Molecular Medicine

, Volume 20, Issue 1, pp 601–611 | Cite as

Brain Region-Specific Alterations in the Gene Expression of Cytokines, Immune Cell Markers and Cholinergic System Components during Peripheral Endotoxin-Induced Inflammation

  • Harold A. Silverman
  • Meghan Dancho
  • Angelique Regnier-Golanov
  • Mansoor Nasim
  • Mahendar Ochani
  • Peder S. Olofsson
  • Mohamed Ahmed
  • Edmund J. Miller
  • Sangeeta S. Chavan
  • Eugene Golanov
  • Christine N. Metz
  • Kevin J. Tracey
  • Valentin A. Pavlov
Research Article


Inflammatory conditions characterized by excessive peripheral immune responses are associated with diverse alterations in brain function, and brain-derived neural pathways regulate peripheral inflammation. Important aspects of this bidirectional peripheral immune-brain communication, including the impact of peripheral inflammation on brain region-specific cytokine responses, and brain cholinergic signaling (which plays a role in controlling peripheral cytokine levels), remain unclear. To provide insight, we studied gene expression of cytokines, immune cell markers and brain cholinergic system components in the cortex, cerebellum, brainstem, hippocampus, hypothalamus, striatum and thalamus in mice after an intraperitoneal lipopolysaccharide injection. Endotoxemia was accompanied by elevated serum levels of interleukin (IL)-1β, IL-6 and other cytokines and brain region-specific increases in Il1b (the highest increase, relative to basal level, was in cortex; the lowest increase was in cerebellum) and Il6 (highest increase in cerebellum; lowest increase in striatum) mRNA expression. Gene expression of brain Gfap (astrocyte marker) was also differentially increased. However, Iba1 (microglia marker) mRNA expression was decreased in the cortex, hippocampus and other brain regions in parallel with morphological changes, indicating microglia activation. Brain choline acetyltransferase (Chat) mRNA expression was decreased in the striatum, acetylcholinesterase (Ache) mRNA expression was decreased in the cortex and increased in the hippocampus, and M1 muscarinic acetylcholine receptor (Chrm1) mRNA expression was decreased in the cortex and the brainstem. These results reveal a previously unrecognized regional specificity in brain immunoregulatory and cholinergic system gene expression in the context of peripheral inflammation and are of interest for designing future antiinflammatory approaches.



This work was supported by the following grants from the National Institute of General Medical Sciences, National Institutes of Health: R01GM057226 (to KJ Tracey) and R01GM089807 (to VA Pavlov).

Supplementary material

10020_2014_2001601_MOESM1_ESM.pdf (447 kb)
Supplementary material, approximately 447 KB.


