The role of developmental transcription factors in maintenance of neuronal properties and in disease remains poorly understood. Lmx1a and Lmx1b are key transcription factors required for the early specification of ventral midbrain dopamine (mDA) neurons. Here we show that conditional ablation of Lmx1a and Lmx1b after mDA neuron specification resulted in abnormalities that show striking resemblance to early cellular abnormalities seen in Parkinson's disease. We found that Lmx1b was required for the normal execution of the autophagic-lysosomal pathway and for the integrity of dopaminergic nerve terminals and long-term mDA neuronal survival. Notably, human LMX1B expression was decreased in mDA neurons in brain tissue affected by Parkinson's disease. Thus, these results reveal a sustained and essential requirement of Lmx1b for the function of midbrain mDA neurons and suggest that its dysfunction is associated with Parkinson's disease pathogenesis.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1.

    & Dopamine neuron systems in the brain: an update. Trends Neurosci. 30, 194–202 (2007).

  2. 2.

    , & Clinical progression in Parkinson disease and the neurobiology of axons. Ann. Neurol. 67, 715–725 (2010).

  3. 3.

    & Maintenance of postmitotic neuronal cell identity. Nat. Neurosci. 17, 899–907 (2014).

  4. 4.

    & Maintaining differentiated cellular identity. Nat. Rev. Genet. 13, 429–439 (2012).

  5. 5.

    et al. Transcription factor Nurr1 maintains fiber integrity and nuclear-encoded mitochondrial gene expression in dopamine neurons. Proc. Natl. Acad. Sci. USA 110, 2360–2365 (2013).

  6. 6.

    et al. Foxa1 and foxa2 are required for the maintenance of dopaminergic properties in ventral midbrain neurons at late embryonic stages. J. Neurosci. 33, 8022–8034 (2013).

  7. 7.

    et al. Otx2 controls neuron subtype identity in ventral tegmental area and antagonizes vulnerability to MPTP. Nat. Neurosci. 13, 1481–1488 (2010).

  8. 8.

    et al. Do polymorphisms in transcription factors LMX1A and LMX1B influence the risk for Parkinson's disease? J. Neural Transm. 116, 333–338 (2009).

  9. 9.

    et al. PITX3 polymorphism is associated with early onset Parkinson's disease. Neurobiol. Aging 31, 114–117 (2010).

  10. 10.

    et al. The transcription factor PITX3 is associated with sporadic Parkinson's disease. Neurobiol. Aging 30, 731–738 (2009).

  11. 11.

    et al. Association of transcription factor polymorphisms PITX3 and EN1 with Parkinson's disease. Neurobiol. Aging 32, 302–307 (2011).

  12. 12.

    et al. Mutations in NR4A2 associated with familial Parkinson disease. Nat. Genet. 33, 85–89 (2003).

  13. 13.

    et al. Characterisation of a novel NR4A2 mutation in Parkinson's disease brain. Neurosci. Lett. 457, 75–79 (2009).

  14. 14.

    et al. Specific and integrated roles of Lmx1a, Lmx1b and Phox2a in ventral midbrain development. Development 138, 3399–3408 (2011).

  15. 15.

    , , , & Lmx1a and lmx1b function cooperatively to regulate proliferation, specification, and differentiation of midbrain dopaminergic progenitors. J. Neurosci. 31, 12413–12425 (2011).

  16. 16.

    et al. Identification of intrinsic determinants of midbrain dopamine neurons. Cell 124, 393–405 (2006).

  17. 17.

    et al. Wnt1-lmx1a forms a novel autoregulatory loop and controls midbrain dopaminergic differentiation synergistically with the SHH-FoxA2 pathway. Cell Stem Cell 5, 646–658 (2009).

  18. 18.

    et al. Efficient production of mesencephalic dopamine neurons by Lmx1a expression in embryonic stem cells. Proc. Natl. Acad. Sci. USA 106, 7613–7618 (2009).

