Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

A preoptic neuronal population controls fever and appetite during sickness

Abstract

During infection, animals exhibit adaptive changes in physiology and behaviour aimed at increasing survival. Although many causes of infection exist, they trigger similar stereotyped symptoms such as fever, warmth-seeking, loss of appetite and fatigue1,2. Yet exactly how the nervous system alters body temperature and triggers sickness behaviours to coordinate responses to infection remains unknown. Here we identify a previously uncharacterized population of neurons in the ventral medial preoptic area (VMPO) of the hypothalamus that are activated after sickness induced by lipopolysaccharide (LPS) or polyinosinic:polycytidylic acid. These neurons are crucial for generating a fever response and other sickness symptoms such as warmth-seeking and loss of appetite. Single-nucleus RNA-sequencing and multiplexed error-robust fluorescence in situ hybridization uncovered the identity and distribution of LPS-activated VMPO (VMPOLPS) neurons and non-neuronal cells. Gene expression and electrophysiological measurements implicate a paracrine mechanism in which the release of immune signals by non-neuronal cells during infection activates nearby VMPOLPS neurons. Finally, we show that VMPOLPS neurons exert a broad influence on the activity of brain areas associated with behavioural and homeostatic functions and are synaptically and functionally connected to circuit nodes controlling body temperature and appetite. Together, these results uncover VMPOLPS neurons as a control hub that integrates immune signals to orchestrate multiple sickness symptoms in response to infection.

Your institute does not have access to this article

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Fig. 1: Activation of a specific VMPO neuronal population after LPS administration.
Fig. 2: Effect of CCL2, PGE2 and IL-1β on intrinsic and synaptic properties of VMPOLPS neurons.
Fig. 3: VMPOLPS neurons drive LPS-induced generation of fever, warmth-seeking behaviour and appetite suppression.
Fig. 4: VMPOLPS neurons regulate body temperature and appetite through direct and indirect synaptic connections.
Fig. 5: Model of the control of body temperature and appetite by VMPOLPS neurons.

Data availability

Raw and processed data have been deposited in the Gene Expression Omnibus and are available under accession number GSE197547.

Code availability

The pipeline used to perform MERFISH analyses is freely available at https://github.com/ZhuangLab/MERlin. Other custom code is available upon request.

References

  1. Konsman, J. P., Parnet, P. & Dantzer, R. Cytokine-induced sickness behaviour: mechanisms and implications. Trends Neurosci. 25, 154–159 (2002).

    CAS  Article  Google Scholar 

  2. McCusker, R. H. & Kelley, K. W. Immune–neural connections: how the immune system’s response to infectious agents influences behavior. J. Exp. Biol. 216, 84–98 (2013).

    CAS  Article  Google Scholar 

  3. Evans, S. S., Repasky, E. A. & Fisher, D. T. Fever and the thermal regulation of immunity: the immune system feels the heat. Nat. Rev. Immunol. 15, 335–349 (2015).

    CAS  Article  Google Scholar 

  4. Quan, N. & Banks, W. A. Brain–immune communication pathways. Brain Behav. Immun. 21, 727–735 (2007).

    CAS  Article  Google Scholar 

  5. Nakamori, T. et al. Organum vasculosum laminae terminalis (OVLT) is a brain site to produce interleukin-1β during fever. Brain Res. 618, 155–159 (1993).

    ADS  CAS  Article  Google Scholar 

  6. Tan, C. L. et al. Warm-sensitive neurons that control body temperature. Cell 167, 47–59.e15 (2016).

    CAS  Article  Google Scholar 

  7. Zhang, Y. et al. Leptin-receptor-expressing neurons in the dorsomedial hypothalamus and median preoptic area regulate sympathetic brown adipose tissue circuits. J. Neurosci. 31, 1873–1884 (2011).

    CAS  Article  Google Scholar 

  8. Zhao, Z. D. et al. A hypothalamic circuit that controls body temperature. Proc. Natl Acad. Sci. USA 114, 2042–2047 (2017).

