Generalization, the process of applying knowledge acquired in one context to other contexts, often drives the expression of similar behaviors in related situations. At the cellular level, generalization is thought to depend on the activity of overlapping neurons that represent shared features between contexts (general representations). Using contextual fear conditioning in mice, we demonstrate that generalization can also occur in response to stress and result from reactivation of specific, rather than general context representations. We found that generalization emerges during memory retrieval, along with stress-induced abnormalities of septohippocampal oscillatory activity and acetylcholine release, which are typically found in negative affective states. In hippocampal neurons that represent aversive memories and drive generalization, cholinergic septohippocampal afferents contributed to a unique reactivation pattern of cFos, Npas4, and repressor element-1 silencing transcription factor (REST). Together, these findings suggest that generalization can be triggered by perceptually dissimilar but valence-congruent memories of specific aversive experiences. Through promoting the reactivation of such memories and their interference with ongoing behavior, abnormal cholinergic signaling could underlie maladaptive cognitive and behavioral generalization linked to negative affective states.
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The data that support the findings of this study and the analysis code are available from the authors on reasonable request.
Banich MT, Dukes P, Caccamise D. Generalization of knowledge: multidisciplinary perspectives. Psychology Press; New York, 2010.
Ono M, Devilly GJ, Shum DH. A meta-analytic review of overgeneral memory: the role of trauma history, mood, and the presence of posttraumatic stress disorder. Psychol Trauma. 2016;8:157–64.
Barry TJ, Chiu CPY, Raes F, Ricarte J, Lau H. The neurobiology of reduced autobiographical memory specificity. Trends Cogn Sci. 2018;22:1038–49.
King MJ, MacDougall AG, Ferris SM, Levine B, MacQueen GM, McKinnon MC. A review of factors that moderate autobiographical memory performance in patients with major depressive disorder. J Clin Exp Neuropsychol. 2010;32:1122–44.
Bennett M, Vervoort E, Boddez Y, Hermans D, Baeyens F. Perceptual and conceptual similarities facilitate the generalization of instructed fear. J Behav Ther Exp Psychiatry. 2015;48:149–55.
Shohamy D, Wagner AD. Integrating memories in the human brain: hippocampal-midbrain encoding of overlapping events. Neuron. 2008;60:378–89.
Berens SC, Bird CM. The role of the hippocampus in generalizing configural relationships. Hippocampus. 2017;27:223–8.
Ren LY, Meyer MAA, Grayson VS, Gao P, Guedea AL, Radulovic J. Stress-induced generalization of negative memories is mediated by an extended hippocampal circuit. Neuropsychopharmacology. 2022;47:516–23.
Kumaran D, McClelland JL. Generalization through the recurrent interaction of episodic memories: a model of the hippocampal system. Psychol Rev. 2012;119:573–616.
Yu JY, Liu DF, Loback A, Grossrubatscher I, Frank LM. Specific hippocampal representations are linked to generalized cortical representations in memory. Nat Commun. 2018;9:2209.
Sun X, Bernstein MJ, Meng M, Rao S, Sorensen AT, Yao L, et al. Functionally distinct neuronal ensembles within the memory engram. Cell. 2020;181:410–23 e17.
Rossi J, Balthasar N, Olson D, Scott M, Berglund E, Lee CE, et al. Melanocortin-4 receptors expressed by cholinergic neurons regulate energy balance and glucose homeostasis. Cell Metab. 2011;13:195–204.
Corcoran KA, Donnan MD, Tronson NC, Guzman YF, Gao C, Jovasevic V, et al. NMDA receptors in retrosplenial cortex are necessary for retrieval of recent and remote context fear memory. J Neurosci. 2011;31:11655–9.
Golden SA, Covington HE 3rd, Berton O, Russo SJ. A standardized protocol for repeated social defeat stress in mice. Nat Protoc. 2011;6:1183–91.
Goni-Balentziaga O, Perez-Tejada J, Renteria-Dominguez A, Lebena A, Labaka A. Social instability in female rodents as a model of stress related disorders: a systematic review. Physiol Behav. 2018;196:190–9.
Labaka A, Gomez-Lazaro E, Vegas O, Perez-Tejada J, Arregi A, Garmendia L. Reduced hippocampal IL-10 expression, altered monoaminergic activity and anxiety and depressive-like behavior in female mice subjected to chronic social instability stress. Behav Brain Res. 2017;335:8–18.
Shin G, Gomez AM, Al-Hasani R, Jeong YR, Kim J, Xie Z, et al. Flexible near-field wireless optoelectronics as subdermal implants for broad applications in optogenetics. Neuron. 2017;93:509–21.
Jeong JW, McCall JG, Shin G, Zhang Y, Al-Hasani R, Kim M, et al. Wireless optofluidic systems for programmable in vivo pharmacology and optogenetics. Cell. 2015;162:662–74.
Corcoran KA, Frick BJ, Radulovic J, Kay LM. Analysis of coherent activity between retrosplenial cortex, hippocampus, thalamus, and anterior cingulate cortex during retrieval of recent and remote context fear memory. Neurobiol Learn Mem. 2016;127:93–101.
