Development Genes and Evolution

, Volume 217, Issue 8, pp 593–601

Nodal regulates neural tube formation in the Ciona intestinalis embryo

Original Article

Abstract

Overexpression of a lefty orthologue, Ci-lefty, caused a failure of neural tube closure in the protochordate ascidian Ciona intestinalis. The body bent dorsally, and anterior–posterior elongation was inhibited. A similar phenotype was observed in embryos treated with SB431542, an inhibitor of Nodal receptors, suggesting that Ci-Lefty antagonized Nodal signaling as reported in other deuterostome species. Overexpression of Ci-nodal also resulted in a similar phenotype, suggesting that a correct quantity and/or a spatial restriction of Nodal signaling are important for the neural tube to form. In addition to known Ci-Nodal target genes, orthologues of Zic (Ci-ZicL) and cdx (Ci-cdx) were activated by Ci-Nodal. Expression of a dominant negative Ci-cdx caused defects in neural tube formation similar to those obtained on treatment with SB431542 or overexpression of Ci-lefty. A regulatory cascade composed of Ci-Nodal, Ci-ZicL, and Ci-Cdx may play an important role in neural tube formation in the Ciona embryo.

Keywords

Ascidian Neural tube formation Lefty Nodal Cdx ZicL 

Supplementary material

427_2007_168_Fig6_ESM.gif (42 kb)
Supplementary Fig. S1

Characterization of a lefty/antivin orthologue in C. intestinalis. (a) The amino acid sequence predicted from the Ci-lefty cDNA is aligned with other Lefty protein sequences obtained from mouse (m), zebrafish (z), and the sea urchin Paracentrotus lividus (Pl). The putative signal peptide region is indicated by a bar. Cysteine residues conserved among Lefty proteins are shown by green shade. An arrowhead indicates the position where cysteine is conserved in other TGF-β superfamily proteins but not in Lefty proteins. Putative protein product of Ci-lefty consists of 385 amino acids. A characteristic configuration of seven cysteine residues, called a cysteine knot, is conserved among TGF-β superfamily members (Kingsley 1994). Among the family, only the Lefty/Antivin group lacks the fourth cysteine residue (Thisse and Thisse 1999; Juan and Hamada 2001). A predicted amino acid sequence of Ci-Lefty showed the Lefty/Antivin-type cysteine knot. The N-terminal signal peptide sequence was observed in the Lefty proteins, including Ci-Lefty. (b) A phylogenetic tree of TGF-β superfamily proteins constructed by the maximum likelihood method (Felsenstein 1981), using full-length amino acid sequences. Zebrafish and mouse TGF-β proteins were defined as an outgroup. Ci, C. intestinalis; m, mouse; Pl, P. lividus; z, zebrafish. Orthologues of lefty formed a monophyletic cluster. Within the cluster, the branching pattern was consistent with widely accepted phylogenetic relationships of deuterostome groups. Paralogous subgroups in vertebrates (e.g. Lefty1 and Lefty2 in the mouse) diverged within the vertebrate clade. (GIF 43 kb)

