Explanted Fish Neural Tubes Give Rise to Differentiating Neural Crest Cells. (neural crest cell culture/pigment cells/Xiphophorus/medaka/fish)

To permit a more detailed analysis of neural crest cell migratory pathways in the chick embryo, neural crest cells were labelled with a nondeleterious membrane intercalating vital dye, DiI. All neural tube cells with endfeet in contact with the lumen, including premigratory neural crest cells, were labelled by pressure injecting a solution of DiI into the lumen of the neural tube. When assayed one to three days later, migrating neural crest cells, motor axons, and ventral root cells were the only cells types external to the neural tube labelled with DiI. During the neural crest cell migratory phase, distinctly labelled cells were found along: (1) a dorsolateral pathway, under the epidermis, as well adjacent to and intercalating through the dermamyotome; and (2) a ventral pathway, through the rostral portion of each sclerotome and around the dorsal aorta as described previously. In contrast to those cells migrating through the sclerotome, labelled cells on the dorsolateral pathway were not segmentally arranged along the rostrocaudal axis. DiI-labelled cells were observed in all truncal neural crest derivatives, including subepidermal presumptive pigment cells, dorsal root ganglia, and sympathetic ganglia. By varying the stage at which the injection was performed, neural crest cell emigration at the level of the wing bud was shown to occur from stage 13 through stage 22. In addition, neural crest cells were found to populate their derivatives in a ventral-to-dorsal order, with the latest emigrating cells migrating exclusively along the dorsolateral pathway.

Download Full-text

The distribution of fibronectin and tenascin along migratory pathways of the neural crest in the trunk of amphibian embryos

Development ◽

10.1242/dev.103.4.743 ◽

1988 ◽

Vol 103 (4) ◽

pp. 743-756 ◽

Cited By ~ 2

Author(s):

H.H. Epperlein ◽

W. Halfter ◽

R.P. Tucker

Keyword(s):

Cell Migration ◽

Neural Crest ◽

Neural Tube ◽

Neural Crest Cell ◽

Neural Crest Cells ◽

Pigment Cells ◽

Dorsal Fin ◽

Amphibian Embryos ◽

Neural Crest Cell Migration ◽

Crest Cell

It is generally assumed that in amphibian embryos neural crest cells migrate dorsally, where they form the mesenchyme of the dorsal fin, laterally (between somites and epidermis), where they give rise to pigment cells, and ventromedially (between somites and neural tube), where they form the elements of the peripheral nervous system. While there is agreement about the crest migratory routes in the axolotl (Ambystoma mexicanum), different opinions exist about the lateral pathway in Xenopus. We investigated neural crest cell migration in Xenopus (stages 23, 32, 35/36 and 41) using the X. laevis-X. borealis nuclear marker system and could not find evidence for cells migrating laterally. We have also used immunohistochemistry to study the distribution of the extracellular matrix (ECM) glycoproteins fibronectin (FN) and tenascin (TN), which have been implicated in directing neural crest cells during their migrations in avian and mammalian embryos, in the neural crest migratory pathways of Xenopus and the axolotl. In premigratory stages of the crest, both in Xenopus (stage 22) and the axolotl (stage 25), FN was found subepidermally and in extracellular spaces around the neural tube, notochord and somites. The staining was particularly intense in the dorsal part of the embryo, but it was also present along the visceral and parietal layers of the lateral plate mesoderm. TN, in contrast, was found only in the anterior trunk mesoderm in Xenopus; in the axolotl, it was absent. During neural crest cell migration in Xenopus (stages 25–33) and the axolotl (stages 28–35), anti-FN stained the ECM throughout the embryo, whereas anti-TN staining was limited to dorsal regions. There it was particularly intense medially, i.e. in the dorsal fin, around the neural tube, notochord, dorsal aorta and at the medial surface of the somites (stage 35 in both species). During postmigratory stages in Xenopus (stage 40), anti-FN staining was less intense than anti-TN staining. In culture, axolotl neural crest cells spread differently on FN- and TN-coated substrata. On TN, the onset of cellular outgrowth was delayed for about 1 day, but after 3 days the extent of outgrowth was indistinguishable from cultures grown on FN. However, neural crest cells in 3-day-old cultures were much more flattened on FN than on TN. We conclude that both FN and TN are present in the ECM that lines the neural crest migratory pathways of amphibian embryos at the time when the neural crest cells are actively migrating. FN is present in the embryonic ECM before the onset of neural crest migration.(ABSTRACT TRUNCATED AT 400 WORDS)