  1. 1.
    Tracey KJ. (2002) The inflammatory reflex. Nature. 420:853–9.CrossRefGoogle Scholar
  2. 2.
    Pavlov VA, Tracey KJ. (2012) The vagus nerve and the inflammatory reflex: linking immunity and metabolism. Nat. Rev. Endocrinol. 8:743–54.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    McInnes IB, Schett G. (2007) Cytokines in the pathogenesis of rheumatoid arthritis. Nat. Rev. Immunol. 7:429–42.CrossRefPubMedGoogle Scholar
  4. 4.
    Diamond B, Volpe BT. (2012) A model for lupus brain disease. Immunol. Rev. 248:56–67.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Chavan SS, et al. (2012) HMGB1 mediates cognitive impairment in sepsis survivors. Mol. Med. 18:930–7.PubMedPubMedCentralGoogle Scholar
  6. 6.
    Sonneville R, et al. (2013) Understanding brain dysfunction in sepsis. Ann. Intensive Care. 3:15.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Strachan MW, Reynolds RM, Marioni RE, Price JF. (2011) Cognitive function, dementia and type 2 diabetes mellitus in the elderly. Nat. Rev. Endocrinol. 7:108–14.CrossRefPubMedGoogle Scholar
  8. 8.
    Samaras K, Sachdev PS. (2012) Diabetes and the elderly brain: sweet memories? Ther. Adv. Endocrinol. Metab. 3:189–96.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Lampa J, et al. (2012) Peripheral inflammatory disease associated with centrally activated IL-1 system in humans and mice. Proc. Natl. Acad. Sci. U. S. A. 109:12728–33.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Hess A, et al. (2011) Blockade of TNF-alpha rapidly inhibits pain responses in the central nervous system. Proc. Natl. Acad. Sci. U. S. A. 108:3731–6.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Diamond B, Tracey KJ. (2011) Mapping the immunological homunculus. Proc. Natl. Acad. Sci. U. S. A. 108:3461–2.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Diamond B, Huerta PT, Mina-Osorio P, Kowal C, Volpe BT. (2009) Losing your nerves? Maybe it’s the antibodies. Nat. Rev. Immunol. 9:449–56.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Thaler JP, et al. (2012) Obesity is associated with hypothalamic injury in rodents and humans. J. Clin. Invest. 122:153–62.CrossRefPubMedGoogle Scholar
  14. 14.
    Holmes C, et al. (2009) Systemic inflammation and disease progression in Alzheimer disease. Neurology. 73:768–74.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Holmes C, Cunningham C, Zotova E, Culliford D, Perry VH. (2011) Proinflammatory cytokines, sickness behavior, and Alzheimer disease. Neurology. 77:212–8.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Ferrari CC, Tarelli R. (2011) Parkinson’s disease and systemic inflammation. Parkinsons Dis. 2011:436813.PubMedPubMedCentralGoogle Scholar
  17. 17.
    Meyer U, Schwarz MJ, Muller N. (2011) Inflammatory processes in schizophrenia: a promising neuroimmunological target for the treatment of negative/cognitive symptoms and beyond. Pharmacol. Ther. 132:96–110.CrossRefPubMedGoogle Scholar
  18. 18.
    Andersson U, Tracey KJ. (2012) Reflex principles of immunological homeostasis. Annu. Rev. Immunol. 30:313–35.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Guarini S, et al. (2004) Adrenocorticotropin reverses hemorrhagic shock in anesthetized rats through the rapid activation of a vagal anti-inflammatory pathway. Cardiovasc. Res. 63:357–65.CrossRefPubMedGoogle Scholar
  20. 20.
    Pavlov VA, et al. (2006) Central muscarinic cholinergic regulation of the systemic inflammatory response during endotoxemia. Proc. Natl. Acad. Sci. U. S. A. 103:5219–23.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Pavlov VA, et al. (2009) Brain acetylcholinesterase activity controls systemic cytokine levels through the cholinergic anti-inflammatory pathway. Brain Behav. Immun. 23:41–5.CrossRefPubMedGoogle Scholar
  22. 22.
    Ji H, et al. (2014) Central cholinergic activation of a vagus nerve-to-spleen circuit alleviates experimental colitis. Mucosal. Immunol. 7:335–47.CrossRefPubMedGoogle Scholar
  23. 23.
    Lee ST, et al. (2010) Cholinergic anti-inflammatory pathway in intracerebral hemorrhage. Brain Res. 1309:164–71.CrossRefPubMedGoogle Scholar
  24. 24.
