Avian neural crest cells can migrate in the dorsolateral path only if they are specified as melanocytes

Neural crest cells are conventionally believed to migrate arbitrarily into various pathways and to differentiate according to the environmental cues that they encounter. We present data consistent with the notion that melanocytes are directed, by virtue of their phenotype, into the dorsolateral path, whereas other neural crest derivatives are excluded. In the avian embryo, trunk neural crest cells that migrate ventrally differentiate largely into neurons and glial cells of the peripheral nervous system. Neural crest cells that migrate into the dorsolateral path become melanocytes, the pigment cells of the skin. Neural crest cells destined for the dorsolateral path are delayed in their migration until at least 24 hours after migration commences ventrally. Previous studies have suggested that invasion into the dorsolateral path is dependent upon a change in the migratory environment. A complementary possibility is that as neural crest cells differentiate into melanocytes they acquire the ability to take this pathway. When quail neural crest cells that have been grown in culture for 12 hours are labeled with Fluoro-gold and then grafted into the early migratory pathway at the thoracic level, they migrate only ventrally and are coincident with the host neural crest. When fully differentiated melanocytes (96 hours old) are back-grafted under identical conditions, however, they enter the dorsolateral path and invade the ectoderm at least one day prior to the host neural crest. Likewise, neural crest cells that have been cultured for at least 20 hours and are enriched in melanoblasts immediately migrate in the dorsolateral path, in addition to the ventral path, when back-grafted into the thoracic level. A population of neural crest cells depleted of melanoblasts--crest cells derived from the branchial arches--are not able to invade the dorsolateral path, suggesting that only pigment cells or their precursors are able to take this migratory route. These results suggest that as neural crest cells differentiate into melanocytes they can exploit the dorsolateral path immediately. Even when 12-hour crest cells are grafted into stage 19–21 embryos at an axial level where host crest are invading the dorsolateral path, these young neural crest cells do not migrate dorsolaterally. Conversely, melanoblasts or melanocytes grafted under the same circumstances are found in the ectoderm. These latter results suggest that during normal development neural crest cells must be specified, if not already beginning to differentiate, as melanocytes in order to take this path.(ABSTRACT TRUNCATED AT 400 WORDS)

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Jaw and branchial arch mutants in zebrafish I: branchial arches

Development ◽

10.1242/dev.123.1.329 ◽

1996 ◽

Vol 123 (1) ◽

pp. 329-344 ◽

Cited By ~ 21

Author(s):

T.F. Schilling ◽

T. Piotrowski ◽

H. Grandel ◽

M. Brand ◽

C.P. Heisenberg ◽

...

Keyword(s):

Neural Crest ◽

Zebrafish Embryo ◽

Neural Crest Cells ◽

Branchial Arch ◽

Pigment Cells ◽

Pharyngeal Arches ◽

Pectoral Fins ◽

Branchial Arches ◽

Group Members ◽

The Brain

Jaws and branchial arches together are a basic, segmented feature of the vertebrate head. Seven arches develop in the zebrafish embryo (Danio rerio), derived largely from neural crest cells that form the cartilaginous skeleton. In this and the following paper we describe the phenotypes of 109 arch mutants, focusing here on three classes that affect the posterior pharyngeal arches, including the hyoid and five gill-bearing arches. In lockjaw, the hyoid arch is strongly reduced and subsets of branchial arches do not develop. Mutants of a large second class, designated the flathead group, lack several adjacent branchial arches and their associated cartilages. Five alleles at the flathead locus all lead to larvae that lack arches 4–6. Among 34 other flathead group members complementation tests are incomplete, but at least six unique phenotypes can be distinguished. These all delete continuous stretches of adjacent branchial arches and unpaired cartilages in the ventral midline. Many show cell death in the midbrain, from which some neural crest precursors of the arches originate. lockjaw and a few mutants in the flathead group, including pistachio, affect both jaw cartilage and pigmentation, reflecting essential functions of these genes in at least two neural crest lineages. Mutants of a third class, including boxer, dackel and pincher, affect pectoral fins and axonal trajectories in the brain, as well as the arches. Their skeletal phenotypes suggest that they disrupt cartilage morphogenesis in all arches. Our results suggest that there are sets of genes that: (1) specify neural crest cells in groups of adjacent head segments, and (2) function in common genetic pathways in a variety of tissues including the brain, pectoral fins and pigment cells as well as pharyngeal arches.