Download Full-text

The role of cell-extracellular matrix interactions in neural crest cell migration

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100156195 ◽

1989 ◽

Vol 47 ◽

pp. 842-843

Author(s):

Marianne Bronner-Fraser

Keyword(s):

Extracellular Matrix ◽

Cell Migration ◽

Neural Crest ◽

Cell Movement ◽

Neural Crest Cell ◽

Neural Crest Cells ◽

Pigment Cells ◽

Surrounding Matrix ◽

Adrenal Chromaffin ◽

Crest Cell

The formation of the embryo involves intricate cell movements, cell proliferation, and differentiation. The neural crest has long served as a model for the study of these processes because these cells: 1. migrate extensively along characteristic pathways during embryogenesis. 2. give rise to diverse and numerous derivatives, including pigment cells, adrenal chromaffin cells, and the ganglia of the peripheral nervous system; and 3. are accessible to surgical, immunological, and biochemical manipulations during both initial and certain later stages in their development. We are in the process of identifying factors that influence cell migration and differentiation in the neural crest system.Neural crest cells follow two primary migratory pathways in the trunk: a dorsolateral route underneath the skin, and a ventral route through the somite. Within the somites, neural crest cells preferentially migrate through the rostral half of each sclerotome but are absent from the caudal sclerotome. The regions through which neural crest cells migrate are lined with extracellular matrix (ECM) molecules. Because of the intimate relationship between neural crest cells and the surrounding matrix, it has been proposed that the ECM plays an important role in the initiation, guidance, and cessation of neural crest cell movement.

Download Full-text

Role of FGF signalling in neural crest cell migration during early chick embryo development

Zygote ◽

10.1017/s096719941800045x ◽

2018 ◽

Vol 26 (6) ◽

pp. 457-464 ◽

Cited By ~ 1

Author(s):

Xiao-tan Zhang ◽

Guang Wang ◽

Yan Li ◽

Manli Chuai ◽

Kenneth Ka Ho Lee ◽

...

Keyword(s):

Neural Crest ◽

Neural Tube ◽

Neural Crest Cell ◽

Dominant Negative ◽

Neural Tubes ◽

Dorsal Neural Tube ◽

Neural Crest Cell Migration ◽

Fgf Signalling ◽

Crest Cell

SummaryFibroblast growth factor (FGF) signalling acts as one of modulators that control neural crest cell (NCC) migration, but how this is achieved is still unclear. In this study, we investigated the effects of FGF signalling on NCC migration by blocking this process. Constructs that were capable of inducing Sprouty2 (Spry2) or dominant-negative FGFR1 (Dn-FGFR1) expression were transfected into the cells making up the neural tubes. Our results revealed that blocking FGF signalling at stage HH10 (neurulation stage) could enhance NCC migration at both the cranial and trunk levels in the developing embryos. It was established that FGF-mediated NCC migration was not due to altering the expression of N-cadherin in the neural tube. Instead, we determined that cyclin D1 was overexpressed in the cranial and trunk levels when Sprouty2 was upregulated in the dorsal neural tube. These results imply that the cell cycle was a target of FGF signalling through which it regulates NCC migration at the neurulation stage.