  19. 19.

    et al. Efficient generation of A9 midbrain dopaminergic neurons by lentiviral delivery of LMX1A in human embryonic stem cells and induced pluripotent stem cells. Hum. Gene Ther. 23, 56–69 (2012).

  20. 20.

    , , , & CNS expression pattern of Lmx1b and coexpression with ptx genes suggest functional cooperativity in the development of forebrain motor control systems. Mol. Cell. Neurosci. 21, 410–420 (2002).

  21. 21.

    , , , & Postnatal ontogeny of the transcription factor Lmx1b in the mouse central nervous system. J. Comp. Neurol. 509, 341–355 (2008).

  22. 22.

    et al. Expression of the LIM-homeodomain gene Lmx1a in the postnatal mouse central nervous system. Brain Res. Bull. 78, 306–312 (2009).

  23. 23.

    et al. Progressive parkinsonism in mice with respiratory-chain-deficient dopamine neurons. Proc. Natl. Acad. Sci. USA 104, 1325–1330 (2007).

  24. 24.

    et al. Nurr1 is required for maintenance of maturing and adult midbrain dopamine neurons. J. Neurosci. 29, 15923–15932 (2009).

  25. 25.

    et al. 3,4-dihydroxyphenylalanine reverses the motor deficits in Pitx3-deficient aphakia mice: behavioral characterization of a novel genetic model of Parkinson's disease. J. Neurosci. 25, 2132–2137 (2005).

  26. 26.

    & The hippocampal-VTA loop: controlling the entry of information into long-term memory. Neuron 46, 703–713 (2005).

  27. 27.

    & A synaptic model of memory: long-term potentiation in the hippocampus. Nature 361, 31–39 (1993).

  28. 28.

    et al. Disrupted autophagy leads to dopaminergic axon and dendrite degeneration and promotes presynaptic accumulation of alpha-synuclein and LRRK2 in the brain. J. Neurosci. 32, 7585–7593 (2012).

  29. 29.

    et al. Regulation of presynaptic neurotransmission by macroautophagy. Neuron 74, 277–284 (2012).

  30. 30.

    , & Fighting neurodegeneration with rapamycin: mechanistic insights. Nat. Rev. Neurosci. 12, 437–452 (2011).

  31. 31.

    et al. Pathogenic lysosomal depletion in Parkinson's disease. J. Neurosci. 30, 12535–12544 (2010).

  32. 32.

    & Axon degeneration in Parkinson's disease. Exp. Neurol. 246, 72–83 (2013).

  33. 33.

    et al. Decreased NURR1 gene expression in patients with Parkinson's disease. J. Neurol. Sci. 273, 29–33 (2008).

  34. 34.

    et al. Decreased NURR1 and PITX3 gene expression in Chinese patients with Parkinson's disease. Eur. J. Neurol. 19, 870–875 (2012).

  35. 35.

    , , , & Alterations in lysosomal and proteasomal markers in Parkinson's disease: relationship to alpha-synuclein inclusions. Neurobiol. Dis. 35, 385–398 (2009).

  36. 36.

    et al. Loss of P-type ATPase ATP13A2/PARK9 function induces general lysosomal deficiency and leads to Parkinson disease neurodegeneration. Proc. Natl. Acad. Sci. USA 109, 9611–9616 (2012).

  37. 37.

    et al. Disease-specific phenotypes in dopamine neurons from human iPS-based models of genetic and sporadic Parkinson's disease. EMBO Mol. Med. 4, 380–395 (2012).

  38. 38.

    , , , & The role of autophagy in Parkinson's disease. Cold Spring Harb. Perspect. Med. 2, a009357 (2012).

  39. 39.

    et al. TFEB-mediated autophagy rescues midbrain dopamine neurons from alpha-synuclein toxicity. Proc. Natl. Acsad. Sci. USA 110, E1817–E1826 (2013).

  40. 40.

