Abstract

The unusually high demand for metals in the brain, along with insufficient understanding of how their dysregulation contributes to neurological diseases, motivates the study of how inorganic chemistry influences neural circuitry. We now report that the transition metal copper is essential for regulating rest–activity cycles and arousal. Copper imaging and gene expression analysis in zebrafish identifies the locus coeruleus–norepinephrine (LC-NE) system, a vertebrate-specific neuromodulatory circuit critical for regulating sleep, arousal, attention, memory and emotion, as a copper-enriched unit with high levels of copper transporters CTR1 and ATP7A and the copper enzyme dopamine β-hydroxylase (DBH) that produces NE. Copper deficiency induced by genetic disruption of ATP7A, which loads copper into DBH, lowers NE levels and hinders LC function as manifested by disruption in rest–activity modulation. Moreover, LC dysfunction caused by copper deficiency from ATP7A disruption can be rescued by restoring synaptic levels of NE, establishing a molecular CTR1–ATP7A–DBH–NE axis for copper-dependent LC function.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from $8.99

All prices are NET prices.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

References

  1. 1.

    Bush, A. I. Metals and neuroscience. Curr. Opin. Chem. Biol. 4, 184–191 (2000).

  2. 2.

    Que, E. L., Domaille, D. W. & Chang, C. J. Metals in neurobiology: probing their chemistry and biology with molecular imaging. Chem. Rev. 108, 1517–1549 (2008).

  3. 3.

    Barnham, K. J., Masters, C. L. & Bush, A. I. Neurodegenerative diseases and oxidative stress. Nat. Rev. Drug Discov. 3, 205–214 (2004).

  4. 4.

    Kaler, S. G. ATP7A-related copper transport diseases-emerging concepts and future trends. Nat. Rev. Neurol. 7, 15–29 (2011).

  5. 5.

    Madsen, E. & Gitlin, J. D. Copper and iron disorders of the brain. Annu. Rev. Neurosci. 30, 317–337 (2007).

  6. 6.

    Zlatic, S., Comstra, H. S., Gokhale, A., Petris, M. J. & Faundez, V. Molecular basis of neurodegeneration and neurodevelopmental defects in Menkes disease. Neurobiol. Dis. 81, 154–161 (2015).

  7. 7.

    Duncan, C. & White, A. R. Copper complexes as therapeutic agents. Metallomics 4, 127–138 (2012).

  8. 8.

    Lutsenko, S., Bhattacharjee, A. & Hubbard, A. L. Copper handling machinery of the brain. Metallomics 2, 596–608 (2010).

  9. 9.

    Prohaska, J. R. Functions of trace elements in brain metabolism. Physiol. Rev. 67, 858–901 (1987).

  10. 10.

    Warren, P. J., Earl, C. J. & Thompson, R. H. The distribution of copper in human brain. Brain 83, 709–717 (1960).

  11. 11.

    Zecca, L. et al. The role of iron and copper molecules in the neuronal vulnerability of locus coeruleus and substantia nigra during aging. Proc. Natl Acad. Sci. USA 101, 9843–9848 (2004).

  12. 12.

    German, D. C. et al. Disease-specific patterns of locus coeruleus cell loss. Ann. Neurol. 32, 667–676 (1992).

  13. 13.

    Braak, H. & Del Tredici, K. Where, when, and in what form does sporadic Alzheimer’s disease begin? Curr. Opin. Neurol. 25, 708–714 (2012).

  14. 14.

    Cotruvo, J. A. Jr, Aron, A. T., Ramos-Torres, K. M. & Chang, C. J. Synthetic fluorescent probes for studying copper in biological systems. Chem. Soc. Rev. 44, 4400–4414 (2015).

  15. 15.

    Nevitt, T., Ohrvik, H. & Thiele, D. J. Charting the travels of copper in eukaryotes from yeast to mammals. Biochim. Biophys. Acta 1823, 1580–1593 (2012).

  16. 16.

    Robinson, N. J. & Winge, D. R. Copper metallochaperones. Annu. Rev. Biochem. 79, 537–562 (2010).

  17. 17.

    O’Halloran, T. V. & Culotta, V. C. Metallochaperones, an intracellular shuttle service for metal ions. J. Biol. Chem. 275, 25057–25060 (2000).

  18. 18.

    Banci, L. et al. Affinity gradients drive copper to cellular destinations. Nature 465, 645–648 (2010).

  19. 19.

    Lutsenko, S., Barnes, N. L., Bartee, M. Y. & Dmitriev, O. Y. Function and regulation of human copper-transporting ATPases. Physiol. Rev. 87, 1011–1046 (2007).

  20. 20.

    Aston-Jones, G. & Waterhouse, B. Locus coeruleus: from global projection system to adaptive regulation of behavior. Brain Res. 1645, 75–78 (2016).

  21. 21.

    Sara, S. J. The locus coeruleus and noradrenergic modulation of cognition. Nat. Rev. Neurosci. 10, 211–223 (2009).

  22. 22.

    Moret, C. & Briley, M. The importance of norepinephrine in depression. Neuropsychiatr. Dis. Treat. 7, 9–13 (2011). (Suppl. 1).

