Abstract
Early growth response gene-1 (Egr-1) is up-regulated by hypoxia-ischemia (HI) and reactive oxygen species (ROS) in adult animals, functioning as a master switch in inflammation and thrombogenesis. We hypothesized that resuscitation from HI with 100% O2 would result in greater Egr-1 expression, ROS, and cell death (CD) in the brains of newborn piglets than 21% O2. Two control groups breathed 21% O2 for 1 h followed by 21% or 100% O2 for 1 h. Two HI groups underwent carotid artery occlusion and breathed 8–12% O2 for 1 h followed by occlusion release and 21% or 100% O2 for 1 h. Brain Egr-1 mRNA and protein were analyzed via quantitative PCR and Western blot. CD and ROS were measured by fluorescence microscopy. Egr-1 mRNA expression increased throughout the brain in response to HI with regional heterogeneity, but protein levels did not. Resuscitation with 100% oxygen did not cause any additional Egr-1 mRNA, Egr-1 protein, CD, or ROS production as compared with 21% oxygen. There was no difference in physiologic recovery after HI with room air compared with 100% O2 resuscitation. However, 100% O2 administration was associated with increased CD in the brainstem independent of HI. Therefore, 100% O2 may have been toxic to some brainstem cells and potentially have significance in long-term neurologic sequelae seen after neonatal HI/resuscitation. Egr-1 protein levels may be tightly regulated in an attempt to diminish neurotoxicity or to enhance plasticity at this stage of development.
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Abbreviations
- aCSF:
-
artificial cerebrospinal fluid
- CD:
-
cell death
- DCFA:
-
5-chloromethyl-2,7-dichlorodihydrofluorescein diacetate
- Egr-1:
-
early growth response gene-1
- HI:
-
hypoxia-ischemia
- HI/R:
-
hypoxia-ischemia and resuscitation
- MAP:
-
mean arterial blood pressure
- PI:
-
propidium iodide
- ROS:
-
reactive oxygen species
References
Datta R, Taneja N, Sukhatme VP, Qureshi SA, Weichselbaum R, Kufe DW 1993 Reactive oxygen intermediates target CC(A/T)6GG sequences to mediate activation of the early growth response 1 transcription factor gene by ionizing radiation. Proc Natl Acad Sci U S A 90: 2419–2422
Honkaniemi J, States BA, Weinstein PR, Espinoza J, Sharp FR 1997 Expression of zinc finger immediate early genes in rat brain after permanent middle cerebral artery occlusion. J Cereb Blood Flow Metab 17: 636–646
Honkaniemi J, Sharp FR 1996 Global ischemia induces immediate-early genes encoding zinc finger transcription factors. J Cereb Blood Flow Metab 16: 557–565
Gashler A, Sukhatme VP 1995 Early growth response protein 1 (Egr-1): prototype of a zinc-finger family of transcription factors. Prog Nucleic Acid Res Mol Biol 50: 191–224
Yan SF, Fujita T, Lu J, Okada K, Shan Zou Y, Mackman N, Pinsky DJ, Stern DM 2000 Egr-1, a master switch coordinating upregulation of divergent gene families underlying ischemic stress. Nat Med 6: 1355–1361
Ramji S, Ahuja S, Thirupuram S, Rootwelt T, Rooth G, Saugstad OD 1993 Resuscitation of asphyxic newborn infants with room air or 100% oxygen. Pediatr Res 34: 809–812
Reddy GR, Zawia NH 2000 Lead exposure alters Egr-1 DNA-binding in the neonatal rat brain. Int J Dev Neurosci 18: 791–795
Jung HY, Kang UG, Joo YH, Cho SC, Jeon SH, Park JB, Kim YS 1998 Electroconvulsive shock does not induce c-fos and junB, but TIS1 and TIS8/zif-268, in neonatal rat hippocampus. Brain Res Dev Brain Res 108: 303–306
Snyder-Keller A, Chandra R, Lin Y, Mitchell ES 2002 Basal EGR-1 (zif268, NGFI-A, Krox-24) expression in developing striatal patches: role of dopamine and glutamate. Brain Res 958: 297–304
