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Projection-specific roles of basolateral amygdala Thy1 neurons in alcohol-induced place preference

Molecular Psychiatry (2025)Cite this article

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Abstract

Alcohol seeking during abstinence is mediated in part by strong associations between the pharmacological effects of alcohol and the environment within which alcohol is administered. The amygdala, particularly the basolateral amygdala (BLA), is a key neural substrate of environmental cue and reward associations since it is involved in associative learning and memory recall. However, we still lack a clear understanding of how alcohol affects the activity of BLA neurons, which may encode information that drives environmental cue-dependent, alcohol-related behaviors. We previously demonstrated that a subset of BLA neurons which express the calcium/calmodulin-dependent protein kinase II (CaMKII) and thymus cell antigen 1 (Thy1) markers project preferentially to the nucleus accumbens (NAcc), rather than the central amygdala; and these neurons mediate fear inhibition rather than fear acquisition or expression, suggesting a specific role in positive valence processing. We now demonstrate that Pavlovian conditioning with alcohol administration increases the activity of these Thy1-expressing (Thy1+) excitatory neurons in mouse BLA, which is necessary for the conditioned appetitive response. In vivo calcium imaging indicates that the temporal activity profile of these neurons is also correlated with alcohol-induced motivated behavior in response to environmental cues. Optogenetic inhibition of BLA Thy1+ neuronal activity at cell body disrupts both the formation and expression of alcohol-induced conditioned place preference. Furthermore, selective axonal inhibition of BLA-Thy1+ efferents reveals that the activity of their NAcc and prefrontal cortex (PFC) projections are differentially necessary for alcohol cue association vs. recall, respectively. Together, these findings provide insights into a molecularly distinct subset of BLA neurons that regulates environmental cue-reward associations and drives alcohol-induced motivated behaviors in a projection-specific manner.

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Fig. 1: Ethanol (EtOH)-induced place preference.
Fig. 2: Fiber photometry of neuronal dynamics during CPP Conditioning and Test.
Fig. 3: Thy1+ neuronal activity at cell bodies in the BLA during Conditioning and Test is necessary for CPP formation and recall, respectively.
Fig. 4: Inhibition of BLA Thy1+ neuronal activity did not disrupt the formation and recall of CPP with natural reward, milk.
Fig. 5: BLA Thy1+ neuronal activity at axons in the NAcc and PFC during Conditioning and Test is necessary for CPP formation and recall, respectively.

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Data availability

The datasets used and analyzed during the current study are available from the corresponding author on reasonable request.

References

  1. Hasin DS, Stinson FS, Ogburn E, Grant BF. Prevalence, correlates, disability, and comorbidity of DSM-IV alcohol abuse and dependence in the United States: results from the National Epidemiologic Survey on Alcohol and Related Conditions. Arch Gen Psychiatry. 2007;64:830–42.

    Article  PubMed  Google Scholar 

  2. Rehm J, Mathers C, Popova S, Thavorncharoensap M, Teerawattananon Y, Patra J. Global burden of disease and injury and economic cost attributable to alcohol use and alcohol-use disorders. Lancet. 2009;373:2223–33.

    Article  PubMed  Google Scholar 

  3. Robinson TE, Berridge KC. The neural basis of drug craving: an incentive-sensitization theory of addiction. Brain Res Brain Res Rev. 1993;18:247–91.

    Article  CAS  PubMed  Google Scholar 

  4. Hyman SE. Addiction: a disease of learning and memory. Am J Psychiatry. 2005;162:1414–22.

    Article  PubMed  Google Scholar 

  5. Heinz A, Beck A, Grusser SM, Grace AA, Wrase J. Identifying the neural circuitry of alcohol craving and relapse vulnerability. Addict Biol. 2009;14:108–18.

