Amygdala circuitry mediating reversible and bidirectional control of anxiety (original) (raw)

Nature volume 471, pages 358–362 (2011)Cite this article

Subjects

Abstract

Anxiety—a sustained state of heightened apprehension in the absence of immediate threat—becomes severely debilitating in disease states1. Anxiety disorders represent the most common of psychiatric diseases (28% lifetime prevalence)2 and contribute to the aetiology of major depression and substance abuse3,4. Although it has been proposed that the amygdala, a brain region important for emotional processing5,6,7,8, has a role in anxiety9,10,11,12,13, the neural mechanisms that control anxiety remain unclear. Here we explore the neural circuits underlying anxiety-related behaviours by using optogenetics with two-photon microscopy, anxiety assays in freely moving mice, and electrophysiology. With the capability of optogenetics14,15,16 to control not only cell types but also specific connections between cells, we observed that temporally precise optogenetic stimulation of basolateral amygdala (BLA) terminals in the central nucleus of the amygdala (CeA)—achieved by viral transduction of the BLA with a codon-optimized channelrhodopsin followed by restricted illumination in the downstream CeA—exerted an acute, reversible anxiolytic effect. Conversely, selective optogenetic inhibition of the same projection with a third-generation halorhodopsin15 (eNpHR3.0) increased anxiety-related behaviours. Importantly, these effects were not observed with direct optogenetic control of BLA somata, possibly owing to recruitment of antagonistic downstream structures. Together, these results implicate specific BLA–CeA projections as critical circuit elements for acute anxiety control in the mammalian brain, and demonstrate the importance of optogenetically targeting defined projections, beyond simply targeting cell types, in the study of circuit function relevant to neuropsychiatric disease.

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

Access options

Subscribe to this journal

Receive 51 print issues and online access

$199.00 per year

only $3.90 per issue

Buy this article

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

Additional access options:

