Skip to main content

her11 is involved in the somitogenesis clock in zebrafish

Abstract

Somitogenesis requires an intricate process of pre-patterning, which is driven by an oscillator mechanism consisting of the Delta-Notch pathway and hairy- (h) and Enhancer of split- [E(spl)] related genes. With the aim of unravelling the complex mechanism of somite pre-patterning, we have conducted an extensive search for h/E(spl)-related genes in the third release of the Danio rerio genomic sequence. We identified 14 new h/E(spl) genes and analysed them by in situ hybridisation for their potential role in the somitogenesis process. We describe here the functional analysis of one of these genes, which we have named her11. her11 is a paralogue of her1 and, similar to her1, is arranged in a head to head fashion with another her gene, namely the previously described her5. It shares an expression in the midbrain-hindbrain boundary with her5, but is in addition cyclically expressed in patterns overlapping those of her1 and her7 and complementary to those of hey1. Furthermore it is expressed in the anterior half of the most caudally formed somites. We show that Delta-Notch pathway genes and fused somites (fss) are necessary for the control of her11 expression. However, some aspects of the her11 regulation suggest that at least one additional as yet unknown gene of the Delta-Notch cascade is required to explain its expression. Morpholino-oligonucleotide-mediated knockdown of her11 shows that it is involved in the zebrafish somitogenesis clock via an interaction with her1 and her7. We have also studied the role of hey1 by morpholino injection, but could not find a direct function for this gene, suggesting that it reflects the output of the clock rather than being a core component of the mechanism.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3a–i
Fig. 4a–j
Fig. 5a–h
Fig. 6a–h
Fig. 7a–i
Fig. 8

References

  • Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    CAS  PubMed  Google Scholar 

  • Bessho Y, Kageyama R (2003) Oscillations, clocks and segmentation. Curr Opin Genet Dev 13:379–384

    Article  CAS  PubMed  Google Scholar 

  • Bessho Y, Miyoshi G, Sakata R, Kageyama R (2001a) Hes7: a bHLH-type repressor gene regulated by Notch and expressed in the presomitic mesoderm. Genes Cells 6:175–185

    Article  CAS  PubMed  Google Scholar 

  • Bessho Y, Sakata R, Komatsu S, Shiota K, Yamada S, Kageyama R (2001b) Dynamic expression and essential functions of Hes7 in somite formation. Genes Dev 15:2642–2647

    Article  CAS  PubMed  Google Scholar 

  • Bessho Y, Hirata H, Masamizu Y, Kageyama R (2003) Periodic repression by the bHLH factor Hes7 is an essential mechanism for the somite segmentation clock. Genes Dev 17:1451–1456

    Article  CAS  PubMed  Google Scholar 

  • van Eeden FJ, Granato M, Schach U, Brand M, Furutani-Seiki M, Haffter P, Hammerschmidt M, Heisenberg CP, Jiang Y-J, Kane DA, Kelsh RN, Mullins MC, Odenthal J, Warga RM, Allende ML, Weinberg ES, Nüsslein-Volhard C (1996) Mutations affecting somite formation and patterning in the zebrafish, Danio rerio. Development 123:153–164

    PubMed  Google Scholar 

  • Fischer A, Leimeister C, Winkler C, Schumacher N, Klamt B, Elmasri A, Steidl C, Maier M, Knobeloch KP, Amann K, Helisch A, Sendtner M, Gessler M (2002) Hey bHLH factors in cardiovascular development. Cold Spring Harb Symp Quant Biol 67:63–70

    CAS  PubMed  Google Scholar 

  • Force A, Lynch M, Pickett FB, Amores A, Yan YL, Postlethwait J (1999) Preservation of duplicate genes by complementary, degenerative mutations. Genetics 151:1531–1545

    CAS  PubMed  Google Scholar 

  • Gajewski M, Voolstra C (2002) Comparative analysis of somitogenesis related genes of the hairy/Enhancer of split class in Fugu and zebrafish. BMC Genomics 3:21