References

  1. Anno C, Satou A, Fujiwara S (2006) Transcriptional regulation of ZicL in the Ciona intestinalis embryo. Dev Genes Evol 216:597–605PubMedCrossRefGoogle Scholar
  2. Boorman CJ, Shimeld SM (2002) The evolution of left–right asymmetry in chordates. Bioessays 24:1004–1011PubMedCrossRefGoogle Scholar
  3. Chea HK, Wright C, Swalla BJ (2005) Nodal signaling and the evolution of deuterostome gastrulation. Dev Dyn 234:269–278PubMedCrossRefGoogle Scholar
  4. Corbo JC, Erives A, Di Gregorio A, Chang A, Levine M (1997a) Dorsoventral patterning of the vertebrate neural tube is conserved in a protochordate. Development 124:2335–2344PubMedGoogle Scholar
  5. Corbo JC, Levine M, Zeller RW (1997b) Characterization of a notochord-specific enhancer from the Brachyury promoter region of the ascidian, Ciona intestinalis. Development 124:589–602PubMedGoogle Scholar
  6. Dehal P, Satou Y, Campbell RK, Chapman J, Degnan B, De Tomaso A, Davidson B, Di Gregorio A, Gelpke M, Goodstein DM, Harafuji N, Hastings KEM, Ho I, Hotta K, Huang W, Kawashima T, Lemaire P, Martinez D, Meinertzhagen IA, Necula S, Nonaka M, Putnam N, Rash S, Saiga H, Satake M, Terry A, Yamada L, Wang H-G, Awazu S, Azumi K, Boore J, Branno M, Chin-bow S, De Santis R, Doyle S, Francino P, Keys DN, Haga S, Hayashi H, Hino K, Imai KS, Inaba K, Kano S, Kobayashi K, Kobayashi M, Lee B-I, Makabe KW, Manohar C, Matassi G, Medina M, Mochizuki Y, Mount S, Morishita T, Miura S, Nakayama A, Nishizaka S, Nomoto H, Ohta F, Oishi K, Rigoutsos I, Sano M, Sasaki A, Sasakura Y, Shoguchi E, Shin-i T, Spagnuolo A, Stainier D, Suzuki MM, Tassy O, Takatori N, Tokuoka M, Yagi K, Yoshizaki F, Wada S, Zhang C, Hyatt PD, Larimer F, Detter C, Doggett N, Glavina T, Hawkins T, Richardson P, Lucas S, Kohara Y, Levine M, Satoh N, Rokhsar DS (2002) The draft genome of Ciona intestinalis: insights into chordate and vertebrate origins. Science 298:2157–2167PubMedCrossRefGoogle Scholar
  7. Duboc V, Röttinger E, Besnardeau L, Lepage T (2004) Nodal and BMP2/4 signaling organizes the oral–aboral axis of the sea urchin embryo. Dev Cell 6:397–410PubMedCrossRefGoogle Scholar
  8. Duboc V, Röttinger E, Lapraz F, Besnardeau L, Lepage T (2005) Left–right asymmetry in the sea urchin embryo is regulated by Nodal signaling on the right side. Dev Cell 9:147–158PubMedCrossRefGoogle Scholar
  9. Epstein M, Pillemer G, Yelin R, Yisraeli JK, Fainsod A (1997) Patterning of the embryo along the anterior–posterior axis: the role of the caudal genes. Development 124:3805–3814PubMedGoogle Scholar
  10. Felsenstein J (1981) Evolutionary trees from DNA sequences: a maximum likelihood approach. J Mol Evol 17:368–376PubMedCrossRefGoogle Scholar
  11. Flowers VL, Courteau GR, Poustka AJ, Weng W, Venuti JM (2004) Nodal/activin signaling establishes oral–aboral polarity in the early sea urchin embryo. Dev Dyn 231:727–740PubMedCrossRefGoogle Scholar
  12. Fujiwara S, Maeda Y, Shin-i T, Kohara Y, Takatori N, Satou Y, Satoh N (2002) Gene expression profiles in Ciona intestinalis cleavage-stage embryos. Mech Dev 112:115–127PubMedCrossRefGoogle Scholar
  13. Hino K, Satou Y, Yagi K, Satoh N (2003) A genomewide survey of developmentally relevant genes in Ciona intestinalis. VI. Genes for Wnt, TGFβ, Hedgehog and JAK/STAT signaling pathways. Dev Genes Evol 213:264–272PubMedCrossRefGoogle Scholar
  14. Hudson C, Yasuo H (2005) Patterning across the ascidian neural plate by lateral Nodal signalling sources. Development 132:1199–1210PubMedCrossRefGoogle Scholar
  15. Hudson C, Yasuo H (2006) A signalling relay involving Nodal and Delta ligands acts during secondary notochord induction in Ciona embryos. Development 133:2855–2864PubMedCrossRefGoogle Scholar
  16. Hudson C, Darras S, Caillol D, Yasuo H, Lemaire P (2003) A conserved role for the MEK signalling pathway in neural tissue specification and posteriorisation in the invertebrate chordate, the ascidian Ciona intestinalis. Development 130:147–159PubMedCrossRefGoogle Scholar
  17. Imai KS (2003) Isolation and characterization of β-catenin downstream genes in early embryos of the ascidian Ciona savignyi. Differentiation 71:346–360PubMedCrossRefGoogle Scholar
  18. Imai KS, Satoh N, Satou Y (2002) An essential role of a FoxD gene in notochord induction in Ciona embryos. Development 129:3441–3453PubMedGoogle Scholar
  19. Imai KS, Hino K, Yagi K, Satoh N, Satou Y (2004) Gene expression profiles of transcription factors and signaling molecules in the ascidian embryo: towards a comprehensive understanding of gene networks. Development 131:4047–4058PubMedCrossRefGoogle Scholar
  20. Imai KS, Levine M, Satoh N, Satou Y (2006) Regulatory blueprint for a chordate embryo. Science 312:1183–1187PubMedCrossRefGoogle Scholar