Froen JF, Amerio G, Stray-Pedersen B, Saugstad OD 2002 Detrimental effects of nicotine and endotoxin in the newborn piglet brain during severe hypoxemia. Biol Neonate 82: 188–196
Vannucci RC, Brucklacher RM, Vannucci SJ 1999 CSF glutamate during hypoxia-ischemia in the immature rat. Brain Res Dev Brain Res 118: 147–151
Kiessling M, Stumm G, Xie Y, Herdegen T, Aguzzi A, Bravo R, Gass P 1993 Differential transcription and translation of immediate early genes in the gerbil hippocampus after transient global ischemia. J Cereb Blood Flow Metab 13: 914–924
Johansson IM, Wester P, Hakova M, Gu W, Seckl JR, Olsson T 2000 Early and delayed induction of immediate early gene expression in a novel focal cerebral ischemia model in the rat. Eur J Neurosci 12: 3615–3625
Paschen W, Linden T, Doutheil J 1998 Effects of transient cerebral ischemia on hsp40 mRNA levels in rat brain. Brain Res Mol Brain Res 55: 341–344
Herdegen T, Leah JD 1998 Inducible and constitutive transcription factors in the mammalian nervous system: control of gene expression by Jun, Fos and Krox, and CREB/ATF proteins. Brain Res Brain Res Rev 28: 370–490
DeGracia DJ, Montie HL 2004 Cerebral ischemia and the unfolded protein response. J Neurochem 91: 1–8
Martin LJ, Brambrink AM, Price AC, Kaiser A, Agnew DM, Ichord RN, Traystman RJ 2000 Neuronal death in newborn striatum after hypoxia-ischemia is necrosis and evolves with oxidative stress. Neurobiol Dis 7: 169–191
Munkeby BH, Borke WB, Bjornland K, Sikkeland LI, Borge GI, Halvorsen B, Saugstad OD 2004 Resuscitation with 100% O2 increases cerebral injury in hypoxemic piglets. Pediatr Res 56: 783–790
Vento M, Asensi M, Sastre J, Garcia-Sala F, Pallardo FV, Vina J 2001 Resuscitation with room air instead of 100% oxygen prevents oxidative stress in moderately asphyxiated term neonates. Pediatrics 107: 642–647
Goplerud JM, Kim S, Delivoria-Papadopoulos M 1995 The effect of post-asphyxial reoxygenation with 21% vs. 100% oxygen on Na+, K(+)-ATPase activity in striatum of newborn piglets. Brain Res 696: 161–164
Temesvari P, Karg E, Bodi I, Nemeth I, Pinter S, Lazics K, Domoki F, Bari F 2001 Impaired early neurologic outcome in newborn piglets reoxygenated with 100% oxygen compared with room air after pneumothorax-induced asphyxia. Pediatr Res 49: 812–819
Acknowledgements
The authors thank Ajay Verma, M.D., Ph.D., for his critical advice and the use of his laboratory facilities; David Jacobowitz, Ph.D., for his guidance and assistance with protein analysis; and Jozsef Czege, Ph.D., and Tom Baginski for their assistance with fluorescence microscopy and digital image analysis.
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The opinions or assertions contained herein are the private views of the authors and are not to be construed as official or to reflect the opinions of the Uniformed Services University of the Health Sciences, the Air Force, or Department of Defense.
Supported by a 2002 Young Investigator Award Research Grant, American Academy of Pediatrics, Neonatal Resuscitation Program, and a research grant from the National Naval Medical Center, Bethesda MD.
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Tyree, M., Dalgard, C. & O'Neill, J. Impact of Room Air Resuscitation on Early Growth Response Gene-1 in a Neonatal Piglet Model of Cerebral Hypoxic Ischemia. Pediatr Res 59, 423–427 (2006). https://doi.org/10.1203/01.pdr.0000199908.30751.ef
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DOI: https://doi.org/10.1203/01.pdr.0000199908.30751.ef
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