Kay LM, Freeman WJ. Bidirectional processing in the olfactory-limbic axis during olfactory behavior. Behav Neurosci. 1998;112:541–53.
Rojas-Libano D, Frederick DE, Egana JI, Kay LM. The olfactory bulb theta rhythm follows all frequencies of diaphragmatic respiration in the freely behaving rat. Front Behav Neurosci. 2014;8:214.
Tadel F, Baillet S, Mosher JC, Pantazis D, Leahy RM. Brainstorm: a user-friendly application for MEG/EEG analysis. Comput Intell Neurosci. 2011;2011:879716.
Lopes G, Bonacchi N, Frazao J, Neto JP, Atallah BV, Soares S, et al. Bonsai: an event-based framework for processing and controlling data streams. Front Neuroinform. 2015;9:7.
Proulx CD, Aronson S, Milivojevic D, Molina C, Loi A, Monk B, et al. A neural pathway controlling motivation to exert effort. Proc Natl Acad Sci USA. 2018;115:5792–7.
Martianova E, Aronson S, Proulx CD. Multi-fiber photometry to record neural activity in freely-moving animals. J Vis Exp. 2019;152:e60278.
Jovasevic V, Corcoran KA, Leaderbrand K, Yamawaki N, Guedea AL, Chen HJ, et al. GABAergic mechanisms regulated by miR-33 encode state-dependent fear. Nat Neurosci. 2015;18:1265–71.
Lin Y, Bloodgood BL, Hauser JL, Lapan AD, Koon AC, Kim TK, et al. Activity-dependent regulation of inhibitory synapse development by Npas4. Nature. 2008;455:1198–204.
Meyer MAA, Anstotz M, Ren LY, Fiske MP, Guedea AL, Grayson VS, et al. Stress-related memories disrupt sociability and associated patterning of hippocampal activity: a role of hilar oxytocin receptor-positive interneurons. Transl Psychiatry. 2020;10:428.
Duzel E, Penny WD, Burgess N. Brain oscillations and memory. Curr Opin Neurobiol. 2010;20:143–9.
Betterton RT, Broad LM, Tsaneva-Atanasova K, Mellor JR. Acetylcholine modulates gamma frequency oscillations in the hippocampus by activation of muscarinic M1 receptors. Eur J Neurosci. 2017;45:1570–85.
Fisahn A, Pike FG, Buhl EH, Paulsen O. Cholinergic induction of network oscillations at 40 Hz in the hippocampus in vitro. Nature. 1998;394:186–9.
Jing M, Zhang P, Wang G, Feng J, Mesik L, Zeng J, et al. A genetically encoded fluorescent acetylcholine indicator for in vitro and in vivo studies. Nat Biotechnol. 2018;36:726–37.
Fadel JR. Regulation of cortical acetylcholine release: insights from in vivo microdialysis studies. Behav Brain Res. 2011;221:527–36.
Nail-Boucherie K, Dourmap N, Jaffard R, Costentin J. Contextual fear conditioning is associated with an increase of acetylcholine release in the hippocampus of rat. Brain Res Cogn Brain Res. 2000;9:193–7.
Zhang H, Lin SC, Nicolelis MA. Spatiotemporal coupling between hippocampal acetylcholine release and theta oscillations in vivo. J Neurosci. 2010;30:13431–40.
Radiske A, Gonzalez MC, Conde-Ocazionez S, Rossato JI, Kohler CA, Cammarota M. Cross-frequency phase-amplitude coupling between hippocampal theta and gamma oscillations during recall destabilizes memory and renders it susceptible to reconsolidation disruption. J Neurosci. 2020;40:6398–408.
Hersh LB, Shimojo M. Regulation of cholinergic gene expression by the neuron restrictive silencer factor/repressor element-1 silencing transcription factor. Life Sci. 2003;72:2021–8.
Srivas S, Thakur MK. Transcriptional co-repressor SIN3A silencing rescues decline in memory consolidation during scopolamine-induced amnesia. J Neurochem. 2018;145:204–16.
Sauer JF, Struber M, Bartos M. Impaired fast-spiking interneuron function in a genetic mouse model of depression. Elife. 2015;4:e04979.
Voget M, Rummel J, Avchalumov Y, Sohr R, Haumesser JK, Rea E, et al. Altered local field potential activity and serotonergic neurotransmission are further characteristics of the Flinders sensitive line rat model of depression. Behav Brain Res. 2015;291:299–305.
Fitzgerald PJ, Watson BO. Gamma oscillations as a biomarker for major depression: an emerging topic. Transl Psychiatry. 2018;8:177.
Rathouz MM, Vijayaraghavan S, Berg DK. Acetylcholine differentially affects intracellular calcium via nicotinic and muscarinic receptors on the same population of neurons. J Biol Chem. 1995;270:14366–75.
Bell LA, Bell KA, McQuiston AR. Activation of muscarinic receptors by ACh release in hippocampal CA1 depolarizes VIP but has varying effects on parvalbumin-expressing basket cells. J Physiol. 2015;593:197–215.
