This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kim, G. W. Articles by Chan, P. H. Search for Related Content PubMed PubMed Citation Articles by Kim, G. W. Articles by Chan, P. H. Related Collections Apoptosis Genetically altered mice Acute Cerebral Infarction Oxidant stress

(Stroke. 2002;33:809.)
© 2002 American Heart Association, Inc.

Original Contributions

Manganese Superoxide Dismutase Deficiency Exacerbates Cerebral Infarction After Focal Cerebral Ischemia/Reperfusion in Mice

Implications for the Production and Role of Superoxide Radicals

Gyung W. Kim, MD, PhD^*; Takeo Kondo, MD, PhD^*; Nobuo Noshita, MD Pak H. Chan, PhD

From the Department of Neurosurgery, Department of Neurology & Neurological Sciences, and Program in Neurosciences, Stanford University School of Medicine, Stanford, Calif.

Correspondence to Pak H. Chan, PhD, Neurosurgical Labs, Stanford University, 1201 Welch Rd., MSLS #P304, Stanford, CA 94305-5487. E-mail phchan{at}leland.stanford.edu

Background and Purpose— Superoxide anion radicals (O₂^·-) are implicated in ischemia/reperfusion injury, although a direct relationship has not been elucidated. Recently, a specific method of hydroethidine (HEt) oxidation by O₂^·- was developed to detect O₂^·- production in a variety of experimental brain injury models. To clarify the role of O₂^·- in the mechanism of ischemia/reperfusion, we investigated O₂^·- production after ischemia/reperfusion and ischemia/reperfusion injury in mutant mice deficient in mitochondrial manganese superoxide dismutase (MnSOD) and in wild-type littermates.

Methods— Ischemia/reperfusion was performed for 60 minutes using intraluminal suture blockade of the middle cerebral artery in the mutant or wild-type mice. We evaluated fluorescent kinetics of HEt or ethidium, the oxidized form of HEt, in brains after an intravenous injection of HEt, followed by measurement of cellular O₂^·- production using specific HEt oxidation by O₂^·- before and after ischemia/reperfusion. Furthermore, we compared O₂^·- production and subsequent infarct volume in the mice using triphenyltetrazolium chloride after ischemia/reperfusion.

Results— HEt oxidation to ethidium is primarily a result of mitochondrially produced O₂^·- under physiological conditions. Cerebral ischemia/reperfusion produced O₂^·- prominently in neurons shortly after reperfusion, followed by a delayed increase in endothelial cells. A deficiency in MnSOD in mutant mice increased mitochondrial O₂^·- production and exacerbated cerebral infarction, worsening neurological deficits after ischemia/reperfusion.

Conclusion— These results suggest that mitochondrial O₂^·- production may be a critical step underlying the mechanism of ischemia/reperfusion injury and that MnSOD may protect against ongoing oxidative cell death after ischemia/reperfusion.

Key Words: cerebral ischemia, transient • oxidative stress • superoxide dismutase • mice, transgenic

This article has been cited by other articles:

				G. S. Kim, J. E. Jung, K. Niizuma, and P. H. Chan CK2 Is a Novel Negative Regulator of NADPH Oxidase and a Neuroprotectant in Mice after Cerebral Ischemia J. Neurosci., November 25, 2009; 29(47): 14779 - 14789. [Abstract] [Full Text] [PDF]

				J. E. Jung, G. S. Kim, P. Narasimhan, Y. S. Song, and P. H. Chan Regulation of Mn-Superoxide Dismutase Activity and Neuroprotection by STAT3 in Mice after Cerebral Ischemia J. Neurosci., May 27, 2009; 29(21): 7003 - 7014. [Abstract] [Full Text] [PDF]

				L. Llacuna, M. Mari, J. M. Lluis, C. Garcia-Ruiz, J. C. Fernandez-Checa, and A. Morales Reactive Oxygen Species Mediate Liver Injury Through Parenchymal Nuclear Factor-{kappa}B Inactivation in Prolonged Ischemia/Reperfusion Am. J. Pathol., May 1, 2009; 174(5): 1776 - 1785. [Abstract] [Full Text] [PDF]

				N. Ooe, K. Motonaga, K. Kobayashi, K. Saito, and H. Kaneko Functional Characterization of Basic Helix-Loop-Helix-PAS Type Transcription Factor NXF in Vivo: PUTATIVE INVOLVEMENT IN AN "ON DEMAND" NEUROPROTECTION SYSTEM J. Biol. Chem., January 9, 2009; 284(2): 1057 - 1063. [Abstract] [Full Text] [PDF]

				V. M. Miller and S. P. Duckles Vascular Actions of Estrogens: Functional Implications Pharmacol. Rev., June 1, 2008; 60(2): 210 - 241. [Abstract] [Full Text] [PDF]

				K. A. Brown, S. P. Didion, J. J. Andresen, and F. M. Faraci Effect of Aging, MnSOD Deficiency, and Genetic Background on Endothelial Function: Evidence for MnSOD Haploinsufficiency Arterioscler Thromb Vasc Biol, September 1, 2007; 27(9): 1941 - 1946. [Abstract] [Full Text] [PDF]

				F. G. De Felice, P. T. Velasco, M. P. Lambert, K. Viola, S. J. Fernandez, S. T. Ferreira, and W. L. Klein Abeta Oligomers Induce Neuronal Oxidative Stress through an N-Methyl-D-aspartate Receptor-dependent Mechanism That Is Blocked by the Alzheimer Drug Memantine J. Biol. Chem., April 13, 2007; 282(15): 11590 - 11601. [Abstract] [Full Text] [PDF]