    CAS  Article  Google Scholar 

  9. Elmquist, J. K., Scammell, T. E., Jacobson, C. D. & Saper, C. B. Distribution of Fos-like immunoreactivity in the rat brain following intravenous lipopolysaccharide administration. J. Comp. Neurol. 371, 85–103 (1996).

    CAS  Article  Google Scholar 

  10. Oka, T. et al. Relationship of EP1-4 prostaglandin receptors with rat hypothalamic cell groups involved in lipopolysaccharide fever responses. J. Comp. Neurol. 428, 20–32 (2000).

    CAS  Article  Google Scholar 

  11. Lazarus, M. et al. EP3 prostaglandin receptors in the median preoptic nucleus are critical for fever responses. Nat. Neurosci. 10, 1131–1133 (2007).

    CAS  Article  Google Scholar 

  12. Machado, N. L. S., Bandaru, S. S., Abbott, S. B. G. & Saper, C. B. EP3R-expressing glutamatergic preoptic neurons mediate inflammatory fever. J. Neurosci. 40, 2573–2588 (2020).

    CAS  Article  Google Scholar 

  13. Moffitt, J. R. et al. Molecular, spatial, and functional single-cell profiling of the hypothalamic preoptic region. Science 362, eaau5324 (2018).

    ADS  Article  Google Scholar 

  14. Chen, K. H., Boettiger, A. N., Moffitt, J. R., Wang, S. & Zhuang, X. RNA imaging. Spatially resolved, highly multiplexed RNA profiling in single cells. Science 348, aaa6090 (2015).

    Article  Google Scholar 

  15. Butler, A., Hoffman, P., Smibert, P., Papalexi, E. & Satija, R. Integrating single-cell transcriptomic data across different conditions, technologies, and species. Nat. Biotechnol. 36, 411–420 (2018).

    CAS  Article  Google Scholar 

  16. Konsman, J. P., Tridon, V. & Dantzer, R. Diffusion and action of intracerebroventricularly injected interleukin-1 in the CNS. Neuroscience 101, 957–967 (2000).

    CAS  Article  Google Scholar 

  17. Kis, B. et al. Effects of LPS stimulation on the expression of prostaglandin carriers in the cells of the blood–brain and blood–cerebrospinal fluid barriers. J. Appl. Physiol. 100, 1392–1399 (2006).

    CAS  Article  Google Scholar 

  18. Zywitza, V., Misios, A., Bunatyan, L., Willnow, T. E. & Rajewsky, N. Single-cell transcriptomics characterizes cell types in the subventricular zone and uncovers molecular defects impairing adult neurogenesis. Cell Rep. 25, 2457–2469.e8 (2018).

    CAS  Article  Google Scholar 

  19. Roessmann, U., Velasco, M. E., Sindely, S. D. & Gambetti, P. Glial fibrillary acidic protein (GFAP) in ependymal cells during development. An immunocytochemical study. Brain Res. 200, 13–21 (1980).

    CAS  Article  Google Scholar 

  20. Duan, L. et al. PDGFRβ cells rapidly relay inflammatory signal from the circulatory system to neurons via chemokine CCL2. Neuron 100, 183–200.e8 (2018).

    CAS  Article  Google Scholar 

  21. Quan, N., Stern, E. L., Whiteside, M. B. & Herkenham, M. Induction of pro-inflammatory cytokine mRNAs in the brain after peripheral injection of subseptic doses of lipopolysaccharide in the rat. J. Neuroimmunol. 93, 72–80 (1999).

    CAS  Article  Google Scholar 

  22. Wilhelms, D. B. et al. Deletion of prostaglandin E2 synthesizing enzymes in brain endothelial cells attenuates inflammatory fever. J. Neurosci. 34, 11684–11690 (2014).

    CAS  Article  Google Scholar 

  23. Hojen, J. F. et al. IL-1R3 blockade broadly attenuates the functions of six members of the IL-1 family, revealing their contribution to models of disease. Nat. Immunol. 20, 1138–1149 (2019).

