Development, Vol 127, Issue 13 2917-2931, Copyright © 2000 by Company of Biologists

JOURNAL ARTICLES

Endogenous patterns of TGFbeta superfamily signaling during early Xenopus development

S Faure, MA Lee, T Keller, P ten Dijke and M Whitman
Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA. mwhitman@hms.harvard.edu

Transforming growth factor beta (TGFbeta) superfamily signaling has been implicated in patterning of the early Xenopus embryo. Upon ligand stimulation, TGFbeta receptors phosphorylate Smad proteins at carboxy-terminal SS(V/M)S consensus motifs. Smads 1/5/8, activated by bone morphogenetic protein (BMP) signaling, induce ventral mesoderm whereas Smad2, activated by activin-like ligands, induces dorsal mesoderm. Although ectopic expression studies are consistent with roles for TGFbeta signals in early Xenopus embryogenesis, when and where BMP and activin-like signaling pathways are active endogenously has not been directly examined. In this study, we investigate the temporal and spatial activation of TGFbeta superfamily signaling in early Xenopus development by using antibodies specific for the type I receptor-phosphorylated forms of Smad1/5/8 and Smad2. We find that Smad1/5/8 and two distinct isoforms of Smad2, full-length Smad2 and Smad2(delta)exon3, are phosphorylated in early embryos. Both Smad1/5/8 and Smad2/Smad2(delta)exon3 are activated after, but not before, the mid-blastula transition (MBT). Endogenous activation of Smad2/Smad2(delta)exon3 requires zygotic transcription, while Smad1/5/8 activation at MBT appears to involve transcription-independent regulation. We also find that the competence of embryonic cells to respond to TGF(delta) superfamily ligands is temporally regulated and may be a determinant of early patterning. Levels of phospho-Smad1/5/8 and of phospho-Smad2/Smad2(delta)exon3 are asymmetrically distributed across both the animal-vegetal and dorsoventral axes. The timing of the development of these asymmetries differs for phospho-Smad1/5/8 and for phospho-Smad2/Smad2(delta)exon3, and the spatial distribution of phosphorylation of each Smad changes dramatically as gastrulation begins. We discuss the implications of our results for endogenous functions of BMP and activin-like signals as candidate morphogens regulating primary germ layer formation and dorsoventral patterning of the early Xenopus embryo.

This article has been cited by other articles:

				S. B. Anderson, A. L. Goldberg, and M. Whitman Identification of a Novel Pool of Extracellular Pro-myostatin in Skeletal Muscle J. Biol. Chem., March 14, 2008; 283(11): 7027 - 7035. [Abstract] [Full Text] [PDF]

				F. M. Spagnoli and A. H. Brivanlou The Gata5 target, TGIF2, defines the pancreatic region by modulating BMP signals within the endoderm Development, February 1, 2008; 135(3): 451 - 461. [Abstract] [Full Text] [PDF]

				Y. Saka, A. I. Hagemann, O. Piepenburg, and J. C. Smith Nuclear accumulation of Smad complexes occurs only after the midblastula transition in Xenopus Development, December 1, 2007; 134(23): 4209 - 4218. [Abstract] [Full Text] [PDF]

				C. Chang and R. M. Harland Neural induction requires continued suppression of both Smad1 and Smad2 signals during gastrulation Development, November 1, 2007; 134(21): 3861 - 3872. [Abstract] [Full Text] [PDF]

				M. Choi, R. W. Stottmann, Y.-P. Yang, E. N. Meyers, and J. Klingensmith The Bone Morphogenetic Protein Antagonist Noggin Regulates Mammalian Cardiac Morphogenesis Circ. Res., February 2, 2007; 100(2): 220 - 228. [Abstract] [Full Text] [PDF]

				A. B. Steiner, M. J. Engleka, Q. Lu, E. C. Piwarzyk, S. Yaklichkin, J. L. Lefebvre, J. W. Walters, L. Pineda-Salgado, P. A. Labosky, and D. S. Kessler FoxD3 regulation of Nodal in the Spemann organizer is essential for Xenopus dorsal mesoderm development Development, December 15, 2006; 133(24): 4827 - 4838. [Abstract] [Full Text] [PDF]

