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Identification of a rudimentary neural crest in a non-vertebrate chordate

Abstract

Neural crest arises at the neural plate border, expresses a core set of regulatory genes and produces a diverse array of cell types, including ectomesenchyme derivatives that elaborate the vertebrate head1,2. The evolution of neural crest has been proposed to be a key event leading to the appearance of new cell types that fostered the transition from filter feeding to active predation in ancestral vertebrates3. However, the origin of neural crest remains controversial, as homologous cell types have not been unambiguously identified in non-vertebrate chordates1,4. Here we show that the tunicate Ciona intestinalis possesses a cephalic melanocyte lineage (a9.49) similar to neural crest that can be reprogrammed into migrating ‘ectomesenchyme’ by the targeted misexpression of Twist (also known as twist-like 2). Our results suggest that the neural crest melanocyte regulatory network pre-dated the divergence of tunicates and vertebrates. We propose that the co-option of mesenchyme determinants, such as Twist, into the neural plate ectoderm was crucial to the emergence of the vertebrate ‘new head’3.

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Figure 1: Wnt signalling promotes ocellus formation.
Figure 2: FoxD represses Mitf in the ocellus.
Figure 3: Twist reprograms the a9.49 lineage.
Figure 4: Lineage tracing of reprogrammed a9.49 cells.

References

  1. 1

    Bronner, M. E. & Le Douarin, N. M. Evolution and development of the neural crest: An overview. Dev. Biol. 366, 2–9 (2012)

    CAS  Article  Google Scholar 

  2. 2

    Le Douarin, N. M. et al. Neural crest cell plasticity and its limits. Development 131, 4637–4650 (2004)

    CAS  Article  Google Scholar 

  3. 3

    Gans, C. & Northcutt, R. G. Neural crest and the origin of vertebrates: a new head. Science 220, 268–273 (1983)

    ADS  CAS  Article  Google Scholar 

  4. 4

    Yu, J. K., Meulemans, D., McKeown, S. J. & Bronner-Fraser, M. Insights from the amphioxus genome on the origin of vertebrate neural crest. Genome Res. 18, 1127–1132 (2008)

    CAS  Article  Google Scholar 

  5. 5

    Delsuc, F., Brinkmann, H., Chourrout, D. & Philippe, H. Tunicates and not cephalochordates are the closest living relatives of vertebrates. Nature 439, 965–968 (2006)

    ADS  CAS  Article  Google Scholar 

  6. 6

    Jeffery, W. R., Strickler, A. G. & Yamamoto, Y. Migratory neural crest-like cells form body pigmentation in a urochordate embryo. Nature 431, 696–699 (2004)

    ADS  CAS  Article  Google Scholar 

  7. 7

    Jeffery, W. R. Ascidian neural crest-like cells: phylogenetic distribution, relationship to larval complexity, and pigment cell fate. J. Exp. Zool. B 306B, 470–480 (2006)

    CAS  Article  Google Scholar 

  8. 8

    Jeffery, W. R. et al. Trunk lateral cells are neural crest-like cells in the ascidian Ciona intestinalis: Insights into the ancestry and evolution of the neural crest. Dev. Biol. 324, 152–160 (2008)

    CAS  Article  Google Scholar 

  9. 9

    Tassy, O. et al. The ANISEED database: digital representation, formalization, and elucidation of a chordate developmental program. Genome Res. 20, 1459–1468 (2010)

    CAS  Article  Google Scholar 

  10. 10

    Russo, M. T. et al. Regulatory elements controlling Ci-msxb tissue-specific expression during Ciona intestinalis embryonic development. Dev. Biol. 267, 517–528 (2004)

    CAS  Article  Google Scholar 

  11. 11

    Imai, K. S., Levine, M., Satoh, N. & Satou, Y. Regulatory blueprint for a chordate embryo. Science 312, 1183–1187 (2006)

    ADS  CAS  Article  Google Scholar 

  12. 12

    Wada, H. & Makabe, K. Genome duplications of early vertebrates as a possible chronicle of the evolutionary history of the neural crest. Int. J. Biol. Sci. 2, 133–141 (2006)

    CAS  Article  Google Scholar 

  13. 13

    Squarzoni, P., Parveen, F., Zanetti, L., Ristoratore, F. & Spagnuolo, A. FGF/MAPK/Ets signaling renders pigment cell precursors competent to respond to Wnt signal by directly controlling Ci-Tcf transcription. Development 138, 1421–1432 (2011)

    CAS  Article  Google Scholar 

  14. 14

    Curran, K. et al. Interplay between Foxd3 and Mitf regulates cell fate plasticity in the zebrafish neural crest. Dev. Biol. 344, 107–118 (2010)

    CAS  Article  Google Scholar 

  15. 15

    Yajima, I. et al. Cloning and functional analysis of ascidian Mitf in vivo: insights into the origin of vertebrate pigment cells. Mech. Dev. 120, 1489–1504 (2003)

    CAS  Article  Google Scholar 

  16. 16

    Nishida, H. & Satoh, N. Determination and regulation in the pigment cell lineage of the ascidian embryo. Dev. Biol. 132, 355–367 (1989)

    CAS  Article  Google Scholar 

  17. 17

    Dorsky, R. I., Moon, R. T. & Raible, D. W. Control of neural crest cell fate by the Wnt signalling pathway. Nature 396, 370–373 (1998)

