Sevoflurane Exposure Results in Sex-Specific Transgenerational Upregulation of Target IEGs in the Subiculum

  • Shelby E. Chastain-Potts
  • Vesna Tesic
  • Quy L. Tat
  • Omar H. Cabrera
  • Nidia Quillinan
  • Vesna Jevtovic-TodorovicEmail author


Large body of animal work and emerging clinical findings have suggested that early exposure to anesthetics may result in increased risk of learning disabilities and behavioral impairments. Recent studies have begun to investigate anesthesia-induced epigenetic modifications to elucidate their role in behavioral and neurodevelopmental abnormalities. Here we examine sevoflurane-induced transgenerational modifications of subicular neuronal DNA methylation and expression of immediate early genes (IEGs), arc and junB, crucial to synaptic plasticity and normal neuronal development. We show that 6 h sevoflurane exposure in postnatal day 7 rat pups resulted in decreased neuronal 5-methycytosine, indicating reduced DNA methylation. This effect is transgenerationally expressed in offspring born to exposed mothers which is of importance considering that decreased DNA methylation in the brain has been linked with functional decline in learning and memory. We further show that sevoflurane exposure induces upregulation of Arc and JunB mRNA expression, 42.7% and 35.2%, respectively. Transgenerational changes in Arc and JunB mRNA were sexually dimorphic only occurring in males born to exposed females, expressed as upregulation of Arc and JunB mRNA, 71.6% and 74.0%, respectively. We further investigated correlation between altered arc promoter methylation and observed upregulation of Arc mRNA and observed that sevoflurane reduced methylation in the 5-upstream promoter region of females exposed to sevoflurane. Transgenerational hypomethylation and modifications to IEGs crucial to synaptic plasticity, observed following neonatal sevoflurane exposure could contribute to morphological and cognitive deficits known to occur with neonatal sevoflurane exposure.


General anesthesia DNA methylation Immature brain Arc junB 



Generation 0


Generation 1


General anesthesia








Immediate early genes


Funding information

Supported in part by funds from the Department of Anesthesiology at the University of Colorado Anschutz Medical campus and R0144517, R0144517-S, R01 GM118197, R01 GM118197, and R21 HD080281, March of Dimes National Award, CU Medicine Endowment.

Compliance with ethical standards

These experiments were approved by the Animal Use and Care Committee of the University of Colorado Anschutz Medical Campus, the Office of Laboratory Animal Resources (OLAR), Aurora, CO. All experiments were conducted in accordance with Public Health Service’s Policy on Humane Care and Use of Laboratory Animals.