    Article  PubMed  Google Scholar 

  • Gajewski M, Sieger D, Alt B, Leve C, Hans S, Wolff C, Rohr KB, Tautz D (2003) Anterior and posterior waves of cyclic her1 gene expression are differentially regulated in the presomitic mesoderm of zebrafish. Development 130:4269–4278

    Article  CAS  PubMed  Google Scholar 

  • Geling A, Itoh M, Tallafuss A, Chapouton P, Tannhauser B, Kuwada JY, Chitnis AB, Bally-Cuif L (2003) bHLH transcription factor Her5 links patterning to regional inhibition of neurogenesis at the midbrain-hindbrain boundary. Development 130:1591–1604

    Article  CAS  PubMed  Google Scholar 

  • Henry CA, Urban MK, Dill KK, Merlie JP, Page MF, Kimmel CB, Amacher SL (2002) Zebrafish her1 and her7 function together to refine alternating somite boundaries. Development 129:3693–3704

    CAS  PubMed  Google Scholar 

  • Hirata H, Yoshiura S, Ohtsuka T, Bessho Y, Harada T, Yoshikawa K, Kageyama R (2002) Oscillatory expression of the bHLH factor Hes1 regulated by a negative feedback loop. Science 298:840–843

    Article  CAS  PubMed  Google Scholar 

  • Holley SA, Geisler R, Nüsslein-Volhard C (2000) Control of her1 expression during zebrafish somitogenesis by a Delta-dependent oscillator and an independent wave front activity. Genes Dev 14:1678–1690

    CAS  PubMed  Google Scholar 

  • Holley SA, Jülich D, Rauch G-J, Geisler R, Nüsslein-Volhard C (2002) her1 and the Notch pathway function within the oscillator mechanism that regulates zebrafish somitogenesis. Development 129:1175–1183

    CAS  PubMed  Google Scholar 

  • Jiang YJ, Aerne BL, Smithers L, Haddon C, Ish-Horowicz D, Lewis J (2000) Notch signalling and the synchronization of the somite segmentation clock. Nature 408:475–479

    Article  CAS  PubMed  Google Scholar 

  • Kimmel CB, Ballard WW, Kimmel SR, Ullmann B, Schilling TH (1995) Stages of embryonic development of the zebrafish. Dev Dyn 203:253–310

    CAS  PubMed  Google Scholar 

  • Ledent V, Paquet O, Vervoort M (2002) Phylogenetic analysis of the human basic helix-loop-helix proteins. Genome Biol 3:0030.1–0030.18

    Article  Google Scholar 

  • Leve C, Gajewski M, Rohr KB, Tautz D (2001) Homologues of c-hairy1 (her9) and lunatic fringe in zebrafish are expressed in the developing central nervous system, but not in the presomitic mesoderm. Dev Genes Evol 211:493–500

    Article  CAS  PubMed  Google Scholar 

  • Maroto M, Pourquié O (2001) A molecular clock involved in somite segmentation. Curr Top Dev Biol 51:221–248

    Article  CAS  PubMed  Google Scholar 

  • Moore AW, Barbel S, Jan LY, Jan YN (2000) A genomewide survey of basic helix-loop-helix factors in Drosophila. Proc Natl Acad Sci USA 97:10436–10441

    Article  CAS  PubMed  Google Scholar 

  • Müller M, Weizsäcker E, Campos-Ortega JA (1996) Expression domains of a zebrafish homolog of the Drosophila pair-rule gene hairy correspond to the primordia of alternating somites. Development 122:2071–2078

    PubMed  Google Scholar 

  • Nicholas KB, Nicholas HB, Deerfield DW II (1997) GeneDoc: analysis and viualization of genetic variation. EMBNEW News 4:14

    Google Scholar 

  • Nikaido M, Kawakami A, Sawada A, Furutani-Seiki M, Takeda H, Araki K (2002) Tbx24, encoding a T-box protein, is mutated in the zebrafish somite-segmentation mutant fused somites. Nat Genet 31:195–199

    Article  CAS  PubMed  Google Scholar 

  • Oates AC, Ho RK (2002) Hairy/E(spl)-related (Her) genes are central components of the segmentation oscillator and display redundancy with the Delta-Notch signaling pathway in the formation of anterior segmental boundaries in the zebrafish. Development 129:2929–2946