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Pax3 is required for cardiac neural crest migration in the mouse: evidence from the splotch (Sp2H) mutant

Development ◽

10.1242/dev.124.2.505 ◽

1997 ◽

Vol 124 (2) ◽

pp. 505-514 ◽

Cited By ~ 14

Author(s):

S.J. Conway ◽

D.J. Henderson ◽

A.J. Copp

Keyword(s):

Neural Crest ◽

Neural Tube ◽

Neural Crest Cells ◽

Outflow Tract ◽

Avian Embryo ◽

Cardiac Neural Crest ◽

Branchial Arches ◽

Aortic Arches ◽

Cardiac Outflow

Neural crest cells originating in the occipital region of the avian embryo are known to play a vital role in formation of the septum of the cardiac outflow tract and to contribute cells to the aortic arches, thymus, thyroid and parathyroids. This ‘cardiac’ neural crest sub-population is assumed to exist in mammals, but without direct evidence. In this paper we demonstrate, using RT-PCR and in situ hybridisation, that Pax3 expression can serve as a marker of cardiac neural crest cells in the mouse embryo. Cells of this lineage were traced from the occipital neural tube, via branchial arches 3, 4 and 6, into the aortic sac and aorto-pulmonary outflow tract. Confirmation that these Pax3-positive cells are indeed cardiac neural crest is provided by experiments in which hearts were deprived of a source of colonising neural crest, by organ culture in vitro, with consequent lack of up-regulation of Pax3. Occipital neural crest cell outgrowths in vitro were also shown to express Pax3. Mutation of Pax3, as occurs in the splotch (Sp2H) mouse, results in development of conotruncal heart defects including persistent truncus arteriosus. Homozygotes also exhibit defects of the aortic arches, thymus, thyroid and parathyroids. Pax3-positive neural crest cells were found to emigrate from the occipital neural tube of Sp2H/Sp2H embryos in a relatively normal fashion, but there was a marked deficiency or absence of neural crest cells traversing branchial arches 3, 4 and 6, and entering the cardiac outflow tract. This decreased expression of Pax3 in Sp2H/Sp2H embryos was not due to down-regulation of Pax3 in neural crest cells, as use of independent neural crest markers, Hoxa-3, CrabpI, Prx1, Prx2 and c-met also revealed a deficiency of migrating cardiac neural crest cells in homozygous embryos. This work demonstrates the essential role of the cardiac neural crest in formation of the heart and great vessels in the mouse and, furthermore, shows that Pax3 function is required for the cardiac neural crest to complete its migration to the developing heart.

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Migrating neural crest cells in the trunk of the avian embryo are multipotent

Development ◽

10.1242/dev.112.4.913 ◽

1991 ◽

Vol 112 (4) ◽

pp. 913-920 ◽

Cited By ~ 33

Author(s):

S.E. Fraser ◽

M. Bronner-Fraser

Keyword(s):