  1. 1.
    U.S. Food & Drug Administration. (2016) FDA Drug Safety Communication: FDA review results in new warnings about using general anesthetics and sedation drugs in young children and pregnant women. Accessed 01 Dec 2018
  2. 2.
    Fan CH, Peng B, Zhang FC (2018) The postoperative effect of sevoflurane inhalational anesthesia on cognitive function and inflammatory response of pediatric patients. Eur Rev Med Pharmacol Sci 22:3971–3975Google Scholar
  3. 3.
    Wilder RT, Flick RP, Sprung J, Katusic SK, Barbaresi WJ, Mickelson C, Gleich SJ, Schroeder DR et al (2009) Early exposure to anesthesia and learning disabilities in a population-based birth cohort. Anesthesiology 110:796–804. CrossRefGoogle Scholar
  4. 4.
    Flick RP, Katusic SK, Colligan RC, Wilder RT, Voigt RG, Olson MD, Sprung J, Weaver AL et al (2011) Cognitive and behavioral outcomes after early exposure to anesthesia and surgery. Pediatrics 128:273–292. CrossRefGoogle Scholar
  5. 5.
    Shen X, Xiao Y, Li W, Chen K, Yu H (2018) Sevoflurane anesthesia during pregnancy in mice induces hearing impairment in the offspring. Drug Des Devel Ther 12:1827–1836CrossRefGoogle Scholar
  6. 6.
    Ozer A, Ceribasi S, Ceribasi A et al (2017) Effects of sevoflurane on apoptosis, BDNF and cognitive functions in neonatal rats. Bratisl Med J 118:80–84. CrossRefGoogle Scholar
  7. 7.
    Shih J, May L d V, Gonzalez HE et al (2012) Delayed environmental enrichment reverses sevoflurane-induced memory impairment in rats. Anesthesiology 116:586–602. CrossRefGoogle Scholar
  8. 8.
    Satomoto M, Satoh Y, Terui K, Miyao H, Takishima K, Ito M, Imaki J (2009) Neonatal exposure to sevoflurane induces abnormal social behaviors and deficits in fear conditioning in mice. Anesthesiology 110:628–637. CrossRefGoogle Scholar
  9. 9.
    Zhang DX, Jiang S, Yu LN et al (2015) The effect of sevoflurane on the cognitive function of rats and its association with the inhibition of synaptic transmission. Int J Clin Exp Med 8:20853–20860 Google Scholar
  10. 10.
    Shen X, Dong Y, Xu Z, Wang H, Miao C, Soriano SG, Sun D, Baxter MG et al (2013) Selective anesthesia-induced neuroinflammation in developing mouse brain and cognitive impairment. Anesthesiology 118:502–515. CrossRefGoogle Scholar
  11. 11.
    Tao G, Luo Y, Xue Q, Li G, Tan Y, Xiao J, Yu B (2016) Docosahexaenoic acid rescues synaptogenesis impairment and long-term memory deficits caused by postnatal multiple sevoflurane exposures. Biomed Res Int 2016:1–7. Google Scholar
  12. 12.
    Chalon J, Tang CK, Ramanathan S et al (1981) Exposure to halothane and enflurane affects learning function of murine progeny. Anesth Analg 60:794–797. CrossRefGoogle Scholar
  13. 13.
    Massara LD, Osuru HP, Ph D et al (2016) General anesthesia causes epigenetic histone modulation of c-Fos and brain-derived neurotrophic factor, target genes important for neuronal development in the immature rat hippocampus. Am Sciety Anesthesiol 124:1311–1327CrossRefGoogle Scholar
  14. 14.
    Jia M, Ji M, Yang J (2017) Epigenetic regulation of general anesthesia-induced neonatal neurodegeneration. Oncotarget 8:5652–5653Google Scholar
  15. 15.
    L sha J, Jia M, Sun J et al (2016) Hypermethylation of hippocampal synaptic plasticity-related genes is involved in neonatal sevoflurane exposure-induced cognitive impairments in rats. Neurotox Res 29:243–255. CrossRefGoogle Scholar
  16. 16.
    Joksimovic SM, Osuru HP, Oklopcic A, Beenhakker MP, Jevtovic-Todorovic V, Todorovic SM (2018) Histone deacetylase inhibitor entinostat (MS-275) restores anesthesia-induced alteration of inhibitory synaptic transmission in the developing rat hippocampus. Mol Neurobiol 55:222–228. CrossRefGoogle Scholar
  17. 17.
    Oliveira AMM (2016) DNA methylation: a permissive mark in memory formation and maintenance. Learn Mem 23:587–593. CrossRefGoogle Scholar
  18. 18.
    Liu L, van Groen T, Kadish I, Tollefsbol TO (2009) DNA methylation impacts on learning and memory in aging. Neurobiol Aging 30:549–560. CrossRefGoogle Scholar
  19. 19.
    Smeester L, Rager JE, Bailey KA et al (2017) An overview of epigenetic assays. J Neurosci 8:e0163690. Google Scholar
  20. 20.
    Ju LS, Yang JJ, Morey TE, Gravenstein N, Seubert CN, Resnick JL, Zhang J (2018) Role of epigenetic mechanisms in transmitting the effects of neonatal sevoflurane to the next generation of male, but not female, rats. Br J Anaesth 121:406–416CrossRefGoogle Scholar
  21. 21.
    Sanchez V, Feinstein SD, Lunardi N, Joksovic PM, Boscolo A, Todorovic SM, Jevtovic-Todorovic V (2011) General anesthesia causes long-term impairment of mitochondrial morphogenesis and synaptic transmission in developing rat brain. Anesthesiology 115:992–1002. CrossRefGoogle Scholar
  22. 22.
    O’Mara S (2005) The subiculum: what it does, what it might do, and what neuroanatomy has yet to tell us. J Anat 207:271–282. CrossRefGoogle Scholar
  23. 23.
    Behr J, Wozny C, Fidzinski P, Schmitz D (2009) Synaptic plasticity in the subiculum. Prog Neurobiol 89:334–342. CrossRefGoogle Scholar
