[en] ABSTRACT: BACKGROUND: Doublecortin (Dcx), a MAP (Microtubule-Associated Protein), is transiently expressed in migrating and differentiating neurons and thereby characterizes neuronal precursors and neurogenesis in developing and adult neurogenesis. In addition, reduced Dcx expression during development has been related to appearance of brain pathologies. Here, we attempt to unveil the molecular mechanisms controlling Dcx gene expression by studying its transcriptional regulation during neuronal differentiation. RESULTS: To determine and analyze important regulatory sequences of the Dcx promoter, we studied a putative regulatory region upstream from the mouse Dcx coding region (pdcx2kb) and several deletions thereof. These different fragments were used in vitro and in vivo to drive reporter gene expression. We demonstrated, using transient expression experiments, that pdcx2kb is sufficient to control specific reporter gene expression in cerebellar cells and in the developing (E14.5) brain. We determined the temporal profile of Dcx promoter activity during neuronal differentiation of mouse embryonic stem cells (mESC) and found that transcriptional activation of the Dcx gene varies along with neuronal differentiation of mESC. Deletion experiments and sequence comparison of Dcx promoters across rodents, human and chicken revealed the importance of a highly conserved sequence in the proximal region of the promoter required for specific and strong expression in neuronal precursors and young neuronal cells. Further analyses revealed the presence in this short sequence of several conserved, putative transcription factor binding sites: LEF/TCF (Lymphoid Enhancer Factor/T-Cell Factor) which are effectors of the canonical Wnt pathway; HNF6/OC2 (Hepatocyte Nuclear Factor-6/Oncecut-2) members of the ONECUT family; and NF-Y/CAAT (Nuclear Factor-Y). CONCLUSIONS: Studies of Dcx gene regulatory sequences using native, deleted and mutated constructs suggest that fragments located upstream of the Dcx coding sequence are sufficient to induce specific Dcx expression in vitro: in heterogeneous differentiated neurons from mESC, in primary mouse cerebellar neurons (PND3) and in organotypic slices cultures. Furthermore, a region in the 3'-end region of the Dcx promoter is highly conserved across several species and exerts positive control on Dcx transcriptional activation. Together, these results indicate that the proximal 3'-end region of the mouse Dcx regulatory sequence is essential for Dcx gene expression during differentiation of neuronal precursors.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Piens, M.
Muller, Marc ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire
Bodson, Marguerite ; Centre Hospitalier Universitaire de Liège - CHU > HOSPITALISATION - UNITE STERILE ( SI -3C)
Baudouin, Gregory
Plumier, Jean-Christophe ; Université de Liège - ULiège > Département des sciences et gestion de l'environnement > Ecophysiologie et physiologie animale
Language :
English
Title :
A short upstream promoter region mediates transcriptional regulation of the mouse doublecortin gene in differentiating neurons.
Francis F, Koulakoff A, Boucher D, Chafey P, Schaar B, Vinet MC, Friocourt G, McDonnell N, Reiner O, Kahn A, et al. Doublecortin is a developmentally regulated, microtubule-associated protein expressed in migrating and differentiating neurons. Neuron 1999, 23(2):247-256. 10.1016/S0896-6273(00)80777-1, 10399932.
des Portes V, Pinard JM, Billuart P, Vinet MC, Koulakoff A, Carrié A, Gelot A, Dupuis E, Motte J, Berwald-Netter Y, et al. A novel CNS gene required for neuronal migration and involved in X-linked subcortical laminar heterotopia and lissencephaly syndrome. Cell 1998, 92(1):51-61. 10.1016/S0092-8674(00)80898-3, 9489699.
Walker TL, Yasuda T, Adams DJ, Bartlett PF. The doublecortin-expressing population in the developing and adult brain contains multipotential precursors in addition to neuronal-lineage cells. Journal of Neuroscience 2007, 27(14):3734-3742. 10.1523/JNEUROSCI.5060-06.2007, 17409237.
Friocourt G, Koulakoff A, Chafey P, Boucher D, Fauchereau F, Chelly J, Francis F. Doublecortin functions at the extremities of growing neuronal processes. Cereb Cortex 2003, 13(6):620-626. 10.1093/cercor/13.6.620, 12764037.
