[en] The cartilaginous elements forming the pharyngeal arches of the zebrafish derive from cranial neural crest cells. Their proper differentiation and patterning are regulated by reciprocal interactions between neural crest cells and surrounding endodermal, ectodermal and mesodermal tissues. In this study, we show that the endodermal factors Runx3 and Sox9b form a regulatory cascade with Egr1 resulting in transcriptional repression of the fsta gene, encoding a BMP antagonist, in pharyngeal endoderm. Using a transgenic line expressing a dominant negative BMP receptor or a specific BMP inhibitor (dorsomorphin), we show that BMP signaling is indeed required around 30 hpf in the neural crest cells to allow cell differentiation and proper pharyngeal cartilage formation. Runx3, Egr1, Sox9b and BMP signaling are required for expression of runx2b, one of the key regulator of cranial cartilage maturation and bone formation. Finally, we show that egr1 depletion leads to increased expression of fsta and inhibition of BMP signaling in the pharyngeal region.
In conclusion, we show that the successive induction of the transcription factors Runx3, Egr1 and Sox9b constitutes a regulatory cascade that controls expression of Follistatin A in pharyngeal endoderm, the latter modulating BMP signaling in developing cranial cartilage in zebrafish.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Dalcq, Julia ; Université de Liège - ULiège > Dép. d'électric., électron. et informat. (Inst.Montefiore) > Systèmes et modélisation
Pasque, Vincent
Ghaye, Aurélie ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire
Larbuisson, Arnaud ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire
Motte, Patrick ; Université de Liège - ULiège > Département des sciences de la vie > Génomique fonctionnelle et imagerie moléculaire végétale
Martial, Joseph ; Université de Liège - ULiège > Département des sciences de la vie > Département des sciences de la vie
Muller, Marc ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire
Language :
English
Title :
Runx3, Egr1 AND Sox9b form a regulatory cascade required to modulate BMP-signaling during cranial cartilage development in zebrafish.
Publication date :
2012
Journal title :
PLoS ONE
eISSN :
1932-6203
Publisher :
Public Library of Science, San Franscisco, United States - California
Schilling TF, Kimmel CB, (1994) Segment and cell type lineage restrictions during pharyngeal arch development in the zebrafish embryo. Development 120: 483-494.
Knight RD, Javidan Y, Zhang T, Nelson S, Schilling TF, (2005) AP2-dependent signals from the ectoderm regulate craniofacial development in the zebrafish embryo. Development 132: 3127-3138.
Yanagisawa H, Clouthier DE, Richardson JA, Charite J, Olson EN, (2003) Targeted deletion of a branchial arch-specific enhancer reveals a role of dHAND in craniofacial development. Development 130: 1069-1078.
Sperber SM, Saxena V, Hatch G, Ekker M, (2008) Zebrafish dlx2a contributes to hindbrain neural crest survival, is necessary for differentiation of sensory ganglia and functions with dlx1a in maturation of the arch cartilage elements. Dev Biol 314: 59-70.
Yan YL, Willoughby J, Liu D, Crump JG, Wilson C, et al. (2005) A pair of Sox: distinct and overlapping functions of zebrafish sox9 co-orthologs in craniofacial and pectoral fin development. Development 132: 1069-1083.
Yan YL, Miller CT, Nissen RM, Singer A, Liu D, et al. (2002) A zebrafish sox9 gene required for cartilage morphogenesis. Development 129: 5065-5079.
Zhou G, Zheng Q, Engin F, Munivez E, Chen Y, et al. (2006) Dominance of SOX9 function over RUNX2 during skeletogenesis. Proc Natl Acad Sci U S A 103: 19004-19009.
Yamashita S, Andoh M, Ueno-Kudoh H, Sato T, Miyaki S, et al. (2009) Sox9 directly promotes Bapx1 gene expression to repress Runx2 in chondrocytes. Exp Cell Res 315: 2231-2240.
Maeno T, Moriishi T, Yoshida CA, Komori H, Kanatani N, et al. (2011) Early onset of Runx2 expression caused craniosynostosis, ectopic bone formation, and limb defects. Bone 49: 673-682.
David NB, Saint-Etienne L, Tsang M, Schilling TF, Rosa FM, (2002) Requirement for endoderm and FGF3 in ventral head skeleton formation. Development 129: 4457-4468.
Walshe J, Mason I, (2003) Fgf signalling is required for formation of cartilage in the head. Dev Biol 264: 522-536.
Nie X, Luukko K, Kettunen P, (2006) BMP signalling in craniofacial development. Int J Dev Biol 50: 511-521.