    Article  CAS  PubMed  Google Scholar 

  6. Wiers CE, Stelzel C, Gladwin TE, Park SQ, Pawelczack S, Gawron CK, et al. Effects of cognitive bias modification training on neural alcohol cue reactivity in alcohol dependence. Am J Psychiatry. 2015;172:335–43.

    Article  PubMed  Google Scholar 

  7. Koob GF, Le Moal M. Drug addiction, dysregulation of reward, and allostasis. Neuropsychopharmacology. 2001;24:97–129.

    Article  CAS  PubMed  Google Scholar 

  8. Suh J, Ressler KJ. Common biological mechanisms of alcohol use disorder and post-traumatic stress disorder. Alcohol Res. 2018;39:131–45.

    PubMed  PubMed Central  Google Scholar 

  9. Crombag HS, Bossert JM, Koya E, Shaham Y. Review. Context-induced relapse to drug seeking: a review. Philos Trans R Soc Lond B Biol Sci. 2008;363:3233–43.

    Article  PubMed  PubMed Central  Google Scholar 

  10. Fenster RJ, McCullough K, Naumenko S, Thompson A, Klengel C, Rodgers A, et al. Cell-type specific induction of cyclo-oxygenase-2 in layer II/III prefrontal cortical neurons mediates stress-induced anxiety phenotypes in mice. bioRxiv 2021.2010.2010.463815 [Preprint]. 2021 https://www.biorxiv.org/content/10.1101/2021.10.10.463815v1.

  11. Claus ED, Ewing SW, Filbey FM, Sabbineni A, Hutchison KE. Identifying neurobiological phenotypes associated with alcohol use disorder severity. Neuropsychopharmacology. 2011;36:2086–96.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Hill KG, Ryabinin AE, Cunningham CL. FOS expression induced by an ethanol-paired conditioned stimulus. Pharmacol Biochem Behav. 2007;87:208–21.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Dayas CV, Liu X, Simms JA, Weiss F. Distinct patterns of neural activation associated with ethanol seeking: effects of naltrexone. Biol Psychiatry. 2007;61:979–89.

    Article  CAS  PubMed  Google Scholar 

  14. Chaudhri N, Woods CA, Sahuque LL, Gill TM, Janak PH. Unilateral inactivation of the basolateral amygdala attenuates context-induced renewal of Pavlovian-conditioned alcohol-seeking. Eur J Neurosci. 2013;38:2751–61.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Herry C, Ciocchi S, Senn V, Demmou L, Muller C, Luthi A. Switching on and off fear by distinct neuronal circuits. Nature. 2008;454:600–6.

    Article  CAS  PubMed  Google Scholar 

  16. Kim J, Pignatelli M, Xu S, Itohara S, Tonegawa S. Antagonistic negative and positive neurons of the basolateral amygdala. Nat Neurosci. 2016;19:1636–46.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Namburi P, Beyeler A, Yorozu S, Calhoon GG, Halbert SA, Wichmann R, et al. A circuit mechanism for differentiating positive and negative associations. Nature. 2015;520:675–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Zhang X, Guan W, Yang T, Furlan A, Xiao X, Yu K, et al. Genetically identified amygdala-striatal circuits for valence-specific behaviors. Nat Neurosci. 2021;24:1586–600.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Feng G, Mellor RH, Bernstein M, Keller-Peck C, Nguyen QT, Wallace M, et al. Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP. Neuron. 2000;28:41–51.

    Article  CAS  PubMed  Google Scholar 

  20. McCullough KM, Choi D, Guo J, Zimmerman K, Walton J, Rainnie DG, et al. Molecular characterization of Thy1 expressing fear-inhibiting neurons within the basolateral amygdala. Nat Commun. 2016;7:13149.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Jasnow AM, Ehrlich DE, Choi DC, Dabrowska J, Bowers ME, McCullough KM, et al. Thy1-expressing neurons in the basolateral amygdala may mediate fear inhibition. J Neurosci. 2013;33:10396–404.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Cunningham CL, Gremel CM, Groblewski PA. Drug-induced conditioned place preference and aversion in mice. Nat Protoc. 2006;1:1662–70.

