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Pavlovian Conditioning and Extinction Methods for Studying the Neurobiology of Fear Learning in PTSD

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Translational Methods for PTSD Research

Part of the book series: Neuromethods ((NM,volume 198))

Abstract

We describe a human threat conditioning and extinction paradigm used to study posttraumatic stress disorder as well as other psychiatric disorders. This paradigm was designed based on rodent threat conditioning paradigms for its application in translational neuroscience research. It is a passive viewing emotional learning and memory task composed of four phases. The first is the threat conditioning phase, followed by a threat extinction learning phase. Subsequently, memory for extinction learning is evaluated in extinction recall after a time delay (typically 24 hours after extinction learning). The last phase is the renewal of threat responding after extinction. Participants undergo this paradigm while in an fMRI scanner to identify brain correlates during all emotional learning phases. The duration of each experimental phase is approximately 13 minutes. Various time delays can be inserted between phases, depending on the aims of the experiment. Other experimental measures such as skin conductance responses and subjective measures could also be acquired during all experimental phases.

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References

  1. APA (2013) Trauma- and stressor-related disorders. In: Diagnostic and statistical manual of mental disorders, 5th edn. https://doi.org/10.1176/appi.books.9780890425596.dsm07

    Chapter  Google Scholar 

  2. Shvil E, Rusch HL, Sullivan GM, Neria Y (2013) Neural, psychophysiological, and behavioral markers of fear processing in PTSD: a review of the literature. Curr Psychiatry Rep 15(5):358–358. https://doi.org/10.1007/s11920-013-0358-3

    Article  PubMed  PubMed Central  Google Scholar 

  3. Hughes KC, Shin LM (2011) Functional neuroimaging studies of post-traumatic stress disorder. Expert Rev Neurother 11(2):275–285. https://doi.org/10.1586/ern.10.198

    Article  PubMed  PubMed Central  Google Scholar 

  4. Garfinkel SN, Abelson JL, King AP, Sripada RK, Wang X, Gaines LM, Liberzon I (2014) Impaired contextual modulation of memories in PTSD: an fMRI and psychophysiological study of extinction retention and fear renewal. J Neurosci 34(40):13435–13443. https://doi.org/10.1523/jneurosci.4287-13.2014

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Milad MR, Pitman RK, Ellis CB, Gold AL, Shin LM, Lasko NB, Zeidan MA, Handwerger K, Orr SP, Rauch SL (2009) Neurobiological basis of failure to recall extinction memory in posttraumatic stress disorder. Biol Psychiatry 66(12):1075–1082. https://doi.org/10.1016/j.biopsych.2009.06.026

    Article  PubMed  PubMed Central  Google Scholar 

  6. Holt DJ, Coombs G, Zeidan MA, Goff DC, Milad MR (2012) Failure of neural responses to safety cues in schizophrenia. Arch Gen Psychiatry 69(9):893–903. https://doi.org/10.1001/archgenpsychiatry.2011.2310

    Article  PubMed  PubMed Central  Google Scholar 

  7. Lonsdorf TB, Menz MM, Andreatta M, Fullana MA, Golkar A, Haaker J, Heitland I, Hermann A, Kuhn M, Kruse O, Meir Drexler S, Meulders A, Nees F, Pittig A, Richter J, Römer S, Shiban Y, Schmitz A, Straube B, Vervliet B, Wendt J, Baas JMP, Merz CJ (2017) Don’t fear ‘fear conditioning’: methodological considerations for the design and analysis of studies on human fear acquisition, extinction, and return of fear. Neurosci Biobehav Rev 77:247–285. https://doi.org/10.1016/j.neubiorev.2017.02.026

    Article  PubMed  Google Scholar 

  8. LaBar KS, Gatenby JC, Gore JC, LeDoux JE, Phelps EA (1998) Human amygdala activation during conditioned fear acquisition and extinction: a mixed-trial fMRI study. Neuron 20(5):937–945. https://doi.org/10.1016/s0896-6273(00)80475-4

    Article  CAS  PubMed  Google Scholar 

  9. Büchel C, Dolan RJ, Armony JL, Friston KJ (1999) Amygdala-hippocampal involvement in human aversive trace conditioning revealed through event-related functional magnetic resonance imaging. J Neurosci 19(24):10869–10876. https://doi.org/10.1523/jneurosci.19-24-10869.1999