Download Full-text

Avian neural crest cell attachment to laminin: involvement of divalent cation dependent and independent integrins

Development ◽

10.1242/dev.113.4.1069 ◽

1991 ◽

Vol 113 (4) ◽

pp. 1069-1084 ◽

Cited By ~ 2

Author(s):

T. Lallier ◽

M. Bronner-Fraser

Keyword(s):

Neural Crest ◽

Cell Attachment ◽

Divalent Cations ◽

Neural Crest Cell ◽

Neural Crest Cells ◽

Cell Interaction ◽

Cell Binding ◽

Cell Adherence ◽

Beta 1 Integrin ◽

Crest Cell

The mechanisms of neural crest cell interaction with laminin were explored using a quantitative cell attachment assay. With increasing substratum concentrations, an increasing percentage of neural crest cells adhere to laminin. Cell adhesion at all substratum concentrations was inhibited by the CSAT antibody, which recognizes the chick beta 1 subunit of integrin, suggesting that beta 1-integrins mediate neural crest cell interactions with laminin. The HNK-1 antibody, which recognizes a carbohydrate epitope, inhibited neural crest cell attachment to laminin at low coating concentrations (greater than 1 microgram ml-1; Low-LM), but not at high coating concentration of laminin (10 micrograms ml-1; High-LM). Attachment to Low-LM occurred in the absence of divalent cations, whereas attachment to High-LM required greater than 0.1 mM Ca2+ or Mn2+. Neural crest cell adherence to the E8 fragment of laminin, derived from its long arm, was similar to that on intact laminin at high and low coating concentrations, suggesting that this fragment contains the neural crest cell binding site(s). The HNK-1 antibody recognizes a protein of 165,000 Mr which is also found in immunoprecipitates using antibodies against the beta 1 subunit of integrin and is likely to be an integrin alpha subunit or an integrin-associated protein. Our results suggest that the HNK-1 epitope on neural crest cells is present on or associated with a novel or differentially glycosylated form of beta 1-integrin, which recognizes laminin in the apparent absence of divalent cations. We conclude that neural crest cells have at least two functionally independent means of attachment to laminin which are revealed at different substratum concentrations and/or conformations of laminin.

Download Full-text

Evidence for collapsin-1 functioning in the control of neural crest migration in both trunk and hindbrain regions

Development ◽

10.1242/dev.126.10.2181 ◽

1999 ◽

Vol 126 (10) ◽

pp. 2181-2189 ◽

Cited By ~ 7

Author(s):

B.J. Eickholt ◽

S.L. Mackenzie ◽

A. Graham ◽

F.S. Walsh ◽

P. Doherty

Keyword(s):

Cell Migration ◽

Neural Crest ◽

Neural Crest Cell ◽

Axon Growth ◽

Neural Crest Cells ◽

Neural Crest Cell Migration ◽

Neural Crest Migration ◽

Migration Pathways ◽

Crest Cell

Collapsin-1 belongs to the Semaphorin family of molecules, several members of which have been implicated in the co-ordination of axon growth and guidance. Collapsin-1 can function as a selective chemorepellent for sensory neurons, however, its early expression within the somites and the cranial neural tube (Shepherd, I., Luo, Y., Raper, J. A. and Chang, S. (1996) Dev. Biol. 173, 185–199) suggest that it might contribute to the control of additional developmental processes in the chick. We now report a detailed study on the expression of collapsin-1 as well as on the distribution of collapsin-1-binding sites in regions where neural crest cell migration occurs. collapsin-1 expression is detected in regions bordering neural crest migration pathways in both the trunk and hindbrain regions and a receptor for collapsin-1, neuropilin-1, is expressed by migrating crest cells derived from both regions. When added to crest cells in vitro, a collapsin-1-Fc chimeric protein induces morphological changes similar to those seen in neuronal growth cones. In order to test the function of collapsin-1 on the migration of neural crest cells, an in vitro assay was used in which collapsin-1-Fc was immobilised in alternating stripes consisting of collapsin-Fc/fibronectin versus fibronectin alone. Explanted neural crest cells derived from both trunk and hindbrain regions avoided the collapsin-Fc-containing substratum. These results suggest that collapsin-1 signalling can contribute to the patterning of neural crest cell migration in the developing chick.