    Article  CAS  PubMed  Google Scholar 

  23. Cunningham CL, Shields CN. Effects of multi-modal cues on conditioned place preference in C57BL/6J and DBA/2J mice. Psychopharmacology (Berl). 2018;235:3535–43.

    Article  CAS  PubMed  Google Scholar 

  24. O’Neal TJ, Bernstein MX, MacDougall DJ, Ferguson SM. A conditioned place preference for heroin is signaled by increased dopamine and direct pathway activity and decreased indirect pathway activity in the nucleus accumbens. J Neurosci. 2022;42:2011–24.

    Article  PubMed  PubMed Central  Google Scholar 

  25. Janak PH, Tye KM. From circuits to behaviour in the amygdala. Nature. 2015;517:284–92.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Beyeler A, Namburi P, Glober GF, Simonnet C, Calhoon GG, Conyers GF, et al. Divergent routing of positive and negative information from the amygdala during memory retrieval. Neuron. 2016;90:348–61.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Ambroggi F, Ishikawa A, Fields HL, Nicola SM. Basolateral amygdala neurons facilitate reward-seeking behavior by exciting nucleus accumbens neurons. Neuron. 2008;59:648–61.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Stuber GD, Sparta DR, Stamatakis AM, van Leeuwen WA, Hardjoprajitno JE, Cho S, et al. Excitatory transmission from the amygdala to nucleus accumbens facilitates reward seeking. Nature. 2011;475:377–80.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Meinhardt MW, Hansson AC, Perreau-Lenz S, Bauder-Wenz C, Stahlin O, Heilig M, et al. Rescue of infralimbic mGluR2 deficit restores control over drug-seeking behavior in alcohol dependence. J Neurosci. 2013;33:2794–806.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Kroener S, Mulholland PJ, New NN, Gass JT, Becker HC, Chandler LJ. Chronic alcohol exposure alters behavioral and synaptic plasticity of the rodent prefrontal cortex. PLoS One. 2012;7:e37541.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Renteria R, Baltz ET, Gremel CM. Chronic alcohol exposure disrupts top-down control over basal ganglia action selection to produce habits. Nat Commun. 2018;9:211.

    Article  PubMed  PubMed Central  Google Scholar 

  32. Gass JT, Sinclair CM, Cleva RM, Widholm JJ, Olive MF. Alcohol-seeking behavior is associated with increased glutamate transmission in basolateral amygdala and nucleus accumbens as measured by glutamate-oxidase-coated biosensors. Addict Biol. 2011;16:215–28.

    Article  CAS  PubMed  Google Scholar 

  33. Fenster RJ, Lebois LAM, Ressler KJ, Suh J. Brain circuit dysfunction in post-traumatic stress disorder: from mouse to man. Nat Rev Neurosci. 2018;19:535–51.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Liu X, Ramirez S, Redondo RL, Tonegawa S. Identification and manipulation of memory engram cells. Cold Spring Harb Symp Quant Biol. 2014;79:59–65.

    Article  PubMed  Google Scholar 

  35. Redondo RL, Kim J, Arons AL, Ramirez S, Liu X, Tonegawa S. Bidirectional switch of the valence associated with a hippocampal contextual memory engram. Nature. 2014;513:426–30.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Gal E, Amsalem O, Schindel A, London M, Schurmann F, Markram H, et al. The role of hub neurons in modulating cortical dynamics. Front Neural Circuits. 2021;15:718270.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Shinonaga Y, Takada M, Mizuno N. Topographic organization of collateral projections from the basolateral amygdaloid nucleus to both the prefrontal cortex and nucleus accumbens in the rat. Neuroscience. 1994;58:389–97.

