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Essential function of HIPK2 in TGFβ-dependent survival of midbrain dopamine neurons

Nature Neuroscience volume 10, pages 77–86 (2007)Cite this article

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Abstract

Transforming growth factor beta (TGFβ) is a potent trophic factor for midbrain dopamine (DA) neurons, but its in vivo function and signaling mechanisms are not entirely understood. We show that the transcriptional cofactor homeodomain interacting protein kinase 2 (HIPK2) is required for the TGFβ-mediated survival of mouse DA neurons. The targeted deletion of Hipk2 has no deleterious effect on the neurogenesis of DA neurons, but leads to a selective loss of these neurons that is due to increased apoptosis during programmed cell death. As a consequence, Hipk2−/− mutants show an array of psychomotor abnormalities. The function of HIPK2 depends on its interaction with receptor-regulated Smads to activate TGFβ target genes. In support of this notion, DA neurons from Hipk2−/− mutants fail to survive in the presence of TGFβ3 and Tgfβ3−/− mutants show DA neuron abnormalities similar to those seen in Hipk2−/− mutants. These data underscore the importance of the TGFβ-Smad-HIPK2 pathway in the survival of DA neurons and its potential as a therapeutic target for promoting DA neuron survival during neurodegeneration.

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Figure 1: Expression of HIPK2 in midbrain DA neurons during development and in postnatal life.
Figure 2: Loss of DA neurons in the SNpc and VTA of Hipk2−/− mutants.
Figure 3: Motor behavioral abnormalities in Hipk2−/− mutants.
Figure 4: Reduced response to unfamiliar environment and to amphetamine challenge in Hipk2−/− mutants.
Figure 5: Loss of HIPK2 does not affect neurogenesis in DA neurons.
Figure 6: Loss of HIPK2 leads to increased apoptosis of DA neurons during periods of PCD.
Figure 7: HIPK2 is important in the TGFβ-Smad signaling pathway.
Figure 8: Loss of TGFβ3, but not TGFβ1, leads to DA neuron deficiencies similar to those in Hipk2−/− mutants.

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Acknowledgements

We thank L.F. Reichardt and S. Pleasure for comments on the manuscript. Special thanks to J. Siegenthaler for TGFβ1 antibody, D.J. Anderson for Ngn2 antibody, R. Akhurst for Tgfβ1 mutants, T. Doestchman for Tgfβ3 mutants, D. Di Monte for dopamine HPLC assays, T. Wyss-Coray for Smads and SBE-luciferase plasmids, A. Buckley for characterizing HIPK2LacZ expression, S. Ku for mutant HIPK2 plasmids, C. Wong for histology and G. Wei for generating Hipk2 mutants. V.P. was supported in part by a postdoctoral fellowship from the American Parkinson Disease Association. This work was supported by grants from the National Institute of Neurological Disorders and Stroke (NS44223 and NS48393), a Pilot Project Grant from the University of California San Francisco (UCSF) Alzheimer's Disease Research Center (AG23501), Veteran's Administration Merit Review, Presidential Early Career Award for Scientists and Engineers, National Parkinson Foundation, and The Michael J. Fox Foundation for Parkinson's Research to E.J.H., funds from the state of California for medical research on alcohol and drug abuse through UCSF to P.H.J., and grants from the US National Institutes of Health to P.H.J. and L.H.T.

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Author notes

  1. Jiasheng Zhang and Vanee Pho: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pathology, University of California San Francisco, 505 Parnassus Avenue, San Francisco, 94143, CA, USA

    Jiasheng Zhang, Vanee Pho, Amy T Tang, Siuwah Tang & Eric J Huang

  2. Department of Medicine, University of California San Francisco, 505 Parnassus Avenue, San Francisco, 94143, CA, USA

    Stephen J Bonasera

  3. Department of Psychiatry, University of California San Francisco, 505 Parnassus Avenue, San Francisco, 94143,, CA, USA

    Jed Holtzman & Laurence H Tecott

  4. Department of Neurology, University of California San Francisco, 505 Parnassus Avenue, San Francisco, 94143, CA, USA

    Joanna Hellmuth & Patricia H Janak

  5. Pathology Service 113B, VA Medical Center, 4150 Clement Street, San Francisco, 94121, CA, USA

    Jiasheng Zhang, Vanee Pho, Amy T Tang, Siuwah Tang & Eric J Huang

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Contributions

J.Z., V.P., A.T.T. and S.T. performed the cellular, molecular and histological experiments and the mouse genetics. L.H.T. supervised and S.J.B. and J.H. performed the Open Field, Rotorod and D1 agonist tests. P.H.J. supervised and J.H. performed the amphetamine tests. J.Z., V.P. and E.J.H. wrote the manuscript.

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Correspondence to Eric J Huang.

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Supplementary information

Supplementary Fig. 1

Absence of HIPK2 expression in locus ceruleus and striatum. (PDF 889 kb)

Supplementary Fig. 2

No detectable reduction in the number of GABAergic neurons (panel a) or the entire volume (panel b) of substantia nigra pars reticularis (SNpr). (PDF 1547 kb)

Supplementary Fig. 3

Loss of HIPK2 has no negative impact on the expression of cell fate or differentiation markers for DA neurons. (PDF 965 kb)

Supplementary Fig. 4

Co-localization of DA neuron marker Nurr1 and activated caspase 3 in SNpc of P0 Hipk2−/− mutants. (PDF 2153 kb)

Supplementary Fig. 5

A working model for HIPK2 in the downstream signaling pathway of TGFβ. (PDF 262 kb)

Supplementary Fig. 6

Determination of total TuJ1–positive and TH–positive neuron numbers in ventral mesencephalon cultures. (PDF 151 kb)

Supplementary Table 1

Genetic background and the number of midbrain dopamine neurons. (PDF 91 kb)

Supplementary Methods (PDF 149 kb)

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Zhang, J., Pho, V., Bonasera, S. et al. Essential function of HIPK2 in TGFβ-dependent survival of midbrain dopamine neurons. Nat Neurosci 10, 77–86 (2007). https://doi.org/10.1038/nn1816

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