    Article  Google Scholar 

  24. Davis, C. J. et al. The neuron-specific interleukin-1 receptor accessory protein is required for homeostatic sleep and sleep responses to influenza viral challenge in mice. Brain Behav. Immun. 47, 35–43 (2015).

    CAS  Article  Google Scholar 

  25. Liege, S., Laye, S., Li, K. S., Moze, E. & Neveu, P. J. Interleukin 1 receptor accessory protein (IL-1RAcP) is necessary for centrally mediated neuroendocrine and immune responses to IL-1β. J. Neuroimmunol. 110, 134–139 (2000).

    CAS  Article  Google Scholar 

  26. Allen, W. E. et al. Thirst-associated preoptic neurons encode an aversive motivational drive. Science 357, 1149–1155 (2017).

    ADS  CAS  Article  Google Scholar 

  27. DeNardo, L. A. et al. Temporal evolution of cortical ensembles promoting remote memory retrieval. Nat. Neurosci. 22, 460–469 (2019).

    CAS  Article  Google Scholar 

  28. Molina-Holgado, E., Ortiz, S., Molina-Holgado, F. & Guaza, C. Induction of COX-2 and PGE2 biosynthesis by IL-1β is mediated by PKC and mitogen-activated protein kinases in murine astrocytes. Br. J. Pharmacol. 131, 152–159 (2000).

    ADS  CAS  Article  Google Scholar 

  29. Wu, Z., Autry, A. E., Bergan, J. F., Watabe-Uchida, M. & Dulac, C. G. Galanin neurons in the medial preoptic area govern parental behaviour. Nature 509, 325–330 (2014).

    ADS  CAS  Article  Google Scholar 

  30. Kozak, W., Conn, C. A. & Kluger, M. J. Lipopolysaccharide induces fever and depresses locomotor activity in unrestrained mice. Am. J. Physiol. 266, R125–R135 (1994).

    CAS  PubMed  Google Scholar 

  31. Akins, C., Thiessen, D. & Cocke, R. Lipopolysaccharide increases ambient temperature preference in C57BL/6J adult mice. Physiol. Behav. 50, 461–463 (1991).

    CAS  Article  Google Scholar 

  32. Liu, Y. et al. Lipopolysacharide rapidly and completely suppresses AgRP neuron-mediated food intake in male mice. Endocrinology 157, 2380–2392 (2016).

    CAS  Article  Google Scholar 

  33. Yizhar, O. et al. Neocortical excitation/inhibition balance in information processing and social dysfunction. Nature 477, 171–178 (2011).

    ADS  CAS  Article  Google Scholar 

  34. Andermann, M. L. & Lowell, B. B. Toward a wiring diagram understanding of appetite control. Neuron 95, 757–778 (2017).

    CAS  Article  Google Scholar 

  35. Pinol, R. A. et al. Brs3 neurons in the mouse dorsomedial hypothalamus regulate body temperature, energy expenditure, and heart rate, but not food intake. Nat. Neurosci. 21, 1530–1540 (2018).

    CAS  Article  Google Scholar 

  36. Farzi, A. et al. Arcuate nucleus and lateral hypothalamic CART neurons in the mouse brain exert opposing effects on energy expenditure. eLife 7, e36494 (2018).

    Article  Google Scholar 

  37. Millington, G. W. The role of proopiomelanocortin (POMC) neurones in feeding behaviour. Nutr. Metab. 4, 18 (2007).

    Article  Google Scholar 

  38. Kapoor, V., Provost, A. C., Agarwal, P. & Murthy, V. N. Activation of raphe nuclei triggers rapid and distinct effects on parallel olfactory bulb output channels. Nat. Neurosci. 19, 271–282 (2016).

