Abstract
Severe acidosis caused death of cultured cerebellar granule neurons (CGNs). Acidosis was accompanied by a progressive increase of the intracellular zinc ions ([Zn2+]i) and decrease of [Ca2+]i. Zn2+ chelator, N,N,N′,N′-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN), prevented the increase of [Zn2+]i and acidosis-induced neuronal death. However, neuronal death was insensitive to blockade of ASIC1 channels with amiloride, as CGNs display considerably lower expression of ASIC1a than other neurons. The antioxidant trolox and menadione significantly protected neurons from acidotic death. Earlier, we demonstrated that menadione rescues neurons from the deleterious effect of inhibition of mitochondrial complex I (Isaev et al. Neuroreport 15:2227–2231, 2004). We speculate that excessive Zn2+-dependent production of reactive oxygen species by mitochondrial complex I may be a general motive for the induction of cell death in CGNs under acidotic conditions.
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Abbreviations
- CGNs:
-
Cerebellar granule neurons
- TPEN:
-
N,N,N′,N′-tetrakis(2-pyridylmethyl)ethylenediamine
- ASIC1a:
-
Acid sensitive ion channel 1a
References
Baron A, Schaefer L, Lingueglia E, Champigny G, Lazdunski M (2001) Zn2+ and H+ are coactivators of acid-sensing ion channels. J Biol Chem 276:35361–35367
Batandier C, Leverve X, Fontaine E (2004) Opening of the mitochondrial permeability transition pore induces reactive oxygen species production at the level of the respiratory chain complex I. J Biol Chem 279:17197–17204
Brown AM, Kristal BS, Effron MS, Shestopalov AI, Ullucci PA, Sheu KF, Blass JP, Cooper AJ (2000) Zn2+ inhibits alpha-ketoglutarate-stimulated mitochondrial respiration and the isolated alpha-ketoglutarate dehydrogenase complex. J Biol Chem 275:13441–13447
Colvin RA (2002) pH dependence and compartmentalization of zinc transported across plasma membrane of rat cortical neurons. Am J Physiol Cell Physiol. 282:C317–C329
Dedukhova VI, Kirillova GP, Mokhova EN, Rozovskaia IA, Skulachev VP (1986) Effect of menadione and vicasol on mitochondrial energy during inhibition of initiation sites of the respiration chain. Biokhimiia 51:567–573
Dineley KE, Votyakova TV, Reynolds IJ (2003) Zinc inhibition of cellular energy production: implications for mitochondria and neurodegeneration. J Neurochem 85:563–570
Ding D, Moskowitz SI, Li R, Lee SB, Esteban M, Tomaselli K, Chan J, Bergold PJ (2000) Acidosis induces necrosis and apoptosis of cultured hippocampal neurons. Exp Neurol 162:1–12
Dirnagl U, Iadecola C, Moskowitz MA (1999) Pathobiology of ischaemic stroke: an integrated view. Trends Neurosci 22:391–397
Frazzini V, Rapposelli IG, Corona C, Rockabrand E, Canzoniero LMT, Sensi SL (2007) Mild acidosis enhances AMPA receptor-mediated intracellular zinc mobilization in cortical neurons. Mol Med 13:356–361
Galli C, Meucci O, Scorziello A, Werge TM, Calissano P, Schettini G (1995) Apoptosis in cerebellar granule cells is blocked by high KCl, forskolin, and IGF-1 through distinct mechanisms of action: the involvement of intracellular calcium and RNA synthesis. J Neurosci 15:1172–1179
Garcia-Anoveros J, Derfler B, Neville-Golden J, Hyman BT, Corey DP (1997) BNaC1 and BNaC2 constitute a new family of human neuronal sodium channels related to degenerins and epithelial sodium channels. Proc Natl Acad Sci USA 94:1459–1464
Hey JG, Chu XP, Seeds J, Simon RP, Xiong ZG (2007) Extracellular zinc protects against acidosis-induced injury of cells expressing Ca2+-permeable acid-sensing ion channels. Stroke 38:670–673
Isaev NK, Stelmashook EV, Ruscher K, Andreeva NA, Zorov DB (2004) Menadione reduces rotenone-induced cell death in cerebellar granule neurons. Neuroreport 15:2227–2231
Isaev NK, Stelmashook EV, Plotnikov EY, Khryapenkova TG, Lozier ER, Doludin YV, Silachev DN, Zorov DB (2008) Role of acidosis, NMDA receptors, and acid-sensitive ion channel 1a (ASIC1a) in neuronal death induced by ischemia. Biochemistry (Mosc) 73:1171–1175
Kim EY, Koh JY, Kim YH, Sohn S, Joe E, Gwag BJ (1999) Zn2+ entry produces oxidative neuronal necrosis in cortical cell cultures. Eur J Neurosci 11:327–334
Link TA, von Jagow G (1995) Zinc ions inhibit the QP center of bovine heart mitochondrial bc1 complex by blocking a protonatable group. J Biol Chem 270:25001–25006
Lorusso M, Cocco T, Sardanelli AM, Minuto M, Bonomi F, Papa S (1991) Interaction of Zn2+ with the bovine-heart mitochondrial bc1 complex. Eur J Biochem 197:555–561
