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σ1 receptor ligands control a switch between passive and active threat responses

Nature Chemical Biology volume 12, pages 552–558 (2016)Cite this article

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Abstract

Humans and many animals show 'freezing' behavior in response to threatening stimuli. In humans, inappropriate threat responses are fundamental characteristics of several mental illnesses. To identify small molecules that modulate threat responses, we developed a high-throughput behavioral assay in zebrafish (Danio rerio) and evaluated 10,000 compounds for their effects on freezing behavior. We found three classes of compounds that switch the threat response from freezing to escape-like behavior. We then screened these for binding activity across 45 candidate targets. Using target profile clustering, we identified the sigma-1 (σ1) receptor as having a role in the mechanism of behavioral switching and confirmed that known σ1 ligands also disrupt freezing behavior. Furthermore, mutation of the gene encoding σ1 prevented the behavioral effect of escape-inducing compounds. One compound, which we call finazine, potently bound mammalian σ1 and altered threat-response behavior in mice. Thus, pharmacological and genetic interrogation of the freezing response revealed σ1 as a mediator of threat responses in vertebrates.

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Figure 1: High-throughput chemical screen for compounds that disrupt zebrafish freezing behavior.
Figure 2: Chemical structures of finazine, finoxetine and finopidil compounds.
Figure 3: Compound target profiling.
Figure 4: σ1 is a functional target of finazine.
Figure 5: Finazine has neuroactive effects in a mouse freezing assay.

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References

  1. Hartley, C.A. & Phelps, E.A. Anxiety and decision-making. Biol. Psychiatry 72, 113–118 (2012).

    Article  Google Scholar 

  2. Laviolette, S.R. Dopamine modulation of emotional processing in cortical and subcortical neural circuits: evidence for a final common pathway in schizophrenia? Schizophr. Bull. 33, 971–981 (2007).

    Article  Google Scholar 

  3. Williams, L.M. et al. Dysregulation of arousal and amygdala-prefrontal systems in paranoid schizophrenia. Am. J. Psychiatry 161, 480–489 (2004).

    Article  Google Scholar 

  4. Paradiso, S. et al. Emotions in unmedicated patients with schizophrenia during evaluation with positron emission tomography. Am. J. Psychiatry 160, 1775–1783 (2003).

    Article  Google Scholar 

  5. Brewin, C.R. What is it that a neurobiological model of PTSD must explain? Prog. Brain Res. 167, 217–228 (2008).

    Article  Google Scholar 

  6. Hyman, S.E. Revolution stalled. Sci. Transl. Med. 4, 155cm11 (2012).

    Article  Google Scholar 

  7. Rennekamp, A.J. & Peterson, R.T. 15 years of zebrafish chemical screening. Curr. Opin. Chem. Biol. 24, 58–70 (2015).

    Article  CAS  Google Scholar 

  8. Bass, S.L. & Gerlai, R. Zebrafish (Danio rerio) responds differentially to stimulus fish: the effects of sympatric and allopatric predators and harmless fish. Behav. Brain Res. 186, 107–117 (2008).

    Article  Google Scholar 

  9. Gerlai, R., Fernandes, Y. & Pereira, T. Zebrafish (Danio rerio) responds to the animated image of a predator: towards the development of an automated aversive task. Behav. Brain Res. 201, 318–324 (2009).

    Article  Google Scholar 

  10. Speedie, N. & Gerlai, R. Alarm substance induced behavioral responses in zebrafish (Danio rerio). Behav. Brain Res. 188, 168–177 (2008).

    Article  CAS  Google Scholar 

  11. Colwill, R.M. & Creton, R. Imaging escape and avoidance behavior in zebrafish larvae. Rev. Neurosci. 22, 63–73 (2011).

    Article  Google Scholar 

  12. Emran, F., Rihel, J. & Dowling, J.E. A behavioral assay to measure responsiveness of zebrafish to changes in light intensities. J. Vis. Exp. 20, 923 (2008).

    Google Scholar 

  13. Bullen, C.R. & Carlson, T.J. Non-physical fish barrier systems: their development and potential applications to marine ranching. Rev. Fish Biol. Fish. 13, 201–212 (2003).

    Article  Google Scholar 

  14. Yeh, C.M., Glöck, M. & Ryu, S. An optimized whole-body cortisol quantification method for assessing stress levels in larval zebrafish. PLoS One 8, e79406 (2013).

    Article  Google Scholar 

  15. Burgess, H.A. & Granato, M. Modulation of locomotor activity in larval zebrafish during light adaptation. J. Exp. Biol. 210, 2526–2539 (2007).

    Article  Google Scholar 

  16. Backman, T.W., Cao, Y. & Girke, T. ChemMine tools: an online service for analyzing and clustering small molecules. Nucleic Acids Res. 39, W486–W491 (2011).

