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1529-240134422014Oct15The Journal of neuroscience : the official journal of the Society for NeuroscienceJ NeurosciHeterogeneously expressed fezf2 patterns gradient Notch activity in balancing the quiescence, proliferation, and differentiation of adult neural stem cells.139111392313911-2310.1523/JNEUROSCI.1976-14.2014Balancing quiescence, self-renewal, and differentiation in adult stem cells is critical for tissue homeostasis. The underlying mechanisms, however, remain incompletely understood. Here we identify Fezf2 as a novel regulator of fate balance in adult zebrafish dorsal telencephalic neural stem cells (NSCs). Transgenic reporters show intermingled fezf2-GFP(hi) quiescent and fezf2-GFP(lo) proliferative NSCs. Constitutive or conditional impairment of fezf2 activity demonstrates its requirement for maintaining quiescence. Analyses of genetic chimeras reveal a dose-dependent role of fezf2 in NSC activation, suggesting that the difference in fezf2 levels directionally biases fate. Single NSC profiling coupled with genetic analysis further uncovers a fezf2-dependent gradient Notch activity that is high in quiescent and low in proliferative NSCs. Finally, fezf2-GFP(hi) quiescent and fezf2-GFP(lo) proliferative NSCs are observed in postnatal mouse hippocampus, suggesting possible evolutionary conservation. Our results support a model in which fezf2 heterogeneity patterns gradient Notch activity among neighbors that is critical to balance NSC fate.Copyright © 2014 the authors 0270-6474/14/3413911-13$15.00/0.BerberogluMichael AMADepartment of Bioengineering and Therapeutic Sciences, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Graduate Program in Neuroscience, Institute of Human Genetics, Department of Molecular Genetics, Ohio State University, Columbus, Ohio 43210.DongZhiqiangZ0000-0001-6259-915XDepartment of Bioengineering and Therapeutic Sciences, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Institute of Human Genetics.LiGuangnanGEli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Department of Neurology.ZhengJiashunJDepartment of Biochemistry and Biophysics.Trejo MartinezLuz del Carmen GLdel CDepartment of Bioengineering and Therapeutic Sciences, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Institute of Human Genetics, Graduate Program in Cell and Molecular Biology, Department of Biology, San Francisco State University, San Francisco, California 94132.PengJisongJDepartment of Bioengineering and Therapeutic Sciences, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Institute of Human Genetics.WagleMahendraMDepartment of Bioengineering and Therapeutic Sciences, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Institute of Human Genetics.ReichholfBrianBDepartment of Neurological Surgery, and.PetritschClaudiaCDepartment of Neurological Surgery, and Brain Tumor Research Center, University of California, San Francisco, California 94143-2811.LiHaoHDepartment of Biochemistry and Biophysics.PleasureSamuel JSJ0000-0001-8599-1613Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Graduate Program in Neuroscience, Department of Neurology.GuoSuS0000-0002-7342-0108Department of Bioengineering and Therapeutic Sciences, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Graduate Program in Neuroscience, Institute of Human Genetics, State