Nature Neuroscience volume 14, pages 1260–1266 (2011)Cite this article

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Abstract

Mice heterozygous for the homeobox gene Engrailed-1 (En1) display progressive loss of mesencephalic dopaminergic (mDA) neurons. We report that exogenous Engrailed-1 and Engrailed-2 (collectively Engrailed) protect mDA neurons from 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), a mitochondrial complex I toxin used to model Parkinson's disease in animals. Engrailed enhances the translation of nuclearly encoded mRNAs for two key complex I subunits, Ndufs1 and Ndufs3, and increases complex I activity. Accordingly, in vivo protection against MPTP by Engrailed is antagonized by Ndufs1 small interfering RNA. An association between Engrailed and complex I is further confirmed by the reduced expression of Ndufs1 and Ndufs3 in the substantia nigra pars compacta of En1 heterozygous mice. Engrailed also confers in vivo protection against 6-hydroxydopamine and α-synuclein-A30P. Finally, the unilateral infusion of Engrailed into the midbrain increases striatal dopamine content, resulting in contralateral amphetamine-induced turning. Therefore, Engrailed is both a survival factor for adult mDA neurons and a regulator of their physiological activity.

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Figure 1: Engrailed protects against complex I, but not against complex II, inhibition.
Figure 2: Engrailed increases the translation of specific complex I subunits and complex I activity.
Figure 3: Engrailed enhances Ndufs1 and Ndufs3 levels in primary midbrain neuron cultures.
Figure 4: Ndufs1 and Ndufs3 expression is diminished in En1+/−En2+/+ mice.
Figure 5: Engrailed protection against MPTP is Ndufs1 dependent.
Figure 6: Engrailed-driven increases in striatal dopamine and turning behavior after MPTP are Ndufs1 dependent.
Figure 7: Engrailed protects against 6-OHDA and α-synuclein-A30P toxicity.

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Acknowledgements

This work was supported by Agence Nationale pour la Recherche (ANR-06-013-01), the Michael. J. Fox Foundation (MJFF), Fondation de France (Physiopathology of Parkinson disease) and the European Community (FP7 222999, mdDANeurodev project). D.A.-F. was funded by the MJFF, the University Medical Center Giessen and Marburg (UKGM) and the German Academic Exchange Service (DAAD). The CIRB group is part of a Global Research Laboratory (KAIST, South Korea) and a Foundation for Medical Research (FRM) team. J.F. was funded by the German Research Foundation (DFG), the MJFF, FRM and NERF (Neuropôle de Recherche Francilien). A.H. was temporarily supported by INSERM. We thank E. Ipendey for technical help. Confocal imaging and quantification were performed by V. Mignon and B. Saubaméa. We thank the Laboratory of Mass Spectrometry and Proteomics, Institut Curie for assistance. We are indebted to A. Prigent, E. Hirsch, P. Michel and M. Volovitch for intellectual input.

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Author notes

  1. Daniel Alvarez-Fischer and Julia Fuchs: These authors contributed equally to this work.

Authors and Affiliations

  1. Centre de Recherche de l'Institut du Cerveau et de la Moelle Epinière University Pierre and Marie Curie/Institut National de la Santé et de la Recherche Médicale, Unité Mixte de Recherche S 975, Paris, France

    Daniel Alvarez-Fischer & Andreas Hartmann

  2. Collège de France, Center for Interdisciplinary Research in Biology (CIRB)/Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7241/Institut National de la Santé et de la Recherche Médicale U1050, Paris, France

    Daniel Alvarez-Fischer, Julia Fuchs, François Castagner, Olivier Stettler, Olivia Massiani-Beaudoin, Kenneth L Moya, Colette Bouillot, Rajiv L Joshi & Alain Prochiantz

  3. Groupe Hospitalier Pitié-Salpêtrière, Paris, France

    Daniel Alvarez-Fischer & Andreas Hartmann

  4. Department of Neurology, Philipps University, Marburg, Germany

    Daniel Alvarez-Fischer & Wolfgang H Oertel

  5. Labex Memolife, Paris Sciences Lettres Research University,

    Julia Fuchs, François Castagner, Olivier Stettler, Olivia Massiani-Beaudoin, Kenneth L Moya, Colette Bouillot, Rajiv L Joshi & Alain Prochiantz

  6. University Pierre et Marie Curie, Ecole Doctorale 158, Paris, France.,

    Julia Fuchs, François Castagner, Olivia Massiani-Beaudoin, Kenneth L Moya, Colette Bouillot, Rajiv L Joshi & Alain Prochiantz

  7. Université Paris Descartes, Paris, France

    Olivier Stettler

  8. Institut National de la Santé et de la Recherche Médicale U975, Hôpital de La Salpêtrière, Paris, France

    Anne Lombès

  9. Laboratoire de Spectrométrie de Masse, Institut Curie, Paris, France

    Wolfgang Faigle

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Contributions

A.P. proposed the initial hypothesis that Engrailed could be used as a protective protein in animal models of Parkinson's disease. D.A.-F., J.F., F.C., O.M.-B. and R.L.J. performed experiments to study Engrailed survival activity. O.S., C.B., K.L.M., J.F., O.M.-B. and R.L.J. examined Engrailed translational targets. W.F. helped with mass spectroscopy. A.L. measured complex I activity. W.H.O. participated in discussions. J.F., D.A.-F., R.L.J., A.H. and A.P. designed the experiments and interpreted the results. D.A.-F., J.F., R.L.J. and A.P. wrote the manuscript.

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Correspondence to Rajiv L Joshi, Andreas Hartmann or Alain Prochiantz.

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The authors declare no competing financial interests.

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Alvarez-Fischer, D., Fuchs, J., Castagner, F. et al. Engrailed protects mouse midbrain dopaminergic neurons against mitochondrial complex I insults. Nat Neurosci 14, 1260–1266 (2011). https://doi.org/10.1038/nn.2916

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