    Article  CAS  Google Scholar 

  17. de Costa, B.R. et al. Synthesis and evaluation of optically pure [3H]-(+)-pentazocine, a highly potent and selective radioligand for sigma receptors. FEBS Lett. 251, 53–58 (1989).

    Article  CAS  Google Scholar 

  18. Lever, J.R., Gustafson, J.L., Xu, R., Allmon, R.L. & Lever, S.Z. σ1 and σ2 receptor binding affinity and selectivity of SA4503 and fluoroethyl SA4503. Synapse 59, 350–358 (2006).

    Article  CAS  Google Scholar 

  19. Walker, J.M. et al. Sigma receptors: biology and function. Pharmacol. Rev. 42, 355–402 (1990).

    CAS  PubMed  Google Scholar 

  20. Bruni, G. et al. Systematic behavioral profiling identifies antipsychotic-like compounds with multi-target polypharmacology. Nat. Chem. Biol. doi:10.1038/nchembio.2097 (2016).

    Article  CAS  Google Scholar 

  21. Antoniadis, E.A. & McDonald, R.J. Amygdala, hippocampus, and unconditioned fear. Exp. Brain Res. 138, 200–209 (2001).

    Article  CAS  Google Scholar 

  22. Adolphs, R. The biology of fear. Curr. Biol. 23, R79–R93 (2013).

    Article  CAS  Google Scholar 

  23. Rosen, J.B. The neurobiology of conditioned and unconditioned fear: a neurobehavioral system analysis of the amygdala. Behav. Cogn. Neurosci. Rev. 3, 23–41 (2004).

    Article  Google Scholar 

  24. Moritz, C., Berardi, F., Abate, C. & Peri, F. Live imaging reveals a new role for the sigma-1 (σ1) receptor in allowing microglia to leave brain injuries. Neurosci. Lett. 591, 13–18 (2015).

    Article  CAS  Google Scholar 

  25. Kamei, H., Kameyama, T. & Nabeshima, T. (+)-SKF-10,047 and dextromethorphan ameliorate conditioned fear stress through the activation of phenytoin-regulated σ1 sites. Eur. J. Pharmacol. 299, 21–28 (1996).

    Article  CAS  Google Scholar 

  26. Wang, J., Mack, A.L., Coop, A. & Matsumoto, R.R. Novel sigma (σ) receptor agonists produce antidepressant-like effects in mice. Eur. Neuropsychopharmacol. 17, 708–716 (2007).

    Article  CAS  Google Scholar 

  27. Reddy, D.S., Kaur, G. & Kulkarni, S.K. Sigma (σ1) receptor mediated anti-depressant-like effects of neurosteroids in the Porsolt forced swim test. Neuroreport 9, 3069–3073 (1998).

    Article  CAS  Google Scholar 

  28. Urani, A., Roman, F.J., Phan, V.L., Su, T.P. & Maurice, T. The antidepressant-like effect induced by σ1-receptor agonists and neuroactive steroids in mice submitted to the forced swimming test. J. Pharmacol. Exp. Ther. 298, 1269–1279 (2001).

    CAS  PubMed  Google Scholar 

  29. Matsuno, K., Kobayashi, T., Tanaka, M.K. & Mita, S. σ1 receptor subtype is involved in the relief of behavioral despair in the mouse forced swimming test. Eur. J. Pharmacol. 312, 267–271 (1996).

    Article  CAS  Google Scholar 

  30. Sabino, V., Cottone, P., Parylak, S.L., Steardo, L. & Zorrilla, E.P. Sigma-1 receptor knockout mice display a depressive-like phenotype. Behav. Brain Res. 198, 472–476 (2009).

    Article  CAS  Google Scholar 

  31. Chevallier, N., Keller, E. & Maurice, T. Behavioural phenotyping of knockout mice for the sigma-1 (σ1) chaperone protein revealed gender-related anxiety, depressive-like and memory alterations. J. Psychopharmacol. 25, 960–975 (2011).

    Article  CAS  Google Scholar 

  32. Fishback, J.A., Robson, M.J., Xu, Y.T. & Matsumoto, R.R. Sigma receptors: potential targets for a new class of antidepressant drug. Pharmacol. Ther. 127, 271–282 (2010).

    Article  CAS  Google Scholar 

  33. Kishimoto, A., Kaneko, M., Gotoh, Y. & Hashimoto, K. Ifenprodil for the treatment of flashbacks in female posttraumatic stress disorder patients with a history of childhood sexual abuse. Biol. Psychiatry 71, e7–e8 (2012).

    Article  Google Scholar 

  34. Sasaki, T. et al. Ifenprodil for the treatment of flashbacks in adolescent female posttraumatic stress disorder patients with a history of abuse. Psychother. Psychosom. 82, 344–345 (2013).