Mergenthaler P, Dirnagl U, Meisel A (2004) Pathophysiology of stroke: lessons from animal models. Meta Brain Dis 19:151–167
Musleh W, Bruce A, Malfroy B, Baudry M (1994) Effects of EUK-8, a synthetic catalytic superoxide scavenger, on hypoxia- and acidosis-induced damage in hippocampal slices. Neuropharmacology 33:929–934
Oy-Yang Y, Kristian T, Mellergarg P, Siejo BK (1994) The influence of pH on glutamate- and depolarization-induced increases of intracellular calcium concentration in cortical neurons in primary culture. Brain Res 646:65–72
Pruss H, Prass K, Ghaeni L, Milosevic M, Muselmann C, Freyer D, Royl G, Reuter U, Baeva N, Dirnagl U, Meisel A, Priller J (2008) Inducible nitric oxide synthase does not mediate brain damage after transient focal cerebral ischemia in mice. J Cereb Blood Flow Metab 28:526–539
Rehncrona S, Hauge H, Siejo BK (1989) Enhancement of iron-catalyzed free radical formation by acidosis in brain homogenates: differences in effect by lactic acid and CO2. J Cereb Blood Flow Metab 9:65–70
Ruscher K, Freyer D, Karsch M, Isaev N, Megow D, Sawitzki B, Priller J, Dirnagl U, Meisel A (2002) Erythropoietin is a paracrine mediator of ischemic tolerance in the brain: evidence from an in vitro model. J Neurosci 22:10291–10301
Sensi SL, Ton-That D, Sullivan PG, Jonas EA, Gee KR, Kaczmarek LK, Weiss JH (2003) Modulation of mitochondrial function by endogenous Zn2+ pools. Proc Natl Acad Sci USA 100:6157–6162
Sensi SL, Paoletti P, Bush AI, Sekler I (2009) Zinc in the physiology and pathology of the CNS. Nat Rev Neurosci 10:780–791
Sharpley MS, Hirst J (2006) The inhibition of mitochondrial complex I (NADH:ubiquinone oxidoreductase) by Zn2+. J Biol Chem 281:34803–34809
Shen H, Chan J, Kass IS, Bergold PJ (1995) Transient acidosis induces delayed neurotoxicity in cultured hippocampal slices. Neurosci Lett 185:115–118
Sipos I, Tretter L, Adam-Vizi V (2003) Quantitative relationship between inhibition of respiratory complexes and formation of reactive oxygen species in isolated nerve terminals. J Neurochem 84:112–118
Stelmashuk EV, Belyaeva EA, Isaev NK (2007) Effect of acidosis, oxidative stress, and glutamate toxicity on the survival of mature and immature cultured cerebellar granule cells. Neurochem J 1:66–69
Tombaugh GC, Sapolsky RM (1990) Mechanistic distinctions between excitotoxic and acidotic hippocampal damage in an in vitro model of ischemia. J Cereb Blood Flow Metab 10:527–535
Tombaugh GC, Somjen GG (1996) Effects of extracellular pH on voltage-gated Na+, K+ and Ca2+ currents in isolated rat CA1 neurons. J Physiol 493:719–732
Wijburg FA, Feller N, de Groot CJ, Wanders RJ (1990) Menadione partially restores NADH-oxidation and ATP-synthesis in complex I deficient fibroblasts. Biochem Int 22:303–309
Xiong ZG, Zhu XM, Chu XP, Minami M, Hey J, Wei WL, MacDonald JF, Wemmie JA, Price MP, Welsh MJ, Simon RP (2004) Neuroprotection in ischemia: blocking calcium-permeable acid-sensing ion channels. Cell 118:665–666
Xiong Z-G, Chu X-P, Simon RP (2006) Ca2+-permeable acid-sensing ion channels and ischemic brain injury. J Membr Biol 209:59–68
Yermolaieva O, Leonard AS, Schnizler MK, Abboud FM, Welsh MJ (2004) Extracellular acidosis increases neuronal cell calcium by activating acid-sensing ion channel 1a. Proc Natl Acad Sci USA 101:6752–6757
Ying W, Han S-K, Miller JW, Swanson RA (1999) FAS-mediated apoptosis and its relation to intrinsic pathway activation in an experimental model of retinal detachment. J Neurochem 73:1549–1556
Zorov DB, Filburn CR, Klotz LO, Zweier JL, Sollott SJ (2000) Reactive oxygen species (ROS)-induced ROS release: a new phenomenon accompanying induction of the mitochondrial permeability transition in cardiac myocytes. J Exp Med 192:1001–1014
Acknowledgments
The authors would like to acknowledge Katharina Stohlmann and Ingo Przesdzing for excellent experimental support. This study was supported by RFBR grants 08-04-00762-a, 08-04-01667-a, 09-04-01096-a and the Deutsche Forschungsgemeinschaft (ME 3603/1-1 to PM).
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Isaev, N.K., Stelmashook, E.V., Lukin, S.V. et al. Acidosis-Induced Zinc-Dependent Death of Cultured Cerebellar Granule Neurons. Cell Mol Neurobiol 30, 877–883 (2010). https://doi.org/10.1007/s10571-010-9516-x
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DOI: https://doi.org/10.1007/s10571-010-9516-x