Abstract
Ischemic preconditioning (IPC) represents an important adaptation of CNS to sub-lethal ischemia, which results in increased tolerance of CNS to the lethal ischemia. Ischemia-induced mitochondrial apoptosis is considered to be an important event leading to neuronal cell death after cerebral blood flow arrest. In presented study, we have determined the effect of IPC on ischemia/reperfusion-induced mitochondrial apoptosis. Global brain ischemia was induced by permanent occlusion of vertebral arteries and temporal occlusion of carotid arteries for 15 min. Rats were preconditioned by 5 min of sub-lethal ischemia and 2 days later 15 min of lethal ischemia was induced. With respect to mitochondrial apoptosis initiation, translocation of p53 to mitochondria was observed in hippocampus but not in cerebral cortex. However, level of both apoptotic bax and anti-apoptotic bcl-xl in both hippocampal and cortical mitochondria was unchanged after global brain ischemia. Detection of genomic DNA fragmentation as well as Fluoro-Jade C staining showed that ischemia induces apoptosis in vulnerable CA1 layer of rat hippocampus. IPC abolished completely ischemia-induced translocation of p53 to mitochondria and had significant protective effect on ischemia-induced DNA fragmentation. In addition, significant decrease of Fluoro-Jade C positive cells was observed as well. Our results indicate that IPC abolished almost completely both initiation and execution of mitochondrial apoptosis induced by global brain ischemia.
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Beere HM, Wolf BB, Cain K, Mosser DD, Mahboubi A, Kuwana T, Tailor P, Morimoto RI, Cohen GM, Green DR (2000) Heat-shock protein 70 inhibits apoptosis by preventing recruitment of procaspase-9 to the Apaf-1 apoptosome. Nat Cell Biol 2:469–475. doi:10.1038/35019501
Billen LP, Kokoski CL, Lovell JF, Leber B, Andrews DW (2008) Bcl-XL inhibits membrane permeabilization by competing with Bax. PLoS Biol 6:1268–1280. doi:10.1371/journal.pbio.0060147
Brunet A, Datta SR, Greenberg ME (2001) Transcription-dependent and -independent control of neuronal survival by the PI3 K-Akt signaling pathway. Curr Opin Neurobiol 11:297–305. doi:10.1016/S0959-4388(00)00211-7
Buss RR, Sun W, Oppenheim RW (2006) Adaptive roles of programmed cell death during nervous system development. Annu Rev Neurosci 29:1–35. doi:10.1146/annurev.neuro.29.051605.112800
Dirnagl U, Simon RP, Hallenbeck JM (2003) Ischemic tolerance and endogenous neuroprotection. Trends Neurosci 26:248–254. doi:10.1016/S0166-2236(03)00071-7
Enari M, Sakahira H, Yokoyama H, Okawa K, Iwamatsu A, Nagata S (1998) A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391:43–50
Endo H, Kamada H, Nito C, Nishi T, Chan PH (2006a) Mitochondrial translocation of p53 mediates release of cytochrome c and hippocampal CA1 neuronal death after transient global cerebral ischemia in rats. J Neurosci 26:7974–7983. doi:10.1523/JNEUROSCI.0897-06.2006
Endo H, Nito C, Kamada H, Nishi T, Chan PH (2006b) Activation of the Akt/GSK3beta signaling pathway mediates survival of vulnerable hippocampal neurons after transient global cerebral ischemia in rats. J Cereb Blood Flow Metab 26:1479–1489. doi:10.1038/sj.jcbfm.9600303
Gidday JM (2006) Cerebral preconditioning and ischaemic tolerance. Nat Rev Neurosci 7:437–448. doi:10.1038/nrn1927