Brown JP, Couillard-Després S, Cooper-Kuhn CM, Winkler J, Aigner L, Kuhn HG. Transient expression of doublecortin during adult neurogenesis. J Comp Neurol 2003, 467(1):1-10. 10.1002/cne.10874, 14574675.
Mattar P, Britz O, Johannes C, Nieto M, Ma L, Rebeyka A, Klenin N, Polleux F, Guillemot F, Schuurmans C. A screen for downstream effectors of Neurogenin2 in the embryonic neocortex. Developmental Biology 2004, 273(2):373-389. 10.1016/j.ydbio.2004.06.013, 15328020.
Couillard-Despres S, Winner B, Schaubeck S, Aigner R, Vroemen M, Weidner N, Bogdahn U, Winkler J, Kuhn H, Aigner L. Doublecortin expression levels in adult brain reflect neurogenesis. Eur J Neurosci 2005, 21(1):1-14. 10.1111/j.1460-9568.2004.03813.x, 15654838.
Gleeson JG, Allen KM, Fox JW, Lamperti ED, Berkovic S, Scheffer I, Cooper EC, Dobyns WB, Minnerath SR, Ross ME, et al. Doublecortin, a brain-specific gene mutated in human X-linked lissencephaly and double cortex syndrome, encodes a putative signaling protein. Cell 1998, 92(1):63-72. 10.1016/S0092-8674(00)80899-5, 9489700.
Corbo JC, Deuel TA, Long JM, LaPorte P, Tsai E, Wynshaw-Boris A, Walsh C. Doublecortin is required in mice for lamination of the hippocampus but not the neocortex. Journal of Neuroscience 2002, 22(17):7548-7557.
Ramos R. Heterotopia Formation in Rat but Not Mouse Neocortex after RNA Interference Knockdown of DCX. Cerebral Cortex 2005, 16(9):1323-1331. 10.1093/cercor/bhj074, 16292002.
Bai J, Ramos R, Ackman J, Thomas A, Lee R, Loturco J. RNAi reveals doublecortin is required for radial migration in rat neocortex. Nat Neurosci 2003, 6(12):1277-1283. 10.1038/nn1153, 14625554.
Loturco J, Bai J. The multipolar stage and disruptions in neuronal migration. Trends in Neurosciences 2006, 29(7):407-413. 10.1016/j.tins.2006.05.006, 16713637.
Friocourt G, Liu JS, Antypa M, Rakic S, Walsh C, Parnavelas JG. Both doublecortin and doublecortin-like kinase play a role in cortical interneuron migration. Journal of Neuroscience 2007, 27(14):3875-3883. 10.1523/JNEUROSCI.4530-06.2007, 17409252.
Deuel TA, Liu JS, Corbo JC, Yoo SY, Rorke-Adams LB, Walsh C. Genetic interactions between doublecortin and doublecortin-like kinase in neuronal migration and axon outgrowth. Neuron 2006, 49(1):41-53. 10.1016/j.neuron.2005.10.038, 16387638.
Zhou Q, Anderson DJ. The bHLH transcription factors OLIG2 and OLIG1 couple neuronal and glial subtype specification. Cell 2002, 109(1):61-73. 10.1016/S0092-8674(02)00677-3, 11955447.
Paratore C, Hagedorn L, Floris J, Hari L, Kléber M, Suter U, Sommer L. Cell-intrinsic and cell-extrinsic cues regulating lineage decisions in multipotent neural crest-derived progenitor cells. Int J Dev Biol 2002, 46(1):193-200.
Kintner C. Neurogenesis in embryos and in adult neural stem cells. J Neurosci 2002, 22(3):639-643.
Gleeson JG, Lin PT, Flanagan LA, Walsh CA. Doublecortin is a microtubule-associated protein and is expressed widely by migrating neurons. Neuron 1999, 23(2):257-271. 10.1016/S0896-6273(00)80778-3, 10399933.
Nadarajah B, Alifragis P, Wong RO, Parnavelas JG. Neuronal migration in the developing cerebral cortex: observations based on real-time imaging. Cereb Cortex 2003, 13(6):607-611. 10.1093/cercor/13.6.607, 12764035.
