Hydrogen peroxide (H2O2) is a reactive oxygen species that is involved in immunity and neuroinflammation. Here, we investigated whether and how pathophysiological levels of H2O2 influenced the differentiation of neural progenitor cells (NPCs). H2O2 levels within the range measured at neuroinflammatory events were applied to rat primary NPC cultures during 24 h, and effects were assessed directly after exposure or in NPCs that were differentiated for 7 days after H2O2 removal. Exposed differentiated NPCs showed significantly increased numbers of neurons and oligodendrocytes compared with unexposed controls. To identify the possible origin of this differentiation result, we characterized the undifferentiated culture and found a significant increase in both OLIG2+ cells and proliferative ASCL1+ C cells that could contribute to both more neurons and oligodendrocytes. In addition, H2O2-induced neurogenesis was supported by western blot and paralleled by gene expression analyses, which revealed an increased expression of the proneural gene Ngn2 and the neuronally expressed gene β-III tubulin. To investigate potential mechanisms for the observed effects on NPC differentiation, we performed gene expression profile analyses for oxidative stress and antioxidant-related and chromatin modification genes where the expression of several important genes was affected by the exposure. Increased oligodendrocyte numbers correlated with increased expression of the chromatin modification enzyme Sirt2, suggesting the involvement of Sirt2 in oligodendrocyte differentiation. Our results suggest a modulatory effect on the differentiation potential of NPCs by H2O2. Our findings indicate that H2O2 exposure has significant effects on NPC proliferation, differentiation, and vulnerability. These results have implications for regeneration after any neuroinflammatory event.
Get full access to this article
View all access options for this article.
References
1.
van HorssenJ, WitteME, SchreibeltG and de VriesHE. (2011). Radical changes in multiple sclerosis pathogenesis. Biochim Biophys Acta, 1812:141–150.
2.
NiethammerP, GrabherC, LookAT and MitchisonTJ. (2009). A tissue-scale gradient of hydrogen peroxide mediates rapid wound detection in zebrafish. Nature, 459:996–999.
3.
AlbertsBJA, LewisJ, RaffM, RobertsK and WalterP. (2002). Molecular Biology of the Cell. Garland Science, a member of the Taylor & Francis Group.
4.
JuarezJC, ManuiaM, BurnettME, BetancourtO, BoivinB, ShawDE, TonksNK, MazarAP and DonateF. (2008). Superoxide dismutase 1 (SOD1) is essential for H2O2-mediated oxidation and inactivation of phosphatases in growth factor signaling. Proc Natl Acad Sci U S A, 105:7147–7152.
5.
ZhuH, SantoA and LiY. (2012). The antioxidant enzyme peroxiredoxin and its protective role in neurological disorders. Exp Biol Med, 237:143–149.
6.
StoneJR and YangS. (2006). Hydrogen peroxide: a signaling messenger. Antioxid Redox Signal, 8:243–270.
7.
RenshawSA, LoynesCA, ElworthyS, InghamPW and WhyteMK. (2007). Modeling inflammation in the zebrafish: how a fish can help us understand lung disease. Exp Lung Res, 33:549–554.
8.
NikicI, MerklerD, SorbaraC, BrinkoetterM, KreutzfeldtM, BareyreFM, BruckW, BishopD, MisgeldT and KerschensteinerM. (2011). A reversible form of axon damage in experimental autoimmune encephalomyelitis and multiple sclerosis. Nat Med, 17:495–499.
9.
TrachoothamD, AlexandreJ and HuangP. (2009). Targeting cancer cells by ROS-mediated mechanisms: a radical therapeutic approach?. Nat Rev Drug Discov, 8:579–591.
10.
CadetJ, DoukiT, GasparuttoD and RavanatJL. (2003). Oxidative damage to DNA: formation, measurement and biochemical features. Mutat Res, 531:5–23.
11.
ChenQM. (2000). Replicative senescence and oxidant-induced premature senescence. Beyond the control of cell cycle checkpoints. Ann N Y Acad Sci, 908:111–125.
12.
JohanssonCB, MommaS, ClarkeDL, RislingM, LendahlU and FrisenJ. (1999). Identification of a neural stem cell in the adult mammalian central nervous system. Cell, 96:25–34.
13.