				L. Parker, J. E. Ellis, M. Q. Nguyen, and K. Arora The divergent TGF-{beta} ligand Dawdle utilizes an activin pathway to influence axon guidance in Drosophila. Development, December 1, 2006; 133(24): 4981 - 4991. [Abstract] [Full Text] [PDF]

				M. Schmerer, I. Torregroza, A. Pascal, M. Umbhauer, and T. Evans STAT5 acts as a repressor to regulate early embryonic erythropoiesis Blood, November 1, 2006; 108(9): 2989 - 2997. [Abstract] [Full Text] [PDF]

				M. Knockaert, G. Sapkota, C. Alarcon, J. Massague, and A. H. Brivanlou Unique players in the BMP pathway: Small C-terminal domain phosphatases dephosphorylate Smad1 to attenuate BMP signaling PNAS, August 8, 2006; 103(32): 11940 - 11945. [Abstract] [Full Text] [PDF]

				J. Heasman Patterning the early Xenopus embryo. Development, April 1, 2006; 133(7): 1205 - 1217. [Abstract] [Full Text] [PDF]

				D. Sakai, T. Suzuki, N. Osumi, and Y. Wakamatsu Cooperative action of Sox9, Snail2 and PKA signaling in early neural crest development Development, April 1, 2006; 133(7): 1323 - 1333. [Abstract] [Full Text] [PDF]

				Y. Onuma, C.-Y. Yeo, and M. Whitman XCR2, one of three Xenopus EGF-CFC genes, has a distinct role in the regulation of left-right patterning Development, January 15, 2006; 133(2): 237 - 250. [Abstract] [Full Text] [PDF]

				B. Reversade, H. Kuroda, H. Lee, A. Mays, and E. M. De Robertis Depletion of Bmp2, Bmp4, Bmp7 and Spemann organizer signals induces massive brain formation in Xenopus embryos Development, August 1, 2005; 132(15): 3381 - 3392. [Abstract] [Full Text] [PDF]

				N. R. Dunn, C. H. Koonce, D. C. Anderson, A. Islam, E. K. Bikoff, and E. J. Robertson Mice exclusively expressing the short isoform of Smad2 develop normally and are viable and fertile Genes & Dev., January 1, 2005; 19(1): 152 - 163. [Abstract] [Full Text] [PDF]

				R. L. C. Carvalho, L. Jonker, M.-J. Goumans, J. Larsson, P. Bouwman, S. Karlsson, P. t. Dijke, H. M. Arthur, and C. L. Mummery Defective paracrine signalling by TGF{beta} in yolk sac vasculature of endoglin mutant mice: a paradigm for hereditary haemorrhagic telangiectasia Development, December 15, 2004; 131(24): 6237 - 6247. [Abstract] [Full Text] [PDF]

				M. Daniels, K. Shimizu, A. M. Zorn, and S.-i. Ohnuma Negative regulation of Smad2 by PIASy is required for proper Xenopus mesoderm formation Development, November 15, 2004; 131(22): 5613 - 5626. [Abstract] [Full Text] [PDF]

				R. M. Monteiro, S. M. C. de Sousa Lopes, O. Korchynskyi, P. t. Dijke, and C. L. Mummery Spatio-temporal activation of Smad1 and Smad5 in vivo: monitoring transcriptional activity of Smad proteins J. Cell Sci., September 15, 2004; 117(20): 4653 - 4663. [Abstract] [Full Text] [PDF]

				F. J. Nicolas, K. De Bosscher, B. Schmierer, and C. S. Hill Analysis of Smad nucleocytoplasmic shuttling in living cells J. Cell Sci., August 15, 2004; 117(18): 4113 - 4125. [Abstract] [Full Text] [PDF]

