Ischemia-induced release of cytochrome c from mitochondria and up-regulation of Bcl-2 expression in rat hippocampus

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Journal Title:
ACTA PHYSIOLOGICA SINICA
Issue:
Volume 56, Issue 02, 2004
DOI:
Key Word:
brain ischemia;cytochrome c;Bcl-2;N-methyl-D-aspartate receptor;L-type voltage-gated Ca2+ channel

Abstract: To evaluate the effects of different antagonists on the release of cytochrome c from mitochondria to cytosol and the expression of Bcl-2 in mitochondria in rat hippocampus after ischemia, we examined Bcl-2 and cytochrome c expression by immunoblotting using 4-vessel occlusion (4-VO) as brain ischemia model. The results showed that after 24 h ischemia/reperfusion (I/R) cytochrome c decreased markedly in mitochondria, which was correspondingly increased in the cytosolic fraction. Bcl-2 expression was time-dependent, reaching its peak level after 6 h I/R. In all those samples, there were no alterations in the subcellular distribution of cytochrome oxidase, a mitochondrial respiratory chain protein. The decreases in Bcl-2 and cytochrome c in mitochondria were restored by pretreatment with non-competitive NMDA receptor antagonist ketamine or L-type voltage-gated Ca2+ channel (L-VGCC) antagonist nifedipine at 20 min prior to ischemia. The results demonstrate that the release of cytochrome c from mitochondria to cytosol and the up-regulation of Bcl-2are possibly mediated by NMDA receptors or L-VGCC following brain ischemia. Cytochrome c release may be injurious while Bcl-2 upregulation may be protective to ischemic hippocampus.