DoetschF, CailleI, LimDA, Garcia-VerdugoJM and Alvarez-BuyllaA. (1999). Subventricular zone astrocytes are neural stem cells in the adult mammalian brain. Cell, 97:703–716.
14.
KuhnHG, Dickinson-AnsonH and GageFH. (1996). Neurogenesis in the dentate gyrus of the adult rat: age-related decrease of neuronal progenitor proliferation. J Neurosci, 16:2027–2033.
15.
ReynoldsBA and WeissS. (1992). Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science, 255:1707–1710.
16.
ArvidssonL, FagerlundM, JaffN, OssoinakA, JanssonK, HagerstrandA, JohanssonCB, BrundinL and SvenssonM. (2011). Distribution and characterization of progenitor cells within the human filum terminale. PloS One, 6:e27393
17.
JhaRM, LiuX, ChrenekR, MadsenJR and CardozoDL. (2013). The postnatal human filum terminale is a source of autologous multipotent neurospheres capable of generating motor neurons. Neurosurgery, 72:118–129; discussion 129.
18.
DanilovAI, CovacuR, MoeMC, LangmoenIA, JohanssonCB, OlssonT and BrundinL. (2006). Neurogenesis in the adult spinal cord in an experimental model of multiple sclerosis. Eur J Neurosci, 23:394–400.
19.
Nait-OumesmarB, Picard-RieraN, KerninonC, DeckerL, SeilheanD, HoglingerGU, HirschEC, ReynoldsR and Baron-Van EvercoorenA. (2007). Activation of the subventricular zone in multiple sclerosis: evidence for early glial progenitors. Proc Natl Acad Sci U S A, 104:4694–4699.
20.
ImitolaJ, RaddassiK, ParkKI, MuellerFJ, NietoM, TengYD, FrenkelD, LiJ, SidmanRL, et al. (2004). Directed migration of neural stem cells to sites of CNS injury by the stromal cell-derived factor 1alpha/CXC chemokine receptor 4 pathway. Proc Natl Acad Sci U S A, 101:18117–18122.
21.
KyritsisN, KizilC and BrandM. (2013). Neuroinflammation and central nervous system regeneration in vertebrates. Trends Cell Biol, 24:128–135.
22.
D'AutreauxB and ToledanoMB. (2007). ROS as signalling molecules: mechanisms that generate specificity in ROS homeostasis. Nat Rev Mol Cell Biol, 8:813–824.
23.
HalliwellB and GutteridgeJ. (2007). Free Radicals in Biology And Medicine. Oxford University Press, UK.
24.
RheeSG. (2006). Cell signaling. H2O2, a necessary evil for cell signaling. Science, 312:1882–1883.
25.
VealEA, DayAM and MorganBA. (2007). Hydrogen peroxide sensing and signaling. Mol Cell, 26:1–14.
26.
Le BelleJE, OrozcoNM, PaucarAA, SaxeJP, MottahedehJ, PyleAD, WuH and KornblumHI. (2011). Proliferative neural stem cells have high endogenous ROS levels that regulate self-renewal and neurogenesis in a PI3K/Akt-dependant manner. Cell Stem Cell, 8:59–71.
27.
DickinsonBC, PeltierJ, StoneD, SchafferDV and ChangCJ. (2011). Nox2 redox signaling maintains essential cell populations in the brain. Nat Chem Biol, 7:106–112.
28.
GutteridgeBH. (2007). Free Radicals in Biology and Medicine. Oxford University Press, New York.
29.
BlockML, ZeccaL and HongJS. (2007). Microglia-mediated neurotoxicity: uncovering the molecular mechanisms. Nat Rev Neurosci, 8:57–69.
30.
DuttaR, McDonoughJ, YinX, PetersonJ, ChangA, TorresT, GudzT, MacklinWB, LewisDA, et al. (2006). Mitochondrial dysfunction as a cause of axonal degeneration in multiple sclerosis patients. Ann Neurol, 59:478–489.
31.
CovacuR, DanilovAI, RasmussenBS, HallenK, MoeMC, LobellA, JohanssonCB, SvenssonMA, OlssonT and BrundinL. (2006). Nitric oxide exposure diverts neural stem cell fate from neurogenesis towards astrogliogenesis. Stem Cells, 24:2792–2800.
32.
Huang daW, ShermanBT and LempickiRA. (2009). Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc, 4:44–57.