Ma X, Zhang Y, Wang L, Li N, Barkai E, Zhang X, et al. The firing of theta state-related septal cholinergic neurons disrupt hippocampal ripple oscillations via muscarinic receptors. J Neurosci. 2020;40:3591–603.
Brazhnik ES, Fox SE. Action potentials and relations to the theta rhythm of medial septal neurons in vivo. Exp Brain Res. 1999;127:244–58.
Salani M, Anelli T, Tocco GA, Lucarini E, Mozzetta C, Poiana G, et al. Acetylcholine-induced neuronal differentiation: muscarinic receptor activation regulates EGR-1 and REST expression in neuroblastoma cells. J Neurochem. 2009;108:821–34.
Mitsushima D. Sex differences in the septo-hippocampal cholinergic system in rats: behavioral consequences. Curr Top Behav Neurosci. 2011;8:57–71.
Giacobini E, Pepeu G. Sex and gender differences in the brain cholinergic system and in the response to therapy of Alzheimer disease with cholinesterase inhibitors. Curr Alzheimer Res. 2018;15:1077–84.
Mineur YS, Mose TN, Vanopdenbosch L, Etherington IM, Ogbejesi C, Islam A, et al. Hippocampal acetylcholine modulates stress-related behaviors independent of specific cholinergic inputs. Mol Psychiatry. 2022. (in press).
Dunsmoor JE, Paz R. Fear generalization and anxiety: behavioral and neural mechanisms. Biol Psychiatry. 2015;78:336–43.
Tulving E. Cue-dependent forgetting: When we forget something we once knew, it does not necessarily mean that the memory trace has been lost; it may only be inaccessible. American Scientist. 1974;62:74–82.
Mineur YS, Picciotto MR. The role of acetylcholine in negative encoding bias: too much of a good thing? Eur J Neurosci. 2021;53:114–25.
Weng FJ, Garcia RI, Lutzu S, Alvina K, Zhang Y, Dushko M, et al. Npas4 is a critical regulator of learning-induced plasticity at mossy fiber-CA3 synapses during contextual memory formation. Neuron. 2018;97:1137–52 e5.
Sun X, Lin Y. Npas4: linking neuronal activity to memory. Trends Neurosci. 2016;39:264–75.
Ramamoorthi K, Fropf R, Belfort GM, Fitzmaurice HL, McKinney RM, Neve RL, et al. Npas4 regulates a transcriptional program in CA3 required for contextual memory formation. Science. 2011;334:1669–75.
Alberini CM, Milekic MH, Tronel S. Mechanisms of memory stabilization and de-stabilization. Cell Mol Life Sci. 2006;63:999–1008.
Stein M, Rohde KB, Henke K. Focus on emotion as a catalyst of memory updating during reconsolidation. Behav Brain Sci. 2015;38:e27.
Besnard A, Sahay A. Adult hippocampal neurogenesis, fear generalization, and stress. Neuropsychopharmacology. 2016;41:24–44.
Lissek S, Kaczkurkin AN, Rabin S, Geraci M, Pine DS, Grillon C. Generalized anxiety disorder is associated with overgeneralization of classically conditioned fear. Biol Psychiatry. 2014;75:909–15.
Lissek S. Toward an account of clinical anxiety predicated on basic, neurally mapped mechanisms of Pavlovian fear-learning: the case for conditioned overgeneralization. Depress Anxiety. 2012;29:257–63.
We thank Gail Mandel (Oregon Health & Science University) for providing the REST antibody, Ryan Drenan (Wake Forest School of Medicine) for providing advice with behavioral analyses of Chat-Cre mice, John A. Kessler (Northwestern University) for helping us finalize the immunohistochemistry studies in his lab, and Gordon Shepherd (Northwestern University) for discussions and feedback on the circuit approaches.
LYR performed the behavioral, chemogenetic, and RAM experiments and data analysis and helped writing the manuscript. AC performed the optogenetic and fiber photometry experiments and analyzed the data, HZ performed the LFP experiments and data analyses, MAAM helped with the behavioral experiments and histochemical analyses, PG helped with the virus injection and expression analyses, ZP performed the circuit manipulations and RAM/REST studies, XS and YL provided all of the RAM constructs and shared key expertise in experimental design with RAM manipulations, JR designed the overall study, helped with data analysis, and wrote the manuscript.
This work was funded by NIMH grants MH078064 and MH108837 and Lundbeck Foundation grant R310-2018-3611 to JR, F30MH122130, and T32MH067564 to LR, and NS115543 to YL.
The authors declare no competing interests.
All animal procedures used in this study were approved by the Northwestern University IACUC and Albert Einstein Medical College IACUC and complied with federal regulations set forth by the National Institutes of Health.
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The original online version of this article was revised: In this article the author name Mariah AA Meyer was incorrectly written as Mariah M. A. A. Meyer.
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Ren, L.Y., Cicvaric, A., Zhang, H. et al. Stress-induced changes of the cholinergic circuitry promote retrieval-based generalization of aversive memories. Mol Psychiatry 27, 3795–3805 (2022). https://doi.org/10.1038/s41380-022-01610-x