Key Laboratory of Genetic Engineering and the Institute of Genetics, School of Life Sciences, Fudan University, Shanghai, China, and su.guo@ucsf.edu.engR01 NS080619NSNINDS NIH HHSUnited StatesR01 NS042626NSNINDS NIH HHSUnited StatesDA035680DANIDA NIH HHSUnited StatesR01 CA164746CANCI NIH HHSUnited StatesR01 MH077694MHNIMH NIH HHSUnited StatesNS042626NSNINDS NIH HHSUnited StatesR01 MH105360MHNIMH NIH HHSUnited StatesR01 DA035680DANIDA NIH HHSUnited StatesT32 NS077984NSNINDS NIH HHSUnited StatesJournal ArticleResearch Support, N.I.H., Extramural
United StatesJ Neurosci81021400270-64740DNA-Binding Proteins0Nerve Tissue Proteins0Receptors, Notch0Zfp312 protein, mouseIMAdult Stem CellsmetabolismAnimalsAnimals, Genetically ModifiedCell DifferentiationphysiologyCell ProliferationphysiologyDNA-Binding ProteinsbiosynthesisFemaleGene Expression Regulation, DevelopmentalMaleMiceMice, TransgenicNerve Tissue ProteinsbiosynthesisNeural Stem CellsmetabolismNeurogenesisphysiologyReceptors, NotchmetabolismZebrafishadult neurogenesishippocampusradial gliaself-renewalsingle-cell analysisvivo morpholino20141017602014101760201527602015415ppublish25319688PMC419853710.1523/JNEUROSCI.1976-14.201434/42/13911Adolf B, Chapouton P, Lam CS, Topp S, Tannhäuser B, Strähle U, Götz M, Bally-Cuif L. Conserved and acquired features of adult neurogenesis in the zebrafish telencephalon. Dev Biol. 2006;295:278–293. doi: 10.1016/j.ydbio.2006.03.023.10.1016/j.ydbio.2006.03.02316828638Alunni A, Krecsmarik M, Bosco A, Galant S, Pan L, Moens CB, Bally-Cuif L. Notch3 signaling gates cell cycle entry and limits neural stem cell amplification in the adult pallium. Development. 2013;140:3335–3347. doi: 10.1242/dev.095018.10.1242/dev.095018PMC373771623863484Bae YK, Kani S, Shimizu T, Tanabe K, Nojima H, Kimura Y, Higashijima S, Hibi M. Anatomy of zebrafish cerebellum and screen for mutations affecting its development. Dev Biol. 2009;330:406–426. doi: 10.1016/j.ydbio.2009.04.013.10.1016/j.ydbio.2009.04.01319371731Baumgart EV, Barbosa JS, Bally-Cuif L, Götz M, Ninkovic J. Stab wound injury of the zebrafish telencephalon: a model for comparative analysis of reactive gliosis. Glia. 2012;60:343–357. doi: 10.1002/glia.22269.10.1002/glia.2226922105794Berberoglu MA, Dong Z, Mueller T, Guo S. fezf2 expression delineates cells with proliferative potential and expressing markers of neural stem cells in the adult zebrafish brain. Gene Expr Patterns. 2009;9:411–422. doi: 10.1016/j.gep.2009.06.002.10.1016/j.gep.2009.06.002PMC274671319524703Blanpain C, Fuchs E. Epidermal homeostasis: a balancing act of stem cells in the skin. Nat Rev Mol Cell Biol. 2009;10:207–217. doi: 10.1038/nrm2636.10.1038/nrm2636PMC276021819209183Bonaguidi MA, Wheeler MA, Shapiro JS, Stadel RP, Sun GJ, Ming GL, Song H. In vivo clonal analysis reveals self-renewing and multipotent adult neural stem cell characteristics. Cell. 2011;145:1142–1155. doi: 10.1016/j.cell.2011.05.024.10.1016/j.cell.2011.05.024PMC312456221664664Carlén M, Meletis K, Göritz C, Darsalia V, Evergren E, Tanigaki K, Amendola M, Barnabé-Heider F, Yeung MS, Naldini L, Honjo T, Kokaia Z, Shupliakov O, Cassidy RM, Lindvall O, Frisén J. Forebrain ependymal cells are Notch-dependent and generate neuroblasts and astrocytes after stroke. Nat Neurosci. 