33.
ScholzenT and GerdesJ. (2000). The Ki-67 protein: from the known and the unknown. J Cell Physiol, 182:311–322.
34.
KriegsteinA and Alvarez-BuyllaA. (2009). The glial nature of embryonic and adult neural stem cells. Annu Rev Neurosci, 32:149–184.
35.
HackMA, SaghatelyanA, de ChevignyA, PfeiferA, Ashery-PadanR, LledoPM and GotzM. (2005). Neuronal fate determinants of adult olfactory bulb neurogenesis. Nat Neurosci, 8:865–872.
36.
IshibashiM, MoriyoshiK, SasaiY, ShiotaK, NakanishiS and KageyamaR. (1994). Persistent expression of helix-loop-helix factor HES-1 prevents mammalian neural differentiation in the central nervous system. EMBO J, 13:1799–1805.
BertrandN, CastroDS and GuillemotF. (2002). Proneural genes and the specification of neural cell types. Nat Rev Neurosci, 3:517–530.
39.
SunY, Nadal-VicensM, MisonoS, LinMZ, ZubiagaA, HuaX, FanG and GreenbergME. (2001). Neurogenin promotes neurogenesis and inhibits glial differentiation by independent mechanisms. Cell, 104:365–376.
40.
GangemiRM, PereraM and CorteG. (2004). Regulatory genes controlling cell fate choice in embryonic and adult neural stem cells. J Neurochem, 89:286–306.
41.
HsiehJ, NakashimaK, KuwabaraT, MejiaE and GageFH. (2004). Histone deacetylase inhibition-mediated neuronal differentiation of multipotent adult neural progenitor cells. Proc Natl Acad Sci U S A, 101:16659–16664.
42.
JiS, DoucetteJR and NazaraliAJ. (2011). Sirt2 is a novel in vivo downstream target of Nkx2.2 and enhances oligodendroglial cell differentiation. J Mol Cell Biol, 3:351–359.
43.
FridovichI. (1986). Biological effects of the superoxide radical. Arch Biochem Biophys, 247:1–11.
44.
WittkoIM, SchanzerA, KuzmichevA, SchneiderFT, ShibuyaM, RaabS and PlateKH. (2009). VEGFR-1 regulates adult olfactory bulb neurogenesis and migration of neural progenitors in the rostral migratory stream in vivo. J Neurosci, 29:8704–8714.
45.
ByrneAM, Bouchier-HayesDJ and HarmeyJH. (2005). Angiogenic and cell survival functions of vascular endothelial growth factor (VEGF). J Cell Mol Med, 9:777–794.
46.
KirkSL and KarlikSJ. (2003). VEGF and vascular changes in chronic neuroinflammation. J Autoimmun, 21:353–363.
47.
KimEJ, AblesJL, DickelLK, EischAJ and JohnsonJE. (2011). Ascl1 (Mash1) defines cells with long-term neurogenic potential in subgranular and subventricular zones in adult mouse brain. PloS One, 6:e18472
48.
ImayoshiI, IsomuraA, HarimaY, KawaguchiK, KoriH, MiyachiH, FujiwaraT, IshidateF and KageyamaR. (2013). Oscillatory control of factors determining multipotency and fate in mouse neural progenitors. Science, 342:1203–1208.
49.
FuentealbaLC, ObernierK and Alvarez-BuyllaA. (2012). Adult neural stem cells bridge their niche. Cell Stem Cell, 10:698–708.
50.
WangX and MichaelisEK. (2010). Selective neuronal vulnerability to oxidative stress in the brain. Front Aging Neurosci, 2:12
51.
SteinbeckMJ, KimJK, TrudeauMJ, HauschkaPV and KarnovskyMJ. (1998). Involvement of hydrogen peroxide in the differentiation of clonal HD-11EM cells into osteoclast-like cells. J Cell Physiol, 176:574–587.
52.
ZhangL, QinX, ZhaoY, FastL, ZhuangS, LiuP, ChengG and ZhaoTC. (2012). Inhibition of histone deacetylases preserves myocardial performance and prevents cardiac remodeling through stimulation of endogenous angiomyogenesis. J Pharmacol Exp Therapeut, 341:285–293.
53.