    Kelley KW, McCusker RH. (2014) Getting nervous about immunity. Semin. Immunol. 26:389–93.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Steinman L. (2012) Lessons learned at the intersection of immunology and neuroscience. J. Clin. Invest. 122:1146–8.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Capuron L, Miller AH. (2011) Immune system to brain signaling: neuropsychopharmacological implications. Pharmacol. Ther. 130:226–38.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Bonaz B. (2013) Inflammatory bowel diseases: a dysfunction of brain-gut interactions? Minerva Gastroenterol. Dietol. 59:241–59.PubMedGoogle Scholar
  28. 28.
    Maier SF, Watkins LR. (2003) Immune-to-central nervous system communication and its role in modulating pain and cognition: implications for cancer and cancer treatment. Brain Behav. Immun. 17 (Suppl. 1):S125–31.CrossRefPubMedGoogle Scholar
  29. 29.
    Doeuvre L, Plawinski L, Toti F, Angles-Cano E. (2009) Cell-derived microparticles: a new challenge in neuroscience. J. Neurochem. 110:457–68.CrossRefPubMedGoogle Scholar
  30. 30.
    Farina C, Aloisi F, Meinl E. (2007) Astrocytes are active players in cerebral innate immunity. Trends Immunol. 28:138–45.CrossRefPubMedGoogle Scholar
  31. 31.
    Lampron A, Elali A, Rivest S. (2013) Innate immunity in the CNS: redefining the relationship between the CNS and Its environment. Neuron. 78:214–32.CrossRefPubMedGoogle Scholar
  32. 32.
    Garden GA, Moller T. (2006) Microglia biology in health and disease. J. Neuroimmune Pharmacol. 1:127–37.CrossRefPubMedGoogle Scholar
  33. 33.
    Hanisch UK, Kettenmann H. (2007) Microglia: active sensor and versatile effector cells in the normal and pathologic brain. Nat. Neurosci. 10:1387–94.CrossRefPubMedGoogle Scholar
  34. 34.
    Ransohoff RM, Perry VH. (2009) Microglial physiology: unique stimuli, specialized responses. Annu. Rev. Immunol. 27:119–45.CrossRefPubMedGoogle Scholar
  35. 35.
    Yirmiya R, Goshen I. (2011) Immune modulation of learning, memory, neural plasticity and neurogenesis. Brain Behav. Immun. 25:181–213.CrossRefPubMedGoogle Scholar
  36. 36.
    Committee for the Update of the Guide for the Care and Use of Laboratory Animals, Institute for Laboratory Animal Research, Division on Earth and Life Studies, National Research Council of the National Academies. (2011) Guide for the Care and Use of Laboratory Animals. 8th edition. Washington (DC): National Academies Press.Google Scholar
  37. 37.
    Glowinski J, Iversen LL. (1966) Regional studies of catecholamines in the rat brain. I. The disposition of [3H]norepinephrine, [3H]dopamine and [3H]dopa in various regions of the brain. J. Neurochem. 13:655–69.CrossRefGoogle Scholar
  38. 38.
    Cikos S, Bukovska A, Koppel J. (2007) Relative quantification of mRNA: comparison of methods currently used for real-time PCR data analysis. BMC Mol. Biol. 8:113.CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Steinman L. (2013) Inflammatory cytokines at the summits of pathological signal cascades in brain diseases. Sci. Signal. 6:e3.CrossRefGoogle Scholar
  40. 40.
    Chen PC, et al. (2009) Nrf2-mediated neuroprotection in the MPTP mouse model of Parkinson’s disease: critical role for the astrocyte. Proc. Natl. Acad. Sci. U. S. A. 106:2933–8.CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Moss A, et al. (2007) Spinal microglia and neuropathic pain in young rats. Pain. 128:215–24.CrossRefPubMedGoogle Scholar
  42. 42.
    Satapathy SK, et al. (2011) Galantamine alleviates inflammation and other obesity-associated complications in high-fat diet-fed mice. Mol. Med. 17:599–606.CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Buttini M, Boddeke H. (1995) Peripheral lipopolysaccharide stimulation induces interleukin-1 beta messenger RNA in rat brain microglial cells. Neuroscience. 65:523–30.CrossRefPubMedGoogle Scholar
  44. 44.
    Wolff S, et al. (2009) Endotoxin-induced gene expression differences in the brain and effects of iNOS inhibition and norepinephrine. Intensive Care Med. 35:730–9.CrossRefPubMedGoogle Scholar
  45. 45.
    Thaler JP, et al. (2009) Atypical protein kinase C activity in the hypothalamus is required for lipopolysaccharide-mediated sickness responses. Endocrinology. 150:5362–72.CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Madore C, et al. (2013) Early morphofunctional plasticity of microglia in response to acute lipopolysaccharide. Brain Behav. Immun. 34:151–8.CrossRefPubMedGoogle Scholar
  47. 47.
