Sfrp5 belongs to the family of secreted frizzled related proteins (Sfrp), secreted inhibitors of Wingless-MMTV Integration Site (Wnt) signaling, which play an important role in cancer and development. We selected sfrp5 because of its compelling expression profile in the developing endoderm in zebrafish, Danio rerio. In this study, overexpression of sfrp5 in embryos results in defects in both convergent extension (CE) by inhibition of non-canonical Wnt signaling and defects in dorsoventral patterning by inhibition of Tolloid-mediated proteolysis of the BMP inhibitor Chordin. From 25 hours post fertilization (hpf) to 3 days post fertilization (dpf), both overexpression and knockdown of Sfrp5 decrease the size of the endoderm, significantly reducing liver cell number. At 3 dpf, insulin-positive endodermal cells fail to coalesce into a single pancreatic islet. We show that Sfrp5 inhibits both canonical and non-canonical Wnt signaling during embryonic and endodermal development, resulting in endodermal abnormalities.
References
[1]
Polakis P (2012) Wnt signaling in cancer. Cold Spring Harbor perspectives in biology 4.
[2]
Nusse R (2012) Wnt signaling. Cold Spring Harbor perspectives in biology 4.
[3]
Verzi MP, Shivdasani RA (2008) Wnt signaling in gut organogenesis. Organogenesis 4: 87–91.
[4]
Lade AG, Monga SP (2011) Beta-catenin signaling in hepatic development and progenitors: which way does the WNT blow? Developmental dynamics: an official publication of the American Association of Anatomists 240: 486–500.
[5]
Verkade H, Heath JK (2009) Wnt signaling mediates diverse developmental processes in zebrafish. Methods Mol Biol 469: 225–251.
[6]
Schepers A, Clevers H (2012) Wnt signaling, stem cells, and cancer of the gastrointestinal tract. Cold Spring Harbor perspectives in biology 4: a007989.
[7]
White BD, Chien AJ, Dawson DW (2012) Dysregulation of Wnt/beta-catenin signaling in gastrointestinal cancers. Gastroenterology 142: 219–232.
[8]
Clevers H, Nusse R (2012) Wnt/beta-catenin signaling and disease. Cell 149: 1192–1205.
[9]
Lee HX, Ambrosio AL, Reversade B, De Robertis EM (2006) Embryonic dorsal-ventral signaling: secreted frizzled-related proteins as inhibitors of tolloid proteinases. Cell 124: 147–159.
[10]
Piccolo S, Agius E, Lu B, Goodman S, Dale L, et al. (1997) Cleavage of Chordin by Xolloid metalloprotease suggests a role for proteolytic processing in the regulation of Spemann organizer activity. Cell 91: 407–416.
[11]
Muraoka O, Shimizu T, Yabe T, Nojima H, Bae Y-K, et al. (2006) Sizzled controls dorso-ventral polarity by repressing cleavage of the Chordin protein. Nat Cell Biol 8: 329–338.
[12]
Ploper D, Lee HX, De Robertis EM (2011) Dorsal-Ventral patterning: crescent is a dorsally secreted Frizzled-related protein that competitively inhibits Tolloid proteases. Dev Biol 352: 317–328.
[13]
Houart C, Caneparo L, Heisenberg C, Barth K, Take-Uchi M, et al. (2002) Establishment of the telencephalon during gastrulation by local antagonism of Wnt signaling. Neuron 35: 255–265.
[14]
Ruiz JM, Rodríguez J, Bovolenta P (2009) Growth and differentiation of the retina and the optic tectum in the medaka fish requires olSfrp5. Dev Neurobiol 69: 617–632.
[15]
Kong W, Yang Y, Zhang T, Shi DL, Zhang Y (2012) Characterization of sFRP2-like in amphioxus: insights into the evolutionary conservation of Wnt antagonizing function. Evolution & development 14: 168–177.
[16]
Satoh W, Matsuyama M, Takemura H, Aizawa S, Shimono A (2008) Sfrp1, Sfrp2, and Sfrp5 regulate the Wnt/beta-catenin and the planar cell polarity pathways during early trunk formation in mouse. Genesis 46: 92–103.