Download Full-text

Neural crest cell migration: requirements for exogenous fibronectin and high cell density.

The Journal of Cell Biology ◽

10.1083/jcb.96.2.462 ◽

1983 ◽

Vol 96 (2) ◽

pp. 462-473 ◽

Cited By ~ 258

Author(s):

R A Rovasio ◽

A Delouvee ◽

K M Yamada ◽

R Timpl ◽

J P Thiery

Keyword(s):

Cell Adhesion ◽

Neural Crest ◽

Type I Collagen ◽

Neural Crest Cell ◽

Neural Crest Cells ◽

Type I ◽

Cell Adhesion And Migration ◽

And Migration ◽

Crest Cell

Cells of the neural crest participate in a major class of cell migratory events during embryonic development. From indirect evidence, it has been suggested that fibronectin (FN) might be involved in these events. We have directly tested the role of FN in neural crest cell adhesion and migration using several in vitro model systems. Avian trunk neural crest cells adhered readily to purified plasma FN substrates and to extracellular matrices containing cellular FN. Their adhesion was inhibited by antibodies to a cell-binding fragment of FN. In contrast, these cells did not adhere to glass, type I collagen, or to bovine serum albumin in the absence of FN. Neural crest cell adhesion to laminin (LN) was significantly less than to FN; however, culturing of crest cells under conditions producing an epithelioid phenotype resulted in cells that could bind equally as well to LN as to FN. The migration of neural crest cells appeared to depend on both the substrate and the extent of cell interactions. Cells migrated substantially more rapidly on FN than on LN or type I collagen substrates; if provided a choice between stripes of FN and glass or LN, cells migrated preferentially on the FN. Migration was inhibited by antibodies against the cell-binding region of FN, and the inhibition could be reversed by a subsequent addition of exogenous FN. However, the migration on FN was random and displayed little persistence of direction unless cells were at high densities that permitted frequent contacts. The in vitro rate of migration of cells on FN-containing matrices was 50 microns/h, similar to their migration rates along the narrow regions of FN-containing extracellular matrix in migratory pathways in vivo. These results indicate that FN is important for neural crest cell adhesion and migration and that the high cell densities of neural crest cells in the transient, narrow migratory pathways found in the embryo are necessary for effective directional migration.

Download Full-text

Modulation of mouse neural crest cell motility by N-cadherin and connexin 43 gap junctions

The Journal of Cell Biology ◽

10.1083/jcb.200105047 ◽

2001 ◽

Vol 154 (1) ◽

pp. 217-230 ◽

Cited By ~ 163

Author(s):

X. Xu ◽

W.E.I. Li ◽

G.Y. Huang ◽

R. Meyer ◽

T. Chen ◽

...

Keyword(s):

Gap Junctions ◽

Neural Crest ◽

Cell Motility ◽

Gap Junction ◽

Connexin 43 ◽

Neural Crest Cell ◽

Neural Crest Cells ◽

Functional Coupling ◽

Dye Coupling ◽

Crest Cell

Connexin 43 (Cx43α1) gap junction has been shown to have an essential role in mediating functional coupling of neural crest cells and in modulating neural crest cell migration. Here, we showed that N-cadherin and wnt1 are required for efficient dye coupling but not for the expression of Cx43α1 gap junctions in neural crest cells. Cell motility was found to be altered in the N-cadherin–deficient neural crest cells, but the alterations were different from that elicited by Cx43α1 deficiency. In contrast, wnt1-deficient neural crest cells showed no discernible change in cell motility. These observations suggest that dye coupling may not be a good measure of gap junction communication relevant to motility. Alternatively, Cx43α1 may serve a novel function in motility. We observed that p120 catenin (p120ctn), an Armadillo protein known to modulate cell motility, is colocalized not only with N-cadherin but also with Cx43α1. Moreover, the subcellular distribution of p120ctn was altered with N-cadherin or Cx43α1 deficiency. Based on these findings, we propose a model in which Cx43α1 and N-cadherin may modulate neural crest cell motility by engaging in a dynamic cross-talk with the cell's locomotory apparatus through p120ctn signaling.