2009;12:259–267. doi: 10.1038/nn.2268.10.1038/nn.226819234458Chapouton P, Adolf B, Leucht C, Tannhäuser B, Ryu S, Driever W, Bally-Cuif L. her5 expression reveals a pool of neural stem cells in the adult zebrafish midbrain. Development. 2006;133:4293–4303. doi: 10.1242/dev.02573.10.1242/dev.0257317038515Chapouton P, Skupien P, Hesl B, Coolen M, Moore JC, Madelaine R, Kremmer E, Faus-Kessler T, Blader P, Lawson ND, Bally-Cuif L. Notch activity levels control the balance between quiescence and recruitment of adult neural stem cells. J Neurosci. 2010;30:7961–7974. doi: 10.1523/JNEUROSCI.6170-09.2010.10.1523/JNEUROSCI.6170-09.2010PMC663267820534844Chen B, Schaevitz LR, McConnell SK. Fezl regulates the differentiation and axon targeting of layer 5 subcortical projection neurons in cerebral cortex. Proc Natl Acad Sci U S A. 2005a;102:17184–17189. doi: 10.1073/pnas.0508732102.10.1073/pnas.0508732102PMC128256916284245Chen JG, Rasin MR, Kwan KY, Sestan N. Zfp312 is required for subcortical axonal projections and dendritic morphology of deep-layer pyramidal neurons of the cerebral cortex. Proc Natl Acad Sci U S A. 2005b;102:17792–17797. doi: 10.1073/pnas.0509032102.10.1073/pnas.0509032102PMC130892816314561Chen L, Zheng J, Yang N, Li H, Guo S. Genomic selection identifies vertebrate transcription factor Fezf2 binding sites and target genes. J Biol Chem. 2011;286:18641–18649. doi: 10.1074/jbc.M111.236471.10.1074/jbc.M111.236471PMC309968021471212Collins SR, Schuldiner M, Krogan NJ, Weissman JS. A strategy for extracting and analyzing large-scale quantitative epistatic interaction data. Genome Biol. 2006;7:R63. doi: 10.1186/gb-2006-7-7-r63.10.1186/gb-2006-7-7-r63PMC177956816859555Del Bene F, Wehman AM, Link BA, Baier H. Regulation of neurogenesis by interkinetic nuclear migration through an apical-basal notch gradient. Cell. 2008;134:1055–1065. doi: 10.1016/j.cell.2008.07.017.10.1016/j.cell.2008.07.017PMC262848718805097Dong Z, Yang N, Yeo SY, Chitnis A, Guo S. Intra-lineage directional Notch signaling regulates self-renewal and differentiation of asymmetrically dividing radial glia. Neuron. 2012;74:65–78. doi: 10.1016/j.neuron.2012.01.031.10.1016/j.neuron.2012.01.031PMC346611422500631Eckler MJ, Chen B. Fez family transcription factors: controlling neurogenesis and cell fate in the developing mammalian nervous system. Bioessays. 2014;36:788–797. doi: 10.1002/bies.201400039.10.1002/bies.201400039PMC547247624913420Eisen MB, Spellman PT, Brown PO, Botstein D. Cluster analysis and display of genome-wide expression patterns. Proc Natl Acad Sci U S A. 1998;95:14863–14868. doi: 10.1073/pnas.95.25.14863.10.1073/pnas.95.25.14863PMC245419843981Fuchs E. The tortoise and the hair: slow-cycling cells in the stem cell race. Cell. 2009;137:811–819. doi: 10.1016/j.cell.2009.05.002.10.1016/j.cell.2009.05.002PMC271612219490891Gage FH. Mammalian neural stem cells. Science. 2000;287:1433–1438. doi: 10.1126/science.287.5457.1433.10.1126/science.287.5457.143310688783Gaiano N, Fishell G. The role of notch in promoting glial and neural stem cell fates. Annu Rev Neurosci. 2002;25:471–490. doi: 10.1146/annurev.neuro.25.030702.130823.10.1146/annurev.neuro.25.030702.13082312052917Grandel H, Kaslin J, Ganz J, Wenzel I, Brand M. Neural stem cells and neurogenesis in the adult zebrafish brain: origin, proliferation dynamics, migration and cell fate. Dev Biol. 2006;295:263–277. doi: 10.1016/j.ydbio.2006.03.040.10.1016/j.ydbio.2006.03.04016682018Guo C, Eckler MJ, McKenna WL, McKinsey GL, Rubenstein JL, Chen B. Fezf2 expression identifies a multipotent progenitor for neocortical projection neurons, astrocytes, and oligodendrocytes. Neuron. 