Early segregation of a neuronal precursor cell line in the neural crest as revealed by culture in a chemically defined medium

doi:10.1016/0092-8674(83)90482-8

Cell

Volume 32, Issue 2, February 1983, Pages 627-638

https://doi.org/10.1016/0092-8674(83)90482-8 Get rights and content

Abstract

This article addresses the problem of the segregation of cell lines during the development of peripheral nervous system components from the neural crest. We show here that committed precursors of peripheral neurons are present in the crest before the migration of its cells has started. If cultured in a serum-deprived medium, a subpopulation of the crest cells readily differentiates into neurons without dividing. Neuronal markers such as neurofilament proteins and receptor sites for tetanus toxin are not expressed in the committed neuronal precursors, but appear after a few hours in culture. They are coexpressed in neurons with the mesenchymal intermediate filament protein, vimentin, which is common to all neural crest cells regardless of their prospective fate. A strong inhibitory effect of serum factor(s) on neurite outgrowth is demonstrated. We show also that conditions stimulating proliferation of crest cells are incompatible with promotion of neuronal differentiation and viceversa.

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Cited by (162)

The issue of the multipotency of the neural crest cells
2018, Developmental Biology
Citation Excerpt :
One is an adaptation from the classical technique described above for trunk NCC, using quail mesencephalon and rhombencephalon (Sieber-Blum et al., 1993; Trentin et al., 2004; Calloni et al., 2007, 2009). Another is to remove mesencephalic NCC in the course of their early migration under the superficial ectoderm (Ziller et al., 1983; Baroffio et al., 1988). Finally, some authors surgically remove the cranial neural folds; in that case, the migratory NCC are produced in vitro (e.g., Abzhanov et al., 2003; McGonnell and Graham, 2002; Ito and Morita, 1995).
In the neural primordium of vertebrate embryos, the neural crest (NC) displays a unique character: the capacity of its component cells to leave the neural primordium, migrate along definite (and, for long, not identified) routes in the developing embryo and invade virtually all tissues and organs, while producing a large array of differentiated cell types. The most striking diversity of the NC derivatives is found in its cephalic domain that produces, not only melanocytes and peripheral nerves and ganglia, but also various mesenchymal derivatives (connective tissues, bones, cartilages…) which, in other parts of the body, are mesoderm-derived. The aim of this article was to review the large amount of work that has been devoted to solving the problem of the differentiation capacities of individual NC cells (NCC) arising from both the cephalic and trunk levels of the neural axis. A variety of experimental designs applied to NCC either in vivo or in vitro are evaluated, including the possibility to culture them in crestospheres, a technique previously designed for cells of the CNS, and which reinforces the notion, previously put forward, of the existence of NC stem cells. At the trunk level, the developmental potentialities of the NCC are more restricted than in their cephalic counterparts, but, in addition to the neural-melanocytic fate that they exclusively express in vivo, it was clearly shown that they harbor mesenchymal capacities that can be revealed in vitro. Finally, a large amount of evidence has been obtained that, during the migration process, most of the NCC are multipotent with a variable array of potentialities among the cells considered. Investigations carried out in adults have shown that multipotent NC stem cells persist in the various sites of the body occupied by NCC. Enlightening new developments concerning the invasive capacity of NCC, the growing peripheral nerves were revealed as migration routes for NCC travelling to distant ventrolateral regions of the body. Designated “Schwann cell precursors” in the mouse embryo, these NCC can leave the nerves and are able to convert to a novel fate. The convertibility of the NC-derived cells, particularly evident in the Schwann cell-melanocyte lineage transition, has also been demonstrated for neuroendocrine cells of the adult carotid body and for the differentiation of parasympathetic neurons of ganglia distant from their origin, the NC. All these new developments attest the vitality of the research on the NC, a field that characterizes vertebrate development and for which the interest has constantly increased during the last decades.
Quo vadis: tracing the fate of neural crest cells
2017, Current Opinion in Neurobiology
Citation Excerpt :
However, growth factors have been identified that at least in vitro selectively promote expansion of neural crest-derived cells adopting particular fates [3]. Moreover, several reports suggested that neural crest cultures contain substantial numbers of lineage-restricted cells [4–7]. On the other hand, clonal analysis of neural crest cells in culture also provided strong evidence for multipotency of a considerable fraction of neural crest cells and for several fate-determining factors acting in an instructive rather than selective manner during neural crest development [3].