    Article  Google Scholar 

  35. Emran, F., Rihel, J., Adolph, A.R. & Dowling, J.E. Zebrafish larvae lose vision at night. Proc. Natl. Acad. Sci. USA 107, 6034–6039 (2010).

    Article  CAS  Google Scholar 

  36. Hwang, W.Y. et al. Targeted mutagenesis in zebrafish using CRISPR RNA-guided nucleases. Methods Mol. Biol. 1311, 317–334 (2015).

    Article  Google Scholar 

  37. Besnard, J. et al. Automated design of ligands to polypharmacological profiles. Nature 492, 215–220 (2012).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank C. Brady, J.J. Colund, I.T. Helenius, Y.N. Jin and A. Nath for helpful comments. Receptor binding profiles and Ki data were provided by the US National Institutes of Health (NIH) Psychoactive Drug Screening Program, contract HHSN-271-2008-025C. This work was supported by NIH grants T32 HL 007208 (A.J.R.), R01 MH 086867 (R.T.P.), R01 GM 088040 (R.T.P.) and U01 MH 105027 (D.K. and J.-R.J.Y.); the Charles and Ann Sanders MGH Research Scholar Award (R.T.P.); and the Harvard NeuroDiscovery Center.

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Authors and Affiliations

  1. Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, Massachusetts, USA

    Andrew J Rennekamp, You Wang, Samir Patel, Lindsay Cade, Andrew P W Gonzales, Jing-Ruey Joanna Yeh & Randall T Peterson

  2. Department of Medicine, Massachusetts General Hospital, Charlestown, Massachusetts, USA

    Andrew J Rennekamp, You Wang, Samir Patel, Lindsay Cade, Andrew P W Gonzales, Jing-Ruey Joanna Yeh & Randall T Peterson

  3. Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA

    Andrew J Rennekamp, You Wang, Samir Patel, Lindsay Cade, Andrew P W Gonzales, Jing-Ruey Joanna Yeh & Randall T Peterson

  4. Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA

    Andrew J Rennekamp, You Wang, Samir Patel, Lindsay Cade, Andrew P W Gonzales & Randall T Peterson

  5. Broad Institute, Cambridge, Massachusetts, USA

    Andrew J Rennekamp, You Wang, Samir Patel, Lindsay Cade, Andrew P W Gonzales & Randall T Peterson

  6. National Institute of Mental Health Psychoactive Drug Screening Program, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA

    Xi-Ping Huang & Bryan L Roth

  7. Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA

    Xi-Ping Huang & Bryan L Roth

  8. NeuroBehavior Laboratory, Harvard NeuroDiscovery Center, Brigham and Women's Hospital, Boston, Massachusetts, USA

    Paul J Lorello & Barbara J Caldarone

  9. Department of Neurology, Brigham and Women's Hospital, Boston, Massachusetts, USA

    Paul J Lorello & Barbara J Caldarone

  10. Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA

    Bryan L Roth

  11. Department of Physiology, University of California, San Francisco, San Francisco, California, USA.,

    David Kokel

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Contributions

A.J.R. designed the experiments and performed the zebrafish behavioral and assays (including the screen), created the zebrafish mutants, analyzed the data and wrote the manuscript with R.T.P. S.P. and L.C. assisted A.J.R. with zebrafish assays. A.P.W.G. assisted with zebrafish mutagenesis. J.-R.J.Y. designed the zebrafish mutagenesis strategies. X.-P.H. performed the in vitro binding assays, which were designed by B.L.R. Y.W. performed the chemical synthesis and purity analyses. P.J.L. performed the mouse experiments and analyzed the data with B.J.C. B.J.C., D.K. and B.L.R. contributed reagents and to research design. All authors contributed to data interpretation and commented on the manuscript.

Corresponding author

Correspondence to Randall T Peterson.

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Competing interests

A.J.R., D.K., Y.W. and R.T.P. declare competing financial interests; they are inventors on a pending patent application, PCT/US2015/037755, covering the σ1 ligands described in this manuscript.

Supplementary information

Supplementary Text and Figures

Supplementary Results, Supplementary Figures 1–21, Supplementary Tables 1–3 and Supplementary Note 1. (PDF 6244 kb)

Strobe-induced freezing response in zebrafish larvae at low magnification. (AVI 40075 kb)

41589_2016_BFnchembio2089_MOESM611_ESM.avi

Strobe-induced freezing response in zebrafish larvae treated with tenfold increasing concentrations of finazine. (AVI 56946 kb)

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Rennekamp, A., Huang, XP., Wang, Y. et al. σ1 receptor ligands control a switch between passive and active threat responses. Nat Chem Biol 12, 552–558 (2016). https://doi.org/10.1038/nchembio.2089

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