Abstract
Both the medial prefrontal cortex (mPFC) and thalamus have been implicated in pain regulation. However, the roles of the mPFC-thalamus connection in pain and how the mPFC modulates nociceptive processing remain unclear. Here, we show that the mPFC neurons projecting to thalamus, marked by Foxp2 expression, are deactivated in both acute and chronic pain in male mice. Persistent inactivation of the mPFC Foxp2+ neurons enhances nociceptive sensitivity, while their activation alleviates multiple aspects of pain. Circuit-specific manipulations revealed that the projections to parataenial nucleus, mediodorsal and ventromedial thalamus differentially modulate sensory and affective pain. Additionally, the mPFC Foxp2+ neurons receive cholinergic input from the basal forebrain, particularly the horizontal diagonal band (HDB). Notably, activation of the α4β2-containing nicotinic acetylcholine receptor in mPFC exerts antinociceptive effects in Foxp2+ neuron-dependent manner. Together, our study defines an HDB→mPFCFoxp2→thalamus circuit essential for sensory and affective pain modulation and underscores the therapeutic potential of targeting mPFC cholinergic signaling in chronic pain management.
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All data necessary to evaluate the conclusions of this study are available in the main text or the Supplementary Information files. Source data are provided with this paper.
References
Raja, S. N. et al. The revised International Association for the Study of Pain definition of pain: concepts, challenges, and compromises. Pain 161, 1976–1982 (2020).
Treede, R. D. et al. Chronic pain as a symptom or a disease: the IASP Classification of Chronic Pain for the International Classification of Diseases (ICD-11). Pain 160, 19–27 (2019).
Kang, Y., Trewern, L., Jackman, J., McCartney, D. & Soni, A. Chronic pain: definitions and diagnosis. BMJ 381, e076036 (2023).
Cohen, S. P., Vase, L. & Hooten, W. M. Chronic pain: an update on burden, best practices, and new advances. Lancet 397, 2082–2097 (2021).
Kuner, R. & Kuner, T. Cellular circuits in the brain and their modulation in acute and chronic pain. Physiol. Rev. 101, 213–258 (2021).
Descalzi, G. et al. Epigenetic mechanisms of chronic pain. Trends Neurosci. 38, 237–246 (2015).
Mercer Lindsay, N., Chen, C., Gilam, G., Mackey, S. & Scherrer, G. Brain circuits for pain and its treatment. Sci. Transl. Med. 13, eabj7360 (2021).
Tan, L. L. & Kuner, R. Neocortical circuits in pain and pain relief. Nat. Rev. Neurosci. 22, 458–471 (2021).
Kummer, K. K., Mitric, M., Kalpachidou, T. & Kress, M. The medial prefrontal cortex as a central hub for mental comorbidities associated with chronic pain. Int. J. Mol. Sci. 21, 3440 (2020).
Ong, W. Y., Stohler, C. S. & Herr, D. R. Role of the prefrontal cortex in pain processing. Mol. Neurobiol. 56, 1137–1166 (2019).
Kuner, R. & Flor, H. Structural plasticity and reorganisation in chronic pain. Nat. Rev. Neurosci. 18, 20–30 (2016).
Apkarian, A. V. et al. Chronic back pain is associated with decreased prefrontal and thalamic gray matter density. J. Neurosci. 24, 10410–10415 (2004).
Seminowicz, D. A. & Moayedi, M. The dorsolateral prefrontal cortex in acute and chronic pain. J. Pain 18, 1027–1035 (2017).
Moisset, X. & Bouhassira, D. Brain imaging of neuropathic pain. Neuroimage 37, S80–S88 (2007).
Ji, G. & Neugebauer, V. Pain-related deactivation of medial prefrontal cortical neurons involves mGluR1 and GABA(A) receptors. J. Neurophysiol. 106, 2642–2652 (2011).
Wang, G. Q. et al. Deactivation of excitatory neurons in the prelimbic cortex via Cdk5 promotes pain sensation and anxiety. Nat. Commun. 6, 7660 (2015).
Huang, J. et al. A neuronal circuit for activating descending modulation of neuropathic pain. Nat. Neurosci. 22, 1659–1668 (2019).
Cheriyan, J. & Sheets, P. L. Altered excitability and local connectivity of mPFC-PAG neurons in a mouse model of neuropathic pain. J. Neurosci. 38, 4829–4839 (2018).
Anastasiades, P. G. & Carter, A. G. Circuit organization of the rodent medial prefrontal cortex. Trends Neurosci. 44, 550–563 (2021).
Ma, Q. A functional subdivision within the somatosensory system and its implications for pain research. Neuron 110, 749–769 (2022).
Gustin, S. M. et al. Different pain, different brain: thalamic anatomy in neuropathic and non-neuropathic chronic pain syndromes. J. Neurosci. 31, 5956–5964 (2011).
Gan, Z. et al. Layer-specific pain relief pathways originating from primary motor cortex. Science 378, 1336–1343 (2022).
Meda, K. S. et al. Microcircuit mechanisms through which mediodorsal thalamic input to anterior cingulate cortex exacerbates pain-related aversion. Neuron 102, 944–959.e943 (2019).