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank K. Prichett-Corning and other veterinary staff at the Harvard Office of Animal Resources for advice on working with sick mice; R. Hellmiss and staff at MCB Graphics for help with illustrations; A. Emery and J. Emery for assistance with histology and RNAscope experiments; members of the Dulac laboratory for helpful advice on experiments, analysis and the manuscript; and F. Engert and R. Losick for comments on the manuscript. This work was supported by NIH awards K99NS114107 to J.A.O. and F31MH120911 to E.V., NIH grant R01NS050835 to L.L, and NIH grant R01NS112399 and Simons Foundation Award 572189 to C.D. C.D., X.Z. and L.L. are investigators at the Howard Hughes Medical Institute.

Author information

Authors and Affiliations

Authors

Contributions

J.A.O. and C.D. conceived and designed the study. J.A.O. performed and analysed tracing, chemogenetic and ablation experiments and associated behavioural assays. J.A.O., C.D., S.W.E. and X.Z. designed the MERFISH experiments and analysis. S.W.E. performed and analysed all MERFISH experiments. J.A.O. and D. Lee. generated cDNA libraries for the snRNA-seq experiments. D. Lee and E.V. performed the snRNA-seq analysis. V.K. performed electrophysiological experiments and analysis. J.A.O. and D. Liu performed the optogenetic experiments. J.D.M. wrote the Matlab program for Fos expression quantification. J.A.O. and D. Lee performed and analysed the immunohistochemistry and in situ hybridization experiments. L.A.D. and L.L. shared the unpublished TRAP2 transgenic mouse line. J.A.O. and C.D. wrote the manuscript with input from all authors.

Corresponding author

Correspondence to Catherine Dulac.

Ethics declarations

Competing interests

X.Z. is an inventor on patents applied for by Harvard University related to MERFISH.

Peer review

Peer review information

Nature thanks Jan Siemens and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data figures and tables

Extended Data Fig. 1 Specificity and identity of VMPOLPS neurons during inflammation.

(a) Mean body temperature 2 h following injection of saline (n = 13), LPS (n = 16) or Poly(I:C) (n = 10). (b-c) Representative images of Fos expression in brain areas displaying significant increases in number of Fos+ cells following LPS administration (b) or saline (c), scale bar for all panels = 200μm. (d) mRNA expression of inhibitory neuronal marker Vgat (green) and Fos (magenta) in the VMPO after LPS injection. (e) Fraction of Fos+ cells that express Vgat or Vglut2 (n = 4). (f) Quantification of the fraction of Fos+ cells within individual snRNA-seq clusters in LPS-injected sample. (g) Dotplot of average expression of marker genes and genes with immunological significance in each cluster: dot size indicates percent of cells in cluster with measurable expression and color indicates average expression levels; ependymal cluster cluster #5 and neuronal cluster #19, found significantly activated after LPS injection are highlighted in red. (h) Quantification of the fraction of Fos+ cells in individual MERFISH neuronal clusters, with statically significant enrichment for Fos+ cells indicated in red, n = 3. (i) mRNA expression of markers for VMPOLPS neurons (calcR, gal and amigo2) in LPS-injected mice. (j) Mean of overlap of markers for VMPOLPS neurons and for warm-sensitive neurons (adcyap1) with LPS-mediated Fos expression in the VMPO, n = 3 mice/experiment. All scale bars = 200μm. All error bars = SEM.

Extended Data Fig. 2 Spatial distribution of activated neuronal and non-neuronal cell type classes in the preoptic area during inflammation.

(a) Cumulative distribution of Fos+ (purple) and Fos- (blue) VMPOGal/Amigo2 neurons as a function of the distance to the bottom of the section (top) or the midline (bottom), n = 3 replicates. (b) MERFISH analysis indicating the fraction of Fos+ cells in major cell type classes in samples from mice injected with LPS versus mice displaying other behaviors: aggression, mating or parenting. Significantly activated populations are indicated in red. (c) Spatial distribution of major cell type classes in MERFISH analysis, cell type (green), Fos+ cell (red), Fos+ cells in the indicated cell type (purple). (d) Cumulative distribution of Fos+ (purple) and Fos- (green) cells in major cell type classes as a function of the distance to the bottom of the section (top) or the midline (bottom), n = 3 mice. All scale bars = 200μm. All error bars = SEM.