The neural crest is a transient structure in vertebrate embryos that produces migratory cells with an astonishing developmental potential. While neural crest fate maps have originally been established through interspecies transplantation assays, dye labeling, and retroviral infection, more recent methods rely on approaches involving transgenesis and genome editing. These technologies allowed the identification of minor neural crest-derived cell populations in tissues of non-neural crest origin. Furthermore, in vivo multipotency at the single cell level and stage-dependent fate acquisitions were demonstrated using genetic technologies. Finally, recent reports indicate that neural crest-derived cells become activated in response to injury to secrete factors supporting tissue repair. Thus, neural crest-derived cells apparently contribute to tissue formation and regeneration by cell autonomous and non-autonomous mechanisms.
The third wave: Intermediate filaments in the maturing nervous system
2017, Molecular and Cellular Neuroscience
Intermediate filaments are critical for the extreme structural specialisations of neurons, providing integrity in dynamic environments and efficient communication along axons a metre or more in length. As neurons mature, an initial expression of nestin and vimentin gives way to the neurofilament triplet proteins and α-internexin, substituted by peripherin in axons outside the CNS, which physically consolidate axons as they elongate and find their targets. Once connection is established, these proteins are transported, assembled, stabilised and modified, structurally transforming axons and dendrites as they acquire their full function. The interaction between these neurons and myelinating glial cells optimises the structure of axons for peak functional efficiency, a property retained across their lifespan. This finely calibrated structural regulation allows the nervous system to maintain timing precision and efficient control across large distances throughout somatic growth and, in maturity, as a plasticity mechanism allowing functional adaptation.
Molecular control of the neural crest and peripheral nervous system development
2015, Current Topics in Developmental Biology
Citation Excerpt :
A tightly regulated balance between extrinsically derived cues and intrinsic regulators is required for the appropriate specification, growth, and function of NCCs during PNS formation. Evidence suggests that the early NCC population is comprised of both fate-restricted and multipotent progenitors (Bronner-Fraser & Fraser, 1988; Coelho-Aguiar, Le Douarin, & Dupin, 2013; Crane & Trainor, 2006; Fraser & Bronner-Fraser, 1991; Greenwood, Turner, & Anderson, 1999; Krispin, Nitzan, & Kalcheim, 2010; Le Douarin & Dupin, 2003; Ziller, Dupin, Brazeau, Paulin, & Le Douarin, 1983). During the course of development in vivo most NCCs undergo progressive fate restriction.
A transient and unique population of multipotent stem cells, known as neural crest cells (NCCs), generate a bewildering array of cell types during vertebrate development. An attractive model among developmental biologists, the study of NCC biology has provided a wealth of knowledge regarding the cellular and molecular mechanisms important for embryogenesis. Studies in numerous species have defined how distinct phases of NCC specification, proliferation, migration, and survival contribute to the formation of multiple functionally distinct organ systems. NCC contributions to the peripheral nervous system (PNS) are well known. Critical developmental processes have been defined that provide outstanding models for understanding how extracellular stimuli, cell–cell interactions, and transcriptional networks cooperate to direct cellular diversification and PNS morphogenesis. Dissecting the complex extracellular and intracellular mechanisms that mediate the formation of the PNS from NCCs may have important therapeutic implications for neurocristopathies, neuropathies, and certain forms of cancer.
Bone morphogenetic proteins regulate enteric gliogenesis by modulating ErbB3 signaling
2011, Developmental Biology
The neural crest-derived cell population that colonizes the bowel (ENCDC) contains proliferating neural/glial progenitors. We tested the hypothesis that bone morphogenetic proteins (BMPs 2 and 4), which are known to promote enteric neuronal differentiation at the expense of proliferation, function similarly in gliogenesis. Enteric gliogenesis was analyzed in mice that overexpress the BMP antagonist, noggin, or BMP4 in the primordial ENS. Noggin-induced loss-of-function decreased, while BMP4-induced gain-of-function increased the glial density and glia/neuron ratio. When added to immunoisolated ENCDC, BMPs provoked nuclear translocation of phosphorylated SMAD proteins and enhanced both glial differentiation and expression of the neuregulin receptor ErbB3. ErbB3 transcripts were detected in E12 rat gut, before glial markers are expressed; moreover, expression of the ErbB3 ligand, glial growth factor 2 (GGF2) escalated rapidly after its first detection at E14. ErbB3-immunoreactive cells were located in the ENS of fetal and adult mice. GGF2 stimulated gliogenesis and proliferation and inhibited glial cell derived neurotrophic factor (GDNF)-promoted neurogenesis. Enhanced glial apoptosis occurred following GGF2 withdrawal; BMPs intensified this GGF2-dependence and reduced GGF2-stimulated proliferation. These observations support the hypotheses that BMPs are required for enteric gliogenesis and act by promoting responsiveness of ENCDC to ErbB3 ligands such as GGF2.
Neural crest progenitors and stem cells
2007, Comptes Rendus - Biologies
Citation Excerpt :
