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Wnt antagonism of Shh facilitates midbrain floor plate neurogenesis

Nature Neuroscience volume 12, pages 125–131 (2009)Cite this article

Abstract

The floor plate, an essential ventral midline organizing center that produces the morphogen Shh, has distinct properties along the neuraxis. The neurogenic potential of the floor plate and its underlying mechanisms remain unknown. Using Shh as a driver for lineage analysis, we found that the mouse midbrain, but not the hindbrain, floor plate is neurogenic, giving rise to dopamine (DA) neurons. Distinct spatiotemporal Shh and Wnt expression may distinguish the neurogenetic potential of these structures. We discovered an inhibitory role for Shh: removal of Shh resulted in neurogenesis from the hindbrain midline and, conversely, high doses of Shh inhibited proliferation and DA neuron production in midbrain cultures. We found that Wnt/beta-catenin signaling is necessary and sufficient for antagonizing Shh, DA progenitor marker induction and promotion of dopaminergic neurogenesis. These findings demonstrate how the dynamic interplay of canonical Wnt/beta-catenin signaling and Shh may orchestrate floor plate neurogenesis or a lack thereof.

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Figure 1: Midline Shh expression inversely correlates with neurogenesis.
Figure 2: Hindbrain midline neurogenesis in Shh::cre;Shh cKO embryos.
Figure 3: Reduced Shh correlates with increased proliferation and Ngn2 expression.
Figure 4: Reduction of dopaminergic neurons results from defective midbrain floor plate neurogenesis in Shh::cre;Ctnnb1 cKO embryos.
Figure 5: Canonical Wnt/beta-catenin signaling is required for Otx2 and Lmx1a, but not Lmx1b and Foxa2, maintenance.
Figure 6: Canonical Wnt/beta-catenin signaling is sufficient to repress Shh and induce a DA progenitor profile.

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Acknowledgements

We thank C. Tabin for Shh::cre mice and Shh cDNA; M. German for antibody to Lmx1a; T. Muller and C. Birchmeier for antibody to Lmx1b; D. Anderson for antibody to Ngn2; T. Edlund for antibodies to Gbx2; and A. Klar for the Spon1l cDNA. We thank A. Chenn for supplying Ctnnb1 mouse strains. We thank J. Kessler for suggestions. R.B.A was supported by the Dana Foundation and the American Parkinson's Disease Association. M.J. was supported by the Parkinson's Disease Foundation.

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Authors and Affiliations

  1. Department of Neurology and Center for Genetic Medicine, Northwestern University Feinberg Medical School, 7-113 Lurie Bldg, 303 E Superior Street, Chicago, 60611, Illinois, USA

    Milan Joksimovic, Beth A Yun, Angela M Anderegg & Rajeshwar B Awatramani

  2. Laboratory of Molecular Biology, National Institute of Neurological Disorders and Stroke, Porter Neuroscience Research Center, Building 35, Room 3A-201, 35 Convent Drive, MSC 3703, Bethesda, 20892-4157, Maryland, USA

    Raja Kittappa, Wendy W Chang & Ronald D G McKay

  3. Department of Pharmacology, Graduate School of Medicine, Kyoto University, Yoshida-Konoé-cho, Sakyo, 606-8501, Kyoto, Japan

    Makoto M Taketo

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  1. Milan Joksimovic
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Contributions

M.J. designed and conducted the experiments, prepared all of the figures and participated in writing the manuscript. B.A.Y. and A.M.A. helped to characterize various conditional mutants. R.K., W.W.C. and R.D.G.M. performed the in vitro analyses and provided useful discussions. M.M.T. provided the stabilized Ctnnb1 strain. R.B.A. supervised the study and wrote the manuscript.

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Correspondence to Rajeshwar B Awatramani.

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Joksimovic, M., Yun, B., Kittappa, R. et al. Wnt antagonism of Shh facilitates midbrain floor plate neurogenesis. Nat Neurosci 12, 125–131 (2009). https://doi.org/10.1038/nn.2243

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