[17]
Satoh W, Gotoh T, Tsunematsu Y, Aizawa S, Shimono A (2006) Sfrp1 and Sfrp2 regulate anteroposterior axis elongation and somite segmentation during mouse embryogenesis. Development 133: 989–999.
[18]
Matsuyama M, Aizawa S, Shimono A (2009) Sfrp controls apicobasal polarity and oriented cell division in developing gut epithelium. PLoS Genet 5: e1000427.
[19]
Mii Y, Taira M (2009) Secreted Frizzled-related proteins enhance the diffusion of Wnt ligands and expand their signalling range. Development 136: 4083–4088.
[20]
Adamska M, Larroux C, Adamski M, Green K, Lovas E, et al. (2010) Structure and expression of conserved Wnt pathway components in the demosponge Amphimedon queenslandica. Evolution & development 12: 494–518.
[21]
Kawakami K, Yamamura S, Hirata H, Ueno K, Saini S, et al. (2011) Secreted frizzled-related protein-5 is epigenetically downregulated and functions as a tumor suppressor in kidney cancer. International journal of cancer Journal international du cancer 128: 541–550.
[22]
Qi J, Zhu Y-Q, Luo J, Tao W-H (2006) Hypermethylation and expression regulation of secreted frizzled-related protein genes in colorectal tumor. World J Gastroenterol 12: 7113–7117.
[23]
Nojima M, Suzuki H, Toyota M, Watanabe Y, Maruyama R, et al. (2007) Frequent epigenetic inactivation of SFRP genes and constitutive activation of Wnt signaling in gastric cancer. Oncogene 26: 4699–4713.
[24]
Bovolenta P, Esteve P, Ruiz JM, Cisneros E, Lopez-Rios J (2008) Beyond Wnt inhibition: new functions of secreted Frizzled-related proteins in development and disease. J Cell Sci 121: 737–746.
[25]
Tendeng C, Houart C (2006) Cloning and embryonic expression of five distinct sfrp genes in the zebrafish Danio rerio. Gene Expr Patterns 6: 761–771.
[26]
Chong JM, Uren A, Rubin JS, Speicher DW (2002) Disulfide bond assignments of secreted Frizzled-related protein-1 provide insights about Frizzled homology and netrin modules. J Biol Chem 277: 5134–5144.
[27]
Uren A, Reichsman F, Anest V, Taylor WG, Muraiso K, et al. (2000) Secreted frizzled-related protein-1 binds directly to Wingless and is a biphasic modulator of Wnt signaling. J Biol Chem 275: 4374–4382.
[28]
Bhat RA, Stauffer B, Komm BS, Bodine PVN (2007) Structure-function analysis of secreted frizzled-related protein-1 for its Wnt antagonist function. J Cell Biochem 102: 1519–1528.
[29]
Wawrzak D, Metioui M, Willems E, Hendrickx M, de Genst E, et al. (2007) Wnt3a binds to several sFRPs in the nanomolar range. Biochemical and biophysical research communications 357: 1119–1123.
[30]
Li Y, Rankin SA, Sinner D, Kenny AP, Krieg PA, et al. (2008) Sfrp5 coordinates foregut specification and morphogenesis by antagonizing both canonical and noncanonical Wnt11 signaling. Genes Dev 22: 3050–3063.
[31]
Wang S, Krinks M, Moos M (1997) Frzb-1, an antagonist of Wnt-1 and Wnt-8, does not block signaling by Wnts -3A, -5A, or -11. Biochem Biophys Res Commun 236: 502–504.
[32]
Mii Y, Taira M (2011) Secreted Wnt “inhibitors” are not just inhibitors: regulation of extracellular Wnt by secreted Frizzled-related proteins. Development, growth & differentiation 53: 911–923.
[33]
Leyns L, Bouwmeester T, Kim SH, Piccolo S, De Robertis EM (1997) Frzb-1 is a secreted antagonist of Wnt signaling expressed in the Spemann organizer. Cell 88: 747–756.