Héron A, Pollard H, Dessi F, Moreau J, Lasbennes F, Ben-Ari Y, Charriaut-Marlangue C (1993) Regional variability in DNA fragmentation after global ischemia evidenced by combined histological and gel electrophoresis observations in the rat brain. J Neurochem 61:1973–1976. doi:10.1111/j.1471-4159.1993.tb09843.x
Hetz C, Vitte PA, Bombrun A, Rostovtseva TK, Montessuit S, Hiver A, Schwarz MK, Church DJ, Korsmeyer SJ, Martinou JC, Antonsson B (2005) Bax channel inhibitors prevent mitochondrion-mediated apoptosis and protect neurons in a model of global brain ischemia. J Biol Chem 280:42960–42970. doi:10.1074/jbc.M505843200
Jin K, Chen J, Nagayama T, Chen M, Sinclair J, Graham SH, Simon RP (1999) In situ detection of neuronal DNA strand breaks using the Klenow fragment of DNA polymerase I reveals different mechanisms of neuron death after global cerebral ischemia. J Neurochem 72:1204–1214. doi:10.1046/j.1471-4159.1999.0721204.x
Jin K, Graham SH, Mao X, Nagayama T, Simon RP, Greenberg DA (2001) Fas (CD95) may mediate delayed cell death in hippocampal CA1 sector after global cerebral ischemia. J Cereb Blood Flow Metab 21:1411–1421. doi:10.1097/00004647-200112000-00005
Kirino T (2002) Ischemic tolerance. J Cereb Blood Flow Metab 22:1283–1296. doi:10.1097/00004647-200211000-00001
Kirino T, Tsujita Y, Tamura A (1991) Induced tolerance to ischemia in gerbil hippocampal neurons. J Cereb Blood Flow Metab 11:299–307
Kitagawa K, Matsumoto M, Kuwabara K, Tagaya M, Ohtsuki T, Hata R, Ueda H, Handa N, Kimura K, Kamada T (1991) “Ischemic tolerance” phenomenon detected in various brain regions. Brain Res 561:203–211. doi:10.1016/0006-8993(91)91596-S
Krajewski S, Krajewska M, Ellerby LM, Welsh K, Xie Y, Deveraux OL, Salvesen GS, Bredesen DE, Rosenthal RE, Fiskum G, Reed JC (1999) Release of caspase-9 from mitochondria during neuronal apoptosis and cerebral ischemia. Proc Natl Acad Sci USA 96:5752–5757. doi:10.1073/pnas.96.10.5752
Liu Y, Kato H, Nakata N, Kogure K (1992) Protection of rat hippocampus against ischemic neuronal damage by pretreatment with sub-lethal ischemia. Brain Res 586:121–124. doi:10.1016/0006-8993(92)91380-W
Liu Y, Kato H, Nakata N, Kogure K (1993) Temporal profile of heat shock protein 70 synthesis in ischemic tolerance induced by preconditioning ischemia in rat hippocampus. Neuroscience 56:921–927. doi:10.1016/0306-4522(93)90138-6
MacManus JP, Buchan AM, Hill IE, Rasquinha I, Preston E (1993) Global ischemia can cause DNA fragmentation indicative of apoptosis in rat brain. Neurosci Lett 164:89–92. doi:10.1016/0304-3940(93)90864-H
Miyawaki T, Mashiko T, Ofengeim D, Flannery RJ, Noh KM, Fujisawa S, Bonanni L, Bennett MV, Zukin RS, Jonas EA (2008) Ischemic preconditioning blocks BAD translocation, Bcl-xL cleavage, and large channel activity in mitochondria of postischemic hippocampal neurons. Proc Natl Acad Sci USA 105:4892–4897. doi:10.1073/pnas.0800628105
Nishi S, Taki W, Uemura Y, Higashi T, Kikuchi H, Kudoh H, Satoh M, Nagata K (1993) Ischemic tolerance due to the induction of HSP70 in a rat ischemic recirculation model. Brain Res 615:281–288. doi:10.1016/0006-8993(93)90039-P
Obrenovitch TP (2008) Molecular physiology of preconditioning-induced brain tolerance to ischemia. Physiol Rev 88:211–247. doi:10.1152/physrev.00039.2006
Perez-Pinzon MA, Born JG (1999) Rapid preconditioning neuroprotection following anoxia in hippocampal slices: role of the K+ ATP channel and protein kinase C. Neuroscience 89:453–459. doi:10.1016/S0306-4522(98)00560-0