    , & Age-related accumulation of Marinesco bodies and lipofuscin in rhesus monkey midbrain dopamine neurons: relevance to selective neuronal vulnerability. J. Comp. Neurol. 502, 683–700 (2007).

  41. 41.

    et al. Syndecan-3-deficient mice exhibit enhanced LTP and impaired hippocampus-dependent memory. Mol. Cell. Neurosci. 21, 158–172 (2002).

  42. 42.

    et al. Enhanced long-term potentiation and impaired learning in mice with mutant postsynaptic density-95 protein. Nature 396, 433–439 (1998).

  43. 43.

    , & Impaired long-term spatial and recognition memory and enhanced CA1 hippocampal LTP in the dystrophin-deficient Dmd(mdx) mouse. Neurobiol. Dis. 17, 10–20 (2004).

  44. 44.

    et al. Disease duration and the integrity of the nigrostriatal system in Parkinson's disease. Brain 136, 2419–2431 (2013).

  45. 45.

    , , , & Autophagy in axonal and dendritic degeneration. Trends Neurosci. 36, 418–428 (2013).

  46. 46.

    et al. Nonmotor symptoms of Parkinson's disease revealed in an animal model with reduced monoamine storage capacity. J. Neurosci. 29, 8103–8113 (2009).

  47. 47.

    Olfaction in Parkinson's disease and related disorders. Neurobiol. Dis. 46, 527–552 (2012).

  48. 48.

    et al. Glutamate receptors on dopamine neurons control the persistence of cocaine seeking. Neuron 59, 497–508 (2008).

  49. 49.

    et al. Lmx1b is required for maintenance of central serotonergic neurons and mice lacking central serotonergic system exhibit normal locomotor activity. J. Neurosci. 26, 12781–12788 (2006).

  50. 50.

    et al. The protein kinase DYRK1A regulates caspase-9-mediated apoptosis during retina development. Dev. Cell 15, 841–853 (2008).

  51. 51.

    et al. Bidirectional regulation of emotional memory by 5–HT1B receptors involves hippocampal p11. Mol. Psychiatry 18, 1096–1105 (2013).

  52. 52.

    et al. PPAR-gamma-mediated neuroprotection in a chronic mouse model of Parkinson's disease. Eur. J. Neurosci. 29, 954–963 (2009).

  53. 53.

    et al. Olfactory discrimination deficits in mice lacking the dopamine transporter or the D2 dopamine receptor. Behav. Brain Res. 172, 97–105 (2006).

  54. 54.

    & T-maze alternation in the rodent. Nat. Protoc. 1, 7–12 (2006).

  55. 55.

    et al. Molecular adaptations underlying susceptibility and resistance to social defeat in brain reward regions. Cell 131, 391–404 (2007).

  56. 56.

    et al. Use of dopamine-beta-hydroxylase-deficient mice to determine the role of norepinephrine in the mechanism of action of antidepressant drugs. J. Pharmacol. Exp. Ther. 298, 651–657 (2001).

  57. 57.

    , , , & Derivatization chemistries for determination of serotonin, norepinephrine and dopamine in brain microdialysis samples by liquid chromatography with fluorescence detection. Biomed. Chromatogr. 20, 267–281 (2006).

  58. 58.

    , , & Isolation and culture of ventral mesencephalic precursor cells and dopaminergic neurons from rodent brains. Curr. Protoc. Stem Cell Biol. Ch. 2, unit 2D 5 (2009).

  59. 59.

    et al. Essential role for DNA-PK-mediated phosphorylation of NR4A nuclear orphan receptors in DNA double-strand break repair. Genes Dev. 25, 2031–2040 (2011).

  60. 60.

    et al. The Sun Health Research Institute Brain Donation Program: description and experience, 1987–2007. Cell Tissue Bank 9, 229–245 (2008).