  23. 23.

    Kaufman, S. & Friedman, S. Dopamine-β-hydroxylase. Pharmacol. Rev. 17, 71–100 (1965).

  24. 24.

    Mangold, J. B. & Klinman, J. P. Mechanism-based inactivation of dopamine β-monooxygenase by β-chlorophenethylamine. J. Biol. Chem. 259, 7772–7779 (1984).

  25. 25.

    Ash, D. E., Papadopoulos, N. J., Colombo, G. & Villafranca, J. J. Kinetic and spectroscopic studies of the interaction of copper with dopamine β-hydroxylase. J. Biol. Chem. 259, 3395–3398 (1984).

  26. 26.

    Kim, B. E., Nevitt, T. & Thiele, D. J. Mechanisms for copper acquisition, distribution and regulation. Nat. Chem. Biol. 4, 176–185 (2008).

  27. 27.

    Scatton, B., Javoy-Agid, F., Rouquier, L., Dubois, B. & Agid, Y. Reduction of cortical dopamine, noradrenaline, serotonin and their metabolites in Parkinson’s disease. Brain Res. 275, 321–328 (1983).

  28. 28.

    Hornby, P. J. & Piekut, D. T. Immunoreactive dopamine β-hydroxylase in neuronal groups in the goldfish brain. Brain Behav. Evol. 32, 252–256 (1988).

  29. 29.

    Ekström, P., Reschke, M., Steinbusch, H. & van Veen, T. Distribution of noradrenaline in the brain of the teleost Gasterosteus aculeatus L.: an immunohistochemical analysis. J. Comp. Neurol. 254, 297–313 (1986).

  30. 30.

    Parent, A. Functional anatomy and evolution of monoaminergic systems. Am. Zool. 24, 783–790 (1984).

  31. 31.

    Hirayama, T., Van de Bittner, G. C., Gray, L. W., Lutsenko, S. & Chang, C. J. Near-infrared fluorescent sensor for in vivo copper imaging in a murine Wilson disease model. Proc. Natl Acad. Sci. USA 109, 2228–2233 (2012).

  32. 32.

    Heffern, M. C. et al. In vivo bioluminescence imaging reveals copper deficiency in a murine model of nonalcoholic fatty liver disease. Proc. Natl Acad. Sci. USA 113, 14219–14224 (2016).

  33. 33.

    Hare, D. J., New, E. J., de Jonge, M. D. & McColl, G. Imaging metals in biology: balancing sensitivity, selectivity and spatial resolution. Chem. Soc. Rev. 44, 5941–5958 (2015).

  34. 34.

    Ackerman, C. M., Lee, S. & Chang, C. J. Analytical methods for imaging metals in biology: from transition metal metabolism to transition metal signaling. Anal. Chem. 89, 22–41 (2017).

  35. 35.

    Colburn, R. W. & Maas, J. W. Adenosine triphosphate–metal–norepinephrine ternary complexes and catecholamine binding. Nature 208, 37–41 (1965).

  36. 36.

    Sato, M., Ohtomo, K., Daimon, T., Sugiyama, T. & Iijima, K. Localization of copper to afferent terminals in rat locus ceruleus, in contrast to mitochondrial copper in cerebellum. J. Histochem. Cytochem. 42, 1585–1591 (1994).

  37. 37.

    Gaier, E. D., Eipper, B. A. & Mains, R. E. Copper signaling in the mammalian nervous system: synaptic effects. J. Neurosci. Res. 91, 2–19 (2013).

  38. 38.

    Madsen, E. C. & Gitlin, J. D. Zebrafish mutants calamity and catastrophe define critical pathways of gene-nutrient interactions in developmental copper metabolism. PLoS Genet. 4, e1000261 (2008).

  39. 39.

    Zimbrean, P. C. & Schilsky, M. L. Psychiatric aspects of Wilson disease: a review. Gen. Hosp. Psychiatry 36, 53–62 (2014).

  40. 40.

    Pantoja, C. et al. Neuromodulatory regulation of behavioral individuality in zebrafish. Neuron 91, 587–601 (2016).

  41. 41.

    Chen, S. et al. Light-dependent regulation of sleep and wake states by prokineticin 2 in zebrafish. Neuron 95, 153–168.e156 (2017).

  42. 42.

    Lein, E. S. et al. Genome-wide atlas of gene expression in the adult mouse brain. Nature 445, 168–176 (2007).

  43. 43.

    Zhu, S. et al. Activated ALK collaborates with MYCN in neuroblastoma pathogenesis. Cancer Cell 21, 362–373 (2012).

  44. 44.

    Deisseroth, K. Circuit dynamics of adaptive and maladaptive behaviour. Nature 505, 309–317 (2014).

  45. 45.

    Caballero, J. & Nahata, M. C. Atomoxetine hydrochloride for the treatment of attention-deficit/hyperactivity disorder. Clin. Ther. 25, 3065–3083 (2003).

  46. 46.

