Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Structural correlates of heterogeneous in vivo activity of midbrain dopaminergic neurons

Nature Neuroscience volume 15, pages 613–619 (2012)Cite this article

Subjects

This article has been updated

Abstract

Dopaminergic neurons of the substantia nigra pars compacta (SNc) exhibit functional heterogeneity that likely underpins their diverse roles in behavior. We examined how the functional diversity of identified dopaminergic neurons in vivo correlates with differences in somato-dendritic architecture and afferent synaptic organization. Stereological analysis of individually recorded and labeled dopaminergic neurons of rat SNc revealed that they received approximately 8,000 synaptic inputs, at least 30% of which were glutamatergic and 40–70% were GABAergic. The latter synapses were proportionally greater in number and denser on dendrites located in the substantia nigra pars reticulata (SNr) than on those located in SNc, revealing the existence of two synaptically distinct and region-specific subcellular domains. We also found that the relative extension of SNc neuron dendrites into the SNr dictated overall GABAergic innervation and predicted inhibition responses to aversive stimuli. We conclude that diverse wiring patterns determine the heterogeneous activities of midbrain dopaminergic neurons in vivo.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Recording and identification of SNc dopaminergic neurons.
Figure 2: Three-dimensional digital reconstructions of identified SNc dopaminergic neurons.
Figure 3: Proportions and localization of glutamatergic and GABAergic synapses made with individual dopaminergic neurons.
Figure 4: Densities of synapses made with different dendritic compartments.
Figure 5: The responsiveness of individual dopaminergic neurons to aversive stimuli is correlated with the proportion of their dendrites located in SNr.

Similar content being viewed by others

Change history

  • 05 March 2012

    In the version of this article initially published online, the large arrowheads were undefined in Figure 3a; the Figure 3b legend stated that the proportion of GABAergic synapses made with dendrites located in the SNr was greater than that of GABAergic and glutamatergic synapses in the SNc, whereas it should have stated that it was greater than that of GABAergic synapses in SNc and glutamatergic synapses in the SNr; the Table 2 legend listed the number of neurons as five, contradicting the correct values given in the table itself; a spurious reference to a Figure 5i should have read Figure 5f; and the units on the y axis in Figure 5e were given as mm × 103 instead of mm. The errors have been corrected for the print, PDF and HTML versions of this article.

References

  1. Wise, R.A. Roles for nigrostriatal–not just mesocorticolimbic–dopamine in reward and addiction. Trends Neurosci. 32, 517–524 (2009).

    Article  CAS  Google Scholar 

  2. Schultz, W. Behavioral dopamine signals. Trends Neurosci. 30, 203–210 (2007).

    Article  CAS  Google Scholar 

  3. Tsai, H.C. et al. Phasic firing in dopaminergic neurons is sufficient for behavioral conditioning. Science 324, 1080–1084 (2009).

    Article  CAS  Google Scholar 

  4. Blythe, S.N., Wokosin, D., Atherton, J.F. & Bevan, M.D. Cellular mechanisms underlying burst firing in substantia nigra dopamine neurons. J. Neurosci. 29, 15531–15541 (2009).

    Article  CAS  Google Scholar 

  5. Kitai, S.T., Shepard, P.D., Callaway, J.C. & Scroggs, R. Afferent modulation of dopamine neuron firing patterns. Curr. Opin. Neurobiol. 9, 690–697 (1999).

    Article  CAS  Google Scholar 

  6. Brischoux, F., Chakraborty, S., Brierley, D.I. & Ungless, M.A. Phasic excitation of dopamine neurons in ventral VTA by noxious stimuli. Proc. Natl. Acad. Sci. USA 106, 4894–4899 (2009).

    Article  CAS  Google Scholar 

  7. Brown, M.T., Henny, P., Bolam, J.P. & Magill, P.J. Activity of neurochemically heterogeneous dopaminergic neurons in the substantia nigra during spontaneous and driven changes in brain state. J. Neurosci. 29, 2915–2925 (2009).

    Article  CAS  Google Scholar 

  8. Bromberg-Martin, E.S., Matsumoto, M. & Hikosaka, O. Dopamine in motivational control: rewarding, aversive and alerting. Neuron 68, 815–834 (2010).

    Article  CAS  Google Scholar 

  9. Matsumoto, M. & Hikosaka, O. Two types of dopamine neuron distinctly convey positive and negative motivational signals. Nature 459, 837–841 (2009).

