Restricted accessResearch articleFirst published online 2018-11
Clozapine attenuates mitochondrial burdens and abnormal behaviors elicited by phencyclidine in mice via inhibition of p47 phox;Possible involvements of phosphoinositide 3-kinase/Akt signaling
Oxidative stress and mitochondrial dysfunction have been implicated in the pathophysiology of schizophrenia.
Aims:
We investigated whether antipsychotic clozapine modulates nicotinamide adenine dinucleotide phosphate oxidase and mitochondrial burdens induced by phencyclidine in mice.
Methods:
We examined the effect of clozapine on nicotinamide adenine dinucleotide phosphate oxidase activation, mitochondrial burdens (i.e. oxidative stress and mitochondrial dysfunction), and activities of enzymatic antioxidant in the prefrontal cortex, and subsequent abnormal behaviors induced by repeated treatment with phencyclidine. p47phox Knockout mice and LY294002, a phosphoinositide 3-kinase inhibitor, were employed to elucidate the pharmacological mechanism of clozapine.
Results:
Phencyclidine treatment resulted in an early increase nicotinamide adenine dinucleotide phosphate oxidase activity, membrane translocation of p47phox, interaction between p-Akt and p47phox, and mitochondrial burdens in wild-type mice. Although these increases returned to near control level four days post-phencyclidine, mitochondrial superoxide dismutase and glutathione peroxidase activities were decreased at that time. Clozapine, LY294002, or p47phox knockout significantly ameliorated social withdrawal and recognition memory deficits produced by phencyclidine. Importantly, LY294002 did not significantly alter the effects of clozapine against abnormal behaviors and the interaction between p-Akt and p47phox induced by phencyclidine. Furthermore, neither LY294002 nor clozapine exhibited any additive effects to the protection afforded by p47phox knockout against phencyclidine insult.
Conclusion:
Our results suggest that p47phox gene mediates phencyclidine-induced mitochondrial burdens and abnormal behaviors, and that the interactive modulation between p47phox and phosphoinositide 3-kinase/Akt is important for the understanding on the pharmacological mechanism of clozapine.
AbramovAYCanevariLDuchenMR (2004) Beta-amyloid peptides induce mitochondrial dysfunction and oxidative stress in astrocytes and death of neurons through activation of NADPH oxidase. J Neurosci24: 565–575.
2.
AndersonKEBoyleKBDavidsonKet al. (2008) CD18-dependent activation of the neutrophil NADPH oxidase during phagocytosis of Escherichia coli or Staphylococcus aureus is regulated by class III but not class I or II PI3Ks. Blood112: 5202–5211.
3.
ArguelloPAGogosJA (2008) A signaling pathway AKTing up in schizophrenia. J Clin Invest118: 2018–2021.
4.
BabiorBM (1999) NADPH oxidase: An update. Blood93: 1464–1476.
5.
BehrensMMAliSSDaoDNet al. (2007) Ketamine-induced loss of phenotype of fast-spiking interneurons is mediated by NADPH-oxidase. Science318: 1645–1647.
6.
BehrensMMAliSSDuganLL (2008) Interleukin-6 mediates the increase in NADPH-oxidase in the ketamine model of schizophrenia. J Neurosci28: 13957–13966.
7.
BhatAHDarKBAneesSet al. (2015) Oxidative stress, mitochondrial dysfunction and neurodegenerative diseases; a mechanistic insight. Biomed Pharmacother74: 101–110.
CabungcalJHCounotteDSLewisEet al. (2014) Juvenile antioxidant treatment prevents adult deficits in a developmental model of schizophrenia. Neuron83: 1073–1084.
10.
CoughlinJMHayesLNTanakaTet al. (2017) Reduced superoxide dismutase-1 (SOD1) in cerebrospinal fluid of patients with early psychosis in association with clinical features. Schizophr Res183: 64–69.
11.
