Title: Transient activation of microglia following acute alcohol exposure in developing mouse neocortex is primarily driven by BAX-dependent neurodegeneration
Abstract: GliaVolume 63, Issue 10 p. 1694-1713 Research Article Transient activation of microglia following acute alcohol exposure in developing mouse neocortex is primarily driven by BAX-dependent neurodegeneration Katelin E. Ahlers, Katelin E. Ahlers Department of Biology, College of Liberal Arts and Sciences, University of Iowa, Iowa City, IowaSearch for more papers by this authorBahri Karaçay, Bahri Karaçay Division of Child Neurology, Department of Pediatrics, University of Iowa, Iowa City, IowaSearch for more papers by this authorLeah Fuller, Leah Fuller Department of Biology, College of Liberal Arts and Sciences, University of Iowa, Iowa City, IowaSearch for more papers by this authorDaniel J. Bonthius, Daniel J. Bonthius Division of Child Neurology, Department of Pediatrics, University of Iowa, Iowa City, Iowa Department of Neurology, the Roy J. Carver College of Medicine, University of Iowa, Iowa City, IowaSearch for more papers by this authorMichael E. Dailey, Corresponding Author Michael E. Dailey Department of Biology, College of Liberal Arts and Sciences, University of Iowa, Iowa City, IowaAddress correspondence to Dr. Michael E. Dailey, Department of Biology, 369 Biology Building, University of Iowa, Iowa City, Iowa 52242-1324. E-mail: [email protected]Search for more papers by this author Katelin E. Ahlers, Katelin E. Ahlers Department of Biology, College of Liberal Arts and Sciences, University of Iowa, Iowa City, IowaSearch for more papers by this authorBahri Karaçay, Bahri Karaçay Division of Child Neurology, Department of Pediatrics, University of Iowa, Iowa City, IowaSearch for more papers by this authorLeah Fuller, Leah Fuller Department of Biology, College of Liberal Arts and Sciences, University of Iowa, Iowa City, IowaSearch for more papers by this authorDaniel J. Bonthius, Daniel J. Bonthius Division of Child Neurology, Department of Pediatrics, University of Iowa, Iowa City, Iowa Department of Neurology, the Roy J. Carver College of Medicine, University of Iowa, Iowa City, IowaSearch for more papers by this authorMichael E. Dailey, Corresponding Author Michael E. Dailey Department of Biology, College of Liberal Arts and Sciences, University of Iowa, Iowa City, IowaAddress correspondence to Dr. Michael E. Dailey, Department of Biology, 369 Biology Building, University of Iowa, Iowa City, Iowa 52242-1324. E-mail: [email protected]Search for more papers by this author First published: 09 April 2015 https://doi.org/10.1002/glia.22835Citations: 63Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat Abstract Fetal alcohol exposure is the most common known cause of preventable mental retardation, yet we know little about how microglia respond to, or are affected by, alcohol in the developing brain in vivo. Using an acute (single day) model of moderate (3 g/kg) to severe (5 g/kg) alcohol exposure in postnatal day (P) 7 or P8 mice, we found that alcohol-induced neuroapoptosis in the neocortex is closely correlated in space and time with the appearance of activated microglia near dead cells. The timing and molecular pattern of microglial activation varied with the level of cell death. Although microglia rapidly mobilized to contact and