    Article  CAS  Google Scholar 

  10. Joshua, M., Adler, A., Mitelman, R., Vaadia, E. & Bergman, H. Midbrain dopaminergic neurons and striatal cholinergic interneurons encode the difference between reward and aversive events at different epochs of probabilistic classical conditioning trials. J. Neurosci. 28, 11673–11684 (2008).

    Article  CAS  Google Scholar 

  11. Tepper, J.M. & Lee, C.R. GABAergic control of substantia nigra dopaminergic neurons. Prog. Brain Res. 160, 189–208 (2007).

    Article  CAS  Google Scholar 

  12. Bolam, J.P., Francis, C.M. & Henderson, Z. Cholinergic input to dopaminergic neurons in the substantia nigra: a double immunocytochemical study. Neuroscience 41, 483–494 (1991).

    Article  CAS  Google Scholar 

  13. Bolam, J.P. & Smith, Y. The GABA and substance P input to dopaminergic neurones in the substantia nigra of the rat. Brain Res. 529, 57–78 (1990).

    Article  CAS  Google Scholar 

  14. Lavoie, B. & Parent, A. Pedunculopontine nucleus in the squirrel monkey: cholinergic and glutamatergic projections to the substantia nigra. J. Comp. Neurol. 344, 232–241 (1994).

    Article  CAS  Google Scholar 

  15. Smith, Y., Charara, A. & Parent, A. Synaptic innervation of midbrain dopaminergic neurons by glutamate-enriched terminals in the squirrel monkey. J. Comp. Neurol. 364, 231–253 (1996).

    Article  CAS  Google Scholar 

  16. Smith, Y., Bevan, M.D., Shink, E. & Bolam, J.P. Microcircuitry of the direct and indirect pathways of the basal ganglia. Neuroscience 86, 353–387 (1998).

    Article  CAS  Google Scholar 

  17. Pinault, D. A novel single-cell staining procedure performed in vivo under electrophysiological control: morpho-functional features of juxtacellularly labeled thalamic cells and other central neurons with biocytin or meurobiotin. J. Neurosci. Methods 65, 113–136 (1996).

    Article  CAS  Google Scholar 

  18. Ascoli, G.A. Mobilizing the base of neuroscience data: the case of neuronal morphologies. Nat. Rev. Neurosci. 7, 318–324 (2006).

    Article  CAS  Google Scholar 

  19. Preston, R.J., McCrea, R.A., Chang, H.T. & Kitai, S.T. Anatomy and physiology of substantia nigra and retrorubral neurons studied by extra- and intracellular recording and by horseradish peroxidase labeling. Neuroscience 6, 331–344 (1981).

    Article  CAS  Google Scholar 

  20. Fallon, J.H. & Loughlin, S.E. Substantia nigra. in The Rat Nervous System (ed. G. Paxinos) 215–238 (Academic Press, London, 1995).

  21. Gerfen, C.R., Baimbridge, K.G. & Thibault, J. The neostriatal mosaic. III. Biochemical and developmental dissociation of patch-matrix mesostriatal systems. J. Neurosci. 7, 3935–3944 (1987).

    Article  CAS  Google Scholar 

  22. West, M.J. Stereological methods for estimating the total number of neurons and synapses: issues of precision and bias. Trends Neurosci. 22, 51–61 (1999).

    Article  CAS  Google Scholar 

  23. Gundersen, H.J. Stereology of arbitrary particles. A review of unbiased number and size estimators and the presentation of some new ones, in memory of William R. Thompson. J. Microsc. 143, 3–45 (1986).

    Article  CAS  Google Scholar 

  24. Naito, A. & Kita, H. The cortico-nigral projection in the rat: an anterograde tracing study with biotinylated dextran amine. Brain Res. 637, 317–322 (1994).

    Article  CAS  Google Scholar 

  25. Omelchenko, N. & Sesack, S.R. Glutamate synaptic inputs to ventral tegmental area neurons in the rat derive primarily from subcortical sources. Neuroscience 146, 1259–1274 (2007).

    Article  CAS  Google Scholar 

  26. Corvaja, N., Doucet, G. & Bolam, J.P. Ultrastructure and synaptic targets of the raphe-nigral projection in the rat. Neuroscience 55, 417–427 (1993).

    Article  CAS  Google Scholar 

  27. Smith, Y., Hazrati, L.N. & Parent, A. Efferent projections of the subthalamic nucleus in the squirrel monkey as studied by the PHA-L anterograde tracing method. J. Comp. Neurol. 294, 306–323 (1990).