  24. 24.
    McNaughton N (2006) The role of the subiculum within the behavioural inhibition system. Behav Brain Res 174:232–250. CrossRefGoogle Scholar
  25. 25.
    Lynch MA (2004) Long-term potentiation and memory. Physiol Rev 84:87–136. CrossRefGoogle Scholar
  26. 26.
    Tischmeyer W, Grimm R (1999) Activation of immediate early genes and memory formation. Cell Mol Life Sci 55:564–574CrossRefGoogle Scholar
  27. 27.
    Sommerlandt FMJ, Brockmann A, Rössler W, Spaethe J (2018) Immediate early genes in social insects: a tool to identify brain regions involved in complex behaviors and molecular processes underlying neuroplasticity. Cell Mol Life Sci 76:1–15. Google Scholar
  28. 28.
    Srivas S, Thakur MK (2017) Epigenetic regulation of neuronal immediate early genes is associated with decline in their expression and memory consolidation in scopolamine-induced amnesic mice. 5107–5119.
  29. 29.
    Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative CT method. Nat Protoc 3:1101–1108. CrossRefGoogle Scholar
  30. 30.
    Penner MR, Roth TL, Chawla MK et al (2011) Age-related changes in Arc transcription and DNA methylation within the hippocampus. Neurobiol Aging 32:2198–2210. CrossRefGoogle Scholar
  31. 31.
    Dyrvig M, Hansen HH, Christiansen SH, Woldbye DPD, Mikkelsen JD, Lichota J (2012) Epigenetic regulation of Arc and c-Fos in the hippocampus after acute electroconvulsive stimulation in the rat. Brain Res Bull 88:507–513. CrossRefGoogle Scholar
  32. 32.
    Zhang L y, Li P l, Wang T z, Zhang X c (2015) Prognostic values of 5-hmC, 5-mC and TET2 in epithelial ovarian cancer. Arch Gynecol Obstet 292:891–897. CrossRefGoogle Scholar
  33. 33.
    Hicks MJ, Hu QP, Macrae E, DeWille J (2015) Mitogen-activated protein kinase signaling controls basal and oncostatin M-mediated JUNB gene expression. Mol Cell Biochem 403:115–124. CrossRefGoogle Scholar
  34. 34.
    Wilkerson JR, Albanesi JP, Huber KM (2018) Roles for Arc in metabotropic glutamate receptor-dependent LTD and synapse elimination: Implications in health and disease. Semin Cell Dev Biol 77:51–62. CrossRefGoogle Scholar
  35. 35.
    Alberini CM (2014) Transcription factors in long-term memory and synaptic plasticity. Physiol Rev 89:1–46. Google Scholar
  36. 36.
    Lister R, Mukamel EA, Nery JR, Urich M, Puddifoot CA, Johnson ND, Lucero J, Huang Y et al (2013) Global epigenomic reconfiguration during mammalian brain development. Science (80- ) 341:1237905. CrossRefGoogle Scholar
  37. 37.
    Bogdanović O, Veenstra GJC (2009) DNA methylation and methyl-CpG binding proteins: developmental requirements and function. Chromosoma 118:549–565. CrossRefGoogle Scholar
  38. 38.
    Ambrosi C, Manzo M, Baubec T (2017) Dynamics and context-dependent roles of DNA methylation. J Mol Biol 429:1459–1475. CrossRefGoogle Scholar
  39. 39.
    Korb E, Finkbeiner S (2011) Arc in synaptic plasticity: from gene to behavior. Trends Neurosci 34:591–598. CrossRefGoogle Scholar
  40. 40.
    Eagle AL, Gajewski PA, Robison AJ (2016) Role of hippocampal activity-induced transcription in memory consolidation. Rev Neurosci 27:559–573. CrossRefGoogle Scholar
  41. 41.
    Clayton DF (2000) The genomic action potential. Neurobiol Learn Mem 74:185–216. CrossRefGoogle Scholar
  42. 42.
    Itoh, Masayuki & Okuno, Hiroyuki & Yamada, Daisuke & Yamashita, Mariko & Abe, Manabu & Natsume, Rie & Kaizuka, Toshie & Sakimura, Kenji & Hoshino, Mikio & Mishina, Masayoshi & Wada, Keiji & Sekiguchi, Masayuki & Hayashi, Takashi. (2018). Perturbed expression pattern of the immediate early gene Arc in the dentate gyrus of GluA1 C‐terminal palmitoylation‐deficient mice. Neuropsychopharmacology Reports. 39.
  43. 43.
    Xiao H, Liu B, Chen Y, Zhang J (2016) Learning, memory and synaptic plasticity in hippocampus in rats exposed to sevoflurane. Int J Dev Neurosci 48:38–49. CrossRefGoogle Scholar
  44. 44.
    Zimering JH, Dong Y, Fang F, Huang L, Zhang Y, Xie Z (2016) Anesthetic sevoflurane causes rho-dependent filopodial shortening in mouse neurons. PLoS One 11:1–15. CrossRefGoogle Scholar
  45. 45.
    Jevtovic-Todorovic V, Hartman RE, Izumi Y, Benshoff ND, Dikranian K, Zorumski CF, Olney JW, Wozniak DF (2003) Early exposure to common anesthetic agents causes widespread neurodegeneration in the developing rat brain and persistent learning deficits. J Neurosci 23:876–882. CrossRefGoogle Scholar
  46. 46.
    Loepke AW, Istaphanous GK, McAuliffe JJ et al (2009) The effects of neonatal isoflurane exposure in mice on brain cell viability, adult behavior, learning, and memory. Anesth Analg 108:90–104. CrossRefGoogle Scholar
  47. 47.
    Paule MG, Li M, Allen RR, Liu F, Zou X, Hotchkiss C, Hanig JP, Patterson TA et al (2011) Ketamine anesthesia during the first week of life can cause long-lasting cognitive deficits in rhesus monkeys. Neurotoxicol Teratol 33:220–230. CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of AnesthesiologyUniversity of Colorado School of MedicineAuroraUSA

Personalised recommendations