    Buttini M, Limonta S, Boddeke HW. (1996) Peripheral administration of lipopolysaccharide induces activation of microglial cells in rat brain. Neurochem. Int. 29:25–35.CrossRefPubMedGoogle Scholar
  48. 48.
    Qin L, et al. (2007) Systemic LPS causes chronic neuroinflammation and progressive neurodegeneration. Glia. 55:453–62.CrossRefPubMedPubMedCentralGoogle Scholar
  49. 49.
    Nadeau S, Rivest S. (1999) Effects of circulating tumor necrosis factor on the neuronal activity and expression of the genes encoding the tumor necrosis factor receptors (p55 and p75) in the rat brain: a view from the blood-brain barrier. Neuroscience. 93:1449–64.CrossRefPubMedGoogle Scholar
  50. 50.
    Goehler LE, et al. (2000) Vagal immune-to-brain communication: a visceral chemosensory pathway. Auton. Neurosci. 85:49–59.CrossRefPubMedGoogle Scholar
  51. 51.
    Pavlov VA, Wang H, Czura CJ, Friedman SG, Tracey KJ. (2003) The cholinergic anti-inflammatory pathway: a missing link in neuroimmunomodulation. Mol. Med. 9:125–34.CrossRefPubMedPubMedCentralGoogle Scholar
  52. 52.
    Tracey KJ. (2009) Reflex control of immunity. Nat. Rev. Immunol. 9:418–28.CrossRefPubMedPubMedCentralGoogle Scholar
  53. 53.
    Pavlov VA, Tracey KJ. (2005) The cholinergic anti-inflammatory pathway. Brain Behav. Immun. 19:493–9.CrossRefPubMedGoogle Scholar
  54. 54.
    Pavlov VA. (2008) Cholinergic modulation of inflammation. Int. J. Clin. Exp. Med. 1:203–12.PubMedPubMedCentralGoogle Scholar
  55. 55.
    Huston JM, et al. (2009) Cholinergic neural signals to the spleen down-regulate leukocyte trafficking via CD11b. J. Immunol. 183:552–9.CrossRefPubMedPubMedCentralGoogle Scholar
  56. 56.
    Metz CN, Tracey KJ. (2005) It takes nerve to dampen inflammation. Nat. Immunol. 6:756–7.CrossRefPubMedGoogle Scholar
  57. 57.
    Olofsson PS, et al. (2012) Alpha7 nicotinic acetylcholine receptor (alpha7nAChR) expression in bone marrow-derived non-T cells is required for the inflammatory reflex. Mol. Med. 18:539–43.CrossRefPubMedGoogle Scholar
  58. 58.
    Woolf NJ. (1991) Cholinergic systems in mammalian brain and spinal cord. Prog. Neurobiol. 37:475–524.CrossRefPubMedGoogle Scholar
  59. 59.
    Mesulam MM, Mufson EJ, Wainer BH, Levey AI. (1983) Central cholinergic pathways in the rat: an overview based on an alternative nomenclature (Ch1-Ch6). Neuroscience. 10:1185–201.CrossRefPubMedGoogle Scholar
  60. 60.
    Hallanger AE, Wainer BH. (1988) Ascending projections from the pedunculopontine tegmental nucleus and the adjacent mesopontine tegmentum in the rat. J. Comp. Neurol. 274:483–515.CrossRefPubMedGoogle Scholar
  61. 61.
    Ibanez CF, Ernfors P, Persson H. (1991) Developmental and regional expression of choline acetyl-transferase mRNA in the rat central nervous system. J. Neurosci. Res 29:163–71.CrossRefPubMedGoogle Scholar
  62. 62.
    Dean B, McLeod M, Keriakous D, McKenzie J, Scarr E. (2002) Decreased muscarinic1 receptors in the dorsolateral prefrontal cortex of subjects with schizophrenia. Mol. Psychiatry. 7:1083–91.CrossRefPubMedGoogle Scholar
  63. 63.
    Mancama D, Arranz MJ, Landau S, Kerwin R. (2003) Reduced expression of the muscarinic 1 receptor cortical subtype in schizophrenia. Am. J. Med. Genet. B. Neuropsychiatr. Genet. 119B:2–6.CrossRefPubMedGoogle Scholar
  64. 64.
    Scarr E, et al. (2013) Decreased cortical muscarinic M1 receptors in schizophrenia are associated with changes in gene promoter methylation, mRNA and gene targeting microRNA. Transl. Psychiatry. 3:e230.CrossRefPubMedPubMedCentralGoogle Scholar
  65. 65.
    Caulfield MP. (1993) Muscarinic receptors: characterization, coupling and function. Pharmacol. Ther. 58:319–79.CrossRefGoogle Scholar
  66. 66.
    Hamilton SE, et al. (1997) Disruption of the m1 receptor gene ablates muscarinic receptor-dependent M current regulation and seizure activity in mice. Proc. Natl. Acad. Sci. U. S. A. 94:13311–6.CrossRefPubMedPubMedCentralGoogle Scholar
  67. 67.
    Ulrich-Lai YM, Herman JP. (2009) Neural regulation of endocrine and autonomic stress responses. Nat. Rev. Neurosci. 10:397–409.CrossRefPubMedPubMedCentralGoogle Scholar
  68. 68.
    Wrona D. (2006) Neural-immune interactions: an integrative view of the bidirectional relationship between the brain and immune systems. J. Neuroimmunol. 172:38–58.CrossRefPubMedGoogle Scholar