    Article  CAS  Google Scholar 

  28. Mann, E.O. & Paulsen, O. Role of GABAergic inhibition in hippocampal network oscillations. Trends Neurosci. 30, 343–349 (2007).

    Article  CAS  Google Scholar 

  29. Fiala, J.C., Spacek, J. & Harris, K.M. Dendrite structure. in Dendrites (eds. G.J. Stuart, N. Spruston & M. Hausser) 1–41 (Oxford University Press, Oxford, 2008).

  30. Magee, J.C. Dendritic integration of excitatory synaptic input. Nat. Rev. Neurosci. 1, 181–190 (2000).

    Article  CAS  Google Scholar 

  31. Comoli, E. et al. A direct projection from superior colliculus to substantia nigra for detecting salient visual events. Nat. Neurosci. 6, 974–980 (2003).

    Article  CAS  Google Scholar 

  32. Borgius, L., Restrepo, C.E., Leao, R.N., Saleh, N. & Kiehn, O. A transgenic mouse line for molecular genetic analysis of excitatory glutamatergic neurons. Mol. Cell. Neurosci. 45, 245–257 (2010).

    Article  CAS  Google Scholar 

  33. Wang, H.L. & Morales, M. Pedunculopontine and laterodorsal tegmental nuclei contain distinct populations of cholinergic, glutamatergic and GABAergic neurons in the rat. Eur. J. Neurosci. 29, 340–358 (2009).

    Article  Google Scholar 

  34. Iribe, Y., Moore, K., Pang, K.C. & Tepper, J.M. Subthalamic stimulation–induced synaptic responses in substantia nigra pars compacta dopaminergic neurons in vitro. J. Neurophysiol. 82, 925–933 (1999).

    Article  CAS  Google Scholar 

  35. Hajós, M. & Greenfield, S.A. Synaptic connections between pars compacta and pars reticulata neurones: electrophysiological evidence for functional modules in the substantia nigra. Brain Res. 660, 216–224 (1994).

    Article  Google Scholar 

  36. Spruston, N. Pyramidal neurons: dendritic structure and synaptic integration. Nat. Rev. Neurosci. 9, 206–221 (2008).

    Article  CAS  Google Scholar 

  37. Megías, M., Emri, Z., Freund, T.F. & Gulyas, A.I. Total number and distribution of inhibitory and excitatory synapses on hippocampal CA1 pyramidal cells. Neuroscience 102, 527–540 (2001).

    Article  Google Scholar 

  38. Grace, A.A. & Bunney, B.S. Paradoxical GABA excitation of nigral dopaminergic cells: indirect mediation through reticulata inhibitory neurons. Eur. J. Pharmacol. 59, 211–218 (1979).

    Article  CAS  Google Scholar 

  39. Gulácsi, A. et al. Cell type–specific differences in chloride-regulatory mechanisms and GABA(A) receptor–mediated inhibition in rat substantia nigra. J. Neurosci. 23, 8237–8246 (2003).

    Article  Google Scholar 

  40. Paladini, C.A., Celada, P. & Tepper, J.M. Striatal, pallidal and pars reticulata evoked inhibition of nigrostriatal dopaminergic neurons is mediated by GABA(A) receptors in vivo. Neuroscience 89, 799–812 (1999).

    Article  CAS  Google Scholar 

  41. Jhou, T.C., Fields, H.L., Baxter, M.G., Saper, C.B. & Holland, P.C. The rostromedial tegmental nucleus (RMTg), a GABAergic afferent to midbrain dopamine neurons, encodes aversive stimuli and inhibits motor responses. Neuron 61, 786–800 (2009).

    Article  CAS  Google Scholar 

  42. Mailly, P., Charpier, S., Menetrey, A. & Deniau, J.M. Three-dimensional organization of the recurrent axon collateral network of the substantia nigra pars reticulata neurons in the rat. J. Neurosci. 23, 5247–5257 (2003).

    Article  CAS  Google Scholar 

  43. Maeda, H. & Mogenson, G.J. Effects of peripheral stimulation on the activity of neurons in the ventral tegmental area, substantia nigra and midbrain reticular formation of rats. Brain Res. Bull. 8, 7–14 (1982).