    Article  PubMed  PubMed Central  Google Scholar 

  10. Phelps EA (2006) Emotion and cognition: insights from studies of the human amygdala. Annu Rev Psychol 57:27–53. https://doi.org/10.1146/annurev.psych.56.091103.070234

    Article  PubMed  Google Scholar 

  11. Milad MR, Orr SP, Lasko NB, Chang Y, Rauch SL, Pitman RK (2008) Presence and acquired origin of reduced recall for fear extinction in PTSD: results of a twin study. J Psychiatr Res 42(7):515–520. https://doi.org/10.1016/j.jpsychires.2008.01.017

    Article  PubMed  PubMed Central  Google Scholar 

  12. Duits P, Cath DC, Lissek S, Hox JJ, Hamm AO, Engelhard IM, van den Hout MA, Baas JM (2015) Updated meta-analysis of classical fear conditioning in the anxiety disorders. Depress Anxiety 32(4):239–253. https://doi.org/10.1002/da.22353

    Article  PubMed  Google Scholar 

  13. Lissek S, Powers AS, McClure EB, Phelps EA, Woldehawariat G, Grillon C, Pine DS (2005) Classical fear conditioning in the anxiety disorders: a meta-analysis. Behav Res Ther 43(11):1391–1424. https://doi.org/10.1016/j.brat.2004.10.007

    Article  PubMed  Google Scholar 

  14. Milad MR, Quirk GJ (2012) Fear extinction as a model for translational neuroscience: ten years of progress. Annu Rev Psychol 63:129–151. https://doi.org/10.1146/annurev.psych.121208.131631

    Article  PubMed  PubMed Central  Google Scholar 

  15. Milad MR, Orr SP, Pitman RK, Rauch SL (2005) Context modulation of memory for fear extinction in humans. Psychophysiology 42(4):456–464. https://doi.org/10.1111/j.1469-8986.2005.00302.x

    Article  PubMed  Google Scholar 

  16. Milad MR, Wright CI, Orr SP, Pitman RK, Quirk GJ, Rauch SL (2007) Recall of fear extinction in humans activates the ventromedial prefrontal cortex and hippocampus in concert. Biol Psychiatry 62(5):446–454. https://doi.org/10.1016/j.biopsych.2006.17.011

    Article  PubMed  Google Scholar 

  17. Hamacher-Dang TC, Merz CJ, Wolf OT (2015) Stress following extinction learning leads to a context-dependent return of fear. Psychophysiology 52(4):489–498. https://doi.org/10.1111/psyp.12384

    Article  PubMed  Google Scholar 

  18. Holt DJ, Lebron-Milad K, Milad MR, Rauch SL, Pitman RK, Orr SP, Cassidy BS, Walsh JP, Goff DC (2009) Extinction memory is impaired in schizophrenia. Biol Psychiatry 65(6):455–463. https://doi.org/10.1016/j.biopsych.2008.09.017

    Article  PubMed  Google Scholar 

  19. Lebron-Milad K, Abbs B, Milad MR, Linnman C, Rougemount-Bucking A, Zeidan MA, Holt DJ, Goldstein JM (2012) Sex differences in the neurobiology of fear conditioning and extinction: a preliminary fMRI study of shared sex differences with stress-arousal circuitry. Biol Mood Anxiety Disord 2:7. https://doi.org/10.1186/2045-5380-2-7

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Linnman C, Zeidan MA, Furtak SC, Pitman RK, Quirk GJ, Milad MR (2012) Resting amygdala and medial prefrontal metabolism predicts functional activation of the fear extinction circuit. Am J Psychiatry 169(4):415–423. https://doi.org/10.1176/appi.ajp.2011.10121780

    Article  PubMed  PubMed Central  Google Scholar 

  21. Pace-Schott EF, Milad MR, Orr SP, Rauch SL, Stickgold R, Pitman RK (2009) Sleep promotes generalization of extinction of conditioned fear. Sleep 32(1):19–26