  21. Inman GJ, Nicolas FJ, Callahan JF, Harling JD, Gaster LM, Reith AD, Laping NJ, Hill CS (2002) SB-431542 is a potent and specific inhibitor of transforming growth factor-β superfamily type I activin receptor-like kinase (ALK) receptors ALK4, ALK5, and ALK7. Mol Pharmacol 62:65–74PubMedCrossRefGoogle Scholar
  22. Isaacs HV, Pownall ME, Slack JMW (1998) Regulation of Hox gene expression and posterior development by the Xenopus caudal homologue Xcad3. EMBO J 17:3413–3427PubMedCrossRefGoogle Scholar
  23. Jaynes JB, O’Farrell PH (1991) Active repression of transcription by the engrailed homeodomain protein. EMBO J 10:1427–1433PubMedGoogle Scholar
  24. Juan H, Hamada H (2001) Roles of nodal-lefty regulatory loops in embryonic patterning of vertebrates. Genes Cells 6:923–930PubMedCrossRefGoogle Scholar
  25. Katsuyama Y, Sato Y, Wada S, Saiga H (1999) Ascidian tail formation requires caudal function. Dev Biol 213:257–268PubMedCrossRefGoogle Scholar
  26. Kingsley DM (1994) The TGF-β superfamily: new members, new receptors, new genetic tests of function in different organisms. Genes Dev 8:133–146PubMedCrossRefGoogle Scholar
  27. Lohnes D (2003) The Cdx1 homeodomain protein: an integrator of posterior signaling in the mouse. Bioessays 25:971–980PubMedCrossRefGoogle Scholar
  28. Macdonald PM, Struhl G (1986) A molecular gradient in early Drosophila embryos and its role in specifying the body pattern. Nature 324:537–545PubMedCrossRefGoogle Scholar
  29. Marom K, Shapira E, Fainsod A (1997) The chicken caudal genes establish an anterior–posterior gradient by partially overlapping temporal and spatial patterns of expression. Mech Dev 64:41–52PubMedCrossRefGoogle Scholar
  30. Morokuma J, Ueno M, Kawanishi H, Saiga H, Nishida H (2002) HrNodal, the ascidian nodal-related gene, is expressed in the left side of the epidermis, and lies upstream of HrPitx. Dev Genes Evol 212:439–446PubMedCrossRefGoogle Scholar
  31. Nagatomo K, Ishibashi T, Satou Y, Satoh N, Fujiwara S (2003) Retinoic acid affects gene expression and morphogenesis without upregulating the retinoic acid receptor in the ascidian Ciona intestinalis. Mech Dev 120:363–372PubMedCrossRefGoogle Scholar
  32. Onuma Y, Takahashi S, Yokota C, Asashima M (2002) Multiple nodal-related genes act coordinately in Xenopus embryogenesis. Dev Biol 241:94–105PubMedCrossRefGoogle Scholar
  33. Osada S, Wright CVE (1999) Xenopus nodal-related signaling is essential for mesendodermal patterning during early embryogenesis. Development 126:3229–3240PubMedGoogle Scholar
  34. Pasini A, Amiel A, Rothbächer U, Roure A, Lemaire P, Darras S (2006) Formation of the ascidian epidermal sensory neurons: insights into the origin of the chordate peripheral nervous system. PLOS Biol 4:1173–1186CrossRefGoogle Scholar
  35. Satou Y, Takatori N, Fujiwara S, Nishikata T, Saiga H, Kusakabe T, Shin-i T, Kohara Y, Satoh N (2002a) Ciona intestinalis cDNA projects: expressed sequence tag analyses and gene expression profiles during embryogenesis. Gene 287:83–96PubMedGoogle Scholar
  36. Satou Y, Yamada L, Mochizuki Y, Takatori N, Kawashima T, Sasaki A, Hamaguchi M, Awazu S, Yagi K, Sasakura Y, Nakayama A, Ishikawa H, Inaba K, Satoh N (2002b) A cDNA resource from the basal chordate Ciona intestinalis. Genesis 33:153–154PubMedCrossRefGoogle Scholar
  37. Schier AF, Shen MM (2000) Nodal signaling in vertebrate development. Nature 403:385–389PubMedCrossRefGoogle Scholar
  38. Thisse C, Thisse B (1999) Antivin, a novel and divergent member of the TGFβ superfamily, negatively regulates mesoderm induction. Development 126:229–240PubMedGoogle Scholar
  39. van den Akker E, Forlani S, Chawengsaksophak K, de Graaff W, Beck F, Meyer BI, Deschamps J (2002) Cdx1 and Cdx2 have overlapping functions in anteroposterior patterning and posterior axis elongation. Development 129:2181–2193PubMedGoogle Scholar
  40. Wallingford JB, Harland RM (2001) Xenopus Dishevelled signaling regulates both neural and mesodermal convergent extension: parallel forces elongating the body axis. Development 128:2581–2592PubMedGoogle Scholar
  41. Whitman M (2001) Nodal signaling in early vertebrate embryos: themes and variations. Dev Cell 1:605–617PubMedCrossRefGoogle Scholar
  42. Yagi K, Satou Y, Satoh N (2004) A zinc finger transcription factor, ZicL, is a direct activator of Brachyury in the notochord specification of Ciona intestinalis. Development 131:1279–1288PubMedCrossRefGoogle Scholar
  43. Yokota C, Kofron M, Zuck M, Houston DW, Isaacs H, Asashima M, Wylie CC, Heasman J (2003) A novel role for a nodal-related protein; Xnr3 regulates convergent extension movements via the FGF receptor. Development 130:2199–2212PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  1. 1.Department of Materials ScienceKochi UniversityKochi-shiJapan

Personalised recommendations