				V. A Fitsanakis, G. Piccola, A. P. Marreilha dos Santos, J. L Aschner, and M. Aschner Putative proteins involved in manganese transport across the blood-brain barr 1ier Human and Experimental Toxicology, April 1, 2007; 26(4): 295 - 302. [Abstract] [PDF]

				P. Pacher, J. S. Beckman, and L. Liaudet Nitric Oxide and Peroxynitrite in Health and Disease Physiol Rev, January 1, 2007; 87(1): 315 - 424. [Abstract] [Full Text] [PDF]

				M. Ohashi, M. S. Runge, F. M. Faraci, and D. D. Heistad MnSOD Deficiency Increases Endothelial Dysfunction in ApoE-Deficient Mice Arterioscler Thromb Vasc Biol, October 1, 2006; 26(10): 2331 - 2336. [Abstract] [Full Text] [PDF]

				L. Esposito, J. Raber, L. Kekonius, F. Yan, G.-Q. Yu, N. Bien-Ly, J. Puolivali, K. Scearce-Levie, E. Masliah, and L. Mucke Reduction in mitochondrial superoxide dismutase modulates Alzheimer's disease-like pathology and accelerates the onset of behavioral changes in human amyloid precursor protein transgenic mice. J. Neurosci., May 10, 2006; 26(19): 5167 - 5179. [Abstract] [Full Text] [PDF]

				B. I. Lee, P. H. Chan, and G. W. Kim Metalloporphyrin-Based Superoxide Dismutase Mimic Attenuates the Nuclear Translocation of Apoptosis-Inducing Factor and the Subsequent DNA Fragmentation After Permanent Focal Cerebral Ischemia in Mice Stroke, December 1, 2005; 36(12): 2712 - 2717. [Abstract] [Full Text] [PDF]

				M. Tribus, J. Galehr, P. Trojer, G. Brosch, P. Loidl, F. Marx, H. Haas, and S. Graessle HdaA, a Major Class 2 Histone Deacetylase of Aspergillus nidulans, Affects Growth under Conditions of Oxidative Stress Eukaryot. Cell, October 1, 2005; 4(10): 1736 - 1745. [Abstract] [Full Text] [PDF]

				H. Xie, P. E. Ray, and B. L. Short NF-{kappa}B Activation Plays a Role in Superoxide-Mediated Cerebral Endothelial Dysfunction After Hypoxia/Reoxygenation Stroke, May 1, 2005; 36(5): 1047 - 1052. [Abstract] [Full Text] [PDF]

				C. Yan, A. Huang, Z. Wu, P. M. Kaminski, M. S. Wolin, T. H. Hintze, G. Kaley, and D. Sun Increased superoxide leads to decreased flow-induced dilation in resistance arteries of Mn-SOD-deficient mice Am J Physiol Heart Circ Physiol, May 1, 2005; 288(5): H2225 - H2231. [Abstract] [Full Text] [PDF]

				H. Zhao, J. Joseph, H. M. Fales, E. A. Sokoloski, R. L. Levine, J. Vasquez-Vivar, and B. Kalyanaraman Detection and characterization of the product of hydroethidine and intracellular superoxide by HPLC and limitations of fluorescence PNAS, April 19, 2005; 102(16): 5727 - 5732. [Abstract] [Full Text] [PDF]

				J. Wang and S. E. Tsirka Tuftsin Fragment 1-3 Is Beneficial When Delivered After the Induction of Intracerebral Hemorrhage Stroke, March 1, 2005; 36(3): 613 - 618. [Abstract] [Full Text] [PDF]

				A. Y. Zhang, E. G. Teggatz, A.-P. Zou, W. B. Campbell, and P.-L. Li Endostatin uncouples NO and Ca2+ response to bradykinin through enhanced O2-{middle dot} production in the intact coronary endothelium Am J Physiol Heart Circ Physiol, February 1, 2005; 288(2): H686 - H694. [Abstract] [Full Text] [PDF]

				W. S. da-Silva, A. Gomez-Puyou, M. T. de Gomez-Puyou, R. Moreno-Sanchez, F. G. De Felice, L. de Meis, M. F. Oliveira, and A. Galina Mitochondrial Bound Hexokinase Activity as a Preventive Antioxidant Defense: STEADY-STATE ADP FORMATION AS A REGULATORY MECHANISM OF MEMBRANE POTENTIAL AND REACTIVE OXYGEN SPECIES GENERATION IN MITOCHONDRIA J. Biol. Chem., September 17, 2004; 279(38): 39846 - 39855. [Abstract] [Full Text] [PDF]

				J. J. Andresen, F. M. Faraci, and D. D. Heistad Vasomotor responses in MnSOD-deficient mice Am J Physiol Heart Circ Physiol, September 1, 2004; 287(3): H1141 - H1148. [Abstract] [Full Text] [PDF]

				D. S. Warner, H. Sheng, and I. Batinic-Haberle Oxidants, antioxidants and the ischemic brain J. Exp. Biol., August 15, 2004; 207(18): 3221 - 3231. [Abstract] [Full Text] [PDF]

				Y. Gursoy-Ozdemir, A. Can, and T. Dalkara Reperfusion-Induced Oxidative/Nitrative Injury to Neurovascular Unit After Focal Cerebral Ischemia Stroke, June 1, 2004; 35(6): 1449 - 1453. [Abstract] [Full Text] [PDF]

				G. W. Kim, Y. Gasche, S. Grzeschik, J.-C. Copin, C. M. Maier, and P. H. Chan Neurodegeneration in Striatum Induced by the Mitochondrial Toxin 3-Nitropropionic Acid: Role of Matrix Metalloproteinase-9 in Early Blood-Brain Barrier Disruption? J. Neurosci., September 24, 2003; 23(25): 8733 - 8742. [Abstract] [Full Text] [PDF]