2013;80:167–174. doi: 10.1016/j.neuron.2013.09.037.10.1016/j.neuron.2013.09.037PMC385758824314728Guo S, Wilson SW, Cooke S, Chitnis AB, Driever W, Rosenthal A. Mutations in the zebrafish unmask shared regulatory pathways controlling the development of catecholaminergic neurons. Dev Biol. 1999;208:473–487. doi: 10.1006/dbio.1999.9204.10.1006/dbio.1999.920410191060Hirata T, Suda Y, Nakao K, Narimatsu M, Hirano T, Hibi M. Zinc finger gene fez-like functions in the formation of subplate neurons and thalamocortical axons. Dev Dyn. 2004;230:546–556. doi: 10.1002/dvdy.20068.10.1002/dvdy.2006815188439Hirata T, Nakazawa M, Muraoka O, Nakayama R, Suda Y, Hibi M. Zinc-finger genes Fez and Fez-like function in the establishment of diencephalon subdivisions. Development. 2006;133:3993–4004. doi: 10.1242/dev.02585.10.1242/dev.0258516971467Imayoshi I, Sakamoto M, Yamaguchi M, Mori K, Kageyama R. Essential roles of Notch signaling in maintenance of neural stem cells in developing and adult brains. J Neurosci. 2010;30:3489–3498. doi: 10.1523/JNEUROSCI.4987-09.2010.10.1523/JNEUROSCI.4987-09.2010PMC663411920203209Jeong JY, Einhorn Z, Mercurio S, Lee S, Lau B, Mione M, Wilson SW, Guo S. Neurogenin1 is a determinant of zebrafish basal forebrain dopaminergic neurons and is regulated by the conserved zinc finger protein Tof/Fezl. Proc Natl Acad Sci U S A. 2006;103:5143–5148. doi: 10.1073/pnas.0600337103.10.1073/pnas.0600337103PMC145880816549779Jeong JY, Einhorn Z, Mathur P, Chen L, Lee S, Kawakami K, Guo S. Patterning the zebrafish diencephalon by the conserved zinc finger protein Fezl. Development. 2007;134:127–136. doi: 10.1242/dev.02705.10.1242/dev.0270517164418Kizil C, Kyritsis N, Dudczig S, Kroehne V, Freudenreich D, Kaslin J, Brand M. Regenerative neurogenesis from neural progenitor cells requires injury-induced expression of Gata3. Dev Cell. 2012;23:1230–1237. doi: 10.1016/j.devcel.2012.10.014.10.1016/j.devcel.2012.10.01423168169Kizil C, Iltzsche A, Kaslin J, Brand M. Micromanipulation of gene expression in the adult zebrafish brain using cerebroventricular microinjection of morpholino oligonucleotides. J Vis Exp. 2013;75:e50415. doi: 10.3791/50415.10.3791/50415PMC371828823728426Kroehne V, Freudenreich D, Hans S, Kaslin J, Brand M. Regeneration of the adult zebrafish brain from neurogenic radial glia-type progenitors. Development. 2011;138:4831–4841. doi: 10.1242/dev.072587.10.1242/dev.07258722007133Levkowitz G, Zeller J, Sirotkin HI, French D, Schilbach S, Hashimoto H, Hibi M, Talbot WS, Rosenthal A. Zinc finger protein too few controls the development of monoaminergic neurons. Nat Neurosci. 2003;6:28–33. doi: 10.1038/nn979.10.1038/nn97912469125Li G, Fang L, Fernández G, Pleasure SJ. The ventral hippocampus is the embryonic origin for adult neural stem cells in the dentate gyrus. Neuron. 2013;78:658–672. doi: 10.1016/j.neuron.2013.03.019.10.1016/j.neuron.2013.03.019PMC366923023643936Li L, Clevers H. Coexistence of quiescent and active adult stem cells in mammals. Science. 2010;327:542–545. doi: 10.1126/science.1180794.10.1126/science.1180794PMC410518220110496Lindsey BW, Darabie A, Tropepe V. The cellular composition of neurogenic periventricular zones in the adult zebrafish forebrain. J Comp Neurol. 