				S. M. C. de Sousa Lopes, B. A.J. Roelen, R. M. Monteiro, R. Emmens, H. Y. Lin, E. Li, K. A. Lawson, and C. L. Mummery BMP signaling mediated by ALK2 in the visceral endoderm is necessary for the generation of primordial germ cells in the mouse embryo Genes & Dev., August 1, 2004; 18(15): 1838 - 1849. [Abstract] [Full Text] [PDF]

				R. A. Randall, M. Howell, C. S. Page, A. Daly, P. A. Bates, and C. S. Hill Recognition of Phosphorylated-Smad2-Containing Complexes by a Novel Smad Interaction Motif Mol. Cell. Biol., February 1, 2004; 24(3): 1106 - 1121. [Abstract] [Full Text] [PDF]

				M. Schmerer and T. Evans Primitive erythropoiesis is regulated by Smad-dependent signaling in postgastrulation mesoderm Blood, November 1, 2003; 102(9): 3196 - 3205. [Abstract] [Full Text] [PDF]

				S.-X. Ying, Z. J. Hussain, and Y. E. Zhang Smurf1 Facilitates Myogenic Differentiation and Antagonizes the Bone Morphogenetic Protein-2-induced Osteoblast Conversion by Targeting Smad5 for Degradation J. Biol. Chem., October 3, 2003; 278(40): 39029 - 39036. [Abstract] [Full Text] [PDF]

				P. Rajan, D. M. Panchision, L. F. Newell, and R. D.G. McKay BMPs signal alternately through a SMAD or FRAP-STAT pathway to regulate fate choice in CNS stem cells J. Cell Biol., June 9, 2003; 161(5): 911 - 921. [Abstract] [Full Text] [PDF]

				S. T. Dougan, R. M. Warga, D. A. Kane, A. F. Schier, and W. S. Talbot The role of the zebrafish nodal-related genes squint and cyclops in patterning of mesendoderm Development, May 1, 2003; 130(9): 1837 - 1851. [Abstract] [Full Text] [PDF]

				S. Sidi, C. Goutel, N. Peyrieras, and F. M. Rosa Maternal induction of ventral fate by zebrafish radar PNAS, March 18, 2003; 100(6): 3315 - 3320. [Abstract] [Full Text] [PDF]

				M. Howell, G. J. Inman, and C. S. Hill A novel Xenopus Smad-interacting forkhead transcription factor (XFast-3) cooperates with XFast-1 in regulating gastrulation movements Development, March 8, 2003; 129(12): 2823 - 2834. [Abstract] [Full Text] [PDF]

				R. K. Moore, F. Otsuka, and S. Shimasaki Molecular Basis of Bone Morphogenetic Protein-15 Signaling in Granulosa Cells J. Biol. Chem., January 3, 2003; 278(1): 304 - 310. [Abstract] [Full Text] [PDF]

				F. M. Rosa Cripto, a Multifunctional Partner in Signaling: Molecular Forms and Activities Sci. Signal., November 12, 2002; 2002(158): pe47 - pe47. [Abstract] [Full Text] [PDF]

				J. B. Xanthos, M. Kofron, Q. Tao, K. Schaible, C. Wylie, and J. Heasman The roles of three signaling pathways in the formation and function of the Spemann Organizer Development, September 1, 2002; 129(17): 4027 - 4043. [Abstract] [Full Text] [PDF]

				A. Rosendahl, E. Pardali, M. Speletas, P. ten Dijke, C.-H. Heldin, and P. Sideras Activation of Bone Morphogenetic Protein/Smad Signaling in Bronchial Epithelial Cells during Airway Inflammation Am. J. Respir. Cell Mol. Biol., August 1, 2002; 27(2): 160 - 169. [Abstract] [Full Text] [PDF]

				G. J. Inman, F. J. Nicolas, J. F. Callahan, J. D. Harling, L. M. Gaster, A. D. Reith, N. J. Laping, and C. S. Hill SB-431542 Is a Potent and Specific Inhibitor of Transforming Growth Factor-beta Superfamily Type I Activin Receptor-Like Kinase (ALK) Receptors ALK4, ALK5, and ALK7 Mol. Pharmacol., July 1, 2002; 62(1): 65 - 74. [Abstract] [Full Text] [PDF]