CoyleJT (2006) Glutamate and schizophrenia: Beyond the dopamine hypothesis. Cell Mol Neurobiol26: 365–384.
DangDKShinEJNamYet al. (2016) Apocynin prevents mitochondrial burdens, microglial activation, and pro-apoptosis induced by a toxic dose of methamphetamine in the striatum of mice via inhibition of p47phox activation by ERK. J Neuroinflammation13: 12.
14.
DangPMStensballeABoussettaTet al. (2006) A specific p47phox -serine phosphorylated by convergent MAPKs mediates neutrophil NADPH oxidase priming at inflammatory sites. J Clin Invest116: 2033–2043.
15.
DikalovS (2011) Cross talk between mitochondria and NADPH oxidases. Free Radic Biol Med51: 1289–1301.
16.
DoKQTrabesingerAHKirsten-KrugerMet al. (2000) Schizophrenia: Glutathione deficit in cerebrospinal fluid and prefrontal cortex in vivo. Eur J Neurosci12: 3721–3728.
17.
DuFCooperAJThidaTet al. (2014) In vivo evidence for cerebral bioenergetic abnormalities in schizophrenia measured using 31P magnetization transfer spectroscopy. JAMA Psychiatry71: 19–27.
18.
El-BennaJDangPMGougerot-PocidaloMA (2008) Priming of the neutrophil NADPH oxidase activation: Role of p47phox phosphorylation and NOX2 mobilization to the plasma membrane. Semin Immunopathol30: 279–289.
19.
EllsonCDGobert-GosseSAndersonKEet al. (2001) PtdIns(3)P regulates the neutrophil oxidase complex by binding to the PX domain of p40(phox). Nat Cell Biol3: 679–682.
20.
EmamianESHallDBirnbaumMJet al. (2004) Convergent evidence for impaired AKT1-GSK3beta signaling in schizophrenia. Nat Genet36: 131–137.
21.
FranklinKBJPaxinosG (2008) The Mouse Brain in Stereotaxic Coordinates. 3rd ed.San Diego: Academic Press.
22.
FreybergZFerrandoSJJavitchJA (2010) Roles of the Akt/GSK-3 and Wnt signaling pathways in schizophrenia and antipsychotic drug action. Am J Psychiatry167: 388–396.
GroempingYRittingerK (2005) Activation and assembly of the NADPH oxidase: A structural perspective. Biochem J386: 401–416.
25.
GubertCStertzLPfaffensellerBet al. (2013) Mitochondrial activity and oxidative stress markers in peripheral blood mononuclear cells of patients with bipolar disorder, schizophrenia, and healthy subjects. J Psychiatr Res47: 1396–1402.
26.
HawkinsPTDavidsonKStephensLR (2007) The role of PI3Ks in the regulation of the neutrophil NADPH oxidase. Biochem Soc Symp. 59–67.
27.
HuXZhouHZhangDet al. (2012) Clozapine protects dopaminergic neurons from inflammation-induced damage by inhibiting microglial overactivation. J Neuroimmune Pharmacol7: 187–201.
28.
JanssenAJTrijbelsFJSengersRCet al. (2007) Spectrophotometric assay for complex I of the respiratory chain in tissue samples and cultured fibroblasts. Clin Chem53: 729–734.
29.
JavittDCZukinSR (1991) Recent advances in the phencyclidine model of schizophrenia. Am J Psychiatry148: 1301–1308.
30.
JentschJDRedmondDEJr.ElsworthJDet al. (1997) Enduring cognitive deficits and cortical dopamine dysfunction in monkeys after long-term administration of phencyclidine. Science277: 953–955.
31.
JentschJDRothRH (1999) The neuropsychopharmacology of phencyclidine: From NMDA receptor hypofunction to the dopamine hypothesis of schizophrenia. Neuropsychopharmacology20: 201–225.
32.