    Article  CAS  Google Scholar 

  44. Coizet, V., Dommett, E.J., Klop, E.M., Redgrave, P. & Overton, P.G. The parabrachial nucleus is a critical link in the transmission of short latency nociceptive information to midbrain dopaminergic neurons. Neuroscience 168, 263–272 (2010).

    Article  CAS  Google Scholar 

  45. Lodge, D.J. The medial prefrontal and orbitofrontal cortices differentially regulate dopamine system function. Neuropsychopharmacology. 36, 1227–1236 (2011).

    Article  CAS  Google Scholar 

  46. Bayer, V.E. & Pickel, V.M. GABA-labeled terminals form proportionally more synapses with dopaminergic neurons containing low densities of tyrosine hydroxylase immunoreactivity in rat ventral tegmental area. Brain Res. 559, 44–55 (1991).

    Article  CAS  Google Scholar 

  47. Paxinos, G. & Watson, C. The Rat Brain in Stereotaxic Coordinates (Elselvier, Amsterdam, 2007).

  48. Bolam, J.P. Experimental Neuroanatomy: a Practical Approach (IRL Press, Oxford, 1992).

  49. Sadek, A.R., Magill, P.J. & Bolam, J.P. A single-cell analysis of intrinsic connectivity in the rat globus pallidus. J. Neurosci. 27, 6352–6362 (2007).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to M. West for providing critical comments during the development of the single-cell stereological protocol. We also thank E. Norman, C. Francis, K. Whitworth, B. Micklem, S. Fernández, J. Mpodozis and G. Marín for expert technical assistance, and C. Morales and M. Quispe for statistical analysis. This work was supported by the Medical Research Council (UK), Parkinson's UK (grant G-0601, to J.P.B., P.J.M. and M.A.U.), and the Comisión Nacional de Investigación Científica y Tecnológica (Fondecyt grant 11100433, Conicyt, Chile to P.H.).

Author information

Author notes

  1. Matthew T C Brown & Mark A Ungless

    Present address: Present addresses: Department of Basic Neurosciences, CMU, Geneva, Switzerland (M.T.C.B.), Medical Research Council Clinical Sciences Centre, Imperial College London, Hammersmith Hospital, London, UK (M.A.U.).,

Authors and Affiliations

  1. Department of Pharmacology, MRC Anatomical Neuropharmacology Unit, University of Oxford, Oxford, UK

    Pablo Henny, Matthew T C Brown, Augustus Northrop, Mark A Ungless, Peter J Magill & J Paul Bolam

  2. Departamento de Anatomía Normal, Escuela de Medicina, Pontificia Universidad Católica de Chile, Santiago, Chile

    Pablo Henny & Macarena Faunes

  3. Centro Interdisciplinario de Neurociencia, NeuroUC, Pontificia Universidad Católica de Chile, Santiago, Chile

    Pablo Henny & Macarena Faunes

  4. Oxford Parkinson's Disease Centre, University of Oxford, Oxford, UK

    Peter J Magill & J Paul Bolam

Authors
  1. Pablo Henny
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Matthew T C Brown
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Augustus Northrop
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Macarena Faunes
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mark A Ungless
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Peter J Magill
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. J Paul Bolam
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

P.H. performed the juxtacellular labeling of some neurons, carried out most of the immunohistochemical procedures, electron microscopic analysis, neuronal reconstructions and stereological analysis, analyzed most of the anatomical and some of the physiological data, prepared the figures, and wrote the manuscript. M.T.C.B. recorded and juxtacellularly labeled most of the neurons that we used, and performed most of the physiological analysis. A.N. carried out neuronal reconstructions in half of the neurons. M.F. performed the light microscopic stereological analysis of immunolabeled varicosities. M.A.U. provided important feedback during the development of the project and gave critical comments during the writing of the manuscript. P.J.M. supervised and contributed to the recording and juxtacellular labeling of neurons and the project in general, and provided insightful comments during the writing of the manuscript. J.P.B. supervised the entire project, provided expertise in immunohistochemistry and tissue processing, helped with ultrastructural analysis, and provided critical and insightful comments during the writing of the manuscript.

Corresponding author

Correspondence to Pablo Henny.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–3 and Supplementary Tables 1 and 2 (PDF 1231 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Henny, P., Brown, M., Northrop, A. et al. Structural correlates of heterogeneous in vivo activity of midbrain dopaminergic neurons. Nat Neurosci 15, 613–619 (2012). https://doi.org/10.1038/nn.3048

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nn.3048

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing