Chordate ancestry of the neural crest: new insights from ascidians

This article reviews new insights from ascidians on the ancestry of vertebrate neural crest (NC) cells. Ascidians have neural crest-like cells (NCLC), which migrate from the dorsal midline, express some of the typical NC markers, and develop into body pigment cells. These characters suggest that primordial NC cells were already present in the common ancestor of the vertebrates and urochordates, which have been recently inferred as sister groups. The primitive role of NCLC may have been in pigment cell dispersal and development. Later, additional functions may have appeared in the vertebrate lineage, resulting in the evolution of definitive NC cells.     

Vertebrate cranial evolution: Contributions and conflict from the fossil record

In modern vertebrates, the craniofacial skeleton is complex, comprising cartilage and bone of the neurocranium, dermatocranium and splanchnocranium (and their derivatives), housing a range of sensory structures such as eyes, nasal and vestibulo-acoustic capsules, with the splanchnocranium including branchial arches, used in respiration and feeding. It is well understood that the skeleton derives from neural crest and mesoderm, while the sensory elements derive from ectodermal thickenings known as placodes. Recent research demonstrates that neural crest and placodes have an evolutionary history outside of vertebrates, while the vertebrate fossil record allows the sequence of the evolution of these various features to be understood. Stem-group vertebrates such as Metaspriggina walcotti (Burgess Shale, Middle Cambrian) possess eyes, paired nasal capsules and well-developed branchial arches, the latter derived from cranial neural crest in extant vertebrates, indicating that placodes and neural crest evolved over 500 million years ago. Since that time the vertebrate craniofacial skeleton has evolved, including different types of bone, of potential neural crest or mesodermal origin. One problematic part of the craniofacial skeleton concerns the evolution of the nasal organs, with evidence for both paired and unpaired nasal sacs being the primitive state for vertebrates.     

The ocular skeleton through the eye of evo-devo

An evolutionary developmental (evo-devo) approach to understanding the evolution, homology, and development of structures has proved important for unraveling complex integrated skeletal systems through the use of modules, or modularity. An ocular skeleton, which consists of cartilage and sometimes bone, is present in many vertebrates; however, the origin of these two components remains elusive. Using both paleontological and developmental data, I propose that the vertebrate ocular skeleton is neural crest derived and that a single cranial neural crest module divided early in vertebrate evolution, possibly during the Ordovician, to give rise to an endoskeletal component and an exoskeletal component within the eye. These two components subsequently became uncoupled with respect to timing, placement within the sclera and inductive epithelia, enabling them to evolve independently and to diversify. In some extant groups, these two modules have become reassociated with one another. Furthermore, the data suggest that the endoskeletal component of the ocular skeleton was likely established and therefore evolved before the exoskeletal component. This study provides important insights into the evolution of the ocular skeleton, a region with a long evolutionary history among vertebrates.     

The Ocular Skeleton Through The Eye of Evo-devo

An evolutionary developmental (evo-devo) approach to understanding the evolution, homology and development of structures has proved important for unraveling complex integrated skeletal systems through the use of modules, or modularity. An ocular skeleton, which consists of cartilage and sometimes bone, is present in many vertebrates; however the origin of these two components remains elusive. Using both palaeontological and developmental data, I propose that the vertebrate ocular skeleton is neural crest derived and that a single cranial neural crest module divided early in vertebrate evolution, possibly during the Ordovician, to give rise to an endoskeletal component and an exoskeletal component within the eye. These two components subsequently became uncoupled with respect to timing, placement within the sclera and inductive epithelia, enabling them to evolve independently and to diversify. In some extant groups, these two modules have become reassociated with one another. Furthermore, the data suggests that the endoskeletal component of the ocular skeleton was likely established and therefore evolved before the exoskeletal component. This study provides important insights into the evolution of the ocular skeleton, a region with a long evolutionary history amongst vertebrates. J. Exp. Zool. (Mol. Dev. Evol.) 9999B: 1-9, 2012. ? 2012 Wiley Periodicals, Inc.     

Chordate origins of the vertebrate central nervous system.

Fine structural, computerized three-dimensional (3D) mapping of cell connectivity in the amphioxus nervous system and comparative molecular genetic studies of amphioxus and tunicates have provided recent insights into the phylogenetic origin of the vertebrate nervous system. The results suggest that several of the genetic mechanisms for establishing and patterning the vertebrate nervous system already operated in the ancestral chordate and that the nerve cord of the proximate invertebrate ancestor of the vertebrates included a diencephalon, midbrain, hindbrain, and spinal cord. In contrast, the telencephalon, a midbrain-hindbrain boundary region with organizer properties, and the definitive neural crest appear to be vertebrate innovations.     

Evolutionary crossroads in developmental biology: cyclostomes (lamprey and hagfish)

Lampreys and hagfish, which together are known as the cyclostomes or 'agnathans', are the only surviving lineages of jawless fish. They diverged early in vertebrate evolution, before the origin of the hinged jaws that are characteristic of gnathostome (jawed) vertebrates and before the evolution of paired appendages. However, they do share numerous characteristics with jawed vertebrates. Studies of cyclostome development can thus help us to understand when, and how, key aspects of the vertebrate body evolved. Here, we summarise the development of cyclostomes, highlighting the key species studied and experimental methods available. We then discuss how studies of cyclostomes have provided important insight into the evolution of fins, jaws, skeleton and neural crest.     

The evolutionary origin of the vertebrate neural crest and its developmental gene regulatory network – insights from amphioxus

The neural crest is an embryonic cell population unique to vertebrates. During vertebrate embryogenesis, neural crest cells are first induced from the neural plate border; subsequently, they delaminate from the dorsal neural tube and migrate to their destination, where they differentiate into a wide variety of derivatives. The emergence of the neural crest is thought to be responsible for the evolution of many complex novel structures of vertebrates that are lacking in invertebrate chordates. Despite its central importance in understanding the origin of vertebrates, the evolutionary origin of the neural crest remains elusive. The basal chordate amphioxus (Branchiostoma floridae) occupies an outgroup position that is useful for investigating this question. In this review, I summarize recent genomic and comparative developmental studies between amphioxus and vertebrates and discuss their implications for the evolutionary origin of neural crest cells. I focus mainly on the origin of the gene regulatory network underlying neural crest development, and suggest several hypotheses regarding how this network could have been assembled during early vertebrate evolution.     

Evolutionary Origins of the Neural Crest and Neural Crest Cells

I evaluate the lines of evidence—cell types, genes, gene pathways, fossils—in putative chordate ancestors—cephalochordates and ascidians—pertaining to the evolutionary origin of the vertebrate neural crest. Given the intimate relationship between the neural crest and the dorsal nervous system during development, I discuss the dorsal nervous system in living (extant) members of the two groups, especially the nature, and genes, and gene regulatory networks of the brain to determine whether any cellular and/or molecular precursors (latent homologues) of the neural may have been present in ancestral cephalochordates or urochordates. I then examine those fossils that have been interpreted as basal chordates or cephalochordates to determine whether they shed any light on the origins of neural crest cell (NCC) derivatives. Do they have, for example, elements of a head skeleton or pharyngeal arches, two fundamental vertebrate characters (synapomorphies)? The third topic recognizes that the origin of the neural crest in the first vertebrates accompanied the evolution of a brain, a muscular pharynx, and paired sensory organs. In a paradigm-breaking hypothesis—often known as the ‘new head hypothesis’—Carl Gans and Glen Northcutt linked these evolutionary innovations to the evolution of the neural crest and ectodermal placodes (Gans and Northcutt Science 220:268-274, 1983. doi:10.1126/science.220.4594.268; Northcutt and Gans The Quarterly Review of Biology 58:1–28, 1983. doi:10.1086/413055). I outline the rationale behind the new head hypothesis before turning to an examination of the pivotal role played by NCCs in the evolution of pharyngeal arches, in the context of the craniofacial skeleton. Integrations between the evolving vertebrate brain, muscular pharynx and paired sensory organs may have necessitated that the pharyngeal arch skeletal system—and subsequently, the skeleton of the jaws and much of the skull (the first vertebrates being jawless)—evolved from NCCs whose developmental connections were to neural ectoderm and neurons rather than to mesoderm and connective tissue; mesoderm produces much of the vertebrate skeleton, including virtually all the skeleton outside the head. The origination of the pharyngeal arch skeleton raises the issue of the group of organisms in which and how cartilage arose as a skeletal tissue. Did cartilage arise in the basal proto-vertebrate from a single germ layer, cell layer or tissue, or were cells and/or genes co-opted from several layers or tissues? Two recent studies utilizing comparative genomics, bioinformatics, molecular fingerprinting, genetic labeling/cell selection, and GeneChip Microarray technologies are introduced as powerful ways to approach the questions that are central to this review.     

Origins and plasticity of neural crest cells and their roles in jaw and craniofacial evolution

The vertebrate head is a complex assemblage of cranial specializations, including the central and peripheral nervous systems, viscero- and neurocranium, musculature and connective tissue. The primary differences that exist between vertebrates and other chordates relate to their craniofacial organization. Therefore, evolution of the head is considered fundamental to the origins of vertebrates (Gans and Northcutt, 1983). The transition from invertebrate to vertebrate chordates was a multistep process, involving the formation and patterning of many new cell types and tissues. The evolution of early vertebrates, such as jawless fish, was accompanied by the emergence of a specialized set of cells, called neural crest cells which have long held a fascination for developmental and evolutionary biologists due to their considerable influence on the complex development of the vertebrate head. Although it has been classically thought that protochordates lacked neural crest counterparts, the recent identification and characterization of amphioxus and ascidian genes homologous to those involved in vertebrate neural crest development challenges this idea. Instead it suggests thatthe neural crest may not be a novel vertebrate cell population, but could have in fact originated from the protochordate dorsal midline epidermis. Consequently, the evolution of the neural crest cells could be reconsidered in terms of the acquisition of new cell properties such as delamination-migration and also multipotency which were key innovations that contributed to craniofacial development. In this review we discuss recent findings concerning the inductive origins of neural crest cells, as well as new insights into the mechanisms patterning this cell population and the subsequent influence this has had on craniofacial evolution.     

Neural crest contributions to the lamprey head

The neural crest is a vertebrate-specific cell population that contributes to the facial skeleton and other derivatives. We have performed focal DiI injection into the cranial neural tube of the developing lamprey in order to follow the migratory pathways of discrete groups of cells from origin to destination and to compare neural crest migratory pathways in a basal vertebrate to those of gnathostomes. The results show that the general pathways of cranial neural crest migration are conserved throughout the vertebrates, with cells migrating in streams analogous to the mandibular and hyoid streams. Caudal branchial neural crest cells migrate ventrally as a sheet of cells from the hindbrain and super-pharyngeal region of the neural tube and form a cylinder surrounding a core of mesoderm in each pharyngeal arch, similar to that seen in zebrafish and axolotl. In addition to these similarities, we also uncovered important differences. Migration into the presumptive caudal branchial arches of the lamprey involves both rostral and caudal movements of neural crest cells that have not been described in gnathostomes, suggesting that barriers that constrain rostrocaudal movement of cranial neural crest cells may have arisen after the agnathan/gnathostome split. Accordingly, neural crest cells from a single axial level contributed to multiple arches and there was extensive mixing between populations. There was no apparent filling of neural crest derivatives in a ventral-to-dorsal order, as has been observed in higher vertebrates, nor did we find evidence of a neural crest contribution to cranial sensory ganglia. These results suggest that migratory constraints and additional neural crest derivatives arose later in gnathostome evolution.     

Consideration of the neural crest and its skeletal derivatives in the context of novelty/innovation

I examine the neural crest and skeletal tissues derived from neural crest cells in the context of novelty/innovation by asking whether the neural crest is a novel tissue and whether the evolutionary origin of the neural crest required innovative developmental processes. As a vertebrate autapomorphy, the neural crest is a novel structure. I equate novelty with innovation and take a hierarchical approach. Some other workers separate the two, using novelty for new structures not found in an ancestor and not homologous with a feature in an ancestor, and innovation for the new processes required to generate the novel structure. While development clearly evolves, I do not separate those processes that result in the production of novel features from those that lead to change in existing structures, whether that change is a transition or transformation from one homologous feature to another (fins-->tetrapod limbs or locomotory appendages-->crustacean maxilliped feeding appendages). The existence of novelties causes us to consider the concept of latent homology. Neural crest cells form cartilage, dentine and bone. Cartilage is found in invertebrates and so is not a vertebrate innovation. No invertebrate cartilage mineralizes in vivo, although some can be induced to mineralize in vitro. Mineralization of cartilage in vivo is a vertebrate innovation. Dentine is a novel tissue that only forms from neural crest cells. Bone is a vertebrate innovation but not one exclusive to the neural crest. The developmental processes responsible for the neural crest and for these skeletal tissues did not arise de novo with the vertebrates. Novelty/innovation results from tinkering with existing processes, from the flexibility that arises from modifications of existing gene networks, and from the selective advantage provided by gene duplications or modifications.     

