Development Genes and Evolution

, Volume 216, Issue 11, pp 709–720 | Cite as

Polarised expression of FoxB and FoxQ2 genes during development of the hydrozoan Clytia hemisphaerica

  • Sandra Chevalier
  • Arnaud Martin
  • Lucas Leclère
  • Aldine Amiel
  • Evelyn HoulistonEmail author
Original Article


We have characterised the expression of four genes coding for Forkhead box-containing (‘Fox’) transcription factors identified from the hydrozoan (Leptomedusa) Clytia hemisphaerica. Phylogenetic analyses including all available non-bilaterian Fox sequences placed these genes in subfamilies B, Q2 (two genes) and O, and indicated that at least 17 Fox subfamilies were present in the common cnidarian/bilaterian ancestor, with multiple subsequent losses in cnidarian lineages. Chordate FoxB and FoxQ2A subfamily genes show polarised expression in early embryos. Correspondingly, Clytia CheFoxB expression was localised around the gastrulation site (future oral pole) at blastula and gastrula stages, with CheFoxQ2a expressed in a complementary aboral domain, maintained through larval development. Distinct later expression domains were observed for CheFoxB in the larval endoderm region, and in the statocyst, gonad and tentacle bulb of the medusa. A second Clytia FoxQ2 gene, CheFoxQ2b, not expressed in the embryo, larva or polyp, was detected uniquely in the gonads of the medusa. In contrast, CheFoxO, whose sequence indicates regulation by the PI3-Kinase/PKB signalling pathway consistent with known roles in bilaterian developmental regulation, was detected throughout the Clytia life cycle. CheFoxO expression was enhanced in regions associated with growth control including larval poles, gonad and the margin of the medusa bell. These results support the idea that an early embryonic patterning system involving FoxB and FoxQ2 family genes has been evolutionary conserved and indicate that Fox family genes have also acquired distinct roles during other phases of the hydrozoan life cycle.


Forkhead Cnidarian Embryonic polarity Statocyst Hydromedusa 



We thank Dany and Claude Carré for encouraging us to develop Clytia as a laboratory model and for giving us advice on its culture. We also thank all the members of our two research groups, in particular, Tsuyoshi Momose, Michaël Manuel, Elsa Denker and Muriel Jager, for useful discussions and for participation in developing tools and techniques. We also thank Mark Martindale for providing a detailed Nematostella in situ hybridisation protocol, and one of the reviewers for alerting us to the Drosophila FoxQ2 expression data.

This research was funded by the CNRS, by joint contracts to our two groups from the GIS “Marine genomics”/ANR, and by ACI, ANR and ARC contracts to EH.

Supplementary material

427_2006_103_MOESM1_ESM.doc (512 kb)
S1 Bayesian analysis of Fox gene phylogeny. Topology of the 50% majority-rule consensus tree resulting from 1,001 trees generated. The posterior probability for each node is indicated above the node (DOC 523 kb)
427_2006_103_MOESM2_ESM.doc (44 kb)
S2 Accession numbers of sequences used in the phylogenetic analysis (DOC 45 kb)
427_2006_103_MOESM3_ESM.doc (44 kb)
S3 Sequence alignment used in the phylogenetic analyses (DOC 44 kb)