Download references


We thank T. Samuelsson, H. Lunden-Miguel and C.-Y. Leung for technical assistance, and members of the Ericson and Perlmann laboratories for discussions. We thank S.-L. Ang (National Institute for Medical Research, London), N.-G. Larsson (Max Planck Institute, Köln) and G. Schütz (DKFZ, Heidelberg) for providing mouse lines. We thank T. Beach at the Banner Sun Health Research Institute, Arizona, USA for providing postmortem human brain samples. This work was supported by funding from the European Union, Seventh Framework Programme under grant agreement mdDANeurodev, NeuroStemCell and Synsys (T.P., J.E. and O.S.), from the Swedish Strategic Research Foundation (SSF; T.P. and P.S.), from the Swedish Research Council, and from Hjärnfonden and Parkinsonfonden (O.S.). The human tissue donation program was supported by the US National Institutes of Health (U24 NS072026 and P30 AG19610), the Arizona Department of Health Services, the Arizona Biomedical Research Commission and the Michael J. Fox Foundation for Parkinson's Research. A.L. was supported by a Marie Curie Intra-European Fellowship for Career Development.

Author information

Author notes

    • Nicoletta Schintu
    •  & André Nobre

    These authors contributed equally to this work.


  1. Ludwig Institute for Cancer Research, Stockholm, Sweden.

    • Ariadna Laguna
    • , André Nobre
    • , Nikolaos Volakakis
    • , Jesper Kjaer Jacobsen
    • , Eliza Joodmardi
    •  & Thomas Perlmann
  2. Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden.

    • Ariadna Laguna
    • , Qiaolin Deng
    • , Johan Ericson
    •  & Thomas Perlmann
  3. Neurodegenerative Diseases Group, Vall d'Hebron Research Institute-CIBERNED, Barcelona, Spain.

    • Ariadna Laguna
  4. Department of Clinical Neuroscience, Center for Molecular Medicine, Karolinska Institutet, Stockholm, Sweden.

    • Nicoletta Schintu
    • , Alexandra Alvarsson
    •  & Per Svenningsson
  5. Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden.

    • Marta Gómez-Galán
    • , Elena Sopova
    •  & Oleg Shupliakov
  6. Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden.

    • Takashi Yoshitake
    •  & Jan Kehr


  1. Search for Ariadna Laguna in:

  2. Search for Nicoletta Schintu in:

  3. Search for André Nobre in:

  4. Search for Alexandra Alvarsson in:

  5. Search for Nikolaos Volakakis in:

  6. Search for Jesper Kjaer Jacobsen in:

  7. Search for Marta Gómez-Galán in:

  8. Search for Elena Sopova in:

  9. Search for Eliza Joodmardi in:

  10. Search for Takashi Yoshitake in:

  11. Search for Qiaolin Deng in:

  12. Search for Jan Kehr in:

  13. Search for Johan Ericson in:

  14. Search for Per Svenningsson in:

  15. Search for Oleg Shupliakov in:

  16. Search for Thomas Perlmann in:


A.L. planned all experiments, performed histological, gene expression and western blot analyses and wrote the manuscript; N.S. performed behavioral analysis and dissection of mouse brain tissue; A.N. performed primary ventral midbrain cultures and gene expression analysis; A.A. performed behavioral analysis; N.V. cloned and produced lentiviruses; J.K.J. performed stereological and western blot analyses; M.G.G. performed electrophysiological analysis; E.S. performed electron microscopy analysis; E.J. performed histological analysis; T.Y. and J.K. performed high-performance liquid chromatography; Q.D. and J.E. generated conditional Lmx1a gene targeted mice and helped with planning; P.S. helped with analysis and planning; O.S. performed electron microscopy analysis, helped with analysis and with writing the manuscript; T.P. together with A.L. planned all experiments and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Ariadna Laguna or Thomas Perlmann.

Integrated supplementary information

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–7 and Supplementary Tables 1 and 2

  2. 2.

    Supplementary Methods Checklist

About this article

Publication history






Further reading