Abstract
Cumulative evidence demonstrates that apoptosis caused by oxidative stress plays a key role in neuronal cell death after transient focal cerebral ischemia. In this study, we investigated exactly the immunohistochemical alterations of neuronal nuclei (NeuN), Cu/Zn-SOD (superoxide dismutase), Mn-SOD, 4-hydroxy-2-nonenal (HNE), and single strand DNA (ssDNA) in the striatum from 3 h up to 15 days after transient focal cerebral ischemia in rats under the same conditions. A conspicuous decrease of NeuN immunoreactive neurons was observed in the ipsilateral striatum from 3 h up to 15 days after focal ischemia. For Cu/Zn-SOD, Mn-SOD and HNE immunostainings, the alteration of Cu/Zn-SOD and HNE immunoreactivity was more pronounced than that of Mn-SOD immunoreactivity in the shrunken or atrophic neurons of ipsilateral striatum 3 h after focal ischemia. Thereafter, a significant increase of HNE immunoreactivity was observed in the shrunken or atrophic neurons of ipsilateral striatum up to 15 days after focal ischemia. In contrast, a significant decrease of Cu/Zn-SOD immunoreactivity was found in the ipsilateral striatum from 3 up to 15 days after focal ischemia. On the other hand, a significant increase of Mn-SOD immunereactivity was observed in the ipsilateral striatum from 1 up to 7 days after focal ischemia. In addition, our Western blot analysis also showed a significant increase of Cu/Zn-SOD and Mn-SOD in the ipsilateral striatum 1 day after focal ischemia, as compared to sham-operated group. In contrast, a significant increase in the number of ssDNA immunoreactive apoptotic neurons was observed in the ipsilateral striatum from 3 h to 3 days after focal cerebral ischemia. The present results also suggest that increased reactive oxygen species (ROS) production during reperfusion may contribute to the induction of the alteration of lipid peroxidation and could thereby lead to apoptosis in neurons of the ipsilateral striatum after transient focal ischemia, because of an insufficient expression of Cu/Zn-SOD and Mn-SOD. Furthermore, our findings demonstrate that the lipid peroxidation against mitochondrial membrane may contribute to apoptosis of striatal neurons after transient focal ischemia. Thus our findings demonstrate that the protection of lipid peroxidation against mitochondrial membrane may offer a novel therapeutic strategy for brain stroke in humans.
Similar content being viewed by others
References
Sugawara T, Fujimura M, Morita-Fujimura Y, Kawase M, Chan PH (1999) Mitochondrial release of cytochrome c corresponds to the selective vulnerability of hippocampal CA1 neurons in rats after transient global cerebral ischemia. J Neurosci 19:RC39: 1–6
Abe K, Araki T, Kogure K (1988) Recovery from edema and of protein synthesis differs between in the cortex and caudate following transient focal cerebral ischemia in rats. J Neurochem 51:1470–1476
Araki T, Kato H, Shuto K, Itoyama Y (1998) Alterations in [3H]L-NG-nitroarginine binding in brain after transient global or transient focal ischemia in gerbils and rats. Eur J Pharmacol 354:153–159
Aronowski J, Strong R, Grotta JC (1997) Reperfusion injury: demonstration of brain damage produced by reperfusion after transient focal ischemia in rats. J Cereb Blood Flow Metab 17:1048–1056
Chan PH (1994) Oxygen radicals in focal cerebral ischemia. Brain Pathol 4:59–65
Chan PH (1996) Role of oxidants in ischemic brain damage. Stroke 27:1124–1129
Chan PH (2001) Reactive oxygen radicals in signaling and damage in the ischemic brain. J Cereb Blood Flow Metab 21:2–14
Chan PH, Kawase M, Murakami K, Chen SF, Li Y, Calagui B, Reola L, Carlson E, Epstein CJ (1998) Overexpression of SOD1 in transgenic rats protects vulnerable neurons against ischemic damage after global cerebral ischemia and reperfusion. J Neurosci 18:8292–8299
Dykens JA (1994) Isolated cerebral and cerebellar mitochondria produce free radicals when exposed to elevated Ca2+ and Na+: implications for neurodegeneration. J Neurochem 63:584–591
Estebauer H, Schaur RJ, Zollner H (1991) Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Radic Biol Med 11:81–128
Fujimura M, Morita-Fujimura Y, Copin JC, Copin JC, Kawase M, Chen PH (1999) Copper-zinc superoxide dismutase prevents the early decrease of apurinic/apyrimidinic endonuclease and subsequent DNA fragmentation after transient focal cerebral ischemia in mice. Stroke 30:2408–2415
Fujimura M, Morita-Fujimura Y, Noshita N, Sugawara T, Kawase M, Chan PH (2000) The cytosolic antioxidant copper/zinc-superoxide dismutase prevents the early release of mitochondrial cytochrome c in ischemic brain after transient cerebral ischemia in mice. J Neurosci 20:2817–2824