Developmental Gene Expression in Amphioxus: New Insights into the Evolutionary Origin of Vertebrate Brain Regions, Neural Crest, and Rostrocaudal Segmentation

SYNOPSIS. Amphioxus is widely held to be the closest invertebraterelative of the vertebrates and the best available stand-infor the proximate ancestor of the vertebrates. The spatiotemporalexpression patterns of developmental genes can help suggestbody part homologies between vertebrates and amphioxus. Thisapproach is illustrated using five homeobox genes (AmphiHoxl,AmphiHox2, AmphiOtx, AmphiDll, and AmphiEri) to provide insightsinto the evolutionary origins of three important vertebratefeatures: the major brain regions, the neural crest, and rostrocaudalsegmentation. During amphioxus development, the neural expressionpatterns of these genes are consistent with the presence ofa forebrain (detailed neuroanatomy indicates that the forebrainis all diencephalon without any telencephalon) and an extensivehindbrain; the possible presence of a midbrain requires additionalstudy. Further, during neurulation, the expression pattern ofAmphiDll as well as migratory cell behavior suggest that theepidermal cells bordering the neural plate may represent a phylogeneticprecursor of the vertebrate neural crest. Finally, when theparaxial mesoderm begins to segment, the earliest expressionof AmphiEn is detected in the posterior part of each nascentand newly formed somite. This pattern recalls the expressionof the segment-polarity gene engrailed during establishmentof the segments of metameric protostomes. Thus, during animalevolution, the role of engrailed in establishing and maintainingmetameric body plans may have arisen in a common segmented ancestorof both the protostomes and deuterostomes.     

Trunk lateral cells are neural crest-like cells in the ascidian Ciona intestinalis: insights into the ancestry and evolution of the neural crest

Neural crest-like cells (NCLC) that express the HNK-1 antigen and form body pigment cells were previously identified in diverse ascidian species. Here we investigate the embryonic origin, migratory activity, and neural crest related gene expression patterns of NCLC in the ascidian Ciona intestinalis. HNK-1 expression first appeared at about the time of larval hatching in dorsal cells of the posterior trunk. In swimming tadpoles, HNK-1 positive cells began to migrate, and after metamorphosis they were localized in the oral and atrial siphons, branchial gill slits, endostyle, and gut. Cleavage arrest experiments showed that NCLC are derived from the A7.6 cells, the precursors of trunk lateral cells (TLC), one of the three types of migratory mesenchymal cells in ascidian embryos. In cleavage arrested embryos, HNK-1 positive TLC were present on the lateral margins of the neural plate and later became localized adjacent to the posterior sensory vesicle, a staging zone for their migration after larval hatching. The Ciona orthologues of seven of sixteen genes that function in the vertebrate neural crest gene regulatory network are expressed in the A7.6/TLC lineage. The vertebrate counterparts of these genes function downstream of neural plate border specification in the regulatory network leading to neural crest development. The results suggest that NCLC and neural crest cells may be homologous cell types originating in the common ancestor of tunicates and vertebrates and support the possibility that a putative regulatory network governing NCLC development was co-opted to produce neural crest cells during vertebrate evolution.     

The serial transformation hypothesis of vertebrate origins: comment on "The new head hypothesis revisited"

In "The New Head Hypothesis Revisited," R.G. Northcutt (2005. J Exp Zool (Mol Dev Evol) 304B:274-297) evaluates the original postulates of this hypothesis (Northcutt and Gans, 1983. Quart Rev Biol 58:1-28). One of these postulates is that the brain-particularly the forebrain-evolved at essentially the same time as many neural crest and neurogenic placode derivatives-including sensory ganglia, dermal skeleton and sensory capsules of the head, and branchial arches. Northcutt's subsequent paper in 1996 concluded with the idea that transitional forms might not have occurred at the origin of vertebrates. Butler proposed a "Serial Transformation" hypothesis in 2000, which disputed the latter idea in that paired eyes and an enlarged brain (but lacking telencephalon) were envisioned to have been gained before elaboration of most neural crest and neurogenic placodal derivatives. In 2003, J. Mallatt and J.-Y. Chen analyzed fossils of the Cambrian animal Haikouella, which strongly support its affinity to craniates and aspects of several hypotheses, including Butler's transformational model, because although branchial bars are present, most other neural crest and placodal derivatives are absent, while paired eyes and an enlarged brain (but probably without telencephalon) are present. A more complete picture of vertebrate origins can be realized when the various hypotheses are constructively reconciled.     