Spellman, T., Svei, M., Kaminsky, J., Manzano-Nieves, G. & Liston, C. Prefrontal deep projection neurons enable cognitive flexibility via persistent feedback monitoring. Cell 184, 2750–2766.e2717 (2021).
Lui, J. H. et al. Differential encoding in prefrontal cortex projection neuron classes across cognitive tasks. Cell 184, 489–506.e426 (2021).
Ballinger, E. C., Ananth, M., Talmage, D. A. & Role, L. W. Basal forebrain cholinergic circuits and signaling in cognition and cognitive decline. Neuron 91, 1199–1218 (2016).
Bannon, A. W. et al. Broad-spectrum, non-opioid analgesic activity by selective modulation of neuronal nicotinic acetylcholine receptors. Science 279, 77–81 (1998).
Pan, Q. et al. Representation and control of pain and itch by distinct prefrontal neural ensembles. Neuron 111, 2414–2431.e2417 (2023).
Li, M., Zhou, H., Teng, S. & Yang, G. Activation of VIP interneurons in the prefrontal cortex ameliorates neuropathic pain aversiveness. Cell Rep. 40, 111333 (2022).
Harris, K. D. & Shepherd, G. M. The neocortical circuit: themes and variations. Nat. Neurosci. 18, 170–181 (2015).
Hodge, R. D. et al. Conserved cell types with divergent features in human versus mouse cortex. Nature 573, 61–68 (2019).
Harris, J. A. et al. Hierarchical organization of cortical and thalamic connectivity. Nature 575, 195–202 (2019).
Sundberg, S. C., Lindstrom, S. H., Sanchez, G. M. & Granseth, B. Cre-expressing neurons in visual cortex of Ntsr1-Cre GN220 mice are corticothalamic and are depolarized by acetylcholine. J. Comp. Neurol. 526, 120–132 (2018).
Olsen, S. R., Bortone, D. S., Adesnik, H. & Scanziani, M. Gain control by layer six in cortical circuits of vision. Nature 483, 47–52 (2012).
Bhattacherjee, A. et al. Cell type-specific transcriptional programs in mouse prefrontal cortex during adolescence and addiction. Nat. Commun. 10, 4169 (2019).
Bhattacherjee, A. et al. Spatial transcriptomics reveals the distinct organization of mouse prefrontal cortex and neuronal subtypes regulating chronic pain. Nat. Neurosci. 26, 1880–1893 (2023).
Tasic, B. et al. Shared and distinct transcriptomic cell types across neocortical areas. Nature 563, 72–78 (2018).
Hisaoka, T., Nakamura, Y., Senba, E. & Morikawa, Y. The forkhead transcription factors, Foxp1 and Foxp2, identify different subpopulations of projection neurons in the mouse cerebral cortex. Neuroscience 166, 551–563 (2010).
Babiczky, A. & Matyas, F. Molecular characteristics and laminar distribution of prefrontal neurons projecting to the mesolimbic system. eLife 11, e78813 (2022).
Kast, R. J., Lanjewar, A. L., Smith, C. D. & Levitt, P. FOXP2 exhibits projection neuron class specific expression, but is not required for multiple aspects of cortical histogenesis. eLife 8, e42012 (2019).
Yao, Z. et al. A transcriptomic and epigenomic cell atlas of the mouse primary motor cortex. Nature 598, 103–110 (2021).
Hunskaar, S. & Hole, K. The formalin test in mice: dissociation between inflammatory and non-inflammatory pain. Pain 30, 103–114 (1987).
Liu, J. et al. Activation of parvalbumin neurons in the rostro-dorsal sector of the thalamic reticular nucleus promotes sensitivity to pain in mice. Neuroscience 366, 113–123 (2017).
Poorthuis, R. B. et al. Layer-specific modulation of the prefrontal cortex by nicotinic acetylcholine receptors. Cereb. Cortex 23, 148–161 (2013).
Guillem, K. et al. Nicotinic acetylcholine receptor β2 subunits in the medial prefrontal cortex control attention. Science 333, 888–891 (2011).
Bloem, B., Poorthuis, R. B. & Mansvelder, H. D. Cholinergic modulation of the medial prefrontal cortex: the role of nicotinic receptors in attention and regulation of neuronal activity. Front. Neural Circuits 8, 17 (2014).
Oswald, M. J. et al. Cholinergic basal forebrain nucleus of Meynert regulates chronic pain-like behavior via modulation of the prelimbic cortex. Nat. Commun. 13, 5014 (2022).
Radzicki, D., Pollema-Mays, S. L., Sanz-Clemente, A. & Martina, M. Loss of M1 receptor dependent cholinergic excitation contributes to mPFC deactivation in neuropathic pain. J. Neurosci. 37, 2292–2304 (2017).
Peron, S. et al. Recurrent interactions in local cortical circuits. Nature 579, 256–259 (2020).
Le Merre, P., Ahrlund-Richter, S. & Carlen, M. The mouse prefrontal cortex: Unity in diversity. Neuron 109, 1925–1944 (2021).
Smith, A. C. W. et al. A master regulator of opioid reward in the ventral prefrontal cortex. Science 384, eadn0886 (2024).
Hadinger, N. et al. Region-selective control of the thalamic reticular nucleus via cortical layer 5 pyramidal cells. Nat. Neurosci. 26, 116–130 (2023).
Sherman, S. M. Thalamus plays a central role in ongoing cortical functioning. Nat. Neurosci. 19, 533–541 (2016).
Collins, D. P., Anastasiades, P. G., Marlin, J. J. & Carter, A. G. Reciprocal circuits linking the prefrontal cortex with dorsal and ventral thalamic nuclei. Neuron 98, 366–379 e364 (2018).
Gao, F. et al. Elevated prelimbic cortex-to-basolateral amygdala circuit activity mediates comorbid anxiety-like behaviors associated with chronic pai. J. Clin. Invest. 133, e166356 (2023).
Zhou, H. et al. Inhibition of the prefrontal projection to the nucleus accumbens enhances pain sensitivity and affect. Front. Cell Neurosci. 12, 240 (2018).
De Ridder, D., Adhia, D. & Vanneste, S. The anatomy of pain and suffering in the brain and its clinical implications. Neurosci. Biobehav. Rev. 130, 125–146 (2021).
Naser, P. V. & Kuner, R. Molecular, cellular and circuit basis of cholinergic modulation of pain. Neuroscience 387, 135–148 (2018).
Sullere, S., Kunczt, A. & McGehee, D. S. A cholinergic circuit that relieves pain despite opioid tolerance. Neuron 111, 3414–3434.e3415 (2023).
Umana, I. C., Daniele, C. A. & McGehee, D. S. Neuronal nicotinic receptors as analgesic targets: it’s a winding road. Biochem. Pharma 86, 1208–1214 (2013).
Nirogi, R., Goura, V., Abraham, R. & Jayarajan, P. alpha4beta2* neuronal nicotinic receptor ligands (agonist, partial agonist and positive allosteric modulators) as therapeutic prospects for pain. Eur. J. Pharmacol. 712, 22–29 (2013).
Mansvelder, H. D., Keath, J. R. & McGehee, D. S. Synaptic mechanisms underlie nicotine-induced excitability of brain reward areas. Neuron 33, 905–919 (2002).
Bailey, C. D., Alves, N. C., Nashmi, R., De Biasi, M. & Lambe, E. K. Nicotinic α5 subunits drive developmental changes in the activation and morphology of prefrontal cortex layer VI neurons. Biol. Psychiatry 71, 120–128 (2012).
Ramirez-Latorre, J. et al. Functional contributions of alpha5 subunit to neuronal acetylcholine receptor channels. Nature 380, 347–351 (1996).
Labrakakis, C. The role of the insular cortex in pain. Int. J. Mol. Sci. 24, 5736 (2023).
Liu, Y. et al. A subset of dopamine receptor-expressing neurons in the nucleus accumbens controls feeding and energy homeostasis. Nat. Metab. 6, 1616–1631 (2024).
Zhao, Z. D. et al. A molecularly defined D1 medium spiny neuron subtype negatively regulates cocaine addiction. Sci. Adv. 8, eabn3552 (2022).
Corder, G. et al. An amygdalar neural ensemble that encodes the unpleasantness of pain. Science 363, 276–281 (2019).
Chaplan, S. R., Bach, F. W., Pogrel, J. W., Chung, J. M. & Yaksh, T. L. Quantitative assessment of tactile allodynia in the rat paw. J. Neurosci. Methods 53, 55–63 (1994).
Acknowledgements
We thank the support of the Mouse Behavior Core of Harvard Medical School and its director, Dr. Barbara Caldarone. We thank Dr. Zhengdong Zhao for the help in using in vivo miniscopic calcium recording. This project was supported by the Open Philanthropy Foundation, the National Institutes of Health (1R01DA050589), and the HHMI. Y.Z. is an investigator of the Howard Hughes Medical Institute. This article is subject to HHMI’s Open Access to Publications policy. HHMI lab heads have previously granted a nonexclusive CC BY 4.0 license to the public and a sublicensable license to HHMI in their research articles. Pursuant to those licenses, the author-accepted manuscript of this article can be made freely available under a CC BY 4.0 license immediately upon publication.
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Y.Z. conceived the project; G.X. designed the experiments; G.X. and Y.L. performed the experiments; X.Q. and G.X. analyzed the calcium imaging results; A.B. acquired the Foxp2-Cre mouse line and conducted preliminary staining and projection mapping; C.Z. analyzed the transcriptomic dataset. All authors are involved in data interpretation. G.X. and Y.Z. wrote the manuscript with input from all authors.
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Xie, G., Liu, Y., Qi, X. et al. A molecularly defined basalo-prefrontal-thalamic circuit regulates sensory and affective dimensions of pain in male mice. Nat Commun (2026). https://doi.org/10.1038/s41467-026-69001-2
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DOI: https://doi.org/10.1038/s41467-026-69001-2