Himeda T, Mizuno K, Kato H, Araki T (2005) Effect of age on immunohistochemical changes in the mouse hippocampus. Mech Ageing Dev 126:673–677
Hudgins WR, Garcia JH (1970) Transorbital approach to the middle cerebral artery of the squirel monkey: a technique for experimental cerebral infarction applicable to ultrastructural studies. Stroke 1:107–111
Kametsu Y, Osuga S, Hakim AM (2003) Apoptosis occurs in the penumbra zone during short-duration focal ischemia in the rat. J Cereb Blood Flow Metab 23:416–422
Keller JN, Kindy MS, Holtsberg FW, St Clair DK, Yen HC, Germeyer A, Steiner SM, Bruce-Keller AJ, Hutchins JB, Mattson MP (1998) Mitochondrial manganese superoxide dismutase prevents neuronal apoptosis and reduces ischemic brain injury: suppression of peroxynitrite production, lipid peroxidation, and mitochondrial dysfunction. J Neurosci 18:687–697
Kinouchi H, Epstein CJ, Mizui T, Carlson E, Chen SF, Chan PH (1991) Attenuation of focal cerebral ischemic injury in transgenic mice overexpressing Cu/Zn superoxide dismutase. Proc Natl Acad Sci USA 88:11158–11162
Komine-Kobayashi M, Chou N, Mochizuki H, Nakano A, Mizuno Y, Urabe T (2004) Dual role of Fcγ receptor in transient focal cerebral ischemia in mice. Stroke 35:958–963
Kontos HA (2001) Oxygen radicals in cerebral ischemia. Stroke 11:2712–2716
Kurosaki R, Muramatsu Y, Kato H, Watanabe Y, Imai Y, Itoyama Y, Araki T (2005) Effect of angiotensin-converting enzyme inhibitor perindopril on interneurons in MPTP-treated mice. Eur Neuropsychopharmacol 15:57–67
Longa EZ, Weinstein PR, Carlson S, Cummins R (1989) Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke 20:84–91
Love S (1999) Oxidative stress in brain ischemia. Brain Pathol 9:119–131
Martin LJ, Al-Abdulla NA, Brambrink AM, Kirsch JR, Sieber FE, Portera-Cailliau C (1998) Neurodegeneration in excitotoxicity, global cerebral ischemia, and target deprivation: a perspective on the contributions of apoptosis and necrosis. Brain Res Bull 46:281–309
Morikawa S, Kurauchi O, Tanaka M, Yoneda M, Uchida K, Itakura A, Furugori K, Mizutani S, Tomoda Y (1997) Increased mitochondrial damage by lipid peroxidation in trophoblast cells of preeclamptic placentas. Biochem Mol Biol Int 41:767–775
Noshita N, Sugawara T, Hayashi T, Lewen A, Omar G, Chan PH (2002) Copper/zinc superoxide dismutase attenuates neuronal cell death by preventing extracellular signal-regulated kinase activation after transient focal cerebral ischemia in mice. J Neurosci 22:7923–7930
O’Brien RG, Waltz AG (1973) Transorbital approach for occluding the middle cerebral artery without craniectomy. Stroke 4:201–206
Oliver CN, Starke-Reed PE, Stadtman ER, Liu GJ, Carney JM, Floyd RA (1990) Oxidative damage to brain proteins, loss of glutamine synthetase activity, and production of free radicals during ischemia/reperfusion-induced injury to gerbil brain. Proc Natl Acad Sci USA 87:5144–5147
Sakuma M, Hayakawa N, Kato H, Araki T (2008) Time dependent changes of striatal interneurons after focal cerebral ischemia. J Neural Transm 115:413–422
Siesjo BK, Agardh CD, Bengtsson F (1989) Free radicals and brain damage. Cerebrovasc Brain Metab Rev 1:165–211
Takagi S, Hayakawa N, Kimoto H, Kato H, Araki T (2007) Damage to oligodendrocytes in the striatum after MPTP neurotoxicity in mice. J Neural Transm 114:1553–1557
White RJ, Reynolds IJ (1996) Mitochondrial depolarization in glutamate-stimulated neurons: an early signal specific to excitotoxin exposure. J Neurosci 16:5688–5697
Yang G, Chan PH, Chen J, Carlson E, Chen SF, Weinstein P, Epstein CJ, Kamii H (1994) Human copper-zinc superoxide dismutase transgenic mice are highly resistant to reperfusion injury after focal cerebral ischemia. Stroke 25:165–170
Yoshino H, Hattori N, Urabe T, Uchida K, Tanaka M, Mizuno Y (1997) Postischemic accumulation of lipid peroxydation products in the rat brain: immunohistochemical detection of 4-hydroxy-2-nonenal modified proteins. Brain Res 767:81–86
Acknowledgments
This study was supported in part by a Grant-in-Aid for Scientific Research (13671095 and 13670627) from the Ministry of Science and Education in Japan. We thank Mio Sakuma for technical assistance.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Matsuda, S., Umeda, M., Uchida, H. et al. Alterations of oxidative stress markers and apoptosis markers in the striatum after transient focal cerebral ischemia in rats. J Neural Transm 116, 395–404 (2009). https://doi.org/10.1007/s00702-009-0194-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00702-009-0194-0