engulf late-stage apoptotic neurons, apoptotic bodies temporarily accumulated in neocortex, suggesting that in severe cases of alcohol toxicity the neurodegeneration rate exceeds the clearance capacity of endogenous microglia. Nevertheless, most dead cells were cleared and microglia began to deactivate within 1–2 days of the initial insult. Coincident with microglial activation and deactivation, there was a transient increase in expression of pro-inflammatory factors, TNFα and IL-1β, after severe (5 g/kg) but not moderate (3 g/kg) EtOH levels. Alcohol-induced microglial activation and pro-inflammatory factor expression were largely abolished in BAX null mice lacking neuroapoptosis, indicating that microglial activation is primarily triggered by apoptosis rather than the alcohol. Therefore, acute alcohol exposure in the developing neocortex causes transient microglial activation and mobilization, promoting clearance of dead cells and tissue recovery. Moreover, cortical microglia show a remarkable capacity to rapidly deactivate following even severe neurodegenerative insults in the developing brain. GLIA 2015;63:1694–1713 Supporting Information Additional Supporting Information may be found in the online version of this article. Filename Description glia22835-sup-0001-suppfig1.tif1.4 MB Supporting Information Figure 1. glia22835-sup-0002-suppfig2.tif1.7 MB Supporting Information Figure 2. glia22835-sup-0003-suppfig3.tif3.1 MB Supporting Information Figure 3. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article. References Adayev T, Estephan R, Meserole S, Mazza B, Yurkow EJ, Banerjee P. 1998. Externalization of phosphatidylserine may not be an early signal of apoptosis in neuronal cells, but only the phosphatidylserine-displaying apoptotic cells are phagocytosed by microglia. J Neurochem 71: 1854–1864. Aguzzi A, Barres BA, Bennett ML. 2013. Microglia: Scapegoat, saboteur, or something else? Science 339: 156–161. Alfonso-Loeches S, Pascual-Lucas M, Blanco AM, Sanchez-Vera I, Guerri C. 2010. Pivotal role of tlr4 receptors in alcohol-induced neuroinflammation and brain damage. J Neurosci 30: 8285–8295. Aroor AR, Baker RC. 1998. Ethanol inhibition of phagocytosis and superoxide anion production by microglia. Alcohol 15: 277–280. Bittigau P, Sifringer M, Genz K, Reith E, Pospischil D, Govindarajalu S, Dzietko M, Pesditschek S, Mai I, Dikranian K, Olney JW, Ikonomidou C. 2002. Antiepileptic drugs and apoptotic neurodegeneration in the developing brain. Proc Natl Acad Sci U S A 99: 15089–15094. Bonthius DJ, Bonthius NE, Li S, Karacay B. 2008. The protective effect of neuronal nitric oxide synthase (nNOS) against alcohol toxicity depends upon the NO-cGMP-PKG pathway and NF-kappaB. Neurotoxicology 29: 1080–1091. Bonthius DJ, Luong T, Bonthius NE, Hostager BS, Karacay B. 2009. Nitric oxide utilizes NF-kappaB to signal its neuroprotective effect against alcohol toxicity. Neuropharmacology 56: 716–731. Bonthius DJ, Tzouras G, Karacay B, Mahoney J, Hutton A, McKim R, Pantazis NJ. 2002. Deficiency of neuronal nitric oxide synthase (nNOS) worsens alcohol-induced microencephaly and neuronal loss in developing mice. Brain Res Dev Brain Res 138: 45–59. Boyadjieva NI, Sarkar DK. 2013. Microglia play a role in ethanol-induced oxidative stress and apoptosis in developing hypothalamic neurons. Alcohol Clin Exp Res 37: 