JevticGNikolicTMircicAet al. (2016) Mitochondrial impairment, apoptosis and autophagy in a rat brain as immediate and long-term effects of perinatal phencyclidine treatment - influence of restraint stress. Prog Neuropsychopharmacol Biol Psychiatry66: 87–96.
33.
JiangLWuXWangSet al. (2016) Clozapine metabolites protect dopaminergic neurons through inhibition of microglial NADPH oxidase. J Neuroinflammation13: 110.
34.
JiangZRompalaGRZhangSet al. (2013) Social isolation exacerbates schizophrenia-like phenotypes via oxidative stress in cortical interneurons. Biol Psychiatry73: 1024–1034.
35.
JohnsonAWJaaro-PeledHShahaniNet al. (2013) Cognitive and motivational deficits together with prefrontal oxidative stress in a mouse model for neuropsychiatric illness. Proc Natl Acad Sci U S A110: 12462–12467.
36.
KameiHNagaiTNakanoHet al. (2006) Repeated methamphetamine treatment impairs recognition memory through a failure of novelty-induced ERK1/2 activation in the prefrontal cortex of mice. Biol Psychiatry59(1): 75–84.
37.
KimSYCohenBMChenXet al. (2017) Redox dysregulation in schizophrenia revealed by in vivo NAD+/NADH measurement. Schizophr Bull43: 197–204.
38.
Kroller-SchonSStevenSKossmannSet al. (2014) Molecular mechanisms of the crosstalk between mitochondria and NADPH oxidase through reactive oxygen species-studies in white blood cells and in animal models. Antioxid Redox Signal20: 247–266.
39.
LahtiACKoffelBLaPorteDet al. (1995) Subanesthetic doses of ketamine stimulate psychosis in schizophrenia. Neuropsychopharmacology13: 9–19.
40.
LahtiACWeilerMATamara MichaelidisBAet al. (2001) Effects of ketamine in normal and schizophrenic volunteers. Neuropsychopharmacology25: 455–467.
LiMHeYZhouZet al. (2017) MicroRNA-223 ameliorates alcoholic liver injury by inhibiting the IL-6-p47phox-oxidative stress pathway in neutrophils. Gut66: 705–715.
43.
LiuXYZhouXYHouJCet al. (2015) Ginsenoside Rd promotes neurogenesis in rat brain after transient focal cerebral ischemia via activation of PI3K/Akt pathway. Acta Pharmacol Sin36(4): 421–428.
44.
LuLMamiyaTLuPet al. (2010) Prenatal exposure to phencyclidine produces abnormal behaviour and NMDA receptor expression in postpubertal mice. Int J Neuropsychopharmacol13: 877–889.
45.
McElneaEMQuillBDochertyNGet al. (2011) Oxidative stress, mitochondrial dysfunction and calcium overload in human lamina cribrosa cells from glaucoma donors. Mol Vis17: 1182–1191.
46.
MalhotraAKPinalsDAWeingartnerHet al. (1996) NMDA receptor function and human cognition: The effects of ketamine in healthy volunteers. Neuropsychopharmacology14: 301–307.
47.
MariMMoralesAColellAet al. (2013) Mitochondrial glutathione: Features, regulation and role in disease. Biochim Biophys Acta1830: 3317–3328.
48.
MeltzerHYRajagopalLHuangMet al. (2013) Translating the N-methyl-D-aspartate receptor antagonist model of schizophrenia to treatments for cognitive impairment in schizophrenia. Int J Neuropsychopharmacol16: 2181–2194.
49.
MichelTMThomeJMartinDet al. (2004) Cu, Zn- and Mn-superoxide dismutase levels in brains of patients with schizophrenic psychosis. J Neural Transm (Vienna)111: 1191–1201.
50.
MiyamotoSMiyakeNJarskogLFet al. (2012) Pharmacological treatment of schizophrenia: A critical review of the pharmacology and clinical effects of current and future therapeutic agents. Mol Psychiatry17: 1206–1227.