Neural Crest ◽

Neural Tube ◽

Sympathetic Neurons ◽

Morphological Characteristics ◽

Neural Crest Cells ◽

Cell Types ◽

Sympathetic Ganglia ◽

Avian Embryo ◽

Developmental Potential ◽

Trunk Neural Crest

Trunk neural crest cells migrate extensively and give rise to diverse cell types, including cells of the sensory and autonomic nervous systems. Previously, we demonstrated that many premigratory trunk neural crest cells give rise to descendants with distinct phenotypes in multiple neural crest derivatives. The results are consistent with the idea that neural crest cells are multipotent prior to their emigration from the neural tube and become restricted in phenotype after leaving the neural tube either during their migration or at their sites of localization. Here, we test the developmental potential of migrating trunk neural crest cells by microinjecting a vital dye, lysinated rhodamine dextran (LRD), into individual cells as they migrate through the somite. By two days after injection, the LRD-labelled clones contained from 2 to 67 cells, which were distributed unilaterally in all embryos. Most clones were confined to a single segment, though a few contributed to sympathetic ganglia over two segments. A majority of the clones gave rise to cells in multiple neural crest derivatives. Individual migrating neural crest cells gave rise to both sensory and sympathetic neurons (neurofilament-positive), as well as cells with the morphological characteristics of Schwann cells, and other non-neuronal cells (both neurofilament-negative). Even those clones contributing to only one neural crest derivative often contained both neurofilament-positive and neurofilament-negative cells. Our data demonstrate that migrating trunk neural crest cells can be multipotent, giving rise to cells in multiple neural crest derivatives, and contributing to both neuronal and non-neuronal elements within a given derivative.(ABSTRACT TRUNCATED AT 250 WORDS)

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Regulative interactions in zebrafish neural crest

Development ◽

10.1242/dev.122.2.501 ◽

1996 ◽

Vol 122 (2) ◽

pp. 501-507 ◽

Cited By ~ 9

Author(s):

D.W. Raible ◽

J.S. Eisen

Keyword(s):

Dorsal Root Ganglion ◽

Environmental Factors ◽

Neural Crest ◽

Dorsal Root ◽

Neural Crest Cells ◽

Dorsal Root Ganglion Neurons ◽

Pigment Cells ◽

Trunk Neural Crest ◽

Ganglion Neurons

Zebrafish trunk neural crest cells that migrate at different times have different fates: early-migrating crest cells produce dorsal root ganglion neurons as well as glia and pigment cells, while late-migrating crest cells produce only non-neuronal derivatives. When presumptive early-migrating crest cells were individually transplanted into hosts such that they migrated late, they retained the ability to generate neurons. In contrast, late-migrating crest cells transplanted under the same conditions never generated neurons. These results suggest that, prior to migration, neural crest cells have intrinsic biases in the types of derivatives they will produce. Transplantation of presumptive early-migrating crest cells does not result in production of dorsal root ganglion neurons under all conditions suggesting that these cells require appropriate environmental factors to express these intrinsic biases. When early-migrating crest cells are ablated, late-migrating crest cells gain the ability to produce neurons, even when they migrate on their normal schedule. Interactions among neural crest cells may thus regulate the types of derivatives neural crest cells produce, by establishing or maintaining intrinsic differences between individual cells.

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Dlx5-augmentation in neural crest cells reveals early development and differentiation potential of mouse apical head mesenchyme

Scientific Reports ◽

10.1038/s41598-021-81434-x ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Tri H. Vu ◽

Masaki Takechi ◽

Miki Shimizu ◽

Taro Kitazawa ◽

Hiroki Higashiyama ◽

...

Keyword(s):

