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Hagfish embryology with reference to the evolution of the neural crest


Hagfish, which lack both jaws and vertebrae, have long been the subject of intense interest owing to their position at a crucial point in the evolutionary transition to a truly vertebrate body plan1,2,3,4. However, unlike the comparatively well characterized vertebrate agnathan lamprey, little is known about hagfish development. The inability to analyse hagfish at early embryonic stages has frustrated attempts to resolve questions with important phylogenetic implications, including fundamental ones relating to the emergence of the neural crest1,5,6. Here we report the obtainment of multiple pharyngula-stage embryos of the hagfish species Eptatretus burgeri and our preliminary analyses of their early development. We present histological evidence of putative neural crest cells, which appear as delaminated cells that migrate along pathways corresponding to neural crest cells in fish and amphibians2,7,8,9,10,11. Molecular cloning studies further revealed the expression of several regulatory genes, including cognates of Pax6, Pax3/7, SoxEa and Sox9, suggesting that the hagfish neural crest is specified by molecular mechanisms that are general to vertebrates. We propose that the neural crest emerged as a population of de-epithelialized migratory cells in a common vertebrate ancestor, and suggest that the possibility of classical and molecular embryology in hagfish opens up new approaches to clarifying the evolutionary history of vertebrates.

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Figure 1: Evolution of vertebrates and neural crest.
Figure 2: Embryos of Eptatretus burgeri.
Figure 3: Neural crest cells in E. burgeri.
Figure 4: Sox9 expression in the neural crest of E. burgeri.


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We thank O. Kakitani for the sample collection, H. Nagashima and Y. K. Ohya for technical advice, and R. Ladher and D. Sipp for critical reading of this manuscript. This work was supported by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

Author Contributions K.G.O. performed sample collection, maintenance of the aquarium tank, molecular cloning and in situ hybridization. S. Kuraku was particularly engaged in isolation of Snail family genes and performed phylogenetic analyses. S. Kuratani operated on the hagfish embryos and conducted histological analysis. K.G.O. and S. Kuratani wrote the manuscript. All of the authors discussed the results and commented on the manuscript.

Sequences for Pax6, Pax3/7, Snail, SoxEa and Sox9 from E. burgeri are deposited in DNA Data Bank of Japan (DDBJ) under accession numbers AB270704, AB270703, AB288229, AB288230 and AB270702, respectively.

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Correspondence to Shigeru Kuratani.

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Sequences for Pax6, Pax3/7, Snail, SoxEa and Sox9 from E. burgeri are deposited in DNA Data Bank of Japan (DDBJ) under accession numbers AB270704, AB270703, AB288229, AB288230 and AB270702, respectively. Reprints and permissions information is available at The authors declare no competing financial interests.

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Ota, K., Kuraku, S. & Kuratani, S. Hagfish embryology with reference to the evolution of the neural crest. Nature 446, 672–675 (2007).

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