2012;520:2275–2316. doi: 10.1002/cne.23065.10.1002/cne.2306522318736März M, Chapouton P, Diotel N, Vaillant C, Hesl B, Takamiya M, Lam CS, Kah O, Bally-Cuif L, Strähle U. Heterogeneity in progenitor cell subtypes in the ventricular zone of the zebrafish adult telencephalon. Glia. 2010;58:870–888. doi: 10.1002/glia.20971.10.1002/glia.2097120155821Mazzone M, Selfors LM, Albeck J, Overholtzer M, Sale S, Carroll DL, Pandya D, Lu Y, Mills GB, Aster JC, Artavanis-Tsakonas S, Brugge JS. Dose-dependent induction of distinct phenotypic responses to Notch pathway activation in mammary epithelial cells. Proc Natl Acad Sci U S A. 2010;107:5012–5017. doi: 10.1073/pnas.1000896107.10.1073/pnas.1000896107PMC284192320194747Merkle FT, Mirzadeh Z, Alvarez-Buylla A. Mosaic organization of neural stem cells in the adult brain. Science. 2007;317:381–384. doi: 10.1126/science.1144914.10.1126/science.114491417615304Miller FD, Gauthier-Fisher A. Home at last: neural stem cell niches defined. Cell Stem Cell. 2009;4:507–510. doi: 10.1016/j.stem.2009.05.008.10.1016/j.stem.2009.05.00819497279Ming GL, Song H. Adult neurogenesis in the mammalian brain: significant answers and significant questions. Neuron. 2011;70:687–702. doi: 10.1016/j.neuron.2011.05.001.10.1016/j.neuron.2011.05.001PMC310610721609825Mizutani K, Yoon K, Dang L, Tokunaga A, Gaiano N. Differential Notch signalling distinguishes neural stem cells from intermediate progenitors. Nature. 2007;449:351–355. doi: 10.1038/nature06090.10.1038/nature0609017721509Molyneaux BJ, Arlotta P, Hirata T, Hibi M, Macklis JD. Fezl is required for the birth and specification of corticospinal motor neurons. Neuron. 2005;47:817–831. doi: 10.1016/j.neuron.2005.08.030.10.1016/j.neuron.2005.08.03016157277Morrison SJ, Kimble J. Asymmetric and symmetric stem-cell divisions in development and cancer. Nature. 2006;441:1068–1074. doi: 10.1038/nature04956.10.1038/nature0495616810241Morrison SJ, Spradling AC. Stem cells and niches: mechanisms that promote stem cell maintenance throughout life. Cell. 2008;132:598–611. doi: 10.1016/j.cell.2008.01.038.10.1016/j.cell.2008.01.038PMC450572818295578Mu Y, Lee SW, Gage FH. Signaling in adult neurogenesis. Curr Opin Neurobiol. 2010;20:416–423. doi: 10.1016/j.conb.2010.04.010.10.1016/j.conb.2010.04.010PMC294296520471243Ninov N, Borius M, Stainier DY. Different levels of Notch signaling regulate quiescence, renewal and differentiation in pancreatic endocrine progenitors. Development. 2012;139:1557–1567. doi: 10.1242/dev.076000.10.1242/dev.076000PMC331796422492351Raj A, van Oudenaarden A. Nature, nurture, or chance: stochastic gene expression and its consequences. Cell. 2008;135:216–226. doi: 10.1016/j.cell.2008.09.050.10.1016/j.cell.2008.09.050PMC311804418957198Rothenaigner I, Krecsmarik M, Hayes JA, Bahn B, Lepier A, Fortin G, Götz M, Jagasia R, Bally-Cuif L. Clonal analysis by distinct viral vectors identifies bona fide neural stem cells in the adult zebrafish telencephalon and characterizes their division properties and fate. Development. 2011;138:459–469. doi: 10.1242/dev.058156.10.1242/dev.05815621367818Rouaux C, Arlotta P. Fezf2 directs the differentiation of corticofugal neurons from striatal progenitors in vivo. Nat Neurosci. 