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Brain endogenous liver X receptor ligands selectively promote midbrain neurogenesis

Nature Chemical Biology volume 9, pages 126–133 (2013)Cite this article

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Abstract

Liver X receptors (Lxrα and Lxrβ) are ligand-dependent nuclear receptors critical for ventral midbrain neurogenesis in vivo. However, no endogenous midbrain Lxr ligand has so far been identified. Here we used LC/MS and functional assays to identify cholic acid as a new Lxr ligand. Moreover, 24(S),25-epoxycholesterol (24,25-EC) was found to be the most potent and abundant Lxr ligand in the developing mouse midbrain. Both Lxr ligands promoted neural development in an Lxr-dependent manner in zebrafish in vivo. Notably, each ligand selectively regulated the development of distinct midbrain neuronal populations. Whereas cholic acid increased survival and neurogenesis of Brn3a-positive red nucleus neurons, 24,25-EC promoted dopaminergic neurogenesis. These results identify an entirely new class of highly selective and cell type–specific regulators of neurogenesis and neuronal survival. Moreover, 24,25-EC promoted dopaminergic differentiation of embryonic stem cells, suggesting that Lxr ligands may thus contribute to the development of cell replacement and regenerative therapies for Parkinson's disease.

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Figure 1: Identification and characterization of Lxr-activating factors derived from the E11.5 mouse ventral midbrain.
Figure 2: CA and 24,25-EC specifically activate Lxrα and Lxrβ by binding to the LBD of Lxr.
Figure 3: 24,25-EC and CA increase transcript levels of dopaminergic neuronal markers and the number of diencephalic Th+ cells in zebrafish.
Figure 4: Differential effects of CA and 24,25-EC on cell survival and differentiation in mouse E11.5 ventral midbrain primary cultures.
Figure 5: 24,25-EC increases dopaminergic neurogenesis, whereas CA increases red nucleus neurogenesis.
Figure 6: 24,25-EC efficiently regulates the dopaminergic differentiation of mESCs.

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Acknowledgements

Work in Karolinska was supported by Hjärnfonden, European Union (Strokemap and NeuroStemCell projects), Swedish Foundation for Strategic Research (SRL Program), Karolinska Institutet (Strategic Research Program and Thematic Center in Stem Cells and Regenerative Medicine) and the Swedish Research Council (DBRM, 2008:2811, 2011-3116 and 3318). The authors are grateful to L. Jansson-Sjöstrand and J. Söderlund for technical assistance, A. Nanni for secretarial help and S. Yang for help with artwork. Work in Swansea was supported by funding from the UK Research Councils Biotechnology and Biological Sciences Research Council (BB/C515771/2, BB/C511356/1 and BB/I001735/1 to W.J.G. and BB/H001018/1 to Y.W.). S.T. was supported by the Swedish Medical Research Council and the Onassis Foundation.

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Author notes

  1. Paola Sacchetti & Kyle M Sousa

    Present address: Present addresses: Department of Biological Sciences, Mount Holyoke College, South Hadley, Massachusetts, USA (P.S.); Department of Gene Regulation and Drug Discovery, City of Hope–Beckman Research Institute, Duarte, California, USA (K.M.S.).,

Authors and Affiliations

  1. Department of Medical Biochemistry and Biophysics, Laboratory of Molecular Neurobiology, Karolinska Institute, Stockholm, Sweden

    Spyridon Theofilopoulos, Satish Srinivas Kitambi, Paola Sacchetti, Kyle M Sousa, Karl Bodin, Carmen Saltó, Magnus Gustafsson, Enrique M Toledo, Patrik Ernfors, Jan Sjövall & Ernest Arenas

  2. Institute of Mass Spectrometry, College of Medicine, Swansea University, Singleton Park, Swansea, UK.,

    Yuqin Wang & William J Griffiths

  3. Waters UK Ltd., Manchester, UK

    Jayne Kirk

  4. The School of Pharmacy, London, UK

    Kersti Karu

  5. Center for Nuclear Receptors and Cell Signaling, University of Houston, Houston, Texas, USA

    Jan-Åke Gustafsson

  6. Department of Biosciences and Nutrition, Center for Biosciences, Stockholm, Sweden

    Knut R Steffensen

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Contributions

S.T. performed most of the experiments. Y.W. performed the HPLC and LC/MSn experiments. S.S.K. performed some of the zebrafish experiments. P.S. contributed to some luciferase assay and primary culture experiments. K.M.S. contributed to some luciferase assay experiments. K.B., J.K., M.G. and K.K. contributed to the HPLC and LC/MSn experiments. C.S. contributed to some of the mESC experiments. E.M.T. contributed to some of the ChIP experiments. J.-Å.G. and K.R.S. provided the Lxrα−/− Lxrβ−/− mice. P.E., J.S. and W.J.G. contributed to draw conclusions from experiments, supervision and provided intellectual input. E.A. contributed to experimental design, supervision, conclusions and coordinated experimental approaches and wrote the manuscript together with S.T.

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Correspondence to Ernest Arenas.

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Theofilopoulos, S., Wang, Y., Kitambi, S. et al. Brain endogenous liver X receptor ligands selectively promote midbrain neurogenesis. Nat Chem Biol 9, 126–133 (2013). https://doi.org/10.1038/nchembio.1156

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