    ADS  CAS  Article  Google Scholar 

  18. 18

    Thomas, A. J. & Erickson, C. A. FOXD3 regulates the lineage switch between neural crest-derived glial cells and pigment cells by repressing MITF through a non-canonical mechanism. Development 136, 1849–1858 (2009)

    CAS  Article  Google Scholar 

  19. 19

    Imai, K. S., Satoh, N. & Satou, Y. An essential role of a FoxD gene in notochord induction in Ciona embryos. Development 129, 3441–3453 (2002)

    CAS  PubMed  Google Scholar 

  20. 20

    Yoshida, T., Vivatbutsiri, P., Morriss-Kay, G., Saga, Y. & Iseki, S. Cell lineage in mammalian craniofacial mesenchyme. Mech. Dev. 125, 797–808 (2008)

    CAS  Article  Google Scholar 

  21. 21

    Bildsoe, H. et al. Requirement for Twist1 in frontonasal and skull vault development in the mouse embryo. Dev. Biol. 331, 176–188 (2009)

    CAS  Article  Google Scholar 

  22. 22

    Soo, K. et al. Twist function is required for the morphogenesis of the cephalic neural tube and the differentiation of the cranial neural crest cells in the mouse embryo. Dev. Biol. 247, 251–270 (2002)

    CAS  Article  Google Scholar 

  23. 23

    Hopwood, N. D., Pluck, A. & Gurdon, J. B. A Xenopus mRNA related to Drosophila twist is expressed in response to induction in the mesoderm and the neural crest. Cell 59, 893–903 (1989)

    CAS  Article  Google Scholar 

  24. 24

    Vincentz, J. W. et al. An absence of Twist1 results in aberrant cardiac neural crest morphogenesis. Dev. Biol. 320, 131–139 (2008)

    CAS  Article  Google Scholar 

  25. 25

    Tokuoka, M., Satoh, N. & Satou, Y. A bHLH transcription factor gene, Twist-like 1, is essential for the formation of mesodermal tissues of Ciona juveniles. Dev. Biol. 288, 387–396 (2005)

    CAS  Article  Google Scholar 

  26. 26

    Vlaeminck-Guillem, V. et al. The Ets family member Erg gene is expressed in mesodermal tissues and neural crests at fundamental steps during mouse embryogenesis. Mech. Dev. 91, 331–335 (2000)

    CAS  Article  Google Scholar 

  27. 27

    Horie, T. et al. Ependymal cells of chordate larvae are stem-like cells that form the adult nervous system. Nature 469, 525–528 (2011)

    ADS  CAS  Article  Google Scholar 

  28. 28

    Shimeld, S. M. & Holland, P. W. H. Vertebrate innovations. Proc. Natl Acad. Sci. USA 97, 4449–4452 (2000)

    ADS  CAS  Article  Google Scholar 

  29. 29

    Shi, W. & Levine, M. Ephrin signaling establishes asymmetric cell fates in an endomesoderm lineage of the Ciona embryo. Development 135, 931–940 (2008)

    CAS  Article  Google Scholar 

  30. 30

    Stolfi, A. & Levine, M. Neuronal subtype specification in the spinal cord of a protovertebrate. Development 138, 995–1004 (2011)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank A. Stolfi for his continued support and guidance, Y. Satou for isolating the Twist enhancer, N. Ellis for cloning Dmbx>Twist and B. Gainous for critical reading of the manuscript. P.B.A is supported by a graduate fellowship from the US National Science Foundation. This work was supported by a grant from the US National Institutes of Health (NS 076542).

Author information

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Authors

Contributions

P.B.A. designed and performed most experiments in consultation with M.L. E.W. isolated the cis-regulatory element for the βγ-crystallin reporter and made the stable β-catenin transgene. I.A.N. examined Mech2 and Erg expression in wild-type and reprogrammed tailbud embryos. P.B.A., M.L. and E.W. wrote the manuscript.

Corresponding author

Correspondence to Michael Levine.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-10 and Supplementary Table 1. (PDF 5463 kb)

Lateral view of a Mitf>Twist expressing larva labeled with Tyr>mCherry

The time-lapse covers a period of about 168 minutes, during which time the protrusive behavior of the a9.49 lineage is seen. The a9.49 lineage misexpressing Twist commonly forms filopodia, which is indicative of cellular reprogramming to mesenchymal fate. (MOV 2329 kb)

Lateral view of a Mitf>Twist expressing early juvenile labeled with Tyr>mCherry

The lateral view of a Mitf>Twist expressing early juvenile labeled with Tyr>mCherry (grey is UV autofluorescence). The time-lapse covers about 6 hours of development, during which time the ectopic a9.49 lineage can be seen migrating around the periphery like the endogenous migrating tunic cells (autofluorescence). The movie loops twice and during the second time the migrating descendants of a9.49 are labeled with asterisks. Other Tyr>mCherry that have been incorporated into the mesenchyme remain stationary. (MOV 19826 kb)

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Abitua, P., Wagner, E., Navarrete, I. et al. Identification of a rudimentary neural crest in a non-vertebrate chordate. Nature 492, 104–107 (2012). https://doi.org/10.1038/nature11589

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