Paiva KB, Silva-Valenzuela MG, Massironi SM, Ko GM, Siqueira FM, et al. (2009) Differential Shh, Bmp and Wnt gene expressions during craniofacial development in mice. Acta Histochem 112: 508-517.
Alexander C, Zuniga E, Blitz IL, Wada N, Le Pabic P, et al. (2011) Combinatorial roles for BMPs and Endothelin 1 in patterning the dorsal-ventral axis of the craniofacial skeleton. Development 138: 5135-5146.
Piotrowski T, Nusslein-Volhard C, (2000) The endoderm plays an important role in patterning the segmented pharyngeal region in zebrafish (Danio rerio). Dev Biol 225: 339-356.
Schilling TF, Piotrowski T, Grandel H, Brand M, Heisenberg CP, et al. (1996) Jaw and branchial arch mutants in zebrafish I: branchial arches. Development 123: 329-344.
Alexander J, Rothenberg M, Henry GL, Stainier DY, (1999) casanova plays an early and essential role in endoderm formation in zebrafish. Dev Biol 215: 343-357.
Dickmeis T, Mourrain P, Saint-Etienne L, Fischer N, Aanstad P, et al. (2001) A crucial component of the endoderm formation pathway, CASANOVA, is encoded by a novel sox-related gene. Genes Dev 15: 1487-1492.
Kikuchi Y, Agathon A, Alexander J, Thisse C, Waldron S, et al. (2001) casanova encodes a novel Sox-related protein necessary and sufficient for early endoderm formation in zebrafish. Genes Dev 15: 1493-1505.
Kikuchi Y, Trinh LA, Reiter JF, Alexander J, Yelon D, et al. (2000) The zebrafish bonnie and clyde gene encodes a Mix family homeodomain protein that regulates the generation of endodermal precursors. Genes Dev 14: 1279-1289.
Piotrowski T, Ahn DG, Schilling TF, Nair S, Ruvinsky I, et al. (2003) The zebrafish van gogh mutation disrupts tbx1, which is involved in the DiGeorge deletion syndrome in humans. Development 130: 5043-5052.
Swirnoff AH, Milbrandt J, (1995) DNA-binding specificity of NGFI-A and related zinc finger transcription factors. Mol Cell Biol 15: 2275-2287.
McMahon AP, Champion JE, McMahon JA, Sukhatme VP, (1990) Developmental expression of the putative transcription factor Egr-1 suggests that Egr-1 and c-fos are coregulated in some tissues. Development 108: 281-287.
Drummond IA, Rohwer-Nutter P, Sukhatme VP, (1994) The zebrafish egr1 gene encodes a highly conserved, zinc-finger transcriptional regulator. DNA Cell Biol 13: 1047-1055.
Close R, Toro S, Martial JA, Muller M, (2002) Expression of the zinc finger Egr1 gene during zebrafish embryonic development. Mech Dev 118: 269-272.
Abdulkadir SA, Qu Z, Garabedian E, Song SK, Peters TJ, et al. (2001) Impaired prostate tumorigenesis in Egr1-deficient mice. Nat Med 7: 101-107.
Eid MA, Kumar MV, Iczkowski KA, Bostwick DG, Tindall DJ, (1998) Expression of early growth response genes in human prostate cancer. Cancer Res 58: 2461-2468.
Huang RP, Fan Y, de Belle I, Niemeyer C, Gottardis MM, et al. (1997) Decreased Egr-1 expression in human, mouse and rat mammary cells and tissues correlates with tumor formation. Int J Cancer 72: 102-109.
Huang RP, Liu C, Fan Y, Mercola D, Adamson ED, (1995) Egr-1 negatively regulates human tumor cell growth via the DNA-binding domain. Cancer Res 55: 5054-5062.
Sukhatme VP, Cao XM, Chang LC, Tsai-Morris CH, Stamenkovich D, et al. (1988) A zinc finger-encoding gene coregulated with c-fos during growth and differentiation, and after cellular depolarization. Cell 53: 37-43.
Nguyen HQ, Hoffman-Liebermann B, Liebermann DA, (1993) The zinc finger transcription factor Egr-1 is essential for and restricts differentiation along the macrophage lineage. Cell 72: 197-209.
Lee SL, Sadovsky Y, Swirnoff AH, Polish JA, Goda P, et al. (1996) Luteinizing hormone deficiency and female infertility in mice lacking the transcription factor NGFI-A (Egr-1). Science 273: 1219-1221.