				D. Y.R. Stainier A glimpse into the molecular entrails of endoderm formation Genes & Dev., April 15, 2002; 16(8): 893 - 907. [Full Text] [PDF]

				Y. Miyanaga, I. Torregroza, and T. Evans A Maternal Smad Protein Regulates Early Embryonic Apoptosis in Xenopus laevis Mol. Cell. Biol., March 1, 2002; 22(5): 1317 - 1328. [Abstract] [Full Text] [PDF]

				P. M. Eimon and R. M. Harland Effects of heterodimerization and proteolytic processing on Derriere and Nodal activity: implications for mesoderm induction in Xenopus Development, January 7, 2002; 129(13): 3089 - 3103. [Abstract] [Full Text] [PDF]

				P.-Y. Bourillot, N. Garrett, and J. B. Gurdon A changing morphogen gradient is interpreted by continuous transduction flow Development, January 5, 2002; 129(9): 2167 - 2180. [Abstract] [Full Text] [PDF]

				A. Schohl and F. Fagotto {beta}-catenin, MAPK and Smad signaling during early Xenopus development Development, January 1, 2002; 129(1): 37 - 52. [Abstract] [Full Text] [PDF]

				Y. Cui, R. Hackenmiller, L. Berg, F. Jean, T. Nakayama, G. Thomas, and J. L. Christian The activity and signaling range of mature BMP-4 is regulated by sequential cleavage at two sites within the prodomain of the precursor Genes & Dev., November 1, 2001; 15(21): 2797 - 2802. [Abstract] [Full Text] [PDF]

				T. S. Yamamoto, C. Takagi, A. C. Hyodo, and N. Ueno Suppression of head formation by Xmsx-1 through the inhibition of intracellular nodal signaling Development, July 15, 2001; 128(14): 2769 - 2779. [Abstract] [Full Text] [PDF]

				K. Shimizu, P.-Y. Bourillot, S. J. Nielsen, A. M. Zorn, and J. B. Gurdon Swift Is a Novel BRCT Domain Coactivator of Smad2 in Transforming Growth Factor {beta} Signaling Mol. Cell. Biol., June 15, 2001; 21(12): 3901 - 3912. [Abstract] [Full Text]

				K. Mintzer, M. Lee, G Runke, J Trout, M Whitman, and M. Mullins Lost-a-fin encodes a type I BMP receptor, Alk8, acting maternally and zygotically in dorsoventral pattern formation Development, January 3, 2001; 128(6): 859 - 869. [Abstract] [PDF]

				J. Xanthos, M Kofron, C Wylie, and J Heasman Maternal VegT is the initiator of a molecular network specifying endoderm in Xenopus laevis Development, January 1, 2001; 128(2): 167 - 180. [Abstract] [PDF]

				J. Esparza-Lopez, J. L. Montiel, M. M. Vilchis-Landeros, T. Okadome, K. Miyazono, and F. Lopez-Casillas Ligand Binding and Functional Properties of Betaglycan, a Co-receptor of the Transforming Growth Factor-beta Superfamily. SPECIALIZED BINDING REGIONS FOR TRANSFORMING GROWTH FACTOR-beta AND INHIBIN A J. Biol. Chem., April 27, 2001; 276(18): 14588 - 14596. [Abstract] [Full Text] [PDF]

				L. Ulloa and S. Tabibzadeh Lefty Inhibits Receptor-regulated Smad Phosphorylation Induced by the Activated Transforming Growth Factor-beta Receptor J. Biol. Chem., June 8, 2001; 276(24): 21397 - 21404. [Abstract] [Full Text] [PDF]

				S. G. Schiffer, S. Foley, A. Kaffashan, X. Hronowski, A. E. Zichittella, C.-Y. Yeo, K. Miatkowski, H. B. Adkins, B. Damon, M. Whitman, et al. Fucosylation of Cripto Is Required for Its Ability to Facilitate Nodal Signaling J. Biol. Chem., October 5, 2001; 276(41): 37769 - 37778. [Abstract] [Full Text] [PDF]