  1. Accili D, Arden KC (2004) FoxOs at the crossroads of cellular metabolism, differentiation, and transformation. Cell 117:421–426PubMedCrossRefGoogle Scholar
  2. Adell T, Muller WE (2004) Isolation and characterization of five Fox (Forkhead) genes from the sponge Suberites domuncula. Gene 334:35–46PubMedCrossRefGoogle Scholar
  3. Anderson MJ, Viars CS, Czekay S, Cavenee WK, Arden KC (1998) Cloning and characterization of three human forkhead genes that comprise an FKHR-like gene subfamily. Genomics 47:187–1991PubMedCrossRefGoogle Scholar
  4. Ang SL, Wierda A, Wong D, Stevens KA, Cascio S, Rossant J, Zaret KS (1993) The formation and maintenance of the definitive endoderm lineage in the mouse: involvement of HNF3/forkhead proteins. Development 119:1301–1315PubMedGoogle Scholar
  5. Ball EE, Hayward DC, Saint R, Miller DJ (2004) A simple plan—cnidarians and the origins of developmental mechanisms. Nat Rev Genet 5:567–577PubMedCrossRefGoogle Scholar
  6. Bodo F, Bouillon J (1968) Etude histologique du développement embryonnaire de quelques Hydroméduses de Roscoff. Cah Biol Mar 9:69–79Google Scholar
  7. Bouillon J (1993) Classe des hydrozoaires. In: Grassé P-P (ed) Traité de zoologie. Masson, Paris pp 29–416Google Scholar
  8. Burgering BM, Kops GJ (2002) Cell cycle and death control: long live forkheads. Trends Biochem Sci 27:352–360PubMedCrossRefGoogle Scholar
  9. Byrum CA (2001) An analysis of hydrozoan gastrulation by unipolar ingression. Dev Biol 240:627–640PubMedCrossRefGoogle Scholar
  10. Carlsson P, Mahlapuu M (2002) Forkhead transcription factors: key players in development and metabolism. Dev Biol 250:1–23PubMedCrossRefGoogle Scholar
  11. Carre D, Carre C (2000) Origin of germ cells, sex determination, and sex inversion in medusae of the genus Clytia (Hydrozoa, leptomedusae): the influence of temperature. J Exp Zool 287:233–242PubMedCrossRefGoogle Scholar
  12. Collins AG (1998) Evaluating multiple alternative hypotheses for the origin of Bilateria: an analysis of 18S rRNA molecular evidence. Proc Natl Acad Sci USA 95:15458–15463PubMedCrossRefGoogle Scholar
  13. Collins AG, Schuchert P, Marques AC, Jankowski T, Medina M, Schierwater B (2006) Cnidarian phylogeny and character evolution clarified by new large and small subunit rDNA data and an assessment of the utility of phylogenetic mixture models. Syst Biol 55:97–110PubMedCrossRefGoogle Scholar
  14. Finnerty JR, Pang K, Burton P, Paulson D, Martindale MQ (2004) Origins of bilateral symmetry: Hox and dpp expression in a sea anemone. Science 304:1335–1337PubMedCrossRefGoogle Scholar
  15. Freeman G (1981) The cleavage initiation site establishes the posterior pole of the hydrozoan embryo. Wilhelm Roux’s Archives 190:123–125CrossRefGoogle Scholar
  16. Freeman G (2005) The effect of larval age on developmental changes in the polyp prepattern of a hydrozoan planula. Zoology 108:55–73PubMedCrossRefGoogle Scholar
  17. Fritzenwanker JH, Saina M, Technau U (2004) Analysis of forkhead and snail expression reveals epithelial–mesenchymal transitions during embryonic and larval development of Nematostella vectensis. Dev Biol 275:389–402PubMedCrossRefGoogle Scholar
  18. Gamse JT, Sive H (2001) Early anteroposterior division of the presumptive neurectoderm in Xenopus. Mech Dev 104:21–36PubMedCrossRefGoogle Scholar
  19. Groger H, Schmid V (2001) Larval development in Cnidaria: a connection to Bilateria? Genesis 29:110–114PubMedCrossRefGoogle Scholar
  20. Guindon S, Lethiec F, Duroux P, Gascuel O (2005) PHYML Online—a web server for fast maximum likelihood-based phylogenetic inference. Nucleic Acids Res 33:W557–W559PubMedCrossRefGoogle Scholar