Topilko P, Schneider-Maunoury S, Levi G, Trembleau A, Gourdji D, et al. (1998) Multiple pituitary and ovarian defects in Krox-24 (NGFI-A, Egr-1)-targeted mice. Mol Endocrinol 12: 107-122.
Slade JP, Carter DA, (2000) Cyclical expression of egr-1/NGFI-A in the rat anterior pituitary: a molecular signal for ovulation? J Neuroendocrinol 12: 671-676.
Suva LJ, Ernst M, Rodan GA, (1991) Retinoic acid increases zif268 early gene expression in rat preosteoblastic cells. Mol Cell Biol 11: 2503-2510.
Granet C, Boutahar N, Vico L, Alexandre C, Lafage-Proust MH, (2001) MAPK and SRC-kinases control EGR-1 and NF-kappa B inductions by changes in mechanical environment in osteoblasts. Biochem Biophys Res Commun 284: 622-631.
Kukita T, Kukita A, Harada H, Iijima T, (1997) Regulation of osteoclastogenesis by antisense oligodeoxynucleotides specific to zinc finger nuclear transcription factors Egr-1 and WT1 in rat bone marrow culture system. Endocrinology 138: 4384-4389.
Srivastava S, Weitzmann MN, Kimble RB, Rizzo M, Zahner M, et al. (1998) Estrogen blocks M-CSF gene expression and osteoclast formation by regulating phosphorylation of Egr-1 and its interaction with Sp-1. J Clin Invest 102: 1850-1859.
Srivastava S, Toraldo G, Weitzmann MN, Cenci S, Ross FP, et al. (2001) Estrogen decreases osteoclast formation by down-regulating receptor activator of NF-kappa B ligand (RANKL)-induced JNK activation. J Biol Chem 276: 8836-8840.
Wang FL, Connor JR, Dodds RA, James IE, Kumar S, et al. (2000) Differential expression of egr-1 in osteoarthritic compared to normal adult human articular cartilage. Osteoarthritis Cartilage 8: 161-169.
Rockel JS, Bernier SM, Leask A, (2009) Egr-1 inhibits the expression of extracellular matrix genes in chondrocytes by TNFalpha-induced MEK/ERK signalling. Arthritis Res Ther 11: R8.
Hu CY, Yang CH, Chen WY, Huang CJ, Huang HY, et al. (2006) Egr1 gene knockdown affects embryonic ocular development in zebrafish. Mol Vis 12: 1250-1258.
Kimmel CB, Ballard WW, Kimmel SR, Ullmann B, Schilling TF, (1995) Stages of embryonic development of the zebrafish. Dev Dyn 203: 253-310.
Pyati UJ, Webb AE, Kimelman D, (2005) Transgenic zebrafish reveal stage-specific roles for Bmp signaling in ventral and posterior mesoderm development. Development 132: 2333-2343.
Flores MV, Lam EY, Crosier P, Crosier K, (2006) A hierarchy of Runx transcription factors modulate the onset of chondrogenesis in craniofacial endochondral bones in zebrafish. Dev Dyn 235: 3166-3176.
Dal-Pra S, Furthauer M, Van-Celst J, Thisse B, Thisse C, (2006) Noggin1 and Follistatin-like2 function redundantly to Chordin to antagonize BMP activity. Dev Biol 298: 514-526.
Hauptmann G, Gerster T, (1994) Two-color whole-mount in situ hybridization to vertebrate and Drosophila embryos. Trends Genet 10: 266.
Lopez M, Nica G, Motte P, Martial JA, Hammerschmidt M, et al. (2006) Expression of the somatolactin beta gene during zebrafish embryonic development. Gene Expr Patterns 6: 156-161.
Quiroz Y, Lopez M, Mavropoulos A, Motte P, Martial JA, et al. (2012) The HMG-Box Transcription Factor Sox4b Is Required for Pituitary Expression of gata2a and Specification of Thyrotrope and Gonadotrope Cells in Zebrafish. Mol Endocrinol 26: 1014-1027.
Dong PD, Munson CA, Norton W, Crosnier C, Pan X, et al. (2007) Fgf10 regulates hepatopancreatic ductal system patterning and differentiation. Nat Genet 39: 397-402.
Akimenko MA, Ekker M, Wegner J, Lin W, Westerfield M, (1994) Combinatorial expression of three zebrafish genes related to distal-less: part of a homeobox gene code for the head. J Neurosci 14: 3475-3486.
Miller CT, Schilling TF, Lee K, Parker J, Kimmel CB, (2000) sucker encodes a zebrafish Endothelin-1 required for ventral pharyngeal arch development. Development 127: 3815-3828.