    Article  CAS  PubMed  Google Scholar 

  38. Huang L, Chen Y, Jin S, Lin L, Duan S, Si K, et al. Organizational principles of amygdalar input-output neuronal circuits. Mol Psychiatry. 2021;26:7118–29.

    Article  PubMed  PubMed Central  Google Scholar 

  39. Cunningham CL, Shields CN. Effects of sex on ethanol conditioned place preference, activity and variability in C57BL/6J and DBA/2J mice. Pharmacol Biochem Behav. 2018;173:84–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Morris R. Thy-1 in developing nervous tissue. Dev Neurosci. 1985;7:133–60.

    Article  CAS  PubMed  Google Scholar 

  41. Barlow JZ, Huntley GW. Developmentally regulated expression of Thy-1 in structures of the mouse sensory-motor system. J Comp Neurol. 2000;421:215–33.

    Article  CAS  PubMed  Google Scholar 

  42. Narendra S, Klengel C, Hamzeh B, Patel D, Otten J, Lardenoije R, et al. Genome-wide transcriptomics of the amygdala reveals similar oligodendrocyte-related responses to acute and chronic alcohol drinking in female mice. Transl Psychiatry. 2022;12:476.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Trouche S, Koren V, Doig NM, Ellender TJ, El-Gaby M, Lopes-Dos-Santos V, et al. A hippocampus-accumbens tripartite neuronal motif guides appetitive memory in space. Cell. 2019;176:1393–406 e1316.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Bruno CA, O’Brien C, Bryant S, Mejaes JI, Estrin DJ, Pizzano C, et al. pMAT: An open-source software suite for the analysis of fiber photometry data. Pharmacol Biochem Behav. 2021;201:173093.

    Article  CAS  PubMed  Google Scholar 

  45. Calipari ES, Bagot RC, Purushothaman I, Davidson TJ, Yorgason JT, Pena CJ, et al. In vivo imaging identifies temporal signature of D1 and D2 medium spiny neurons in cocaine reward. Proc Natl Acad Sci USA. 2016;113:2726–31.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by NIH awards R21-AA027450 (JS and KR), R01-AA030585 (JS), P50-MH115874 (KJR), R01-MH108665 (KJR), and the Frazier Institute at McLean Hospital (KJR).

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Authors and Affiliations

  1. Division of Depression and Anxiety Disorders, McLean Hospital, Department of Psychiatry, Harvard Medical School, Belmont, MA, 02478, USA

    Junghyup Suh, Quilla C. Flanagan-Burt, Byung Kwon Moon, Amanda L. Pasqualini, Maria A. Zambrano & Kerry J. Ressler

  2. Department of Anesthesiology and Pain Medicine, University of Washington, Seattle, WA, 98195, USA

    Amanda L. Pasqualini

  3. Department of Immunology, Tufts University, Boston, MA, 02111, USA

    Maria A. Zambrano

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Contributions

Participated in research design: JS, KJR. Conducted experiments: JS, QCFB, ALP, MAZ. Performed data analysis: JS, BKM. Wrote or contributed to the writing of the manuscript: JS, KJR.

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Correspondence to Junghyup Suh or Kerry J. Ressler.

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Competing interests

The authors declare no competing interests. KJR has received consulting income from Acer, Bionomics, and Jazz Pharma; serves on Scientific Advisory Boards for Sage, Boehringer Ingelheim, Senseye, the Brain and Behavior Research Foundation, and the Brain Research Foundation, and he has received sponsored research support from Alto Neuroscience. None of this work is directly related to the work presented here.

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All the methods in the current study were approved by McLean Hospital’s Institutional Animal Care and Use Committee (#2018N000123) and maintained adherence to the National Research Council’s Guide for the Care and Use of Laboratory Animals.

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Suh, J., Flanagan-Burt, Q.C., Moon, B.K. et al. Projection-specific roles of basolateral amygdala Thy1 neurons in alcohol-induced place preference. Mol Psychiatry (2025). https://doi.org/10.1038/s41380-025-03184-w

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