[34]
Scardigli R, Gargioli C, Tosoni D, Borello U, Sampaolesi M, et al. (2008) Binding of sFRP-3 to EGF in the extra-cellular space affects proliferation, differentiation and morphogenetic events regulated by the two molecules. PLoS ONE 3: e2471.
[35]
Kobayashi K, Luo M, Zhang Y, Wilkes DC, Ge G, et al. (2009) Secreted Frizzled-related protein 2 is a procollagen C proteinase enhancer with a role in fibrosis associated with myocardial infarction. Nat Cell Biol 11: 46–55.
[36]
Su HY, Lai HC, Lin YW, Liu CY, Chen CK, et al. (2010) Epigenetic silencing of SFRP5 is related to malignant phenotype and chemoresistance of ovarian cancer through Wnt signaling pathway. International journal of cancer Journal international du cancer 127: 555–567.
[37]
Peng G, Westerfield M (2006) Lhx5 promotes forebrain development and activates transcription of secreted Wnt antagonists. Development 133: 3191–3200.
[38]
Buchert M, Athineos D, Abud HE, Burke ZD, Faux MC, et al. (2010) Genetic dissection of differential signaling threshold requirements for the Wnt/beta-catenin pathway in vivo. PLoS genetics 6: e1000816.
[39]
Stuckenholz C, Lu L, Thakur P, Kaminski N, Bahary N (2009) FACS-assisted microarray profiling implicates novel genes and pathways in zebrafish gastrointestinal tract development. Gastroenterology 137: 1321–1332.
[40]
Ng ANY, de Jong-Curtain TA, Mawdsley DJ, White SJ, Shin J, et al. (2005) Formation of the digestive system in zebrafish: III. Intestinal epithelium morphogenesis. Dev Biol 286: 114–135.
[41]
Ober EA, Verkade H, Field HA, Stainier DYR (2006) Mesodermal Wnt2b signalling positively regulates liver specification. Nature 442: 688–691.
[42]
Matsui T, Raya A, Kawakami Y, Callol-Massot C, Capdevila J, et al. (2005) Noncanonical Wnt signaling regulates midline convergence of organ primordia during zebrafish development. Genes Dev 19: 164–175.
[43]
Axelrod JD, Miller JR, Shulman JM, Moon RT, Perrimon N (1998) Differential recruitment of Dishevelled provides signaling specificity in the planar cell polarity and Wingless signaling pathways. Genes Dev 12: 2610–2622.
[44]
Du SJ, Purcell SM, Christian JL, McGrew LL, Moon RT (1995) Identification of distinct classes and functional domains of Wnts through expression of wild-type and chimeric proteins in Xenopus embryos. Mol Cell Biol 15: 2625–2634.
[45]
Ku M, Melton DA (1993) Xwnt-11: a maternally expressed Xenopus wnt gene. Development 119: 1161–1173.
[46]
Xie J, Fisher S (2005) Twisted gastrulation enhances BMP signaling through chordin dependent and independent mechanisms. Development 132: 383–391.
[47]
Chung W-S, Shin CH, Stainier DYR (2008) Bmp2 signaling regulates the hepatic versus pancreatic fate decision. Dev Cell 15: 738–748.
[48]
Kwan KM, Fujimoto E, Grabher C, Mangum BD, Hardy ME, et al. (2007) The Tol2kit: a multisite gateway-based construction kit for Tol2 transposon transgenesis constructs. Dev Dyn 236: 3088–3099.
[49]
Zhou Y, Cashman TJ, Nevis KR, Obregon P, Carney SA, et al. (2011) Latent TGF-beta binding protein 3 identifies a second heart field in zebrafish. Nature 474: 645–648.
[50]
Tucker JA, Mintzer KA, Mullins MC (2008) The BMP signaling gradient patterns dorsoventral tissues in a temporally progressive manner along the anteroposterior axis. Dev Cell 14: 108–119.