Bibel M, Richter J, Schrenk K, Tucker K, Staiger V, Korte M, Goetz M, Barde Y. Differentiation of mouse embryonic stem cells into a defined neuronal lineage. Nat Neurosci 2004, 7(9):1003-1009. 10.1038/nn1301, 15332090.
Zhang JQ, Yu XB, Ma BF, Yu WH, Zhang AX, Huang G, Mao FF, Zhang XM, Wang ZC, Li SN, et al. Neural differentiation of embryonic stem cells induced by conditioned medium from neural stem cell. Neuroreport 2006, 17(10):981-986. 10.1097/01.wnr.0000227977.60271.ca, 16791088.
Billon N, Jolicoeur C, Tokumoto Y, Vennström B, Raff M. Normal timing of oligodendrocyte development depends on thyroid hormone receptor alpha 1 (TRalpha1). EMBO J 2002, 21(23):6452-6460. 10.1093/emboj/cdf662, 136965, 12456652.
Lemaigre F. [Structure, function and physiopathological implications of HNF-6, a prototype of a new family of gene expression regulators]. Bull Mem Acad R Med Belg 1999, 154(6 Pt 2):295-301.
Landry C, Clotman F, Hioki T, Oda H, Picard JJ, Lemaigre FP, Rousseau GG. HNF-6 is expressed in endoderm derivatives and nervous system of the mouse embryo and participates to the cross-regulatory network of liver-enriched transcription factors. Developmental Biology 1997, 192(2):247-257. 10.1006/dbio.1997.8757, 9441665.
Jacquemin P, Lannoy VJ, Rousseau GG, Lemaigre FP. OC-2, a novel mammalian member of the ONECUT class of homeodomain transcription factors whose function in liver partially overlaps with that of hepatocyte nuclear factor-6. J Biol Chem 1999, 274(5):2665-2671. 10.1074/jbc.274.5.2665, 9915796.
Willems E, Mateizel I, Kemp C, Cauffman G, Sermon K, Leyns L. Selection of reference genes in mouse embryos and in differentiating human and mouse ES cells. Int J Dev Biol 2006, 50(7):627-635. 10.1387/ijdb.052130ew, 16892176.
Reyes JH, O'Shea KS, Wys NL, Velkey JM, Prieskorn DM, Wesolowski K, Miller JM, Altschuler RA. Glutamatergic neuronal differentiation of mouse embryonic stem cells after transient expression of neurogenin 1 and treatment with BDNF and GDNF: in vitro and in vivo studies. J Neurosci 2008, 28(48):12622-12631. 10.1523/JNEUROSCI.0563-08.2008, 2729437, 19036956.
Abranches E, Silva M, Pradier L, Schulz H, Hummel O, Henrique D, Bekman E. Neural differentiation of embryonic stem cells in vitro: a road map to neurogenesis in the embryo. PLoS ONE 2009, 4(7):e6286. 10.1371/journal.pone.0006286, 2709448, 19621087.
Pereira L, Yi F, Merrill BJ. Repression of Nanog gene transcription by Tcf3 limits embryonic stem cell self-renewal. Mol Cell Biol 2006, 26(20):7479-7491. 10.1128/MCB.00368-06, 1636872, 16894029.
Tóth G, Gáspári Z, Jurka J. Microsatellites in different eukaryotic genomes: survey and analysis. Genome Res 2000, 10(7):967-981. 10.1101/gr.10.7.967, 310925, 10899146.
Katti MV, Ranjekar PK, Gupta VS. Differential distribution of simple sequence repeats in eukaryotic genome sequences. Mol Biol Evol 2001, 18(7):1161-1167.
Huang T-S, Lee C-C, Chang A-C, Lin S, Chao C-C, Jou Y-S, Chu Y-W, Wu C-W, Whang-Peng J. Shortening of microsatellite deoxy(CA) repeats involved in GL331-induced down-regulation of matrix metalloproteinase-9 gene expression. Biochem Biophys Res Commun 2003, 300(4):901-907. 10.1016/S0006-291X(02)02962-5, 12559958.
Gebhardt F, Zänker KS, Brandt B. Modulation of epidermal growth factor receptor gene transcription by a polymorphic dinucleotide repeat in intron 1. J Biol Chem 1999, 274(19):13176-13180. 10.1074/jbc.274.19.13176, 10224073.