252–262. Boyadjieva NI, Sarkar DK. 2010. Role of microglia in ethanol's apoptotic action on hypothalamic neuronal cells in primary cultures. Alcohol Clin Exp Res 34: 1835–1842. Cabal-Hierro L, Lazo PS. 2012. Signal transduction by tumor necrosis factor receptors. Cell Signal 24: 1297–1305. Carloni S, Mazzoni E, and, Balduini W. 2004. Caspase-3 and calpain activities after acute and repeated ethanol administration during the rat brain growth spurt. J Neurochem 89: 197–203. Chastain LG, Sarkar DK. 2014. Role of microglia in regulation of ethanol neurotoxic action. Int Rev Neurobiol 118: 81–103. Chekeni FB, Ravichandran KS. 2011. The role of nucleotides in apoptotic cell clearance: Implications for disease pathogenesis. J Mol Med (Berl) 89: 13–22. Chen WJ, Maier SE, Parnell SE, West JR. 2003. Alcohol and the developing brain: Neuroanatomical studies. Alcohol Res Health 27: 174–180. Coleman LGJ, Oguz I, Lee J, Styner M, Crews FT. 2012. Postnatal day 7 ethanol treatment causes persistent reductions in adult mouse brain volume and cortical neurons with sex specific effects on neurogenesis. Alcohol 46: 603–612. Colton CA, Snell-Callanan J, Chernyshev ON. 1998. Ethanol induced changes in superoxide anion and nitric oxide in cultured microglia. Alcohol Clin Exp Res 22: 710–716. Colton CA, Wilcock DM. 2010. Assessing activation states in microglia. CNS Neurol Disord Drug Targets 9: 174–191. Creeley CE, Olney JW. 2013. Drug-induced apoptosis: Mechanism by which alcohol and many other drugs can disrupt brain development. Brain Sci 3: 1153–1181. Crews F, Nixon K, Kim D, Joseph J, Shukitt-Hale B, Qin L, Zou J. 2006. BHT blocks NF-kB activation and ethanol-induced brain damage. Alcohol Clin Exp Res 30: 1938–1949. Dalmau I, Finsen B, Zimmer J, Gonzalez B, Castellano B. 1998. Development of microglia in the postnatal rat hippocampus. Hippocampus 8: 458–474. Dalmau I, Vela JM, Gonzalez B, Finsen B, Castellano B. 2003. Dynamics of microglia in the developing rat brain. J Comp Neurol 458: 144–157. Damoiseaux JG, Dopp EA, Calame W, Chao D, MacPherson GG, Dijkstra CD. 1994. Rat macrophage lysosomal membrane antigen recognized by monoclonal antibody ed1. Immunology 83: 140–147. Dobbing J, Sands J. 1979. Comparative aspects of the brain growth spurt. Early Hum Dev 3: 79–83. Drew PD, Kane CJ. 2014. Fetal alcohol spectrum disorders and neuroimmune changes. Int Rev Neurobiol 118: 41–80. Eggen BJ, Raj D, Hanisch UK, Boddeke HW. 2013. Microglial phenotype and adaptation. J Neuroimmune Pharmacol 8: 807–823. Elmore S. 2007. Apoptosis: A review of programmed cell death. Toxicol Pathol 35: 495–516. Eyo UB, Dailey ME. 2013. Microglia: Key elements in neural development, plasticity, and pathology. J Neuroimmune Pharmacol 8: 494–509. Fernandez-Lizarbe S, Montesinos J, Guerri C. 2013. Ethanol induces tlr4/tlr2 association, triggering an inflammatory response in microglial cells. J Neurochem 126: 261–273. Fernandez-Lizarbe S, Pascual M, Guerri C. 2009. Critical role of tlr4 response in the activation of microglia induced by ethanol. J Immunol 183: 4733–4744. Ferrer I, Bernet E, Soriano E, Del Rio T, Fonseca M. 1990. Naturally occurring cell death in the cerebral cortex of the rat and removal of dead cells by transitory phagocytes. Neuroscience 39: 451–458. Ferrer I, Soriano E, del Rio JA, Alcantara S, Auladell C. 1992. Cell death and removal in the cerebral