    PubMed  PubMed Central  Google Scholar 

  22. Rougemont-Bücking A, Linnman C, Zeffiro TA, Zeidan MA, Lebron-Milad K, Rodriguez-Romaguera J, Rauch SL, Pitman RK, Milad MR (2011) Altered processing of contextual information during fear extinction in PTSD: an fMRI study. CNS Neurosci Ther 17(4):227–236. https://doi.org/10.1111/j.1755-5949.2010.00152.x

    Article  PubMed  Google Scholar 

  23. Shvil E, Sullivan GM, Schafer S, Markowitz JC, Campeas M, Wager TD, Milad MR, Neria Y (2014) Sex differences in extinction recall in posttraumatic stress disorder: a pilot fMRI study. Neurobiol Learn Mem 113:101–108. https://doi.org/10.1016/j.nlm.2014.02.003

    Article  PubMed  PubMed Central  Google Scholar 

  24. Zeidan MA, Lebron-Milad K, Thompson-Hollands J, Im JJY, Dougherty DD, Holt DJ, Orr SP, Milad MR (2012) Test-retest reliability during fear acquisition and fear extinction in humans. CNS Neurosci Ther 18(4):313–317. https://doi.org/10.1111/j.1755-5949.2011.00238.x

    Article  PubMed  Google Scholar 

  25. van ‘t Wout M, Mariano TY, Garnaat SL, Reddy MK, Rasmussen SA, Greenberg BD (2016) Can transcranial direct current stimulation augment extinction of conditioned fear? Brain Stimul 9(4):529–536. https://doi.org/10.1016/j.brs.2016.03.004

    Article  PubMed  PubMed Central  Google Scholar 

  26. Quirk GJ, Repa JC, LeDoux JE (1995) Fear conditioning enhances short-latency auditory responses of lateral amygdala neurons: parallel recordings in the freely behaving rat. Neuron 15(5):1029–1039. https://doi.org/10.1016/0896-6273(95)90092-6

    Article  CAS  PubMed  Google Scholar 

  27. Quirk GJ, Armony JL, LeDoux JE (1997) Fear conditioning enhances different temporal components of tone-evoked spike trains in auditory cortex and lateral amygdala. Neuron 19(3):613–624. https://doi.org/10.1016/S0896-6273(00)80375-X

    Article  CAS  PubMed  Google Scholar 

  28. Milad MR, Quirk GJ (2002) Neurons in medial prefrontal cortex signal memory for fear extinction. Nature 420(6911):70–74. https://doi.org/10.1038/nature01138

    Article  CAS  PubMed  Google Scholar 

  29. Dawson ME, Schell AM, Filion DL (2007) The electrodermal system. In: Cacioppo JT, Tassinary LG, Berntson GG (eds) Handbook of psychophysiology, 3rd edn. Cambridge University Press, New York, pp 159–181. https://doi.org/10.1017/CBO9780511546396.007

    Chapter  Google Scholar 

  30. Harris JA, Jones ML, Bailey GK, Westbrook RF (2000) Contextual control over conditioned responding in an extinction paradigm. J Exp Psychol Anim Behav Process 26(2):174–185. https://doi.org/10.1037//0097-7403.26.2.174

    Article  CAS  PubMed  Google Scholar 

  31. Bouton ME (2000) A learning theory perspective on lapse, relapse, and the maintenance of behavior change. Health Psychol 19(1s):57–63. https://doi.org/10.1037/0278-6133.19.suppl1.57

    Article  CAS  PubMed  Google Scholar 

  32. Bach DR, Friston KJ, Dolan RJ (2013) An improved algorithm for model-based analysis of evoked skin conductance responses. Biol Psychol 94(3):490–497. https://doi.org/10.1016/j.biopsycho.2013.09.010

    Article  PubMed  PubMed Central  Google Scholar 

  33. Bach DR, Flandin G, Friston KJ, Dolan RJ (2009) Time-series analysis for rapid event-related skin conductance responses. J Neurosci Methods 184(2):224–234. https://doi.org/10.1016/j.jneumeth.2009.08.005

    Article  PubMed  PubMed Central  Google Scholar 

  34. Peri T, Ben-Shakhar G, Orr SP, Shalev AY (2000) Psychophysiologic assessment of aversive conditioning in posttraumatic stress disorder. Biol Psychiatry 47(6):512–519. https://doi.org/10.1016/s0006-3223(99)00144-4

