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 Mori, T. Articles by Asano, T. Search for Related Content PubMed PubMed Citation Articles by Mori, T. Articles by Asano, T. Pubmed/NCBI databases Compound via MeSH Substance via MeSH Hazardous Substances DB IRON MANGANESE COMPOUNDS MANGANESE, ELEMENTAL Related Collections Animal models of human disease Cerebral Aneurysm, AVM, & Subarachnoid hemorrhage Pathology of Stroke

(Stroke. 2001;32:636.)
© 2001 American Heart Association, Inc.

Original Contributions

Intracisternal Increase of Superoxide Anion Production in a Canine Subarachnoid Hemorrhage Model

Takashi Mori, DVM, PhD; Kazuya Nagata, MD, DMSc; Terrence Town, BA; Jun Tan, MD, PhD; Toru Matsui, MD, DMSc Takao Asano, MD, DMSc

From the Institute of Laboratory Animal Science (T.M., T.T., J.T.) and the Department of Neurosurgery (T.M., T.A.), Saitama Medical Center/School, Saitama, and the Department of Neurosurgery (K.N.), Showa General Hospital, Tokyo, Japan.

Correspondence to Takao Asano, MD, Department of Neurosurgery, Saitama Medical Center/School, 1981 Kamoda, Kawagoe, Saitama 350-8550, Japan. E-mail asano{at}ns2.saitama-med.ac.jp

Background and Purpose—Reactive oxygen species (ROS) are thought to be primary in the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage (SAH). However, as direct evidence of ROS has not yet been demonstrated in cerebral vasospasm, we sought to substantiate superoxide anion (·O₂^-) generation in the subarachnoid space after SAH using a modification of Karnovsky’s manganese/diaminobenzidine (Mn²⁺/DAB) technique.

Methods—SAH or sham operation was induced according to a 2-hemorrhage model in a total of 24 beagle dogs. On day 2 or 7 after SAH or sham operation, dogs were intrathecally infused with buffer containing Mn²⁺ and DAB, and the brain stem was prepared for light and electron microscopy. Possible colocalization of ferrous (Fe²⁺) or ferric (Fe³⁺) iron ions with ·O₂^- was also examined with the use of Turnbull blue or Berlin blue staining, respectively.

Results—Light microscopy revealed amorphous, amber deposits within the subarachnoid hematoma, the periarterial space, and the tunica adventitia of the basilar artery on days 2 and 7 after SAH. ·O₂^- deposits were eliminated by addition of superoxide dismutase or exclusion of either Mn²⁺ or DAB from the perfusate, confirming the specificity of the reaction. These deposits were colocalized with blue reaction deposits indicating Fe²⁺ and Fe³⁺. Within the subarachnoid space, ·O₂^- indicating electron-dense fine granules were preferentially located around degenerated erythrocytes and, secondarily, infiltrating macrophages and neutrophils.

Conclusions—We show direct evidence for enhanced production of ·O₂^- and Fe²⁺/Fe³⁺ iron ions in the subarachnoid space after SAH, lending further support to the pathogenic role of ROS in cerebral vasospasm after SAH.

Key Words: reactive oxygen species • subarachnoid hemorrhage • superoxides • vasospasm, intracranial • dogs

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]

				D. Zemke, M. U Farooq, A. Mohammed Yahia, and A. Majid Delayed ischemia after subarachnoid hemorrhage: result of vasospasm alone or a broader vasculopathy? Vascular Medicine, August 1, 2007; 12(3): 243 - 249. [Abstract] [PDF]

				M. Yamaguchi, C. Zhou, D. D. Heistad, Y. Watanabe, and J. H. Zhang Gene Transfer of Extracellular Superoxide Dismutase Failed to Prevent Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage Stroke, November 1, 2004; 35(11): 2512 - 2517. [Abstract] [Full Text] [PDF]

				H. Suzuki, M. Muramatsu, T. Kojima, and W. Taki Intracranial Heme Metabolism and Cerebral Vasospasm After Aneurysmal Subarachnoid Hemorrhage Stroke, December 1, 2003; 34(12): 2796 - 2800. [Abstract] [Full Text] [PDF]

				F. M. Faraci Vascular Protection Stroke, February 1, 2003; 34(2): 327 - 329. [Full Text] [PDF]

				Y. Watanabe, Y. Chu, J. J. Andresen, H. Nakane, F. M. Faraci, and D. D. Heistad Gene Transfer of Extracellular Superoxide Dismutase Reduces Cerebral Vasospasm After Subarachnoid Hemorrhage Stroke, February 1, 2003; 34(2): 434 - 440. [Abstract] [Full Text] [PDF]

				M. J. McGirt, A. Parra, H. Sheng, Y. Higuchi, T. D. Oury, D. T. Laskowitz, R. D. Pearlstein, and D. S. Warner Attenuation of Cerebral Vasospasm After Subarachnoid Hemorrhage in Mice Overexpressing Extracellular Superoxide Dismutase Stroke, September 1, 2002; 33(9): 2317 - 2323. [Abstract] [Full Text] [PDF]

				S. P. Didion, C. A. Hathaway, and F. M. Faraci Superoxide levels and function of cerebral blood vessels after inhibition of CuZn-SOD Am J Physiol Heart Circ Physiol, October 1, 2001; 281(4): H1697 - H1703. [Abstract] [Full Text] [PDF]

				A. Saito, H. Kamii, I. Kato, S. Takasawa, T. Kondo, P. H. Chan, H. Okamoto, T. Yoshimoto, and R. L. Macdonald Transgenic CuZn-Superoxide Dismutase Inhibits NO Synthase Induction in Experimental Subarachnoid Hemorrhage Editorial Comment Stroke, July 1, 2001; 32(7): 1652 - 1657. [Abstract] [Full Text] [PDF]