[51]
Miller-Bertoglio VE, Fisher S, Sánchez A, Mullins MC, Halpern ME (1997) Differential regulation of chordin expression domains in mutant zebrafish. Dev Biol 192: 537–550.
[52]
Stachel SE, Grunwald DJ, Myers PZ (1993) Lithium perturbation and goosecoid expression identify a dorsal specification pathway in the pregastrula zebrafish. Development 117: 1261–1274.
[53]
Schulte-Merker S, Lee KJ, McMahon AP, Hammerschmidt M (1997) The zebrafish organizer requires chordino. Nature. 387: 862–863.
[54]
Joly JS, Joly C, Schulte-Merker S, Boulekbache H, Condamine H (1993) The ventral and posterior expression of the zebrafish homeobox gene eve1 is perturbed in dorsalized and mutant embryos. Development 119: 1261–1275.
[55]
Meani N, Pezzimenti F, Deflorian G, Mione M, Alcalay M (2009) The tumor suppressor PRDM5 regulates Wnt signaling at early stages of zebrafish development. PLoS ONE 4: e4273.
[56]
Tsang M, Kim R, de Caestecker MP, Kudoh T, Roberts AB, et al. (2000) Zebrafish nma is involved in TGFbeta family signaling. Genesis 28: 47–57.
[57]
Dougan ST, Warga RM, Kane DA, Schier AF, Talbot WS (2003) The role of the zebrafish nodal-related genes squint and cyclops in patterning of mesendoderm. Development 130: 1837–1851.
[58]
Lele Z, Bakkers J, Hammerschmidt M (2001) Morpholino phenocopies of the swirl, snailhouse, somitabun, minifin, silberblick, and pipetail mutations. Genesis 30: 190–194.
[59]
Mullins MC, Hammerschmidt M, Kane DA, Odenthal J, Brand M, et al. (1996) Genes establishing dorsoventral pattern formation in the zebrafish embryo: the ventral specifying genes. Development 123: 81–93.
[60]
Müller M, von Weizs？cker E, Campos-Ortega JA (1996) Transcription of a zebrafish gene of the hairy-Enhancer of split family delineates the midbrain anlage in the neural plate. Development Genes and Evolution 206: 153–160.
[61]
Seo J, Asaoka Y, Nagai Y, Hirayama J, Yamasaki T, et al. (2010) Negative regulation of wnt11 expression by Jnk signaling during zebrafish gastrulation. J Cell Biochem 110: 1022–1037.
[62]
Connors SA, Tucker JA, Mullins MC (2006) Temporal and spatial action of tolloid (mini fin) and chordin to pattern tail tissues. Dev Biol 293: 191–202.
[63]
Hammerschmidt M, Pelegri F, Mullins MC, Kane DA, van Eeden FJ, et al. (1996) dino and mercedes, two genes regulating dorsal development in the zebrafish embryo. Development 123: 95–102.
[64]
Blader P, Rastegar S, Fischer N, Str？hle U (1997) Cleavage of the BMP-4 antagonist chordin by zebrafish tolloid. Science 278: 1937–1940.
[65]
Poulain M, Ober EA (2011) Interplay between Wnt2 and Wnt2bb controls multiple steps of early foregut-derived organ development. Development 138: 3557–3568.
[66]
Shin D, Lee Y, Poss KD, Stainier DYR (2011) Restriction of hepatic competence by Fgf signaling. Development 138: 1339–1348.
[67]
Ho CY, Houart C, Wilson SW, Stainier DY (1999) A role for the extraembryonic yolk syncytial layer in patterning the zebrafish embryo suggested by properties of the hex gene. Curr Biol 9: 1131–1134.
[68]
Wallace KN, Yusuff S, Sonntag JM, Chin AJ, Pack M (2001) Zebrafish hhex regulates liver development and digestive organ chirality. Genesis 30: 141–143.
[69]
Ober EA, Field HA, Stainier DYR (2003) From endoderm formation to liver and pancreas development in zebrafish. Mech Dev 120: 5–18.
[70]
Odenthal J, Nüsslein-Volhard C (1998) fork head domain genes in zebrafish. Dev Genes Evol 208: 245–258.