51.
MollerMDu PreezJLViljoenFPet al. (2013) Social isolation rearing induces mitochondrial, immunological, neurochemical and behavioural deficits in rats, and is reversed by clozapine or N-acetyl cysteine. Brain Behav Immun30: 156–167.
52.
MouriAKosekiTNarusawaSet al. (2012) Mouse strain differences in phencyclidine-induced behavioural changes. Int J Neuropsychopharmacol15: 767–779.
53.
MouriANodaYEnomotoTet al. (2007) Phencyclidine animal models of schizophrenia: Approaches from abnormality of glutamatergic neurotransmission and neurodevelopment. Neurochem Int51: 173–184.
54.
NabeshimaTMouriAMuraiRet al. (2006) Animal model of schizophrenia: Dysfunction of NMDA receptor-signaling in mice following withdrawal from repeated administration of phencyclidine. Ann N Y Acad Sci1086: 160–168.
55.
NakajimaAAoyamaYNguyenTTet al. (2013) Nobiletin, a citrus flavonoid, ameliorates cognitive impairment, oxidative burden, and hyperphosphorylation of tau in senescence-accelerated mouse. Behav Brain Res250: 351–360.
NewcomerJWFarberNBJevtovic-TodorovicVet al. (1999) Ketamine-induced NMDA receptor hypofunction as a model of memory impairment and psychosis. Neuropsychopharmacology20: 106–118.
58.
NgFBerkMDeanOet al. (2008) Oxidative stress in psychiatric disorders: Evidence base and therapeutic implications. Int J Neuropsychopharmacol11: 851–876.
59.
NiCWKumarSAnkenyCJet al. (2014) Development of immortalized mouse aortic endothelial cell lines. Vasc Cell6: 7.
60.
NichollsDG (2009) Mitochondrial calcium function and dysfunction in the central nervous system. Biochim Biophys Acta1787: 1416–1424.
61.
NodaANodaYKameiHet al. (2001) Phencyclidine impairs latent learning in mice: Interaction between glutamatergic systems and sigma(1) receptors. Neuropsychopharmacology24: 451–460.
62.
NodaYKameiHMamiyaTet al. (2000) Repeated phencyclidine treatment induces negative symptom-like behavior in forced swimming test in mice: Imbalance of prefrontal serotonergic and dopaminergic functions. Neuropsychopharmacology23: 375–387.
63.
NodaYYamadaKFurukawaHet al. (1995) Enhancement of immobility in a forced swimming test by subacute or repeated treatment with phencyclidine: A new model of schizophrenia. Br J Pharmacol116: 2531–2537.
64.
NodaYYamadaKKomoriYet al. (1996) Role of nitric oxide in the development of tolerance and sensitization to behavioural effects of phencyclidine in mice. Br J Pharmacol117: 1579–1585.
65.
NuciforaFCJrMihaljevicMLeeBJet al. (2017) Clozapine as a Model for Antipsychotic Development. Neurotherapeutics14: 750–761.
PrabakaranSSwattonJERyanMMet al. (2004) Mitochondrial dysfunction in schizophrenia: Evidence for compromised brain metabolism and oxidative stress. Mol Psychiatry9: 684–697, 643.
68.
QianLWeiS-JZhangDet al. (2008) Potent anti-inflammatory and neuroprotective effects of TGFβ1 are mediated through the inhibition of ERK and p47(phox)-Ser345 phosphorylation and translocation in microglia. J Immunol181: 660–668.
69.
QiaoHNodaYKameiHet al. (2001) Clozapine, but not haloperidol, reverses social behavior deficit in mice during withdrawal from chronic phencyclidine treatment. Neuroreport12(1): 11–15.
70.
RadonjicNVJakovcevskiIBumbasirevicVet al. (2013) Perinatal phencyclidine administration decreases the density of cortical interneurons and increases the expression of neuregulin-1. Psychopharmacology (Berl)227: 673–683.