Copyright information

© The Author(s) 2014

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, and provide a link to the Creative Commons license. You do not have permission under this license to share adapted material derived from this article or parts of it.

The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this license, visit (

Authors and Affiliations

  • Harold A. Silverman
    • 1
    • 2
  • Meghan Dancho
    • 1
  • Angelique Regnier-Golanov
    • 3
  • Mansoor Nasim
    • 4
  • Mahendar Ochani
    • 1
  • Peder S. Olofsson
    • 1
  • Mohamed Ahmed
    • 5
    • 6
  • Edmund J. Miller
    • 2
    • 6
  • Sangeeta S. Chavan
    • 1
  • Eugene Golanov
    • 7
  • Christine N. Metz
    • 2
    • 8
  • Kevin J. Tracey
    • 1
    • 2
  • Valentin A. Pavlov
    • 1
    • 2
  1. 1.Laboratory of Biomedical Science, Center for Biomedical ScienceThe Feinstein Institute for Medical ResearchManhassetUSA
  2. 2.Hofstra North Shore-LIJ School of Medicine at Hofstra UniversityHempsteadUSA
  3. 3.Pediatrics-NeurologyBaylor College of MedicineHoustonUSA
  4. 4.Neuropathology-Anatomic PathologyNorth Shore-LIJ Health SystemNew Hyde ParkUSA
  5. 5.Cohen Children’s Medical CenterNorth Shore-LIJ Health SystemNew Hyde ParkUSA
  6. 6.Center for Heart and Lung ResearchThe Feinstein Institute for Medical ResearchManhassetUSA
  7. 7.The Houston Methodist Research InstituteHoustonUSA
  8. 8.Laboratory of Medicinal BiochemistryThe Feinstein Institute for Medical ResearchManhassetUSA

Personalised recommendations