  • [1]Boyer PD, Chance B, Emster L, Mitchell P, Racker E, Slater EC.Oxidative phosphorylation and photophosphorylation. Annu Rev Biochem 1977; 46: 955-1026.
  • [2]Liu X, Kim CN, Yang J, Jemmerson R, Wang X. Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome c. Cell 1996; 86: 147-157.
  • [3]Vaux DL. CED-4-the third horseman of apoptosis. Cell 1997;90: 389-390.
  • [4]Zou H, Henzel WJ, Liu X, Lutschg A, Wang X. Apaf- 1, a human protein homologous to C. elegans CED-4, participates in cytochrome c-dependent activation of caspase-3. Cell 1997; 90: 405-413.
  • [5]Li P, Nijhawan D, Budihardjo I, Srinivasula SM, Ahmad M,Alnemri ES, Wang X. Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 1997; 91: 479-489.
  • [6]Zhang CY, Shen WH, Zhang GY. N-methyl-D-aspartate receptor and L-type voltage-gated Ca2+ channel antagonists suppress the release of cytochrome c and the expression of procaspase-3 in rat hippocampus after global brain ischemia. Neurosci Lett 2002;328: 265-268.
  • [7]Lithgow T, Van-Driel R, Bertram JF, Strasser A. The protein product of the oncogene Bcl-2 is a component of the nuclear envelope, the endoplasmic reticulum, and the outer mitochondrial membrane. Cell 1994; 5: 411-417.
  • [8]Kluck RM, Bossy-Wetzel E, Green DR, Newmeyer DD. The release of cytochrome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis. Science 1997; 275:1132-1136.
  • [9]Yang J, Lin X, Bhalla K, Kim CN, Ibrado AM, Cai J, Peng T, Jones DP, Wang X. Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science 1997; 275:1129-1132.
  • [10]Behl C, Hovey L, Krajewski S, Schubert D, Reed JC. Bcl-2prevents killing of neuronal cells by glutamate but not by amyloid beta protein. Biochem Biophys Res Commun 1993; 197:949-956.
  • [11]Zhong LT, Kane DJ, Bredesent DE. BCL-2 blocks glutamate toxicity in neural cell lines. Mol Brain Res 1993; 19: 353-355.
  • [12]Shen WH, Zhang CY, Zhang GY. Nuclear factor kB activation is mediated by NMDA and non-NMDA receptor and L-type voltage-gated Ca2+ channel following severe global iscbemia in rat hippocampus. Brain Res 2002; 933: 23-30.
  • [13]Maulik D, Numagami Y, Ohnishi ST, Mishra OP, DelivoriaPapadopoulos M. Direct measurement of oxygen free radicals during in utero hypoxia in the fetal guinea pig brain. Brain Res 1998; 798: 166-172.
  • [14]Numagami Y, Zubrow AB, Mishra OP, Delivoria-Papadopoulos M. Lipid free radical generation and brain cell membrane alternation following nitric oxide synthase inhibition during cerebral hypoxia in the newborn piglets. J Neurochem 1997; 69: 1542-1547.
  • [15]Susin SA, Zamzami N, Castedo M, Hirsch T, Marchetti P, Macho A, Daugas E, Geuskens M, Kroemer G. Bcl-2 inhibits the mitochondrial release of an apoptogenic protease. J Exp Med 1996; 184: 1331-1341.
  • [16]Fisher M, Bogousslavsky J. Further evolution toward effective therapy for acute ischemic stroke. JAMA 1998; 279: 1298-1303.
  • [17]Shen WH, Zhang CY, Zhang GY. Antioxidants attenuate reperfusion injury after global brain ischemia through inhibiting nuclear factor-kappa B activity in rats. Acta Pharmacol Sin 2003; 24: 1125-1130.
  • [18]Lowry OH, Rosebrough HJ, Fart AL. Protein measurement with Folin-Phenol reagent. J Biol Chem 1951; 193: 265-275.
  • [19]Sambrook J, Fritsch EF, Maniatis T, eds. Immunoassay of Expressed Protein, Molecular Cloning: A Laboratory Manual.2nd ed. Cold Spring Harbor Laboratory Press, 1989: 880.
  • [20]Shen WH, Zhang CY, Zhang GY. Modulation of Iκ B kinase autophosphorylation and activity following brain ischemia. Acta Pharmacol Sin 2003; 24:311-315.
  • [21]MacManus JP, Linnik MD. Gene expression induced by cerebral ischemia: an apoptotic perspective. J Cereb Blood Flow Metab 1997; 17: 815-832.
  • [22]Merry DE, Korsmeyer SJ. Bcl-2 gene family in the nervous system. Annu Rev Neurosci 1997; 20: 245-267.
  • [23]Gillardon F, Lenz C, Waschke KF, Krajewski S, Reed JC,Zimmermann M, Kuschinsky W. Altered expression of Bcl-2,Bcl-X, Bax, and c-Fos colocalizes with DNA fragmentation and ischemic cell damage following middle cerebral artery occlusion in rots. Brain Res Mol Brain Res 1996; 40: 254-260.
  • [24]Urabe T, Hattori N, Yoshikawa W, Yoshino H, Uchida K, Mizuno Y. Colocalization of Bcl-2 and 4-hydroxynonenal modified proteins in microglial cells and neurons of rat brain following transient focal ischemia. Neurosci Lett 1998; 247: 159-162.
  • [25]Choi DW. Glutamate neurotoxicity and diseases of the nervous system. Neuron 1988; 1:623-634.
  • [26]Choi DW. Glutamate receptors and the induction of excitotoxic neuronal death. Prog Brain Res 1994; 100: 47-51.
  • [27]Budd SL, Tenneti L, Lishnak T, Lipton SA. Mitochondrial and extramitochondrial apoptotic signaling pathways in cerebrocortical neurons. Proc Nail Acad Sci 2000; 97: 6161-6166.
  • [28]Marshall KA, Daniel SE, Cairns N, Jenner P, Halliwell B.Upregulation of the anti-apoptotic protein Bcl-2 may be an early event in neurodegeneration: studies on Parkinson' s and incidental Lewy body disease. Biochem Biophys Res Commun 1997; 240: 84-87.
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