cortex during development. Prog Neurobiol 39: 1–43. Guizzetti M, Zhang X, Goeke C, Gavin DP. 2014. Glia and neurodevelopment: Focus on fetal alcohol spectrum disorders. Front Pediatr 2: 123. Hamerman JA, Jarjoura JR, Humphrey MB, Nakamura MC, Seaman WE, Lanier LL. 2006. Cutting edge: Inhibition of TLR and FcR responses in macrophages by triggering receptor expressed on myeloid cells (TREM)-2 and dap12. J Immunol 177: 2051–2055. Han JY, Joo Y, Kim YS, Lee YK, Kim HJ, Cho GJ, Choi WS, Kang SS. 2005. Ethanol induces cell death by activating caspase-3 in the rat cerebral cortex. Mol Cells 20: 189–195. Hanisch UK. 2002. Microglia as a source and target of cytokines. Glia 40: 140–155. Harry GJ. 2013. Microglia during development and aging. Pharmacol Ther 139: 313–326. Haynes SE, Hollopeter G, Yang G, Kurpius D, Dailey ME, Gan WB, Julius D. 2006. The p2y12 receptor regulates microglial activation by extracellular nucleotides. Nat Neurosci 9: 1512–1519. Ikonomidou C, Bosch F, Miksa M, Bittigau P, Vöckler J, Dikranian K, Tenkova T, Stevoska V, Turski L, Olney JW. 1999. Blockade of NMDA receptors and apoptotic neurodegeneration in the developing brain. Science 283: 70–74. Ikonomidou C, Bittigau P, Ishimaru MJ, Wozniak DF, Koch C, Genz K, Price MT, Stefovska V, Horster F, Tenkova T, Dikranian K, Olney JW. 2000. Ethanol-induced apoptotic neurodegeneration and fetal alcohol syndrome. Science 287: 1056–1060. Istaphanous GK, Howard J, Nan X, Hughes EA, McCann JC, McAuliffe JJ, Danzer SC, Loepke AW. 2011. Comparison of the neuroapoptotic properties of equipotent anesthetic concentrations of desflurane, isoflurane, or sevoflurane in neonatal mice. Anesthesiology 114: 578–587. 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. Jung S, Aliberti J, Graemmel P, Sunshine MJ, Kreutzberg GW, Sher A, Littman DR. 2000. Analysis of fractalkine receptor CX(3)cr1 function by targeted deletion and green fluorescent protein reporter gene insertion. Mol Cell Biol 20: 4106–4114. Kane CJ, Phelan KD, Drew PD. 2012. Neuroimmune mechanisms in fetal alcohol spectrum disorder. Dev Neurobiol 72: 1302–1316. Kane CJ, Phelan KD, Han L, Smith RR, Xie J, Douglas JC, Drew PD. 2011. Protection of neurons and microglia against ethanol in a mouse model of fetal alcohol spectrum disorders by peroxisome proliferator-activated receptor-gamma agonists. Brain Behav Immun 25(Suppl 1): S137–S145. Knudson CM, Tung KS, Tourtellotte WG, Brown GA, Korsmeyer SJ. 1995. Bax-deficient mice with lymphoid hyperplasia and male germ cell death. Science 270: 96–99. Kurpius D, Wilson N, Fuller L, Hoffman A, Dailey ME. 2006. Early activation, motility, and homing of neonatal microglia to injured neurons does not require protein synthesis. Glia 54: 58–70. Ling EA. 1976. Some aspects of amoeboid microglia in the corpus callosum and neighbouring regions of neonatal rats. J Anat 121: 29–45. Marin-Teva JL, Dusart I, Colin C, Gervais A, van Rooijen N, Mallat M. 2004. Microglia promote the death of developing purkinje cells. Neuron 41: 535–547. Marino MD, Aksenov MY, Kelly SJ. 2004. Vitamin E protects against alcohol-induced cell loss and oxidative stress in the neonatal rat hippocampus. Int J Dev Neurosci 22: 363–377. Marshall SA, McClain JA, Kelso ML, Hopkins DM, Pauly JR, Nixon K. 2013. Microglial activation is not equivalent to neuroinflammation in alcohol-induced neurodegeneration: The importance of microglia phenotype. Neurobiol Dis 54: 239–251. May PA, Gossage JP, Kalberg WO, Robinson LK, Buckley D, Manning M, Hoyme HE. 2009. Prevalence and epidemiologic characteristics of FASD from various research methods with an emphasis on recent in-school studies. Dev Disabil Res Rev 15: 176–192. McClain JA, Morris SA, Deeny MA, Marshall SA, Hayes DM, Kiser ZM, Nixon K. 2011. Adolescent binge alcohol exposure induces long-lasting partial activation of microglia. Brain Behav Immun 25(Suppl 1): S120–S128. Napoli I, Neumann H. 2009. Microglial clearance function in health and disease. Neuroscience 158: 1030–1038. Neumann H, Kotter MR, Franklin RJ. 2009. Debris clearance by microglia: An essential link between degeneration and regeneration. Brain 132: 288–295. Neumann H, Takahashi K. 2007. Essential role of the microglial triggering receptor expressed on myeloid cells-2 (trem2) for central nervous tissue immune homeostasis. J Neuroimmunol 184: 92–99. O'Connor JJ. 2013. Targeting tumour necrosis factor-alpha in hypoxia and synaptic signalling. Ir J Med Sci 182: 157–162. Olney JW, Tenkova T, Dikranian K, Qin YQ, Labruyere J, Ikonomidou C. 2002. Ethanol-induced apoptotic neurodegeneration in the developing C57BL/6 mouse brain. Brain Res Dev Brain Res 133: 115–126. Oppenheim RW. 1991. Cell death during development of the nervous system. Annu Rev Neurosci 14: 453–501. Otero GC, Merrill JE. 1994. Cytokine receptors on glial cells. Glia 11: 117–128. Petersen MA, Dailey ME. 2004. Diverse microglial motility behaviors during clearance of dead cells in hippocampal slices. Glia 46: 195–206. Qin L, He J, Hanes RN, Pluzarev O, Hong JS, Crews FT. 2008. Increased systemic and brain cytokine production and neuroinflammation by endotoxin following ethanol treatment. J Neuroinflammation 5: 10. Ramprasad MP, Fischer W, Witztum JL, Sambrano GR, Quehenberger O, Steinberg D. 1995. The 94- to 97-kDa mouse macrophage membrane protein that recognizes oxidized low density lipoprotein and phosphatidylserine-rich liposomes is identical to macrosialin, the mouse homologue of human cd68. Proc Natl Acad Sci U S A 92: 9580–9584. Rice D, Barone SJ. 2000. Critical periods of vulnerability for the developing nervous system: Evidence from humans and animal models. Environ Health Perspect 108(Suppl 3): 511–533. Rizzi S, Carter LB, Ori C, Jevtovic-Todorovic V. 2008. Clinical anesthesia causes permanent damage to the fetal guinea pig brain. Brain Pathol 18: 198–210. Rizzi S, Ori C, Jevtovic-Todorovic V. 2010. Timing versus duration: Determinants of anesthesia-induced developmental apoptosis in the young mammalian brain. Ann N Y Acad Sci 1199: 43–51. Sadrian B, Subbanna S, Wilson DA, Basavarajappa BS, Saito M. 2012. Lithium prevents long-term neural and behavioral pathology induced by early alcohol exposure. Neuroscience 206: 122–135. Saijo K, Glass CK. 2011. Microglial cell origin and phenotypes in health and disease. Nat Rev Immunol 11: 775–787. Saito M, Mao RF, Wang R, Vadasz C, Saito M. 2007a. Effects of gangliosides on ethanol-induced neurodegeneration in the developing mouse brain. Alcohol Clin Exp Res 31: 665–674. Saito M, Chakraborty G, Mao RF, Wang R, Cooper TB, Vadasz C, Saito M. 2007b. Ethanol alters lipid profiles and phosphorylation status of AMP-activated protein kinase in the neonatal mouse brain. J Neurochem 103: 