MontgomeryRL, HsiehJ, BarbosaAC, RichardsonJA and OlsonEN. (2009). Histone deacetylases 1 and 2 control the progression of neural precursors to neurons during brain development. Proc Natl Acad Sci U S A, 106:7876–7881.
54.
ZhouQ, DalgardCL, WynderC and DoughtyML. (2011). Histone deacetylase inhibitors SAHA and sodium butyrate block G1-to-S cell cycle progression in neurosphere formation by adult subventricular cells. BMC Neurosci, 12:50
55.
KaoHY, OrdentlichP, Koyano-NakagawaN, TangZ, DownesM, KintnerCR, EvansRM and KadeschT. (1998). A histone deacetylase corepressor complex regulates the Notch signal transduction pathway. Genes Dev, 12:2269–2277.
56.
AjamianF, SuuronenT, SalminenA and ReebenM. (2003). Upregulation of class II histone deacetylases mRNA during neural differentiation of cultured rat hippocampal progenitor cells. Neurosci Lett, 346:57–60.
57.
PerdizD, MackehR, PousC and BailletA. (2011). The ins and outs of tubulin acetylation: more than just a post-translational modification?. Cell Signal, 23:763–771.
58.
Etienne-MannevilleS. (2010). From signaling pathways to microtubule dynamics: the key players. Curr Opin Cell Biol, 22:104–111.
59.
FriedmanHV, BreslerT, GarnerCC and ZivNE. (2000). Assembly of new individual excitatory synapses: time course and temporal order of synaptic molecule recruitment. Neuron, 27:57–69.
60.
DickO, tom DieckS, AltrockWD, AmmermullerJ, WeilerR, GarnerCC, GundelfingerED and BrandstatterJH. (2003). The presynaptic active zone protein bassoon is essential for photoreceptor ribbon synapse formation in the retina. Neuron, 37:775–786.
61.
BecherA, DrenckhahnA, PahnerI, MargittaiM, JahnR and Ahnert-HilgerG. (1999). The synaptophysin-synaptobrevin complex: a hallmark of synaptic vesicle maturation. J Neurosci, 19:1922–1931.
ZhouQ, ChoiG and AndersonDJ. (2001). The bHLH transcription factor Olig2 promotes oligodendrocyte differentiation in collaboration with Nkx2.2. Neuron, 31:791–807.
64.
ImaiS, ArmstrongCM, KaeberleinM and GuarenteL. (2000). Transcriptional silencing and longevity protein Sir2 is an NAD-dependent histone deacetylase. Nature, 403:795–800.
65.
LiW, ZhangB, TangJ, CaoQ, WuY, WuC, GuoJ, LingEA and LiangF. (2007). Sirtuin 2, a mammalian homolog of yeast silent information regulator-2 longevity regulator, is an oligodendroglial protein that decelerates cell differentiation through deacetylating alpha-tubulin. J Neurosci, 27:2606–2616.
66.
DringenR. (2000). Metabolism and functions of glutathione in brain. Prog Neurobiol, 62:649–671.
67.
FridovichI. (1995). Superoxide radical and superoxide dismutases. Annu Rev Biochem, 64:97–112.
68.
SmithJ, LadiE, Mayer-ProschelM and NobleM. (2000). Redox state is a central modulator of the balance between self-renewal and differentiation in a dividing glial precursor cell. Proc Natl Acad Sci U S A, 97:10032–10037.
69.
ThorburneSK and JuurlinkBH. (1996). Low glutathione and high iron govern the susceptibility of oligodendroglial precursors to oxidative stress. J Neurochem, 67:1014–1022.
70.
KhannaS, RoyS, BagchiD, BagchiM and SenCK. (2001). Upregulation of oxidant-induced VEGF expression in cultured keratinocytes by a grape seed proanthocyanidin extract. Free Radic Biol Med, 31:38–42.
71.
SondellM, LundborgG and KanjeM. (1999). Vascular endothelial growth factor has neurotrophic activity and stimulates axonal outgrowth, enhancing cell survival and Schwann cell proliferation in the peripheral nervous system. J Neurosci, 19:5731–5740.
72.
StoneJ, ItinA, AlonT, Pe'erJ, GnessinH, Chan-LingT and KeshetE. (1995). Development of retinal vasculature is mediated by hypoxia-induced vascular endothelial growth factor (VEGF) expression by neuroglia. J Neurosci, 15:4738–4747.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