71.
RadonjicNVKnezevicIDVilimanovichUet al. (2010) Decreased glutathione levels and altered antioxidant defense in an animal model of schizophrenia: Long-term effects of perinatal phencyclidine administration. Neuropharmacology58: 739–745.
72.
RaffaMAtigFMhallaAet al. (2011) Decreased glutathione levels and impaired antioxidant enzyme activities in drug-naive first-episode schizophrenic patients. BMC Psychiatry11: 124.
73.
RossCAMargolisRLReadingSAet al. (2006) Neurobiology of schizophrenia. Neuron52: 139–153.
74.
ScainiGRochiNMoraisMOet al. (2013) In vitro effect of antipsychotics on brain energy metabolism parameters in the brain of rats. Acta Neuropsychiatr25: 18–26.
75.
SchiavoneSSorceSDubois-DauphinMet al. (2009) Involvement of NOX2 in the development of behavioral and pathologic alterations in isolated rats. Biol Psychiatry66: 384–392.
76.
SelivanovVAVotyakovaTVPivtoraikoVNet al. (2011) Reactive oxygen species production by forward and reverse electron fluxes in the mitochondrial respiratory chain. PLoS Comput Biol7: e1001115.
77.
ShinEJDangDKTranHQet al. (2016a) PKCdelta knockout mice are protected from para-methoxymethamphetamine-induced mitochondrial stress and associated neurotoxicity in the striatum of mice. Neurochem Int100: 146–158.
78.
ShinEJDuongCXNguyenXTet al. (2012) Role of oxidative stress in methamphetamine-induced dopaminergic toxicity mediated by protein kinase Cdelta. Behav Brain Res232: 98–113.
79.
ShinEJJeongJHKimAYet al. (2009) Protection against kainate neurotoxicity by ginsenosides: Attenuation of convulsive behavior, mitochondrial dysfunction, and oxidative stress. J Neurosci Res87: 710–722.
80.
ShinEJKimJMNguyenXKet al. (2011) Effects of gastrodia elata bl on phencyclidine-induced schizophrenia-like psychosis in mice. Curr Neuropharmacol9: 247–250.
81.
ShinEJKoKHKimWKet al. (2008) Role of glutathione peroxidase in the ontogeny of hippocampal oxidative stress and kainate seizure sensitivity in the genetically epilepsy-prone rats. Neurochem Int52: 1134–1147.
82.
ShinEJNamYTuTHet al. (2016b) Protein kinase C delta mediates trimethyltin-induced neurotoxicity in mice in vivo via inhibition of glutathione defense mechanism. Arch Toxicol90: 937–953.
83.
ShinEJShinSWNguyenTTet al. (2014) Ginsenoside Re rescues methamphetamine-induced oxidative damage, mitochondrial dysfunction, microglial activation, and dopaminergic degeneration by inhibiting the protein kinase Cdelta gene. Mol Neurobiol49: 1400–1421.
84.
ShinEJSuhSKLimYKet al. (2005) Ascorbate attenuates trimethyltin-induced oxidative burden and neuronal degeneration in the rat hippocampus by maintaining glutathione homeostasis. Neuroscience133: 715–727.
85.
ShinSYChoiBHKoJet al. (2006) Clozapine, a neuroleptic agent, inhibits Akt by counteracting Ca2+/calmodulin in PTEN-negative U-87MG human glioblastoma cells. Cell Signal18: 1876–1886.
86.
SorceSSchiavoneSTucciPet al. (2010) The NADPH oxidase NOX2 controls glutamate release: A novel mechanism involved in psychosis-like ketamine responses. J Neurosci30: 11317–11325.
87.
StojkovicTRadonjicNVVelimirovicMet al. (2012) Risperidone reverses phencyclidine induced decrease in glutathione levels and alterations of antioxidant defense in rat brain. Prog Neuropsychopharmacol Biol Psychiatry39: 192–199.