Extended Data Fig. 3 Expression of key immune molecules and their receptors in the VMPO.

(a-i) mRNA expression in the VMPO at 2hrs post LPS administration, genes of interest in green, LPS-induced Fos is in magenta. Scale bar for a-i = 200μm (a) Prostaglandin E synthase 2 (ptgs2) and its receptors ep2 (d), ep1 (g), ep3 (h), ep4 (i). (b) interleukin-1β (il1β) and its receptor il1rap (e). (c) chemokine ligand 2 (ccl2) and it’s receptor chemkine receptor 2 (ccr2) (f). (j) Expression of ptgs2 in absence of LPS stimulation and at 60min and 120min post LPS. No or weak expression is found without LPS stimulation, and expression after LPS challenge shows overlap with markers for endothelial cells and microglia. (k) Expression of il1β with no stimulation and at 60min and 120min post LPS, overlap with markers for ependymal cells and microglia. Scale bar for j-l = 50μm (l) Expression of ccl2 with no stimulation and 120 min post LPS inejction, overlap with gfap+ astrocytes. (m) Expression of il1r1 in the VMPO after LPS injection, Scale bar = 200μm.

Extended Data Fig. 4 Effects of PGE-2 and cytokines on the intrinsic properties and synaptic activity of VMPOLPS neurons.

(a) Wide field microscope images depicting tdTomato expression in the VMPO of TRAP2;Ai9 during whole cell patch clamp recording with pipette containing Alexa 488. (b) Changes in rheobase current for VMPOLPS neurons in presence of ACSF (black), CCL2 (pink), PGE2 (green), addition of EP2 antagonist in presence of PGE2 (blue), IL-1β (orange) and further addition of PGE2 (yellow), and (c) IL-1β (orange), addition of COX-2 inhibitor in the presence of IL-1β (light orange), further addition of PGE2 (light green) and further addition of EP2 antagonist (light blue). During whole cell voltage clamp recordings, changes in (d) amplitude of miniature EPSCs, (e) inter event interval for miniature EPSCs, (f) amplitude of miniature IPSCs and (g) inter event interval for miniature IPSCs for VMPOLPS neurons in presence of ACSF (black), CCL2 (pink), PGE2 (green), addition of EP2 antagonist in presence of PGE2 (blue), IL-1β (orange) and further addition of PGE2 (yellow). Changes in (h) amplitude of miniature EPSCs, (i) inter event interval for miniature EPSCs, (j) amplitude of miniature IPSCs and (k) inter event interval for miniature IPSCs for VMPOLPS neurons in the presence of IL-1β and COX-2 inhibitor (light orange), further addition of PGE2 (light green) and further addition of EP2 antagonist (light blue). (l) Cumulative plots showing total charge transferred over 1s for inhibitory events and excitatory events for control (black) and CCL2 (pink). (m, top) Cumulative charge transferred by IPSCs for control (mean value of 24.81 ± 0.14 μC/s) and CCL2 (mean value of 19.06 ± 0.09 μC/s p = 2.0 * 10−03 for). (m, bottom) Cumulative charge transferred by miniature EPSCs for control (mean value of 18.81 ± 0.09 μC/s) and CCL2 (mean value of 23.37 ± 0.11 μC/s p = 9.0 * 10−03). All other values and statistical tests are catalogued in Extended Data Table 2. For violin plots, central white circle depicts the mean value and thick black line depicts the interquartile range. All p values shown are for two-sided Wilcoxon rank sum test with * = p< 0.05, ** = p< 0, *** = p< 0.005 and ns = p > 0.05. All error bars = SEM. See Statistics and Reproducibility section for exact n values.

Extended Data Fig. 5 Specificity of viral injections and cell type identity.