Induction of the neural crest: a multigene process

In the embryo, the neural crest is an important population of cells that gives rise to diverse derivatives, including the peripheral nervous system and the craniofacial skeleton. Evolutionarily, the neural crest is of interest as an important innovation in vertebrates. Experimentally, it represents an excellent system for studying fundamental developmental processes, such as tissue induction. Classical embryologists have identified interactions between tissues that lead to neural crest formation. More recently, geneticists and molecular biologists have identified the genes that are involved in these interactions; this recent work has revealed that induction of the neural crest is a complex multistep process that involves many genes.     

Specification and formation of the neural crest: Perspectives on lineage segregation

The neural crest is a fascinating embryonic population unique to vertebrates that is endowed with remarkable differentiation capacity. Thought to originate from ectodermal tissue, neural crest cells generate neurons and glia of the peripheral nervous system, and melanocytes throughout the body. However, the neural crest also generates many ectomesenchymal derivatives in the cranial region, including cell types considered to be of mesodermal origin such as cartilage, bone, and adipose tissue. These ectomesenchymal derivatives play a critical role in the formation of the vertebrate head, and are thought to be a key attribute at the center of vertebrate evolution and diversity. Further, aberrant neural crest cell development and differentiation is the root cause of many human pathologies, including cancers, rare syndromes, and birth malformations. In this review, we discuss the current findings of neural crest cell ontogeny, and consider tissue, cell, and molecular contributions toward neural crest formation. We further provide current perspectives into the molecular network involved during the segregation of the neural crest lineage.     

Myosin-X is critical for migratory ability of Xenopus cranial neural crest cells

The neural crest is a highly migratory cell population, unique to vertebrates, that forms much of the craniofacial skeleton and peripheral nervous system. In exploring the cell biological basis underlying this behavior, we have identified an unconventional myosin, myosin-X (Myo10) that is required for neural crest migration. Myo10 is highly expressed in both premigratory and migrating cranial neural crest (CNC) cells in Xenopus embryos. Disrupting Myo10 expression using antisense morpholino oligonucleotides leads to impaired neural crest migration and subsequent cartilage formation, but only a slight delay in induction. In vivo grafting experiments reveal that Myo10-depleted CNC cells migrate a shorter distance and fail to segregate into distinct migratory streams. Finally, in vitro cultures and cell dissociation-reaggregation assays suggest that Myo10 may be critical for cell protrusion and cell-cell adhesion. These results demonstrate an essential role for Myo10 in normal cranial neural crest migration and suggest a link to cell-cell interactions and formation of processes.     

The origins and key innovations of vertebrates and arthropods

The Lower Cambrian Maotianshan Shale near Kunming (Southern China) not only retains beautifully preserved and diverse organisms but also documents a missing evolutionary history between living vertebrates and their amphioxus-like ancestor. Presented here are the key novelties both for the evolutionary origins of the chordates and for the evolutionary transition from amphioxus-like ancestor toward the vertebrates. The adaptation for a burrowing life style is considered a key adaptive pressure for generating novel chordate-only anatomical characters including the first axial skeleton, e.g., notochord and myotomes. The transition from amphioxus-like ancestor toward the vertebrates was presumably triggered by the way toward more active life style, resulted in the origination of numerous novel structures including neural crests, a more complex head with upper and lower lips, an active gilled pharyngeal system, a large brain, image-forming paired eyes, and a bony axial skeleton (vertebrae). The diverse limb-bearing organisms from the Lower Cambrian Maotianshan Shale arthropods representing different evolutionary stages shed light on the mysterious field of the evolutionary origins of the arthropods and reveal a grand scenario of how the arthropods paved their way through step-wise evolution from worm-like ancestor toward living crown-lineage arthropods.