    Dehal, P. & Boore, J. L. Two rounds of whole genome duplication in the ancestral vertebrate. PLoS Biol. 3, e314 (2005).

  47. 47.

    Panopoulou, G. & Poustka, A. J. Timing and mechanism of ancient vertebrate genome duplications — the adventure of a hypothesis. Trends Genet. 21, 559–567 (2005).

  48. 48.

    Dmitriev, O. et al. Solution structure of the N-domain of Wilson disease protein: distinct nucleotide-binding environment and effects of disease mutations. Proc. Natl Acad. Sci. USA 103, 5302–5307 (2006).

  49. 49.

    Li, S. B., Jones, J. R. & de Lecea, L. Hypocretins, neural systems, physiology, and psychiatric disorders. Curr. Psychiatry Rep. 18, 7 (2016).

  50. 50.

    Singh, C., Oikonomou, G. & Prober, D. A. Norepinephrine is required to promote wakefulness and for hypocretin-induced arousal in zebrafish. Elife 4, e07000 (2015).

  51. 51.

    Atkinson, M. J. & Bingman, C. Elemental composition of commercial seasalts. J. Aquaricult. Aquat. Sci. VIII, 39–43 (1998).

  52. 52.

    Lister, J. A., Robertson, C. P., Lepage, T., Johnson, S. L. & Raible, D. W. nacre encodes a zebrafish microphthalmia-related protein that regulates neural-crest-derived pigment cell fate. Development 126, 3757–3767 (1999).

  53. 53.

    Ahrens, M. B., Orger, M. B., Robson, D. N., Li, J. M. & Keller, P. J. Whole-brain functional imaging at cellular resolution using light-sheet microscopy. Nat. Methods 10, 413–420 (2013).

  54. 54.

    Xiao, T. & Baier, H. Lamina-specific axonal projections in the zebrafish tectum require the type IV collagen Dragnet. Nat. Neurosci. 10, 1529–1537 (2007).

  55. 55.

    Vilella, A. J. et al. EnsemblCompara GeneTrees: complete, duplication-aware phylogenetic trees in vertebrates. Genome Res. 19, 327–335 (2009).

Download references

Acknowledgements

We thank A. T. Look (Dana-Farber Cancer Institute) and J. Gitlin (Marine Biological Laboratory) for providing plasmids and transgenic fish lines, C. Miller (University of California, Berkeley) and R. Segev (Ben Gurion University of the Negev) for providing fish samples, and R. Feng and R. Fish for assistance with pilot experiments. We thank the NIH (GM79465 to C.J.C. and PN2EY018241 to E.Y.I.) for providing funding for this work. C.J.C. is an Investigator of the Howard Hughes Medical Institute and a CIFAR Senior Fellow. C.M.A. was partially supported by a Hertz Foundation Graduate Fellowship and a Chemical Biology Training Grant from the NIH (T32 GM066698). Experiments at the CRL Molecular Imaging Center were supported by the Helen Wills Neuroscience Institute.

Author information

Author notes

    • Elizabeth C Carroll

    Present address: Department of Imaging Physics, Delft University of Technology, Delft, The Netherlands

Affiliations

  1. Department of Chemistry, University of California, Berkeley, CA, USA

    • Tong Xiao
    • , Cheri M. Ackerman
    • , Shang Jia
    • , Jefferson Chan
    • , Christine S. Liu
    •  & Christopher J. Chang
  2. Howard Hughes Medical Institute, University of California, Berkeley, CA, USA

    • Tong Xiao
    • , Cheri M. Ackerman
    • , Shang Jia
    • , Jefferson Chan
    • , Bao Thai
    • , Christine S. Liu
    •  & Christopher J. Chang
  3. Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA

    • Elizabeth C Carroll
    • , Adam Hoagland
    • , Bao Thai
    • , Ehud Y. Isacoff
    •  & Christopher J. Chang
  4. Helen Wills Neuroscience Institute, University of California, Berkeley, CA, USA

    • Ehud Y. Isacoff
    •  & Christopher J. Chang

Authors

  1. Search for Tong Xiao in:

  2. Search for Cheri M. Ackerman in:

  3. Search for Elizabeth C Carroll in:

  4. Search for Shang Jia in:

  5. Search for Adam Hoagland in:

  6. Search for Jefferson Chan in:

  7. Search for Bao Thai in:

  8. Search for Christine S. Liu in:

  9. Search for Ehud Y. Isacoff in:

  10. Search for Christopher J. Chang in:

Contributions

T.X. and C.J.C. designed research; T.X., C.M.A., E.C.C. and A.H. performed imaging and behavioral assays; T.X., C.M.A. and B.T. performed copper imaging and analysis assays; S.J. and J.C. synthesized and characterized fluorescent copper probes; T.X. and C.S.L. conducted in situ hybridization and IHC assays; T.X., C.M.A., E.C.C. and C.J.C. wrote the manuscript; E.Y.I. provided valuable input on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Christopher J. Chang.

Supplementary information

  1. Supplementary Text and Figures

    Supplementary Table 1, Supplementary Figures 1–32 and Supplementary Note

  2. Reporting Summary

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/s41589-018-0062-z