(a) Example of viral injection specificity that qualified for inclusion in the study. Note viral expression in the VMPO and absence in surrounding brain areas. Injections found to be more broadly dispersed were not included in analysis. (b-d) Chemogenetic activation of Calcr+ and Gal+ neurons in the VMPO. (b) CNO injection elicited strong increase in body temperature in both genetic backgrounds (n = 6 mice/group). (c) Activation of Gal-Cre but not Calcr-Cre increased preferred temperature (n = 7mice/group). (d) Activation of Calcr-Cre (n = 6 mice) but not Gal-Cre (n = 8) decreased chow consumed (saline n = 6 mice). (e) DTA-mediated Ablation of VMPOLPS neurons had no effect on circadian temperatures, dark bar indicates dark phase, WT mice were used as controls. (f-g) Effect of DTA-mediated ablation of VMPO neurons in Saline-TRAP mice had no effect on body temperature or preferred temperature following LPS injection (n = 7 mice/group). (h) Overlap of warm-TRAP reporter expression with markers of warm-sensitive neurons adcyap1 and sncg. (i) Overlap of hunger-TRAP-mediated reporter expression (magenta) with markers of appetite-increasing neurons, agrp, or appetite-decreasing neurons, pomc, (green). Scale bars = 200μm. All error bars = SEM. For all graphs * = p< 0.05, ** = p< 0.01, *** = p< 0.001.

Extended Data Fig. 6 Chemogenetic and optogenetic activation of VMPOLPS neurons.

(a) Widefield microscope images of VMPOLPS neurons in the VMPO of TRAP2 injected with AAV8-hSyn-DIO-hM3D(Gq)-mCherry. (b) Whole cell current clamp recording of a VMPOLPS neuron showing the effects of bath application of 1mM CNO (baseline: black, CNO: pink and washout: green). (c) Effects of CNO application on the firing rate of VMPOLPS neurons; black (n = 5 cells with mean values of 0.20 ± 0.05 Hz for baseline), pink (0.91 ± 0.06 Hz for CNO application and p = 7.0 * 10−03 compared to baseline), green (0.25 ± 0.06 Hz for washout and p = 6.9 * 10−03 compared to CNO application), boxes represent the 25th and 75th percentile, whiskers extend to the minimum and maximum data points (d) Widefield microscope images of VMPOLPS neurons in the VMPO of TRAP2 injected with AAV5-EF1a-DIO-hChR2(C128S/D156A)-EYFP. (e) Whole cell current clamp recording of a VMPOLPS neuron showing activation of hChR2(C128S/D156A) by 20 ms blue light (460 ± 10 nm) light pulse and inactivation by 50 ms green light (525 ± 9 nm) pulse. (f) Firing rate of VMPOLPS neurons (n = 4 cells) in response to activation of hChR2(C128S/D156A), baseline (black, mean firing frequency of 0.21 ± 0.08 Hz), following blue light activation (blue, mean firing frequency of 1.46 ± 0.72 Hz and p = 2.1 * 10−02) and following green light mediated inactivation of hChR2(C128S/D156A) (green, mean firing frequency of 0.12 ± 0.05 Hz and p = 2.0 * 10−02), boxes represent the 25th and 75th percentile, whiskers extend to the minimum and maximum data points. All p values shown are for two-sided Wilcoxon rank sum test with * = p< 0.05, ** = p< 0, *** = p< 0.005 and ns = p > 0.05.

Extended Data Table 1 Effects of prostaglandin E2 and cytokines (Il-1β and CCL2) on the intrinsic properties of VMPOLPS neurons
Extended Data Table 2 Effects of prostaglandin E2 and cytokines (Il-1β and CCL2) on the synaptic inputs to VMPOLPS neurons
Extended Data Table 3 Abbreviations of brain areas
Extended Data Table 4 List of drugs
Extended Data Table 5 Precise animal numbers for Fig. 3
Extended Data Table 6 Precise cell numbers for Fig. 2 and Extended Data Fig. 4

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Osterhout, J.A., Kapoor, V., Eichhorn, S.W. et al. A preoptic neuronal population controls fever and appetite during sickness. Nature 606, 937–944 (2022). https://doi.org/10.1038/s41586-022-04793-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41586-022-04793-z

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing