Advertisement

Development Genes and Evolution

, Volume 226, Issue 2, pp 99–107 | Cite as

vox homeobox gene: a novel regulator of midbrain-hindbrain boundary development in medaka fish?

  • Peter Fabian
  • Chrysoula N. Pantzartzi
  • Iryna Kozmikova
  • Zbynek Kozmik
Short Communication

Abstract

The midbrain-hindbrain boundary (MHB) is one of the key organizing centers of the vertebrate central nervous system (CNS). Its patterning is governed by a well-described gene regulatory network (GRN) involving several transcription factors, namely, pax, gbx, en, and otx, together with signaling molecules of the Wnt and Fgf families. Here, we describe the onset of these markers in Oryzias latipes (medaka) early brain development in comparison to previously known zebrafish expression patterns. Moreover, we show for the first time that vox, a member of the vent gene family, is expressed in the developing neural tube similarly to CNS markers. Overexpression of vox leads to profound changes in the gene expression patterns of individual components of MHB-specific GRN, most notably of fgf8, a crucial organizer molecule of MHB. Our data suggest that genes from the vent family, in addition to their crucial role in body axis formation, may play a role in regionalization of vertebrate CNS.

Keywords

Midbrain-hindbrain boundary vox medaka Heat shock element fgf8 Gene regulatory network 

Notes

Acknowledgments

We are grateful to Jindra Pohorela, Anna Zitova, Ivana Dobiasovska, and Vladimir Soukup for the technical support. We are grateful to Thomas Czerny for providing reagents and to Sarka Takacova for the manuscript proofreading. This work was supported by the Ministry of Education, Youth, and Sports (LO1419).

Supplementary material

427_2016_533_Fig5_ESM.gif (52 kb)
Fig. S1

vox expression is overlapping with otx2 expression in neural plate border and neural keel. (a-e´) Double RNA in situ hybridization for vox (purple) and otx2 (red). Dorsal views at stages 15 (a-d) and 17 (). Lateral view at stage 17 (e). Dashed line indicates position of the shield in the medaka embryo (a-d) or embryonic body (e, ). (b-d) Dissected shield area from (a). Arrowhead marks the edge of vox domain. (GIF 52 kb)

427_2016_533_MOESM1_ESM.tif (2.7 mb)
High Resolution Image (TIF 2769 kb)
427_2016_533_Fig6_ESM.gif (149 kb)
Fig. S2

Knockdown of medaka vox by morpholino (MO). (a) Schematic drawing of the mRNA construct vox_aug_region-eGFP (b) mRNA construct with vox MO. Embryos injected with the synthetic mRNA alone () and with vox MO (). (c-f´) RNA in situ hybridization for otx2, en2, wnt1 and fgf8 genes on embryos injected with control MO (c-f) and vox morpholino (c´-f´). Embryos are shown, anterior to right, in (c-f´ top) lateral view, and (c-f´ bottom) dorsal view. Arrowheads mark developing MHB. (GIF 149 kb)

427_2016_533_MOESM2_ESM.tif (6.4 mb)
High Resolution Image (TIF 6596 kb)
427_2016_533_Fig7_ESM.gif (83 kb)
Fig. S3

The effects of vox overexpression during early development. (a) mCherry:HSE:vox construct. (b) Experimental strategy used in this work. (c, ) overexpression of vox gene (c) and mCherry marker gene () after heat shock. (d-e´) Heat-treated control and mCherry:HSE:vox transgenic line embryos at stage 16. (d, d´) Heat shock does not seem to influence development of control embryos at later developmental stages (stage 30). (e, e´) Overexpression of vox during gastrulation affects development of mCherry:HSE:vox transgenic line embryos. Inset: MHB in detail. (GIF 82 kb)

427_2016_533_MOESM3_ESM.tif (3.7 mb)
High Resolution Image (TIF 3807 kb)

References

  1. Bajoghli B, Aghaallaei N, Heimbucher T, Czerny T (2004) An artificial promoter construct for heat-inducible misexpression during fish embryogenesis. Dev Biol 271:416–430. doi: 10.1016/j.ydbio.2004.04.006 CrossRefPubMedGoogle Scholar
  2. Canning CA, Lee L, Irving C, Mason I, Jones CM (2007) Sustained interactive Wnt and FGF signaling is required to maintain isthmic identity. Dev Biol 305:276–286. doi: 10.1016/j.ydbio.2007.02.009 CrossRefPubMedGoogle Scholar
  3. Dworkin S, Jane SM (2013) Novel mechanisms that pattern and shape the midbrain-hindbrain boundary. Cell Mol Life Sci 70:3365–3374. doi: 10.1007/s00018-012-1240-x CrossRefPubMedGoogle Scholar
  4. Echelard Y, Epstein DJ, St-Jacques B, Shen L, Mohler J, McMahon JA, McMahon AP (1993) Sonic hedgehog, a member of a family of putative signaling molecules, is implicated in the regulation of CNS polarity. Cell 75:1417–1430CrossRefPubMedGoogle Scholar
  5. Echevarria D, Vieira C, Gimeno L, Martinez S (2003) Neuroepithelial secondary organizers and cell fate specification in the developing brain. Brain Res Brain Res Rev 43:179–191CrossRefPubMedGoogle Scholar
  6. Fabian P, Kozmikova I, Kozmik Z, Pantzartzi CN (2015) Pax2/5/8 and Pax6 alternative splicing events in basal chordates and vertebrates: a focus on paired box domain. Front Genet 6:228. doi: 10.3389/fgene.2015.00228 CrossRefPubMedPubMedCentralGoogle Scholar
  7. Gilardelli CN, Pozzoli O, Sordino P, Matassi G, Cotelli F (2004) Functional and hierarchical interactions among zebrafish vox/vent homeobox genes. Dev Dyn 230:494–508. doi: 10.1002/dvdy.20073 CrossRefPubMedGoogle Scholar
  8. Heimbucher T et al (2007) Gbx2 and Otx2 interact with the WD40 domain of Groucho/Tle corepressors. Mol Cell Biol 27:340–351. doi: 10.1128/MCB.00811-06 CrossRefPubMedPubMedCentralGoogle Scholar
  9. Houart C, Westerfield M, Wilson SW (1998) A small population of anterior cells patterns the forebrain during zebrafish gastrulation. Nature 391:788–792. doi: 10.1038/35853 CrossRefPubMedGoogle Scholar
  10. Imai Y, Gates MA, Melby AE, Kimelman D, Schier AF, Talbot WS (2001) The homeobox genes vox and vent are redundant repressors of dorsal fates in zebrafish. Development 128:2407–2420PubMedGoogle Scholar
  11. Iwamatsu T (2004) Stages of normal development in the medaka Oryzias latipes. Mech Dev 121:605–618. doi: 10.1016/j.mod.2004.03.012 CrossRefPubMedGoogle Scholar
  12. Jaszai J, Reifers F, Picker A, Langenberg T, Brand M (2003) Isthmus-to-midbrain transformation in the absence of midbrain-hindbrain organizer activity. Development 130:6611–6623. doi: 10.1242/dev.00899 CrossRefPubMedGoogle Scholar
  13. Jessell TM, Sanes JR (2000) Development. The decade of the developing brain. Curr Opin Neurobiol 10:599–611CrossRefPubMedGoogle Scholar
  14. Joyner AL, Liu A, Millet S (2000) Otx2, Gbx2 and Fgf8 interact to position and maintain a mid-hindbrain organizer. Curr Opin Cell Biol 12:736–741CrossRefPubMedGoogle Scholar
  15. Kawahara A, Wilm T, Solnica-Krezel L, Dawid IB (2000) Antagonistic role of vega1 and bozozok/dharma homeobox genes in organizer formation. Proc Natl Acad Sci U S A 97:12121–12126. doi: 10.1073/pnas.97.22.12121 CrossRefPubMedPubMedCentralGoogle Scholar
  16. Kozmikova I, Candiani S, Fabian P, Gurska D, Kozmik Z (2013) Essential role of Bmp signaling and its positive feedback loop in the early cell fate evolution of chordates. Dev Biol 382:538–554. doi: 10.1016/j.ydbio.2013.07.021 CrossRefPubMedGoogle Scholar
  17. Lee KJ, Jessell TM (1999) The specification of dorsal cell fates in the vertebrate central nervous system. Annu Rev Neurosci 22:261–294. doi: 10.1146/annurev.neuro.22.1.261 CrossRefPubMedGoogle Scholar
  18. Li Y, Allende ML, Finkelstein R, Weinberg ES (1994) Expression of two zebrafish orthodenticle-related genes in the embryonic brain. Mech Dev 48:229–244CrossRefPubMedGoogle Scholar
  19. Liu A, Joyner AL (2001) EN and GBX2 play essential roles downstream of FGF8 in patterning the mouse mid/hindbrain region. Development 128:181–191PubMedGoogle Scholar
  20. Louis A, Nguyen NT, Muffato M, Roest Crollius H (2015) Genomicus update 2015: KaryoView and MatrixView provide a genome-wide perspective to multispecies comparative genomics. Nucleic Acids Res 43:D682–689. doi: 10.1093/nar/gku1112 CrossRefPubMedPubMedCentralGoogle Scholar
  21. Lun K, Brand M (1998) A series of no isthmus (noi) alleles of the zebrafish pax2.1 gene reveals multiple signaling events in development of the midbrain-hindbrain boundary. Development 125:3049–3062PubMedGoogle Scholar
  22. Melby AE, Beach C, Mullins M, Kimelman D (2000) Patterning the early zebrafish by the opposing actions of bozozok and vox/vent. Dev Biol 224:275–285. doi: 10.1006/dbio.2000.9780 CrossRefPubMedGoogle Scholar
  23. Nicholas K, Nicholas H, Deerfield D (1997) GeneDoc: Analysis and visualization of genetic variationGoogle Scholar
  24. Pfeffer PL, Gerster T, Lun K, Brand M, Busslinger M (1998) Characterization of three novel members of the zebrafish Pax2/5/8 family: dependency of Pax5 and Pax8 expression on the Pax2.1 (noi) function. Development 125:3063–3074PubMedGoogle Scholar
  25. Raible F, Brand M (2004) Divide et Impera—the midbrain-hindbrain boundary and its organizer. Trends Neurosci 27:727–734. doi: 10.1016/j.tins.2004.10.003 CrossRefPubMedGoogle Scholar
  26. Reifers F, Bohli H, Walsh EC, Crossley PH, Stainier DY, Brand M (1998) Fgf8 is mutated in zebrafish acerebellar (ace) mutants and is required for maintenance of midbrain-hindbrain boundary development and somitogenesis. Development 125:2381–2395PubMedGoogle Scholar
  27. Rhinn M, Brand M (2001) The midbrain—hindbrain boundary organizer. Curr Opin Neurobiol 11:34–42CrossRefPubMedGoogle Scholar
  28. Rhinn M, Lun K, Amores A, Yan YL, Postlethwait JH, Brand M (2003) Cloning, expression and relationship of zebrafish gbx1 and gbx2 genes to Fgf signaling. Mech Dev 120:919–936CrossRefPubMedGoogle Scholar
  29. Rhinn M, Lun K, Luz M, Werner M, Brand M (2005) Positioning of the midbrain-hindbrain boundary organizer through global posteriorization of the neuroectoderm mediated by Wnt8 signaling. Development 132:1261–1272. doi: 10.1242/dev.01685 CrossRefPubMedGoogle Scholar
  30. Rhinn M, Lun K, Ahrendt R, Geffarth M, Brand M (2009) Zebrafish gbx1 refines the midbrain-hindbrain boundary border and mediates the Wnt8 posteriorization signal. Neural Dev 4:12. doi: 10.1186/1749-8104-4-12 CrossRefPubMedPubMedCentralGoogle Scholar
  31. Ristoratore F et al (1999) The midbrain-hindbrain boundary genetic cascade is activated ectopically in the diencephalon in response to the widespread expression of one of its components, the medaka gene Ol-eng2. Development 126:3769–3779PubMedGoogle Scholar
  32. Sievers F et al (2011) Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol Syst Biol 7:539. doi: 10.1038/msb.2011.75 CrossRefPubMedPubMedCentralGoogle Scholar
  33. Thermes V, Grabher C, Ristoratore F, Bourrat F, Choulika A, Wittbrodt J, Joly JS (2002) I-SceI meganuclease mediates highly efficient transgenesis in fish. Mech Dev 118:91–98CrossRefPubMedGoogle Scholar
  34. Wilson SW, Houart C (2004) Early steps in the development of the forebrain. Dev Cell 6:167–181CrossRefPubMedPubMedCentralGoogle Scholar
  35. Wurst W, Bally-Cuif L (2001) Neural plate patterning: upstream and downstream of the isthmic organizer. Nat Rev Neurosci 2:99–108. doi: 10.1038/35053516 CrossRefPubMedGoogle Scholar
  36. Zhao J, Lambert G, Meijer AH, Rosa FM (2013) The transcription factor Vox represses endoderm development by interacting with Casanova and Pou2. Development 140:1090–1099. doi: 10.1242/dev.082008 CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Peter Fabian
    • 1
  • Chrysoula N. Pantzartzi
    • 1
  • Iryna Kozmikova
    • 1
  • Zbynek Kozmik
    • 1
  1. 1.Institute of Molecular Genetics, Academy of Sciences of the Czech RepublicPrague 4Czech Republic

Personalised recommendations