    Article  CAS  PubMed  Google Scholar 

  35. Orr SP, Metzger LJ, Lasko NB, Macklin ML, Peri T, Pitman RK (2000) De novo conditioning in trauma-exposed individuals with and without posttraumatic stress disorder. J Abnorm Psychol 109(2):290–298

    Article  CAS  PubMed  Google Scholar 

  36. Marin MF, Hammoud MZ, Klumpp H, Simon NM, Milad MR (2020) Multimodal categorical and dimensional approaches to understanding threat conditioning and its extinction in individuals with anxiety disorders. JAMA Psychiatry 77(6):618–627. https://doi.org/10.1001/jamapsychiatry.2019.4833

    Article  PubMed  Google Scholar 

  37. Marin M-F, Zsido RG, Song H, Lasko NB, Killgore WDS, Rauch SL, Simon NM, Milad MR (2017) Skin conductance responses and neural activations during fear conditioning and extinction recall across anxiety disorders. JAMA Psychiatry 74(6):622–631. https://doi.org/10.1001/jamapsychiatry.2017.0329

    Article  PubMed  PubMed Central  Google Scholar 

  38. Homan P, Lin Q, Murrough JW, Soleimani L, Bach DR, Clem RL, Schiller D (2017) Prazosin during threat discrimination boosts memory of the safe stimulus. Learn Mem Cold Spring Harbor 24(11):597–601. https://doi.org/10.1101/lm.045898.117

    Article  Google Scholar 

  39. Lonsdorf T, Kuhn M, Gerlicher A (2021) Navigating the manifold of skin conductance response quantification approaches. PsyArXiv. https://doi.org/10.31234/osf.io/9h2kd

  40. Esteban O, Birman D, Schaer M, Koyejo OO, Poldrack RA, Gorgolewski KJ (2017) MRIQC: advancing the automatic prediction of image quality in MRI from unseen sites. PLoS One 12(9):e0184661. https://doi.org/10.1371/journal.pone.0184661

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Esteban O, Markiewicz CJ, Blair RW, Moodie CA, Isik AI, Erramuzpe A, Kent JD, Goncalves M, DuPre E, Snyder M, Oya H, Ghosh SS, Wright J, Durnez J, Poldrack RA, Gorgolewski KJ (2019) fMRIPrep: a robust preprocessing pipeline for functional MRI. Nat Methods 16(1):111–116. https://doi.org/10.1038/s41592-018-0235-4

    Article  CAS  PubMed  Google Scholar 

  42. Liberzon I, Sripada CS (2008) The functional neuroanatomy of PTSD: a critical review. Prog Brain Res 167:151–169. https://doi.org/10.1016/s0079-6123(07)67011-3

    Article  PubMed  Google Scholar 

  43. Negreira AM, Abdallah CG (2019) A review of fMRI affective processing paradigms used in the neurobiological study of posttraumatic stress disorder. Chronic Stress 3:2470547019829035. https://doi.org/10.1177/2470547019829035

    Article  PubMed  PubMed Central  Google Scholar 

  44. Milad MR, Rosenbaum BL, Simon NM (2014) Neuroscience of fear extinction: implications for assessment and treatment of fear-based and anxiety related disorders. Behav Res Ther 62:17–23. https://doi.org/10.1016/j.brat.2014.08.006

    Article  PubMed  Google Scholar 

  45. Phelps EA, Delgado MR, Nearing KI, LeDoux JE (2004) Extinction learning in humans: role of the amygdala and vmPFC. Neuron 43(6):897–905. https://doi.org/10.1016/j.neuron.2004.08.042

    Article  CAS  PubMed  Google Scholar 

  46. Fullana MA, Albajes-Eizagirre A, Soriano-Mas C, Vervliet B, Cardoner N, Benet O, Radua J, Harrison BJ (2018) Fear extinction in the human brain: a meta-analysis of fMRI studies in healthy participants. Neurosci Biobehav Rev 88:16–25. https://doi.org/10.1016/j.neubiorev.2018.03.002

    Article  PubMed  Google Scholar 

  47. Kalisch R, Korenfeld E, Stephan KE, Weiskopf N, Seymour B, Dolan RJ (2006) Context-dependent human extinction memory is mediated by a ventromedial prefrontal and hippocampal network. J Neurosci 26(37):9503–9511. https://doi.org/10.1523/jneurosci.2021-06.2006

