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miR-7a regulation of Pax6 controls spatial origin of forebrain dopaminergic neurons

Nature Neuroscience volume 15, pages 1120–1126 (2012)Cite this article

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Abstract

In the postnatal and adult mouse forebrain, a mosaic of spatially separated neural stem cells along the lateral wall of the ventricles generates defined types of olfactory bulb neurons. To understand the mechanisms underlying the regionalization of the stem cell pool, we focused on the transcription factor Pax6, a determinant of the dopaminergic phenotype in this system. We found that, although Pax6 mRNA was transcribed widely along the ventricular walls, Pax6 protein was restricted to the dorsal aspect. This dorsal restriction was a result of inhibition of protein expression by miR-7a, a microRNA (miRNA) that was expressed in a gradient opposing Pax6. In vivo inhibition of miR-7a in Pax6-negative regions of the lateral wall induced Pax6 protein expression and increased dopaminergic neurons in the olfactory bulb. These findings establish miRNA-mediated fine-tuning of protein expression as a mechanism for controlling neuronal stem cell diversity and, consequently, neuronal phenotype.

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Figure 1: Discrepancy between expression of Pax6 promoter activity and protein along the lateral wall of the lateral ventricle.
Figure 2: Fate of dorsal and lateral neural progenitors in the olfactory bulb.
Figure 3: Pax6 expression is post-transcriptionally inhibited in a dorso-ventral gradient via its 3′ UTR.
Figure 4: Identification of miR-7a as a potential regulator of Pax6.
Figure 5: miR-7a controls Pax6 expression through its 3′ UTR in vivo.
Figure 6: De-repression of Pax6 protein expression by miR-7a inhibition in the lateral PVZ favors a dopaminergic phenotype in the olfactory bulb.

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Acknowledgements

We thank D. Price (University of Edinburgh) for tissue from PAX6-GFP mice, D. van Vector (Harvard Medical School) for miR-7a sponge plasmid, U. Bissels (Miltenyi Biotec) for advice on luciferase assays, Y. Manhoun for help with deep-sequencing data analyses, A. Barani (Precym platform of Centre d'Océanologie de Marseille) for fluorescence-activated cell sorting, J. Ninkovic for helpful tips on antibody labeling and X. Morin (Ecole Normale Superieure) for plasmids. We acknowledge the expertise of V. Magnone and K. Lebrigand (Functional Genomics Platform of Nice Sophia Antipolis), and R. Waldmann and L.-E. Zaragosi for fruitful discussions. We thank A. Bosio, R. Kelly and M.C. Tiveron for critical reading of the manuscript. This work was supported by funds from the Agence National de la Recherche (ANR, FORDOPA), Foundation pour la Recherche Médicale (Label Equipe FRM), Fondation de France and the European Commission (Marie-Curie: ITN AXREGEN and IAPP DopaNew) to H.C. P.B. was supported by Région PACA, CG06 and by grants from ANR, Vaincre la Mucoviscidose, Association pour la Recherche sur le Cancer and Institut National du Cancer.

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Author notes

  1. Antoine de Chevigny and Nathalie Coré: These authors contributed equally to this work.

Authors and Affiliations

  1. Institut de Biologie de Développement de Marseille-Luminy, Aix-Marseille University, Marseille, France

    Antoine de Chevigny, Nathalie Coré, Philipp Follert, Marion Gaudin, Christophe Béclin & Harold Cremer

  2. CNRS (Centre National de la Recherche Scientifique), UMR 7288, Marseille, France.,

    Antoine de Chevigny, Nathalie Coré, Philipp Follert, Marion Gaudin, Christophe Béclin & Harold Cremer

  3. CNRS and University of Nice Sophia Antipolis, Institut de Pharmacologie Moléculaire et Cellulaire, Sophia Antipolis, France

    Pascal Barbry

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Contributions

A.d.C. and N.C. designed and conducted experiments, and wrote the manuscript. P.F. and P.B. contributed miRNA expression data. M.G. performed molecular cloning. C.B. designed experiments and performed miRNA studies. H.C. designed the research and wrote the manuscript.

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Correspondence to Nathalie Coré or Harold Cremer.

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de Chevigny, A., Coré, N., Follert, P. et al. miR-7a regulation of Pax6 controls spatial origin of forebrain dopaminergic neurons. Nat Neurosci 15, 1120–1126 (2012). https://doi.org/10.1038/nn.3142

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