Craniofacial development of hagfishes and the evolution of vertebrates

Journal name:
Nature
Volume:
493,
Pages:
175–180
Date published:
DOI:
doi:10.1038/nature11794
Received
Accepted
Published online

Abstract

Cyclostomes, the living jawless vertebrates including hagfishes and lampreys, represent the most basal lineage of vertebrates. Although the monophyly of cyclostomes has been supported by recent molecular analyses, the phenotypic traits of hagfishes, especially the lack of some vertebrate-defining features and the reported endodermal origin of the adenohypophysis, have been interpreted as hagfishes exhibiting a more ancestral state than those of all other vertebrates. Furthermore, the adult anatomy of hagfishes cannot be compared easily with that of lampreys. Here we describe the craniofacial development of a series of staged hagfish embryos, which shows that their adenohypophysis arises ectodermally, consistent with the molecular phylogenetic data. This finding also allowed us to identify a pan-cyclostome pattern, one not shared by jawed vertebrates. Comparative analyses indicated that many of the hagfish-specific traits can be explained by changes secondarily introduced into the hagfish lineage. We also propose a possibility that the pan-cyclostome pattern may reflect the ancestral programme for the craniofacial development of all living vertebrates.

At a glance

Figures

  1. Cyclostomes and gnathostomes.
    Figure 1: Cyclostomes and gnathostomes.

    a–c, Eptatretus burgeri (hagfish). a, Dorsal view. b, Ventral view of a hagfish head showing a single nostril (en) and jawless mouth (mo). c, A diagram of the head anatomy. dg, Lethenteron japonicum (lamprey). d, Lateral view. e, Ventral view to show the oral funnel surrounding the mouth. f, Dorsal view to show the dorsal nostril. g, A diagram of the head anatomy. h, i, Scyliorhinus torazame (shark). h, Lateral view. i, Frontal view to show upper and lower jaws (uj and lj), and paired nostrils (en). j, Diagram of the generalized gnathostome head. ah, adenohypophysis; et, ethmoidal region; llp, lower lip; ne, nasal epithelium; ons, oronasohypophyseal septum; T1–4, tentacles 1–4; ulp, upper lip.

  2. Craniofacial development of E. burgeri.
    Figure 2: Craniofacial development of E. burgeri.

    af, Frontal (a, c, e) and lateral (b, d, f) views. The ectoderm is coloured light blue, the endoderm is coloured yellow, and the mesoendoderm is coloured orange. Arrows indicate the SOM. g, h, Sagittal sections at the plane represented in e. ip, Lateral views (i, k, m, o) and sagittal sections (j, l, n, p). Inset in p shows the differentiated adenohypophysis (ah). e, eye; ht, hypothalamus; nhd, nasohypophyseal duct; nhp, nasohypophyseal plate; nt, notochord; oc, oral cavity; onc, oronasohypophyseal cavity; opm, oropharyngeal membrane; pcp, prechordal plate; p1, pharyngeal pouch 1; st, stomodeum. See Fig. 1 for other abbreviations. Scale bars, 100μm. Numbers indicate developmental stages.

  3. Embryonic gene expression in Eptatretus burgeri.
    Figure 3: Embryonic gene expression in Eptatretus burgeri.

    af, Expression of genes involved in the NHP development from stages 26 to 40. g, Dorsal view of three-dimensional-reconstructed image. The blue area shows pan-placodal domain. h, Schematic diagrams summarizing the gene expression in the embryos. In the graph, the hypothalamic domain is characterized by EbHh1, EbNkx2.1 and EbSix3/6 expression. Adenohypophysis-like expression (blue bars) is restricted in the ectodermal domain rostral to and including the oropharyngeal membrane (opm). ch, chiasma; fb, forebrain; hb, hindbrain; mb, midbrain; ot, otic placode. See Figs 1 and 2 for other abbreviations. Scale bars, 100 μm.

  4. Comparison of vertebrate heads.
    Figure 4: Comparison of vertebrate heads.

    ah, Mid-sagittal sections (a, e) and ventral views (bd, fh) of Eptatretus burgeri (a, stage 45; b, stage 40; c, stage 50; d, stage 53; SOM has been removed from bd) and Lethenteron japonicum (e, stage 26; f, stage 23; g, stage 26; h, stage 27). ik, Ventral views of Scyliorhinus torazame (i, stage 20; j, stage 25; k, stage 28). The premandibular domain (asterisk) is comparable to the post-hypophyseal process (php) of cyclostomes. The blue domain (lateral and medial nasal prominences (fnp+lnp)) resembles the anterior nasal process (anp) of cyclostomes (ref. 22). l, Generalized patterns of vertebrate heads. ma, mandibular arch; np, nasal placodes. See Figs 1 and 2 for other abbreviations. Scale bars, 100μm.

  5. Evolution of the vertebrate head.
    Figure 5: Evolution of the vertebrate head.

    A model based on the assumption that the pan-cyclostome pattern (in box) represents a plesiomorphic pattern for the entire vertebrates. The hagfish oronasohypophyseal septum (ons) and the lamprey upper lip (ulp) are both derivatives of the post-hypophyseal process (php). Separation of the NHP into the nasal epithelium (ne) and adenohypophysis (ah) may have led to the pattern of galeaspids42. Further bilateral separation of the nasal epithelia is regarded as a prerequisite for acquisition of the trabecula (tr) and upper and lower jaws (uj and lj) in crown gnathostomes. ph, pharynx. See Fig. 1, 2 and 4 for other abbreviations.

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Author information

Affiliations

  1. Department of Biology, Graduate School of Science, Kobe University, Kobe 657-8501, Japan

    • Yasuhiro Oisi
  2. Laboratory for Evolutionary Morphology, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan

    • Yasuhiro Oisi,
    • Satoko Fujimoto &
    • Shigeru Kuratani
  3. Marine Research Station, Institute of Cellular and Organismic Biology, Academia Sinica, Yilan 26242, Taiwan

    • Kinya G. Ota
  4. Genome Resource and Analysis Unit, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan

    • Shigehiro Kuraku

Contributions

Y.O. performed sample collection, maintenance of aquarium tanks, histological preparation and three-dimensional reconstructions. Y.O. and S.F. performed the molecular cloning of EbPitxA, EbSix3/6A, EbFgf8/17A, EbHh1, EbTbx1/10A, EbSoxB1, EbLhx3/4A and EbNkx2.1 genes and in situ hybridization. S. Kuraku performed the molecular evolutionary analysis. Y.O. and S. Kuratani wrote the first draft of the manuscript. K.G.O., S. Kuraku and S. Kuratani wrote the final version of the manuscript. All of the authors discussed the results and commented on the manuscript.

Competing financial interests

The authors declare no competing financial interests.

Corresponding author

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Sequences for EbPitxA, EbSix3/6A, EbFgf8/17, EbHh1, EbTbx1/10A, EbSoxB1, EbLhx3/4A and EbNkx2.1 from E. burgeri are deposited in DDBJ/GenBank/EMBL under accession numbers AB703678AB703682, AB729075AB729076, and AB747372.

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Supplementary information

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  1. Supplementary Information (12.5M)

    This file contains Supplementary Figures 1-12, which show embryonic morphological data, Supplementary Table 1 and additional references.

Comments

  1. Report this comment #54730

    Peter Gibson said:

    Oisi et al have given an anatomical explanation for the single nasal opening in hagfish and lamprey. This does not appear to take into account the function of this orifice and that of the month. The two are perhaps generally confused. The present nasal opening was possibly the mouth in the early chordates. That is why it is a single opening. Filter feeders such as Amphioxus only needs one opening (the pharynx). Possibly the species evolved from an entirely planktonic stage (e.g. A. palagicus). This was trapped above a thermocline and only later, after destruction of the thermocline, become benthic. The emphasis was then on eating solids or for which a mouth and teeth were necessary. This has remained so to the present. The original opening was possibly used for this purpose in the Ostracodrems which were benthic. Clearly this is unsatisfactory so the present day mouth with teeth was separated by septum (Fig 5, ONS). This probably grew in from the edge of the pharynx leaving a passage to the gut. The original mouth was used for respiration as it still is and for sensing nature of the water (close proximity to the central nerve chord) (Fig 5, ne, ah). As food became larger so did the mouth. In the clycostomes the new mouth was clamped onto prey and could not be used for respiration. The same is still true vertebrates when eating.
    Amphioxus has a single anterior orifice ? the pharynx and atrium. In evolution the gill slits moved caudally (or the preoral region moved rostrally). The atrium becomes the mouth with teeth (adapted scales moving into the new mouth). In the detailed analysis of head of hagfish and lamprey Oisi et al suggest that the nasal tube become separated (shut off) from the evolving mouth and buccal cavity (Fig 3, OMP). To show this convincingly must be difficult. I suggest that the nasal passage has always remained open. Since the function of the duct was for respiration and not feeding there was no need for it to remain as a single opening. Two opening, as with eyes, gives a degree of directional location.
    The larval-like form of Amphioxus and other chordates (as well as invertebrates in general) have a single feeding opening in the side of the body. This is the pharynx. The ciliated band on the body surface and pharynx may have given rise to the gills (ciliated structures) through overgrowth of the sides of the body to give the atrium and its atriopore. The gills in vertebrate evolution gave rise to jaws and ears. The Eurasian tube may be a remnant of the atrium (divided atriopore). This is a demonstration that nothing is lost during evolution: structures are shuffled about the body to be used for other purposes.

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