  21. Hacker U, Grossniklaus U, Gehring WJ, Jackle H (1992) Developmentally regulated Drosophila gene family encoding the fork head domain. Proc Natl Acad Sci USA 89:8754–8758PubMedCrossRefGoogle Scholar
  22. Honegger TG, Achermann J, Littlefield RJ, Baenninger R, Tardent P (1980) Light-controlled spawning in Phialidium hemisphaericum (Leptomedusae). In: Tardent P, Tardent R (eds) Developmental and cellular biology of coelenterates. Proceedings of the IV international coelenterate conference. Elsevier/N. Holland Biomedical, Amsterdam, pp 83–88Google Scholar
  23. Hope IA, Mounsey A, Bauer P, Aslam S (2003) The forkhead gene family of Caenorhabditis elegans. Gene 304:43–55PubMedCrossRefGoogle Scholar
  24. Hosaka T, Biggs WH 3rd, Tieu D, Boyer AD, Varki NM, Cavenee WK, Arden KC (2004) Disruption of forkhead transcription factor (FOXO) family members in mice reveals their functional diversification. Proc Natl Acad Sci USA 101:2975–2980PubMedCrossRefGoogle Scholar
  25. Huelsenbeck JP, Ronquist F (2001) MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics 17:754–755PubMedCrossRefGoogle Scholar
  26. Jager M, Queinnec E, Houliston E, Manuel M (2006) Expansion of the SOX gene family predated the emergence of the Bilateria. Mol Phylogenet Evol 39:468–477PubMedCrossRefGoogle Scholar
  27. Kaestner KH, Knochel W, Martinez DE (2000) Unified nomenclature for the winged helix/forkhead transcription factors. Genes Dev 14:142–146PubMedGoogle Scholar
  28. Kamm K, Schierwater B, Jakob W, Dellaporta SL, Miller DJ (2006) Axial patterning and diversification in the Cnidaria Predate the Hox system. Curr Biol 16:920–926PubMedCrossRefGoogle Scholar
  29. Kortschak RD, Samuel G, Saint R, Miller DJ (2003) EST analysis of the cnidarian Acropora millepora reveals extensive gene loss and rapid sequence divergence in the model invertebrates. Curr Biol 13:2190–2195PubMedCrossRefGoogle Scholar
  30. Kusserow A, Pang K, Sturm C, Hrouda M, Lentfer J, Schmidt HA, Technau U, von Haeseler A, Hobmayer B, Martindale MQ, Holstein TW (2005) Unexpected complexity of the Wnt gene family in a sea anemone. Nature 433:156–160PubMedCrossRefGoogle Scholar
  31. Larroux C, Fahey B, Liubicich D, Hinman VF, Gauthier M, Gongora M, Green K, Worheide G, Leys SP, Degnan BM (2006) Developmental expression of transcription factor genes in a demosponge: insights into the origin of metazoan multicellularity. Evol Dev 8:150–173PubMedCrossRefGoogle Scholar
  32. Lee HH, Frasch M (2004) Survey of forkhead domain encoding genes in the Drosophila genome: classification and embryonic expression patterns. Dev Dyn 229:357–366PubMedCrossRefGoogle Scholar
  33. Magie CR, Pang K, Martindale MQ (2005) Genomic inventory and expression of Sox and Fox genes in the cnidarian Nematostella vectensis. Dev Genes Evol 215:1–13CrossRefGoogle Scholar
  34. Martindale MQ (2005) The evolution of metazoan axial properties. Nat Rev Genet 6:917–927PubMedGoogle Scholar
  35. Martindale MQ, Pang K, Finnerty JR (2004) Investigating the origins of triploblasty: ‘mesodermal’ gene expression in a diploblastic animal, the sea anemone Nematostella vectensis (phylum, Cnidaria; class, Anthozoa). Development 131:2463–2474PubMedCrossRefGoogle Scholar
  36. Martinez DE, Dirksen ML, Bode PM, Jamrich M, Steele RE, Bode HR (1997) Budhead, a fork head/HNF-3 homologue, is expressed during axis formation and head specification in hydra. Dev Biol 192:523–536PubMedCrossRefGoogle Scholar
  37. Mazet F, Shimeld SM (2002) The evolution of chordate neural segmentation. Dev Biol 251:258–270PubMedCrossRefGoogle Scholar
  38. Mazet F, Yu JK, Liberles DA, Holland LZ, Shimeld SM (2003) Phylogenetic relationships of the Fox (Forkhead) gene family in the Bilateria. Gene 316:79–89PubMedCrossRefGoogle Scholar
  39. Miller DJ, Ball EE, Technau U (2005) Cnidarians and ancestral genetic complexity in the animal kingdom. Trends Genet 21:536–539PubMedCrossRefGoogle Scholar
  40. Minokawa T, Rast JP, Arenas-Mena C, Franco CB, Davidson EH (2004) Expression patterns of four different regulatory genes that function during sea urchin development. Gene Expression Patterns 4:449–456PubMedCrossRefGoogle Scholar
  41. Odenthal J, Nusslein-Volhard C (1998) Fork head domain genes in zebrafish. Dev Genes Evol 208:245–258PubMedCrossRefGoogle Scholar
  42. Pohl BS, Knochel S, Dillinger K, Knochel W (2002) Sequence and expression of FoxB2 (XFD-5) and FoxI1c (XFD-10) in Xenopus embryogenesis. Mech Dev 117:283–287PubMedCrossRefGoogle Scholar
  43. Roosen-Runge EC, Szollosi D (1965) On biology and structure of the testis of Philidium Leuckhart (Leptomedusae). Z Zellforsch Mikrosk Anat 68:597–610PubMedCrossRefGoogle Scholar
  44. Schmid B, Schmid V, Tardent P (1974) The umbrellar growth process in the leptomedusa Phialidium hemisphaericum (Syn. Campanularia johnstoni). Experimentia 30:1399–1400CrossRefGoogle Scholar
  45. Seipel K, Schmid V (2005) Evolution of striated muscle: jellyfish and the origin of triploblasty. Dev Biol 282:14–26PubMedCrossRefGoogle Scholar
  46. Seoane J, Le HV, Shen L, Anderson SA, Massague J (2004) Integration of Smad and forkhead pathways in the control of neuroepithelial and glioblastoma cell proliferation. Cell 117:211–223PubMedCrossRefGoogle Scholar
  47. Singla CL (1975) Statocysts of hydromedusae. Cell Tissue Res 158:391–407PubMedCrossRefGoogle Scholar
  48. Spring J, Yanze N, Josch C, Middel AM, Winninger B, Schmid V (2002) Conservation of Brachyury, Mef2, and Snail in the myogenic lineage of jellyfish: a connection to the mesoderm of bilateria. Dev Biol 244:372–384PubMedCrossRefGoogle Scholar
  49. Tan PB, Lackner MR, Kim SK (1998) MAP kinase signaling specificity mediated by the LIN-1 Ets/LIN-31 WH transcription factor complex during C. elegans vulval induction. Cell 93:569–580PubMedCrossRefGoogle Scholar
  50. Technau U, Bode HR (1999) HyBra1, a Brachyury homologue, acts during head formation in Hydra. Development 126:999–1010PubMedGoogle Scholar
  51. Thomas MB, Freeman G, Martin VJ (1987) The embryonic origin of neurosensory cells and the role of nerve cells in metamorphosis in Phialidium gregarium (Cnidaria, Hydrozoa). Int J Invertebr Reprod Dev 11:265–287Google Scholar
  52. Wikramanayake AH, Hong M, Lee PN, Pang K, Byrum CA, Bince JM, Xu R, Martindale MQ (2003) An ancient role for nuclear beta-catenin in the evolution of axial polarity and germ layer segregation. Nature 426:446–450PubMedCrossRefGoogle Scholar
  53. Yamada A, Martindale MQ (2002) Expression of the ctenophore Brain Factor 1 forkhead gene ortholog (ctenoBF-1) mRNA is restricted to the presumptive mouth and feeding apparatus: implications for axial organization in the Metazoa. Dev Genes Evol 212:338–348PubMedCrossRefGoogle Scholar
  54. Yu JK, Holland ND, Holland LZ (2003) AmphiFoxQ2, a novel winged helix/forkhead gene, exclusively marks the anterior end of the amphioxus embryo. Dev Genes Evol 213:102–105PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Sandra Chevalier
    • 1
  • Arnaud Martin
    • 1
  • Lucas Leclère
    • 2
  • Aldine Amiel
    • 1
  • Evelyn Houliston
    • 1
    Email author
  1. 1.“Biologie du Développement”, Observatoire OcéanologiqueUMR7009 CNRS/Université Pierre et Marie Curie (Paris VI)Villefranche-sur-merFrance
  2. 2.“Evolution et Développement”UMR 7138 CNRS UPMC MNHN IRD ENS, Université Pierre et Marie Curie (Paris VI)ParisFrance

Personalised recommendations