[71]
Kim HJ, Schleiffarth JR, Jessurun J, Sumanas S, Petryk A, et al. (2005) Wnt5 signaling in vertebrate pancreas development. BMC Biol 3: 23.
[72]
Shin D, Weidinger G, Moon RT, Stainier DY (2012) Intrinsic and extrinsic modifiers of the regulative capacity of the developing liver. Mechanisms of development 128: 525–535.
[73]
Tada M, Concha ML, Heisenberg CP (2002) Non-canonical Wnt signalling and regulation of gastrulation movements. Semin Cell Dev Biol 13: 251–260.
[74]
Heisenberg CP, Tada M, Rauch GJ, Saúde L, Concha ML, et al. (2000) Silberblick/Wnt11 mediates convergent extension movements during zebrafish gastrulation. Nature 405: 76–81.
[75]
Kilian B, Mansukoski H, Barbosa FC, Ulrich F, Tada M, et al. (2003) The role of Ppt/Wnt5 in regulating cell shape and movement during zebrafish gastrulation. Mech Dev 120: 467–476.
[76]
Zorn AM, Wells JM (2009) Vertebrate endoderm development and organ formation. Annual review of cell and developmental biology 25: 221–251.
[77]
Tremblay KD (2011) Inducing the liver: understanding the signals that promote murine liver budding. Journal of cellular physiology 226: 1727–1731.
[78]
Niu X, Shi H, Peng J (2010) The role of mesodermal signals during liver organogenesis in zebrafish. Science China Life sciences 53: 455–461.
[79]
Zaret KS, Grompe M (2008) Generation and regeneration of cells of the liver and pancreas. Science 322: 1490–1494.
[80]
Goessling W, North TE, Lord AM, Ceol C, Lee S, et al. (2008) APC mutant zebrafish uncover a changing temporal requirement for wnt signaling in liver development. Dev Biol 320: 161–174.
[81]
McLin VA, Rankin SA, Zorn AM (2007) Repression of Wnt/beta-catenin signaling in the anterior endoderm is essential for liver and pancreas development. Development 134: 2207–2217.
[82]
Stoick-Cooper CL, Weidinger G, Riehle KJ, Hubbert C, Major MB, et al. (2007) Distinct Wnt signaling pathways have opposing roles in appendage regeneration. Development 134: 479–489.
[83]
Yuzugullu H, Benhaj K, Ozturk N, Senturk S, Celik E, et al. (2009) Canonical Wnt signaling is antagonized by noncanonical Wnt5a in hepatocellular carcinoma cells. Molecular cancer 8: 90.
[84]
Leaf I, Tennessen J, Mukhopadhyay M, Westphal H, Shawlot W (2006) Sfrp5 is not essential for axis formation in the mouse. Genesis 44: 573–578.
[85]
Hardy KM, Garriock RJ, Yatskievych TA, D’Agostino SL, Antin PB, et al. (2008) Non-canonical Wnt signaling through Wnt5a/b and a novel Wnt11 gene, Wnt11b, regulates cell migration during avian gastrulation. Dev Biol 320: 391–401.
[86]
Langdon YG, Mullins MC (2011) Maternal and zygotic control of zebrafish dorsoventral axial patterning. Annual review of genetics 45: 357–377.
[87]
Pei J, Grishin NV (2012) Cysteine-rich domains related to Frizzled receptors and Hedgehog-interacting proteins. Protein Sci 21: 1172–1184.
[88]
Filmus J, Capurro M, Rast J (2008) Glypicans. Genome biology 9: 224.
[89]
Lin X (2004) Functions of heparan sulfate proteoglycans in cell signaling during development. Development 131: 6009–6021.
[90]
Sawada Y, Yoshikawa T, Nobuoka D, Shirakawa H, Kuronuma T, et al. (2012) Phase I trial of a glypican-3-derived peptide vaccine for advanced hepatocellular carcinoma: immunologic evidence and potential for improving overall survival. Clinical cancer research : an official journal of the American Association for Cancer Research 18: 3686–3696.