Meyer G, Perez-Garcia CG, Gleeson J. Selective expression of doublecortin and LIS1 in developing human cortex suggests unique modes of neuronal movement. Cereb Cortex 2002, 12(12):1225-1236. 10.1093/cercor/12.12.1225, 12427674.
Ying Q, Stavridis M, Griffiths D, Li M, Smith A. Conversion of embryonic stem cells into neuroectodermal precursors in adherent monoculture. Nat Biotech 2003, 21(2):183-186.
Flanagan LA, Rebaza LM, Derzic S, Schwartz PH, Monuki ES. Regulation of human neural precursor cells by laminin and integrins. J Neurosci Res 2006, 83(5):845-856. 10.1002/jnr.20778, 2409144, 16477652.
De Arcangelis A, Georges-Labouesse E. Integrin and ECM functions: roles in vertebrate development. Trends Genet 2000, 16(9):389-395. 10.1016/S0168-9525(00)02074-6, 10973067.
Andressen C, Adrian S, Fässler R, Arnhold S, Addicks K. The contribution of beta1 integrins to neuronal migration and differentiation depends on extracellular matrix molecules. Eur J Cell Biol 2005, 84(12):973-982. 10.1016/j.ejcb.2005.09.017, 16325506.
Hannan AJ, Henke RC, Seeto GS, Capes-Davis A, Dunn J, Jeffrey PL. Expression of doublecortin correlates with neuronal migration and pattern formation in diverse regions of the developing chick brain. J Neurosci Res 1999, 55(5):650-657. 10.1002/(SICI)1097-4547(19990301)55:5<650::AID-JNR12>3.0.CO;2-S, 10082087.
Lee SH, Kim YJ, Lee KM, Ryu S, Yoon BW. Ischemic preconditioning enhances neurogenesis in the subventricular zone. Neuroscience 2007, 146(3):1020-1031. 10.1016/j.neuroscience.2007.02.058, 17434685.
Karl C, Couillard-Despres S, Prang P, Munding M, Kilb W, Brigadski T, Plotz S, Mages W, Luhmann H, Winkler J, et al. Neuronal precursor-specific activity of a human doublecortin regulatory sequence. J Neurochem 2005, 92(2):264-282. 10.1111/j.1471-4159.2004.02879.x, 15663475.
Tang K, Yang J, Gao X, Wang C, Liu L, Kitani H, Atsumi T, Jing N. Wnt-1 promotes neuronal differentiation and inhibits gliogenesis in P19 cells. Biochemical and Biophysical Research Communications 2002, 293(1):167-173. 10.1016/S0006-291X(02)00215-2, 12054580.
Korinek V, Barker N, Willert K, Molenaar M, Roose J, Wagenaar G, Markman M, Lamers W, Destree O, Clevers H. Two members of the Tcf family implicated in Wnt/beta-catenin signaling during embryogenesis in the mouse. Mol Cell Biol 1998, 18(3):1248-1256. 108837, 9488439.
Katoh M. Regulation of WNT signaling molecules by retinoic acid during neuronal differentiation in NT2 cells: threshold model of WNT action (review). Int J Mol Med 2002, 10(6):683-687.
Haegele L, Ingold B, Naumann H, Tabatabai G, Ledermann B, Brandner S. Wnt signalling inhibits neural differentiation of embryonic stem cells by controlling bone morphogenetic protein expression. Mol Cell Neurosci 2003, 24(3):696-708. 10.1016/S1044-7431(03)00232-X, 14664819.
Czyz J, Wobus A. Embryonic stem cell differentiation: the role of extracellular factors. Differentiation 2001, 68(4-5):167-174. 10.1046/j.1432-0436.2001.680404.x, 11776469.
Kielman MF, Rindapää M, Gaspar C, van Poppel N, Breukel C, van Leeuwen S, Taketo MM, Roberts S, Smits R, Fodde R. Apc modulates embryonic stem-cell differentiation by controlling the dosage of beta-catenin signaling. Nat Genet 2002, 32(4):594-605. 10.1038/ng1045, 12426568.
Hirabayashi Y. The Wnt/-catenin pathway directs neuronal differentiation of cortical neural precursor cells. Development 2004, 131(12):2791-2801. 10.1242/dev.01165, 15142975.