Neural Crest ◽

Normal Development ◽

Neural Crest Cells ◽

Pigment Cells ◽

Osteogenic Potential ◽

Heterotopic Bone ◽

Differentiation Potential ◽

Dermal Layer ◽

Development And Differentiation ◽

Dermal Cells

AbstractNeural crest cells (NCCs) give rise to various tissues including neurons, pigment cells, bone and cartilage in the head. Distal-less homeobox 5 (Dlx5) is involved in both jaw patterning and differentiation of NCC-derivatives. In this study, we investigated the differentiation potential of head mesenchyme by forcing Dlx5 to be expressed in mouse NCC (NCCDlx5). In NCCDlx5 mice, differentiation of dermis and pigment cells were enhanced with ectopic cartilage (ec) and heterotopic bone (hb) in different layers at the cranial vertex. The ec and hb were derived from the early migrating mesenchyme (EMM), the non-skeletogenic cell population located above skeletogenic supraorbital mesenchyme (SOM). The ec developed within Foxc1+-dura mater with increased PDGFRα signalling, and the hb formed with upregulation of BMP and WNT/β-catenin signallings in Dermo1+-dermal layer from E11.5. Since dermal cells express Runx2 and Msx2 in the control, osteogenic potential in dermal cells seemed to be inhibited by an anti-osteogenic function of Msx2 in normal context. We propose that, after the non-skeletogenic commitment, the EMM is divided into dermis and meninges by E11.5 in normal development. Two distinct responses of the EMM, chondrogenesis and osteogenesis, to Dlx5-augmentation in the NCCDlx5 strongly support this idea.

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An in vitro assay for neural crest cell migration through the somites

Development ◽

10.1242/dev.98.1.85 ◽

1986 ◽

Vol 98 (1) ◽

pp. 85-97

Author(s):

Georgia Guillory ◽

Marianne Bronner-Fraser

Keyword(s):

Neural Crest ◽

Neural Crest Cell ◽

Neural Crest Cells ◽

Enzymatic Digestion ◽

Pigment Cells ◽

Avian Embryo ◽

Vitro Assay ◽

Neural Crest Cell Migration ◽

Migratory Routes

Neural crest cells in the trunk of the avian embryo come into contact with the somites and neural tube during the course of their migration. However, the relationship between the somites and the early migratory routes followed by these cells is not yet completely understood. Here, we use a tissue culture assay to examine if avian neural crest cells migrate through the somites. Cultures of quail somites were prepared from four adjacent regions along the neural axis in the trunk. Each region had four pairs of consecutive somites with region I being most anterior and region IV containing the last four segments. Within each region, the somites were separated from other tissues by enzymatic digestion and plated onto collagen-coated dishes. Immuno-cytochemical techniques were used to confirm that no neural crest cells, recognized by the HNK-1 antibody, were present on the surface of the somites at the time of explantation. After several days in culture, the explanted somites were screened to identify pigment cells. Because neural crest cells give rise to all of the melanocytes in the trunk, the presence of pigment cells indicated that neural crest precursors were contained within the initial explant. After 5–11 days in vitro, the percentage of somite cultures containing pigment cells in regions I through IV, respectively, was 36%, 51%, 31% and 1%. These results suggest that neural crest cells migrate through the somitic mesenchyme and first enter the somites between 5 to 9 segments rostral to the most recently formed somite.

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T-cadherin expression alternates with migrating neural crest cells in the trunk of the avian embryo

Development ◽

10.1242/dev.111.1.15 ◽

1991 ◽

Vol 111 (1) ◽

pp. 15-22 ◽

Cited By ~ 2

Author(s):

B. Ranscht ◽

M. Bronner-Fraser

Keyword(s):

Cell Migration ◽

Neural Crest ◽

Neural Crest Cell ◽

Neural Crest Cells ◽

Small Population ◽

Avian Embryo ◽

Caudal Portion ◽

Neural Crest Cell Migration ◽

Trunk Neural Crest ◽

Crest Cell

Trunk neural crest cells and motor axons move in a segmental fashion through the rostral (anterior) half of each somitic sclerotome, avoiding the caudal (posterior) half. This metameric migration pattern is thought to be caused by molecular differences between the rostral and caudal portions of the somite. Here, we describe the distribution of T-cadherin (truncated-cadherin) during trunk neural crest cell migration. T-cadherin, a novel member of the cadherin family of cell adhesion molecules was selectively expressed in the caudal half of each sclerotome at all times examined. T-cadherin immunostaining appeared graded along the rostrocaudal axis, with increasing levels of reactivity in the caudal halves of progressively more mature (rostral) somites. The earliest T-cadherin expression was detected in a small population of cells in the caudal portion of the somite three segments rostral to last-formed somite. This initial T-cadherin expression was observed concomitant with the invasion of the first neural crest cells into the rostral portion of the same somite in stage 16 embryos. When neural crest cells were ablated surgically prior to their emigration from the neural tube, the pattern of T-cadherin immunoreactivity was unchanged compared to unoperated embryos, suggesting that the metameric T-cadherin distribution occurs independent of neural crest cell signals. This expression pattern is consistent with the possibility that T-cadherin plays a role in influencing the pattern of neural crest cell migration and in maintaining somite polarity.