Similar content being viewed by others

References

  1. Lieb, R. Anxiety disorders: clinical presentation and epidemiology. Handb. Exp. Pharmacol. 169, 405–432 (2005)
    Article Google Scholar
  2. Kessler, R. C. et al. Lifetime prevalence and age-of-onset distributions of DSM-IV disorders in the National Comorbidity Survey Replication. Arch. Gen. Psychiatry 62, 593–602 (2005)
    Article Google Scholar
  3. Koob, G. F. Brain stress systems in the amygdala and addiction. Brain Res. 1293, 61–75 (2009)
    Article CAS Google Scholar
  4. Ressler, K. J. & Mayberg, H. S. Targeting abnormal neural circuits in mood and anxiety disorders: from the laboratory to the clinic. Nature Neurosci. 10, 1116–1124 (2007)
    Article CAS Google Scholar
  5. LeDoux, J. The emotional brain, fear, and the amygdala. Cell. Mol. Neurobiol. 23, 727–738 (2003)
    Article Google Scholar
  6. Pare, D., Quirk, G. J. & Ledoux, J. E. New vistas on amygdala networks in conditioned fear. J. Neurophysiol. 92, 1–9 (2004)
    Article Google Scholar
  7. Tye, K. M., Stuber, G. D., de Ridder, B., Bonci, A. & Janak, P. H. Rapid strengthening of thalamo-amygdala synapses mediates cue–reward learning. Nature 453, 1253–1257 (2008)
    Article CAS ADS Google Scholar
  8. Weiskrantz, L. Behavioral changes associated with ablation of the amygdaloid complex in monkeys. J. Comp. Physiol. Psychol. 49, 381–391 (1956)
    Article CAS Google Scholar
  9. Kalin, N. H., Shelton, S. E. & Davidson, R. J. The role of the central nucleus of the amygdala in mediating fear and anxiety in the primate. J. Neurosci. 24, 5506–5515 (2004)
    Article CAS Google Scholar
  10. Lesscher, H. M. et al. Amygdala protein kinase C epsilon regulates corticotropin-releasing factor and anxiety-like behavior. Genes Brain Behav. 7, 323–333 (2008)
    Article CAS Google Scholar
  11. Etkin, A., Prater, K. E., Schatzberg, A. F., Menon, V. & Greicius, M. D. Disrupted amygdalar subregion functional connectivity and evidence of a compensatory network in generalized anxiety disorder. Arch. Gen. Psychiatry 66, 1361–1372 (2009)
    Article Google Scholar
  12. Lyons, A. M. & Thiele, T. E. Neuropeptide Y conjugated to saporin alters anxiety-like behavior when injected into the central nucleus of the amygdala or basomedial hypothalamus in BALB/cJ mice. Peptides 31, 2193–2199 (2010)
    Article CAS Google Scholar
  13. Roozendaal, B., McEwen, B. S. & Chattarji, S. Stress, memory and the amygdala. Nature Rev. Neurosci. 10, 423–433 (2009)
    Article CAS Google Scholar
  14. Boyden, E. S., Zhang, F., Bamberg, E., Nagel, G. & Deisseroth, K. Millisecond-timescale, genetically targeted optical control of neural activity. Nature Neurosci. 8, 1263–1268 (2005)
    Article CAS Google Scholar
  15. Gradinaru, V. et al. Molecular and cellular approaches for diversifying and extending optogenetics. Cell 141, 154–165 (2010)
    Article CAS Google Scholar
  16. Deisseroth, K. Optogenetics: controlling the brain with light. Sci. Am. 303, 48–55 (2010)
    Article ADS Google Scholar
  17. Woods, J. H., Katz, J. L. & Winger, G. Benzodiazepines: use, abuse, and consequences. Pharmacol. Rev. 44, 151–347 (1992)
    CAS PubMed Google Scholar
  18. Ciocchi, S. et al. Encoding of conditioned fear in central amygdala inhibitory circuits. Nature 468, 277–282 (2010)
    Article CAS ADS Google Scholar
  19. Haubensak, W. et al. Genetic dissection of an amygdala microcircuit that gates conditioned fear. Nature 468, 270–276 (2010)
    Article CAS ADS Google Scholar
  20. Carlsen, J. Immunocytochemical localization of glutamate decarboxylase in the rat basolateral amygdaloid nucleus, with special reference to GABAergic innervation of amygdalostriatal projection neurons. J. Comp. Neurol. 273, 513–526 (1988)
    Article CAS Google Scholar
  21. Smith, Y. & Pare, D. Intra-amygdaloid projections of the lateral nucleus in the cat: PHA-L anterograde labeling combined with postembedding GABA and glutamate immunocytochemistry. J. Comp. Neurol. 342, 232–248 (1994)
    Article CAS Google Scholar
  22. McDonald, A. J. Cytoarchitecture of the central amygdaloid nucleus of the rat. J. Comp. Neurol. 208, 401–418 (1982)
    Article CAS Google Scholar
  23. Krettek, J. E. & Price, J. L. A description of the amygdaloid complex in the rat and cat with observations on intra-amygdaloid axonal connections. J. Comp. Neurol. 178, 255–279 (1978)
    Article CAS Google Scholar
  24. Krettek, J. E. & Price, J. L. Amygdaloid projections to subcortical structures within the basal forebrain and brainstem in the rat and cat. J. Comp. Neurol. 178, 225–253 (1978)
    Article CAS Google Scholar
  25. Davis, M. in The Amygdala: A Functional Analysis (ed. Aggleton, J. P. ) 213–288 (Oxford Univ. Press, 2000)
    Google Scholar
  26. Pitkanen, A. in The Amygdala: A Functional Analysis (ed. Aggleton, J. P. ) 31–99 (Oxford Univ. Press, 2000)
    Google Scholar
  27. Carola, V., D’Olimpio, F., Brunamonti, E., Mangia, F. & Renzi, P. Evaluation of the elevated plus-maze and open-field tests for the assessment of anxiety-related behaviour in inbred mice. Behav. Brain Res. 134, 49–57 (2002)
    Article Google Scholar
  28. Davis, M., Walker, D. L., Miles, L. & Grillon, C. Phasic vs sustained fear in rats and humans: role of the extended amygdala in fear vs anxiety. Neuropsychopharmacology 35, 105–135 (2010)
    Article Google Scholar
  29. Shin, L. M. & Liberzon, I. The neurocircuitry of fear, stress, and anxiety disorders. Neuropsychopharmacology 35, 169–191 (2010)
    Article Google Scholar
  30. Gray, J. A. & McNaughton, N. The neuropsychology of anxiety: reprise. Nebr. Symp. Motiv. 43, 61–134 (1996)
    CAS PubMed Google Scholar

Download references

Acknowledgements

We would like to thank P. Janak, H. Fields, G. Stuber, E. Thomas, F. Zhang, I. Witten, V. Sohal, T. Davidson and M. Warden as well as J. Mattis, R. Durand, M. Mogri, J. Mirzabekov and E. Steinberg for discussions, and the entire K.D. laboratory for their support. All viruses were packaged at University of North Carolina (UNC) Vector Core. Supported by NIMH (1F32MH088010-01, K.M.T.), NARSAD (K.R.T.), Samsung Scholarship (S.-Y.K.), NSF IGERT Award 0801700 (L.G.) and the Defense Advanced Research Projects Agency Reorganization and Plasticity to Accelerate Injury Recovery (N66001-10-C-2010), the Alice Woo, Albert Yu, Snyder, and McKnight Foundations, as well as NIDA, NIMH and the NIH Pioneer Award (K.D.)