Pinto JP, Conceicao NM, Viegas CS, Leite RB, Hurst LD, et al. (2005) Identification of a new pebp2alphaA2 isoform from zebrafish runx2 capable of inducing osteocalcin gene expression in vitro. J Bone Miner Res 20: 1440-1453.
Flores MV, Tsang VW, Hu W, Kalev-Zylinska M, Postlethwait J, et al. (2004) Duplicate zebrafish runx2 orthologues are expressed in developing skeletal elements. Gene Expr Patterns 4: 573-581.
Bauer H, Meier A, Hild M, Stachel S, Economides A, et al. (1998) Follistatin and noggin are excluded from the zebrafish organizer. Dev Biol 204: 488-507.
Erickson T, French CR, Waskiewicz AJ, (2010) Meis1 specifies positional information in the retina and tectum to organize the zebrafish visual system. Neural Dev 5: 22.
Thisse B, Pfumio S, Fürthauer M, Loppin B, Heyer V, et al. (2001) Expression of the zebrafish genome during embryogenesis. ZFIN on-line publication.
Poulain M, Furthauer M, Thisse B, Thisse C, Lepage T, (2006) Zebrafish endoderm formation is regulated by combinatorial Nodal, FGF and BMP signalling. Development 133: 2189-2200.
Hao J, Daleo MA, Murphy CK, Yu PB, Ho JN, et al. (2008) Dorsomorphin, a selective small molecule inhibitor of BMP signaling, promotes cardiomyogenesis in embryonic stem cells. PLoS One 3: e2904.
Massague J, Gomis RR, (2006) The logic of TGFbeta signaling. FEBS Lett 580: 2811-2820.
Dudas M, Sridurongrit S, Nagy A, Okazaki K, Kaartinen V, (2004) Craniofacial defects in mice lacking BMP type I receptor Alk2 in neural crest cells. Mech Dev 121: 173-182.
Ko SO, Chung IH, Xu X, Oka S, Zhao H, et al. (2007) Smad4 is required to regulate the fate of cranial neural crest cells. Dev Biol 312: 435-447.
Tang S, Snider P, Firulli AB, Conway SJ, (2010) Trigenic neural crest-restricted Smad7 over-expression results in congenital craniofacial and cardiovascular defects. Dev Biol 344: 233-247.
Holzschuh J, Wada N, Wada C, Schaffer A, Javidan Y, et al. (2005) Requirements for endoderm and BMP signaling in sensory neurogenesis in zebrafish. Development 132: 3731-3742.
Martinez-Barbera JP, Toresson H, Da Rocha S, Krauss S, (1997) Cloning and expression of three members of the zebrafish Bmp family: Bmp2a, Bmp2b and Bmp4. Gene 198: 53-59.
Wilson J, Tucker AS, (2004) Fgf and Bmp signals repress the expression of Bapx1 in the mandibular mesenchyme and control the position of the developing jaw joint. Dev Biol 266: 138-150.
Xia Y, Schneyer AL, (2009) The biology of activin: recent advances in structure, regulation and function. J Endocrinol 202: 1-12.
Canalis E, Economides AN, Gazzerro E, (2003) Bone morphogenetic proteins, their antagonists, and the skeleton. Endocr Rev 24: 218-235.
Esch FS, Shimasaki S, Mercado M, Cooksey K, Ling N, et al. (1987) Structural characterization of follistatin: a novel follicle-stimulating hormone release-inhibiting polypeptide from the gonad. Mol Endocrinol 1: 849-855.
de Kretser DM, Hedger MP, Loveland KL, Phillips DJ, (2002) Inhibins, activins and follistatin in reproduction. Hum Reprod Update 8: 529-541.
Gajos-Michniewicz A, Piastowska AW, Russell JA, Ochedalski T, (2010) Follistatin as a potent regulator of bone metabolism. Biomarkers 15: 563-574.
Liu W, Selever J, Murali D, Sun X, Brugger SM, et al. (2005) Threshold-specific requirements for Bmp4 in mandibular development. Dev Biol 283: 282-293.
Chaboissier MC, Kobayashi A, Vidal VI, Lutzkendorf S, van de Kant HJ, et al. (2004) Functional analysis of Sox8 and Sox9 during sex determination in the mouse. Development 131: 1891-1901.
Balczerski B, Matsutani M, Castillo P, Osborne N, Stainier DY, et al. (2011) Analysis of Sphingosine-1-phosphate signaling mutants reveals endodermal requirements for the growth but not dorsoventral patterning of jaw skeletal precursors. Dev Biol 362: 230-241.