    CAS  PubMed  Google Scholar 

  • Page RDM (1996) TREEVIEW: an application to display phylogenetic trees on personal computers. Comp Appl Biosci 12:357–358

    CAS  PubMed  Google Scholar 

  • Pasini A, Henrique D, Wilkinson DG (2001) The zebrafish Hairy/Enhancer-of-split-related gene her6 is segmentally expressed during the early development of hindbrain and somites. Mech Dev 100:317–321

    Article  CAS  PubMed  Google Scholar 

  • Plickert G, Gajewski M, Gehrke G, Gausepohl H, Schlossherr J, Ibrahim H (1999) Automated in situ detection (AISD) of biomolecules. Dev Genes Evol 207:362–367

    Article  Google Scholar 

  • Rida PC, Le Minh N, Jiang YJ (2004) A Notch feeling of somite segmentation and beyond. Dev Biol 265:2–22

    Article  CAS  PubMed  Google Scholar 

  • Saga Y, Takeda H (2001) The making of the somite: molecular events in vertebrate segmentation. Nat Rev 2:835–845

    Article  CAS  PubMed  Google Scholar 

  • Sawada A, Fritz A, Jiang Y-J, Yamamoto A, Yamasu K, Kuroiwa A, Saga Y, Takeda H (2000) Zebrafish Mesp family genes, mesp-a and mesp-b are segmentally expressed in the presomitic mesoderm, and Mesp-b confers the anterior identity to the developing somites. Development 127:1691–1702

    CAS  PubMed  Google Scholar 

  • Sieger D, Tautz D, Gajewski M (2003) The role of Suppressor of Hairless in Notch mediated signalling during zebrafish somitogenesis. Mech Dev 120:1083–1094

    Article  CAS  PubMed  Google Scholar 

  • Takke C, Dornseifer P, von Weizsäcker E, Campos-Ortega JA (1999) her4, a zebrafish homologue of the Drosophila neurogenic gene E(spl), is a target of Notch signalling, Development 126:1811–1821

  • Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The ClustalX Windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 24:4876–4882

    Article  Google Scholar 

  • Weinmaster G, Kintner C (2003) Modulation of notch signaling during somitogenesis. Annu Rev Cell Dev Biol 19:367–395

    Article  CAS  PubMed  Google Scholar 

  • von Weizsäcker E (1994) Molekulargenetische Untersuchungen an sechs Zebrafisch-Genen mit Homologie zur Enhancer of split Gen-Familie von Drosophila. PhD Thesis, Cologne University, Cologne

  • Westin J, Lardelli M (1997) Three novel Notch genes in zebrafish: implications for vertebrate Notch gene evolution and function. Dev Genes Evol 207:51–63

    Article  CAS  Google Scholar 

  • Winkler C, Elmasri H, Klamt B, Volff JN, Gessler M (2003) Characterization of hey bHLH genes in teleost fish. Dev Genes Evol 213:541–553

    Article  CAS  PubMed  Google Scholar 

  • Zhong TP, Rosenberg M, Mohideen MA, Weinstein B, Fishman MC (2000) gridlock, an HLH gene required for assembly of the aorta in zebrafish. Science 287:1820–1824

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We wish to thank Irene Steinfartz and Eva Schetter for excellent technical assistance, Nina Kobs and Bastian Ackermann for fish care. The zebrafish sequence data has been provided freely by the Wellcome Trust Sanger Institute at the Ensembl Genome Browser (http://www.ensembl.org/Danio_rerio/). The work was supported by the Deutsche Forschungsgemeinschaft (SFB 572) and by the Fond der Chemischen Industrie.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Martin Gajewski.

Additional information

Edited by B. Herrmann

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Sieger, D., Tautz, D. & Gajewski, M. her11 is involved in the somitogenesis clock in zebrafish. Dev Genes Evol 214, 393–406 (2004). https://doi.org/10.1007/s00427-004-0427-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00427-004-0427-z

Keywords

  • Somitogenesis
  • bHLH transcription factor
  • her genes
  • Morpholino-oligonucleotide-mediated knockdown