1208–1218. Saito M, Chakraborty G, Mao RF, Paik SM, Vadasz C, Saito M. 2010. Tau phosphorylation and cleavage in ethanol-induced neurodegeneration in the developing mouse brain. Neurochem Res 35: 651–659. Sierra A, Abiega O, Shahraz A, Neumann H. 2013. Janus-faced microglia: Beneficial and detrimental consequences of microglial phagocytosis. Front Cell Neurosci 7: 6. Smith ME, van der Maesen K, Somera FP. 1998. Macrophage and microglial responses to cytokines in vitro: Phagocytic activity, proteolytic enzyme release, and free radical production. J Neurosci Res 54: 68–78. Snigdha S, Smith ED, Prieto GA, Cotman CW. 2012. Caspase-3 activation as a bifurcation point between plasticity and cell death. Neurosci Bull 28: 14–24. Spittau B, Wullkopf L, Zhou X, Rilka J, Pfeifer D, Krieglstein K. 2013. Endogenous transforming growth factor-beta promotes quiescence of primary microglia in vitro. Glia 61: 287–300. Stence N, Waite M, Dailey ME. 2001. Dynamics of microglial activation: A confocal time-lapse analysis in hippocampal slices. Glia 33: 256–266. Subbanna S, Shivakumar M, Umapathy NS, Saito M, Mohan PS, Kumar A, Nixon RA, Verin AD, Psychoyos D, Basavarajappa BS. 2013. G9a-mediated histone methylation regulates ethanol-induced neurodegeneration in the neonatal mouse brain. Neurobiol Dis 54: 475–485. Tiwari V, Chopra K. 2011. Resveratrol prevents alcohol-induced cognitive deficits and brain damage by blocking inflammatory signaling and cell death cascade in neonatal rat brain. J Neurochem 117: 678–690. Turnbull IR, Gilfillan S, Cella M, Aoshi T, Miller M, Piccio L, Hernandez M, Colonna M. 2006. Cutting edge: TREM-2 attenuates macrophage activation. J Immunol 177: 3520–3524. von Zahn J, Möller T, Kettenmann H, Nolte C. 1997. Microglial phagocytosis is modulated by pro- and anti-inflammatory cytokines. Neuroreport 8: 3851–3856. Wilson DA, Peterson J, Basavaraj BS, Saito M 2011. Local and regional network function in behaviorally relevant cortical circuits of adult mice following postnatal alcohol exposure. Alcohol. Clin Exp Res 35: 1974–1984. Witting A, Muller P, Herrmann A, Kettenmann H, Nolte C. 2000. Phagocytic clearance of apoptotic neurons by microglia/brain macrophages in vitro: Involvement of lectin-, integrin-, and phosphatidylserine-mediated recognition. J Neurochem 75: 1060–1070. Wozniak DF, Hartman RE, Boyle MP, Vogt SK, Brooks AR, Tenkova T, Young C, Olney JW, Muglia LJ. 2004. Apoptotic neurodegeneration induced by ethanol in neonatal mice is associated with profound learning/memory deficits in juveniles followed by progressive functional recovery in adults. Neurobiol Dis 17: 403–414. Young C, Klocke BJ, Tenkova T, Choi J, Labruyere J, Qin YQ, Holtzman DM, Roth KA, Olney JW. 2003. Ethanol-induced neuronal apoptosis in vivo requires BAX in the developing mouse brain. Cell Death Differ 10: 1148–1155. Young C, Roth KA, Klocke BJ, West T, Holtzman DM, Labruyere J, Qin YQ, Dikranian K, Olney JW. 2005. Role of caspase-3 in ethanol-induced developmental neurodegeneration. Neurobiol Dis 20: 608–614. Zhao YN, Wang F, Fan YX, Ping GF, Yang JY, Wu CF. (2013). Activated microglia are implicated in cognitive deficits, neuronal death, and successful recovery following intermittent ethanol exposure. Behav Brain Res 2013; 236: 270–282. Citing Literature Volume63, Issue10October 2015Pages 1694-1713 ReferencesRelatedInformation