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Cancer-associated HIF-2α impacts trunk neural crest stemness

10.1101/2020.01.22.915199 ◽

2020 ◽

Author(s):

Sofie Mohlin ◽

Camilla U. Persson ◽

Elina Fredlund ◽

Emanuela Monni ◽

Jessica M. Lindvall ◽

...

Keyword(s):

Neural Crest ◽

Normal Development ◽

Hypoxia Inducible Factor ◽

Neural Crest Cells ◽

Sympathetic Ganglia ◽

Stem Cell Population ◽

Neural Crest Development ◽

Trunk Neural Crest

AbstractThe neural crest is a stem cell population that gives rise to sympathetic ganglia, the cell type of origin of neuroblastoma. Hypoxia Inducible Factor (HIF)-2α is associated with high risk neuroblastoma, however, little is known about its role in normal neural crest development. To address this important question, here we show that HIF-2α is expressed in trunk neural crest cells of human, murine and avian embryos. Modulating HIF-2α in vivo not only causes developmental delays but also induces proliferation and stemness of neural crest cells while altering the number of cells migrating ventrally to sympathoadrenal sites. Transcriptome changes after loss of HIF-2α reflect the in vivo phenotype. The results suggest that expression levels of HIF-2α must be strictly controlled and abnormal levels increase stemness and may promote metastasis. Our findings help elucidate the role of HIF-2α during normal development with implications also in tumor initiation at the onset of neuroblastoma.

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Analysis of cranial neural crest migratory pathways in axolotl using cell markers and transplantation

Development ◽

10.1242/dev.127.12.2751 ◽

2000 ◽

Vol 127 (12) ◽

pp. 2751-2761 ◽

Cited By ~ 1

Author(s):

H. Epperlein ◽

D. Meulemans ◽

M. Bronner-Fraser ◽

H. Steinbeisser ◽

M.A. Selleck

Keyword(s):

Transcription Factor ◽

Neural Crest ◽

Neural Crest Cells ◽

Branchial Arch ◽

Cranial Neural Crest ◽

Branchial Arches ◽

Trunk Neural Crest ◽

Cranial Neural Crest Cells ◽

Random Nature

We have examined the ability of normal and heterotopically transplanted neural crest cells to migrate along cranial neural crest pathways in the axolotl using focal DiI injections and in situ hybridization with the neural crest marker, AP-2. DiI labeling demonstrates that cranial neural crest cells migrate as distinct streams along prescribed pathways to populate the maxillary and mandibular processes of the first branchial arch, the hyoid arch and gill arches 1–4, following migratory pathways similar to those observed in other vertebrates. Another neural crest marker, the transcription factor AP-2, is expressed by premigratory neural crest cells within the neural folds and migrating neural crest cells en route to and within the branchial arches. Rotations of the cranial neural folds suggest that premigratory neural crest cells are not committed to a specific branchial arch fate, but can compensate when displaced short distances from their targets by migrating to a new target arch. In contrast, when cells are displaced far from their original location, they appear unable to respond appropriately to their new milieu such that they fail to migrate or appear to migrate randomly. When trunk neural folds are grafted heterotopically into the head, trunk neural crest cells migrate in a highly disorganized fashion and fail to follow normal cranial neural crest pathways. Importantly, we find incorporation of some trunk cells into branchial arch cartilage despite the random nature of their migration. This is the first demonstration that trunk neural crest cells can form cartilage when transplanted to the head. Our results indicate that, although cranial and trunk neural crest cells have inherent differences in ability to recognize migratory pathways, trunk neural crest can differentiate into cranial cartilage when given proper instructive cues.

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