How the multipotent trunk NC cells integrate these different signals is not yet fully understood [30], although Notch proved to be dominant over BMP2 signalling by triggering irreversible switch from neurogenesis to gliogenesis [31]. Interestingly, these cells were not capable to generate sensory neurones, in support to previous data suggesting an early segregation of sensory and autonomic NC lineages [32–34]. Sensory neurones were found to develop from cultures of the mammalian neural primordium (including the premigratory trunk NC) and arise from determined progenitors that do not respond to BMP2 autonomic cue [35].
In the vertebrate embryo, multiple cell types originate from a common structure, the neural crest (NC), which forms at the dorsal tips of the neural epithelium. The NC gives rise to migratory cells that colonise a wide range of embryonic tissues and later differentiate into neurones and glial cells of the peripheral nervous system (PNS), pigment cells (melanocytes) in the skin and endocrine cells in the adrenal and thyroid glands. In the head and the neck, the NC also yields mesenchymal cells that form craniofacial cartilages, bones, dermis, adipose tissue, and vascular smooth muscle cells. The NC is therefore a model system to study cell diversification during embryogenesis and phenotype maintenance in the adult.
By analysing the developmental potentials of quail NC cells in clonal cultures, we have shown that the migratory NC is a collection of heterogeneous progenitors, including various types of intermediate precursors and highly multipotent cells, some of which being endowed of self-renewal capacity. We also have identified common progenitors for mesenchymal derivatives and neural/melanocytic cells in the cephalic NC. These results are consistent with a hierarchical model of lineage segregation wherein environmental cytokines control the fate of progenitors and stem cells. One of these cytokines, the endothelin3 peptide, promotes the survival, proliferation, and self-renewal capacity of common progenitors for glial cells and melanocytes. At post-migratory stages, when they have already differentiated, NC-derived cells exhibit phenotypic plasticity. Epidermal pigment cells and Schwann cells from peripheral nerves in single-cell culture are able to reverse into multipotent NC-like progenitors endowed with self-renewal.
Therefore, stem cell properties are expressed by a variety of NC progenitors and can be re-acquired by differentiated cells of NC origin, suggesting potential function for repair. To cite this article: E. Dupin et al., C. R. Biologies 330 (2007).
Chez l'embryon de Vertébrés, différents types cellulaires ont une origine commune dans une structure transitoire, la crête neurale, qui se forme aux bords dorsaux de l'épithélium neural. La crête neurale donne naissance à des cellules migratrices, qui colonisent un grand nombre de tissus et, plus tard, se différencient en neurones et cellules gliales dans le système nerveux périphérique, en cellules pigmentaires (mélanocytes) dans la peau et en cellules endocrines dans les glandes surrénales et la thyroïde. Dans la tête, la crête neurale fournit également des cellules mésenchymateuses, qui formeront les cartilages et les os du crâne et de la face, ainsi que du derme, du tissu adipeux et des cellules musculaires de la paroi des vaisseaux. La crête neurale constitue donc un modèle pour l'étude des mécanismes de la diversification cellulaire au cours du développement et du maintien des phénotypes cellulaires chez l'adulte.
En réalisant une analyse in vitro des potentiels de développement des cellules de la crête neurale de caille cultivées en cultures clonales, nous avons montré que les cellules de la crête neurale en migration constituent une collection de progéniteurs, où sont présents des cellules multipotentes, des précurseurs déjà déterminés et différents types de précurseurs oligopotents intermédiaires. La crête neurale céphalique comprend notamment des précurseurs communs aux cellules mésenchymateuses (chondrocytes, cellules musculaires vasculaires) et aux cellules nerveuses et pigmentaires. Certains de ces précurseurs peuvent se propager in vitro après sous-clonages successifs, et possèdent donc la capacité de s'autorenouveler, caractéristique des cellules souches. Ces résultats suggèrent un modèle hiérarchique dans lequel les lignages dérivés de la crête neurale sont ségrégés au cours des restrictions progressives des potentialités des cellules souches et progéniteurs oligopotents, sous l'influence de facteurs environnementaux. L'un de ces facteurs, le peptide endothéline3, est nécessaire au développement des mélanocytes dans la peau, et favorise la survie, la prolifération et l'autorenouvellement in vitro des précurseurs donnant naissance aux cellules pigmentaires et gliales.
Aux stades les plus tardifs du développement, les types cellulaires dérivés de la crête neurale montrent une plasticité phénotypique remarquable. Ainsi, est-il possible, en culture in vitro, d'obtenir l'interconversion de cellules pigmentaires et de cellules gliales différenciées, isolées respectivement de l'épiderme et des nerfs périphériques de l'embryon. Dans le cas des cellules pigmentaires, nous avons montré que, sous l'influence mitogénique de l'endothéline3, la reprogrammation phénotypique s'accomplit en culture après dédifférenciation et retour à l'état de cellule souche multipotente.
Il apparaît donc qu'au cours du développement, des propriétés caractéristiques de cellules souches sont présentes dans une diversité de progéniteurs dérivés de la crête neurale. En outre, ces propriétés peuvent être ré-exprimées par des cellules différenciées, suggérant un potentiel de réparation chez l'adulte. Pour citer cet article : E. Dupin et al., C. R. Biologies 330 (2007).

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Cell

ArticleEarly segregation of a neuronal precursor cell line in the neural crest as revealed by culture in a chemically defined medium

Abstract

Exp. Cell Res.

Regulatory Peptides

Dev. Biol.

Exp. Cell Res.

Cell

Exp. Cell Res.

J. Biol. Chem.

Brain Res.

Exp. Cell Res.

Cell

Cell

Dev. Biol.

Prog. Brain Res.

The staining of paraffin sections of nervous tissues with activated protargol

Growth of a rat neuroblastoma cell line in serum-free supplemented medium

Control of differentiation pathways by the extracellular environment in an embryonal carcinoma cell line

Human monoclonal IgM with autoantibody activity against intermediate filaments

Cholinesterase in embryonic development

Progr. Histochem. Cytochem.

Differentiation of autonomic neuron precursors in vitro: cholinergic and adrenergic traits in cultured neural crest cells

J. Neursci.

Vimentin and 70k neurofilament protein co-exist in embryonic neurones from spinal ganglia

J. Neurochem.

Growth and development of sympathetic neurons in tissue culture

A “direct-colouring” thiocholine method for cholinesterases

J. Histochem. Cytochem.

Cleavage of structural proteins during the assembly of the head of bacteriophage T4

Nature

Article
Early segregation of a neuronal precursor cell line in the neural crest as revealed by culture in a chemically defined medium