88.
TakahataRMoghaddamB (2003) Activation of glutamate neurotransmission in the prefrontal cortex sustains the motoric and dopaminergic effects of phencyclidine. Neuropsychopharmacology28: 1117–1124.
89.
TangXLiuKJRamuJet al. (2010) Inhibition of gp91(phox) contributes towards normobaric hyperoxia afforded neuroprotection in focal cerebral ischemia. Brain Res1348: 174–180.
90.
TianWLiXJStullNDet al. (2008) Fc gamma R-stimulated activation of the NADPH oxidase: Phosphoinositide-binding protein p40phox regulates NADPH oxidase activity after enzyme assembly on the phagosome. Blood112: 3867–3877.
91.
ToriumiKMouriANarusawaSet al. (2012) Prenatal NMDA receptor antagonism impaired proliferation of neuronal progenitor, leading to fewer glutamatergic neurons in the prefrontal cortex. Neuropsychopharmacology37: 1387–1396.
92.
ToriumiKOkiMMutoEet al. (2016) Prenatal phencyclidine treatment induces behavioral deficits through impairment of GABAergic interneurons in the prefrontal cortex. Psychopharmacology (Berl)233: 2373–2381.
93.
TranHQLeeYShinEJet al. (2018) PKCdelta knockout mice are protected from dextromethorphan-induced serotonergic behaviors in mice: Involvements of downregulation of 5-HT1A receptor and upregulation of Nrf2-dependent GSH synthesis. Mol Neurobiol.
94.
TranTVShinEJDangDKet al. (2017a) Ginsenoside Re protects against phencyclidine-induced behavioral changes and mitochondrial dysfunction via interactive modulation of glutathione peroxidase-1 and NADPH oxidase in the dorsolateral cortex of mice. Food Chem Toxicol110: 300–315.
95.
TranTVShinEJJeongJHet al. (2017b) Protective potential of the glutathione peroxidase-1 gene in abnormal behaviors induced by phencyclidine in mice. Mol Neurobiol54: 7042–7062.
VignaisPV (2002) The superoxide-generating NADPH oxidase: Structural aspects and activation mechanism. Cell Mol Life Sci59: 1428–1459.
98.
VolzHRRiehemannSMaurerIet al. (2000) Reduced phosphodiesters and high-energy phosphates in the frontal lobe of schizophrenic patients: A (31)P chemical shift spectroscopic-imaging study. Biol Psychiatry47: 954–961.
99.
WangQChuCHOyarzabalEet al. (2014a) Subpicomolar diphenyleneiodonium inhibits microglial NADPH oxidase with high specificity and shows great potential as a therapeutic agent for neurodegenerative diseases. Glia62: 2034–2043.
100.
WangQChuCHQianLet al. (2014b) Substance P exacerbates dopaminergic neurodegeneration through neurokinin-1 receptor-independent activation of microglial NADPH oxidase. J Neurosci34: 12490–12503.
101.
WeiZBaiORichardsonJSet al. (2003) Olanzapine protects PC12 cells from oxidative stress induced by hydrogen peroxide. J Neurosci Res73: 364–368.
102.
WolfRPaelchenKMatzkeKet al. (2007) Acute or subchronic clozapine treatment does not ameliorate prepulse inhibition (PPI) deficits in CPB-K mice with low levels of hippocampal NMDA receptor density. Psychopharmacology (Berl)194(1): 93–102.
103.
ZhangXYZhouDFQiLYet al. (2009) Superoxide dismutase and cytokines in chronic patients with schizophrenia: Association with psychopathology and response to antipsychotics. Psychopharmacology (Berl)204: 177–184.
104.
ZhuSWangHShiRet al. (2014) Chronic phencyclidine induces inflammatory responses and activates GSK3beta in mice. Neurochem Res39: 2385–2393.
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