Viti J, Gulacsi A, Lillien L. Wnt regulation of progenitor maturation in the cortex depends on Shh or fibroblast growth factor 2. Journal of Neuroscience 2003, 23(13):5919-5927.
Hong SK, Kim CH, Yoo KW, Kim HS, Kudoh T, Dawid IB, Huh TL. Isolation and expression of a novel neuron-specific onecut homeobox gene in zebrafish. Mech Dev 2002, 112(1-2):199-202. 10.1016/S0925-4773(01)00647-5, 11850195.
Sasakura Y, Makabe KW. A gene encoding a new ONECUT class homeodomain protein in the ascidian Halocynthia roretzi functions in the differentiation and specification of neural cells in ascidian embryogenesis. Mech Dev 2001, 104(1-2):37-48. 10.1016/S0925-4773(01)00352-5, 11404078.
Nguyen DN, Rohrbaugh M, Lai Z. The Drosophila homolog of Onecut homeodomain proteins is a neural-specific transcriptional activator with a potential role in regulating neural differentiation. Mech Dev 2000, 97(1-2):57-72. 10.1016/S0925-4773(00)00431-7, 11025207.
Bibel M, Richter J, Schrenk K, Tucker K, Staiger V, Korte M, Goetz M, Barde Y. Differentiation of mouse embryonic stem cells into a defined neuronal lineage. Nat Neurosci 2004, 7(9):1003-1009. 10.1038/nn1301, 15332090.
Matsuo N, Kawamoto S, Matsubara K, Okubo K. Cloning and developmental expression of the murine homolog of doublecortin. Biochemical and Biophysical Research Communications 1998, 252(3):571-576. 10.1006/bbrc.1998.9698, 9837748.
Howe D, Bromidge T. Variation of LEF-1 mRNA expression in low-grade B-cell non-Hodgkin's lymphoma. Leuk Res 2006, 30(1):29-32. 10.1016/j.leukres.2005.06.004, 16054689.
Murakami T, Toda S, Fujimoto M, Ohtsuki M, Byers HR, Etoh T, Nakagawa H. Constitutive activation of Wnt/beta-catenin signaling pathway in migration-active melanoma cells: role of LEF-1 in melanoma with increased metastatic potential. Biochemical and Biophysical Research Communications 2001, 288(1):8-15. 10.1006/bbrc.2001.5719, 11594745.
Pierreux CE, Vanhorenbeeck V, Jacquemin P, Lemaigre FP, Rousseau GG. The transcription factor hepatocyte nuclear factor-6/Onecut-1 controls the expression of its paralog Onecut-3 in developing mouse endoderm. J Biol Chem 2004, 279(49):51298-51304. 10.1074/jbc.M409038200, 15381696.
Jacquemin P, Pierreux CE, Fierens S, van Eyll JM, Lemaigre FP, Rousseau GG. Cloning and embryonic expression pattern of the mouse Onecut transcription factor OC-2. Gene Expr Patterns 2003, 3(5):639-644. 10.1016/S1567-133X(03)00110-8, 12971999.
Tomita T, Kimura S. Regulation of mouse Scgb3a1 gene expression by NF-Y and association of CpG methylation with its tissue-specific expression. BMC Mol Biol 2008, 9:5. 10.1186/1471-2199-9-5, 2266941, 18194566.
Li B, Mackay DR, Dai Q, Li TW, Nair M, Fallahi M, Schonbaum CP, Fantes J, Mahowald AP, Waterman ML, et al. The LEF1/beta -catenin complex activates movo1, a mouse homolog of Drosophila ovo required for epidermal appendage differentiation. Proc Natl Acad Sci USA 2002, 99(9):6064-6069. 10.1073/pnas.092137099, 122902, 11983900.
Desaint S, Hansmannel F, Clémencet MC, Le Jossic-Corcos C, Nicolas-Frances V, Latruffe N, Cherkaoui-Malki M. NFY interacts with the promoter region of two genes involved in the rat peroxisomal fatty acid beta-oxidation: the multifunctional protein type 1 and the 3-ketoacyl-CoA B thiolase. Lipids in health and disease 2004, 3:4. 10.1186/1476-511X-3-4, 400753, 15046640.