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Blechert J, Michael T, Vriends N, Margraf J, Wilhelm FH (2007) Fear conditioning in posttraumatic stress disorder: evidence for delayed extinction of autonomic, experiential, and behavioural responses. Behav Res Ther 45(9):2019–2033. https://doi.org/10.1016/j.brat.2007.02.012

    Article  PubMed  Google Scholar 

  49. Lonsdorf TB, Klingelhöfer-Jens M, Andreatta M, Beckers T, Chalkia A, Gerlicher A, Jentsch VL, Meir Drexler S, Mertens G, Richter J, Sjouwerman R, Wendt J, Merz CJ (2019) Navigating the garden of forking paths for data exclusions in fear conditioning research. elife 8. https://doi.org/10.7554/eLife.52465

  50. Marin M-F, Barbey F, Rosenbaum BL, Hammoud MZ, Orr SP, Milad MR (2020) Absence of conditioned responding in humans: a bad measure or individual differences? Psychophysiology 57(1):e13350. https://doi.org/10.1111/psyp.13350

    Article  PubMed  Google Scholar 

  51. Fullana MA, Harrison BJ, Soriano-Mas C, Vervliet B, Cardoner N, Àvila-Parcet A, Radua J (2016) Neural signatures of human fear conditioning: an updated and extended meta-analysis of fMRI studies. Mol Psychiatry 21(4):500–508. https://doi.org/10.1038/mp.2015.88

    Article  CAS  PubMed  Google Scholar 

  52. Quirk GJ, Armony JL, Repa JC, Li XF, LeDoux JE (1996) Emotional memory: a search for sites of plasticity. Cold Spring Harb Symp Quant Biol 61:247–257

    Article  CAS  PubMed  Google Scholar 

  53. Fullana MA, Albajes-Eizagirre A, Soriano-Mas C, Vervliet B, Cardoner N, Benet O, Radua J, Harrison BJ (2019) Amygdala where art thou? Neurosci Biobehav Rev 102:430–431. https://doi.org/10.1016/j.neubiorev.2018.06.003

    Article  PubMed  Google Scholar 

  54. Milad MR, Rauch SL, Pitman RK, Quirk GJ (2006) Fear extinction in rats: implications for human brain imaging and anxiety disorders. Biol Psychol 73(1):61–71. https://doi.org/10.1016/j.biopsycho.2006.01.008

    Article  PubMed  Google Scholar 

  55. Schiller D, Levy I, Niv Y, LeDoux JE, Phelps EA (2008) From fear to safety and back: reversal of fear in the human brain. J Neurosci 28(45):11517–11525. https://doi.org/10.1523/jneurosci.2265-08.2008

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Zhou F, Geng Y, Xin F, Li J, Feng P, Liu C, Zhao W, Feng T, Guastella AJ, Ebstein RP, Kendrick KM, Becker B (2019) Human extinction learning is accelerated by an angiotensin antagonist via ventromedial prefrontal cortex and its connections with basolateral amygdala. Biol Psychiatry 86(12):910–920. https://doi.org/10.1016/j.biopsych.2019.07.007

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Psychiatry, NYU Grossman School of Medicine, New York, NY, USA

    Dylan B. Miller, Madeleine M. Rassaby, Zhenfu Wen & Mohammed R. Milad

  2. Nathan Kline Institute for Psychiatric Research, Orangeburg, NY, USA

    Mohammed R. Milad

Authors
  1. Dylan B. Miller
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  2. Madeleine M. Rassaby
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  3. Zhenfu Wen
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  4. Mohammed R. Milad
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Correspondence to Mohammed R. Milad .

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Editors and Affiliations

  1. Psychiatric Institute, University of Chicago, Chicago, IL, USA

    Graziano Pinna

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Miller, D.B., Rassaby, M.M., Wen, Z., Milad, M.R. (2023). Pavlovian Conditioning and Extinction Methods for Studying the Neurobiology of Fear Learning in PTSD. In: Pinna, G. (eds) Translational Methods for PTSD Research. Neuromethods, vol 198. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3218-5_5

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  • DOI: https://doi.org/10.1007/978-1-0716-3218-5_5

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