2010;13:1345–1347. doi: 10.1038/nn.2658.10.1038/nn.2658PMC420744220953195Selkoe D, Kopan R. Notch and Presenilin: regulated intramembrane proteolysis links development and degeneration. Annu Rev Neurosci. 2003;26:5697. doi: 10.1146/annurev.neuro.26.041002.131334.10.1146/annurev.neuro.26.041002.13133412730322Shimizu T, Hibi M. Formation and patterning of the forebrain and olfactory system by zinc-finger genes Fezf1 and Fezf2. Dev Growth Differ. 2009;51:221–231. doi: 10.1111/j.1440-169X.2009.01088.x.10.1111/j.1440-169X.2009.01088.x19222525Shimizu T, Nakazawa M, Kani S, Bae YK, Kageyama R, Hibi M. Zinc finger genes Fezf1 and Fezf2 control neuronal differentiation by repressing Hes5 expression in the forebrain. Development. 2010;137:1875–1885. doi: 10.1242/dev.047167.10.1242/dev.04716720431123Simons BD, Clevers H. Strategies for homeostatic stem cell self-renewal in adult tissues. Cell. 2011;145:851–862. doi: 10.1016/j.cell.2011.05.033.10.1016/j.cell.2011.05.03321663791Spurgeon SL, Jones RC, Ramakrishnan R. High throughput gene expression measurement with real time PCR in a microfluidic dynamic array. PLoS One. 2008;3:e1662. doi: 10.1371/journal.pone.0001662.10.1371/journal.pone.0001662PMC224470418301740Sugiarto S, Persson A, Munoz EG, Waldhuber M, Lamagna C, Andor N, Hanecker P, Ayers-Ringler J, Phillips J, Siu J, Lim DA, Vandenberg S, Stallcup W, Berger MS, Bergers G, Weiss WA, Petritsch C. Asymmetry-defective oligodendrocyte progenitors are glioma precursors. Cancer Cell. 2011;20:328–340. doi: 10.1016/j.ccr.2011.08.011.10.1016/j.ccr.2011.08.011PMC329749021907924Takke C, Dornseifer P, v Weizsäcker E, Campos-Ortega JA. her4, a zebrafish homologue of the Drosophila neurogenic gene E (spl), is a target of NOTCH signalling. Development. 1999;126:1811–1821.10101116Temple S, Alvarez-Buylla A. Stem cells in the adult mammalian central nervous system. Curr Opin Neurobiol. 1999;9:135–141. doi: 10.1016/S0959-4388(99)80017-8.10.1016/S0959-4388(99)80017-810072370Weissman IL, Anderson DJ, Gage F. Stem and progenitor cells: origins, phenotypes, lineage commitments, and transdifferentiations. Annu Rev Cell Dev Biol. 2001;17:387–403. doi: 10.1146/annurev.cellbio.17.1.387.10.1146/annurev.cellbio.17.1.38711687494Wullimann MF, Puelles L. Postembryonic neural proliferation in the zebrafish forebrain and its relationship to prosomeric domains. Anat Embryol (Berl) 1999;199:329–348. doi: 10.1007/s004290050232.10.1007/s00429005023210195307Yang J, Chan CY, Jiang B, Yu X, Zhu GZ, Chen Y, Barnard J, Mei W. hnRNP I inhibits Notch signaling and regulates intestinal epithelial homeostasis in the zebrafish. PLoS Genet. 2009;5:e1000363. doi: 10.1371/journal.pgen.1000363.10.1371/journal.pgen.1000363PMC262957719197356Yang N, Dong Z, Guo S. Fezf2 regulates multi-lineage neuronal differentiation through activating bHLH and HD genes in the zebrafish ventral forebrain. J Neurosci. 2012;32:10940–10948. doi: 10.1523/JNEUROSCI.2216-12.2012.10.1523/JNEUROSCI.2216-12.2012PMC347889522875928Zhao C, Deng W, Gage FH. Mechanisms and functional implications of adult neurogenesis. Cell. 2008;132:645–660. doi: 10.1016/j.cell.2008.01.033.10.1016/j.cell.2008.01.03318295581Zupanc GK, Hinsch K, Gage FH. Proliferation, migration, neuronal differentiation, and long-term survival of new cells in the adult zebrafish brain. J Comp Neurol. 2005;488:290–319. doi: 10.1002/cne.20571.10.1002/cne.2057115952170