Perez-Pinzon MA, Xu GP, Born J, Lorenyo J, Busto R, Rosenthal M, Sick TJ (1999) Cytochrome C is released from mitochondria into the cytosol after cerebral anoxia or ischemia. J Cereb Blood Flow Metab 19:39–43. doi:10.1097/00004647-199901000-00004
Polosa PL, Attardi G (1991) Distinctive pattern and translational control of mitochondrial protein synthesis in rat brain synaptic endings. J Biol Chem 266:10011–10017
Polster BM, Fiskum G (2004) Mitochondrial mechanisms of neural cell apoptosis. J Neurochem 90:1281–1289. doi:10.1111/j.1471-4159.2004.02572.x
Pulsinelli WA, Brierley JB, Plum F (1982) Temporal profile of neuronal damage in a model of transient forebrain ischemia. Ann Neurol 11:491–498. doi:10.1002/ana.410110509
Racay P, Tatarkova Z, Drgova A, Kaplan P, Dobrota D (2007) Effect of ischemic preconditioning on mitochondrial dysfunction and mitochondrial p53 translocation after transient global cerebral ischemia in rats. Neurochem Res 32:1823–1832. doi:10.1007/s11064-007-9437-3
Schmued LC, Stowers CC, Scallet AC, Xu L (2005) Fluoro-Jade C results in ultra high resolution and contrast labeling of degenerating neurons. Brain Res 1035:24–31. doi:10.1016/j.brainres.2004.11.054
Simon RP, Niiro M, Gwinn R (1993) Prior ischemic stress protects against experimental stroke. Neurosci Lett 163:135–137. doi:10.1016/0304-3940(93)90364-Q
Slee EA, Adrain C, Martin SJ (2001) Executioner caspase-3, -6, and -7 perform distinct, non-redundant roles during the demolition phase of apoptosis. J Biol Chem 276:7320–7326. doi:10.1074/jbc.M008363200
Stankiewicz AR, Lachapelle G, Foo CP, Radicioni SM, Mosser DD (2005) Hsp70 inhibits heat-induced apoptosis upstream of mitochondria by preventing Bax translocation. J Biol Chem 280:38729–38739. doi:10.1074/jbc.M509497200
Steel R, Doherty JP, Buzzard K, Clemons N, Hawkins CJ, Anderson RL (2004) Hsp72 inhibits apoptosis upstream of the mitochondria and not through interactions with Apaf-1. J Biol Chem 279:51490–51499. doi:10.1074/jbc.M401314200
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 19:1–6
Tanaka H, Yokota H, Jover T, Cappuccio I, Calderone A, Simionescu M, Bennett MV, Zukin RS (2004) Ischemic preconditioning: neuronal survival in the face of caspase-3 activation. J Neurosci 24:2750–2759. doi:10.1523/JNEUROSCI.5475-03.2004
Tsuchiya D, Hong S, Matsumori Y, Shiina H, Kayama T, Swanson RA, Dillman WH, Liu J, Panter SS, Weinstein PR (2003) Overexpression of rat heat shock protein 70 is associated with reduction of early mitochondrial cytochrome c release and subsequent DNA fragmentation after permanent focal ischemia. J Cereb Blood Flow Metab 23:718–727. doi:10.1097/01.WCB.0000054756.97390.F7
Wolf BB, Schuler M, Echeverri F, Green DR (1999) Caspase-3 is the primary activator of apoptotic DNA fragmentation via DNA fragmentation factor-45/inhibitor of caspase-activated DNase inactivation. J Biol Chem 274:30651–30656. doi:10.1074/jbc.274.43.30651
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This work was supported by the Ministry of Education of Slovak Republic (grant VEGA 1/4255/07 to P.R.). Authors are grateful to Zdenka Cetlova for her excellent technical assistance.
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Racay, P., Chomova, M., Tatarkova, Z. et al. Ischemia-Induced Mitochondrial Apoptosis is Significantly Attenuated by Ischemic Preconditioning. Cell Mol Neurobiol 29, 901–908 (2009). https://doi.org/10.1007/s10571-009-9373-7
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DOI: https://doi.org/10.1007/s10571-009-9373-7