Author information

Author notes

  1. Kay M. Tye, Rohit Prakash, Sung-Yon Kim and Lief E. Fenno: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Bioengineering, Stanford University, Stanford, 94305, California, USA
    Kay M. Tye, Rohit Prakash, Sung-Yon Kim, Lief E. Fenno, Logan Grosenick, Hosniya Zarabi, Kimberly R. Thompson, Viviana Gradinaru, Charu Ramakrishnan & Karl Deisseroth
  2. Neurosciences Program, Stanford University, Stanford, 94305, California, USA
    Rohit Prakash, Sung-Yon Kim, Lief E. Fenno, Logan Grosenick, Viviana Gradinaru & Karl Deisseroth
  3. Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, 94305, California, USA
    Karl Deisseroth
  4. Howard Hughes Medical Institute, Stanford University, Stanford, 94305, California, USA
    Karl Deisseroth
  5. CNC Program, Stanford University, Stanford, 94305, California, USA
    Karl Deisseroth

Authors

  1. Kay M. Tye
    You can also search for this author inPubMed Google Scholar
  2. Rohit Prakash
    You can also search for this author inPubMed Google Scholar
  3. Sung-Yon Kim
    You can also search for this author inPubMed Google Scholar
  4. Lief E. Fenno
    You can also search for this author inPubMed Google Scholar
  5. Logan Grosenick
    You can also search for this author inPubMed Google Scholar
  6. Hosniya Zarabi
    You can also search for this author inPubMed Google Scholar
  7. Kimberly R. Thompson
    You can also search for this author inPubMed Google Scholar
  8. Viviana Gradinaru
    You can also search for this author inPubMed Google Scholar
  9. Charu Ramakrishnan
    You can also search for this author inPubMed Google Scholar
  10. Karl Deisseroth
    You can also search for this author inPubMed Google Scholar

Contributions

K.M.T., R.P., S.-Y.K., L.E.F. and K.D. contributed to study design and data interpretation. K.M.T., R.P., S.-Y.K. and L.E.F. contributed to data collection and K.M.T. coordinated data collection and analysis. K.M.T., S.-Y.K., H.Z. and K.R.T. contributed to immunohistochemical processing, fluorescence imaging and quantitative analyses. K.M.T. and L.G. performed the behavioural and ex vivo electrophysiology statistical analyses. V.G. and C.R. contributed to the design of eNpHR3.0. C.R. cloned all constructs and managed viral packaging processes. K.D. supervised all aspects of the work. All authors contributed to writing the paper.

Corresponding author

Correspondence toKarl Deisseroth.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-15 with legends, Supplementary Materials and Methods and additional references. (PDF 11353 kb)

Supplementary Movie 1

This movie shows a representative mouse from the ChR2:BLA-CeA group on elevated plus maze. The 15-minute elevated plus maze session is shown at 5x speed; each 5-min epoch is shown in 1 min and the duration of the light epoch is indicated by the appearance of blue text detailing light stimulation parameters. During the light-on epoch, the mouse increased open arm entry and open arm time. (MP4 6635 kb)

PowerPoint slides

Rights and permissions

About this article

Cite this article

Tye, K., Prakash, R., Kim, SY. et al. Amygdala circuitry mediating reversible and bidirectional control of anxiety.Nature 471, 358–362 (2011). https://doi.org/10.1038/nature09820

Download citation

This article is cited by

Editorial Summary

Neuronal circuitry of anxiety

The amygdala, a brain region important for learning fearful memories, is thought to have a role in generalized anxiety, but the subregions and connections involved in this response are unknown. Now, using optogenetic stimulation of basolateral amygdala terminals in the central nucleus of the amygdala of rats, a specific circuit for natural bidirectional anxiety control has been identified. Stimulating these neurons has a calming effect, whereas blocking the same projection increases anxiety-related behaviours. These findings are consistent with a role for the central nucleus of the amygdala in anxiety, although there may be other circuits working in parallel or downstream of the amygdala.