Download Full-text

Connexin 43 impacts the chick premigratory cranial neural crest cell population without affecting the neural crest cell epithelial-to-mesenchymal transition

10.1101/673921 ◽

2019 ◽

Author(s):

Karyn Jourdeuil ◽

Lisa A. Taneyhill

Keyword(s):

Gap Junctions ◽

Neural Crest ◽

Gap Junction ◽

Neural Crest Cell ◽

Neural Crest Cells ◽

Cranial Neural Crest ◽

Mesenchymal Transition ◽

Junction Protein ◽

Cranial Neural Crest Cells ◽

Crest Cell

ABSTRACTGap junctions are intercellular channels that allow for the diffusion of small ions and solutes between coupled cells. Connexin 43 (Cx43), also known as Gap Junction Protein α1, is the most broadly expressed gap junction protein in vertebrate development. Cx43 is strongly expressed in premigratory cranial neural crest cells and is maintained throughout the neural crest cell epithelial-to-mesenchymal transition (EMT), but its function in these cells is not known. To this end, we have used a combination of in vivo and ex vivo live imaging with confocal microscopy, immunohistochemistry, and functional assays to assess gap junction formation, and Cx43 function, in chick premigratory cranial neural crest cells. Our results demonstrate that gap junctions exist between chick premigratory and migratory cranial neural crest cells, with Cx43 depletion inhibiting the function of gap junctions. While a reduction in Cx43 levels just prior to neural crest cell EMT did not affect EMT and subsequent emigration of neural crest cells from the neural tube, the size of the premigratory neural crest cell domain was decreased in the absence of any changes in cell proliferation or death. Collectively, these data identify a role for Cx43 within the chick premigratory cranial neural crest cell population prior to EMT and migration.

Download Full-text

In ovo time-lapse analysis of chick hindbrain neural crest cell migration shows cell interactions during migration to the branchial arches

Development ◽

10.1242/dev.127.6.1161 ◽

2000 ◽

Vol 127 (6) ◽

pp. 1161-1172 ◽

Cited By ~ 9

Author(s):

P.M. Kulesa ◽

S.E. Fraser

Keyword(s):

Cell Migration ◽

Neural Crest ◽

Neural Crest Cell ◽

Neural Crest Cells ◽

Otic Vesicle ◽

In Ovo ◽

Branchial Arches ◽

Neural Crest Cell Migration ◽

Crest Cell ◽

Cell Cell

Hindbrain neural crest cells were labeled with DiI and followed in ovo using a new approach for long-term time-lapse confocal microscopy. In ovo imaging allowed us to visualize neural crest cell migration 2–3 times longer than in whole embryo explant cultures, providing a more complete picture of the dynamics of cell migration from emergence at the dorsal midline to entry into the branchial arches. There were aspects of the in ovo neural crest cell migration patterning which were new and different. Surprisingly, there was contact between neural crest cell migration streams bound for different branchial arches. This cell-cell contact occurred in the region lateral to the otic vesicle, where neural crest cells within the distinct streams diverted from their migration pathways into the branchial arches and instead migrated around the otic vesicle to establish a contact between streams. Some individual neural crest cells did appear to cross between the streams, but there was no widespread mixing. Analysis of individual cell trajectories showed that neural crest cells emerge from all rhombomeres (r) and sort into distinct exiting streams adjacent to the even-numbered rhombomeres. Neural crest cell migration behaviors resembled the wide diversity seen in whole embryo chick explants, including chain-like cell arrangements; however, average in ovo cell speeds are as much as 70% faster. To test to what extent neural crest cells from adjoining rhombomeres mix along migration routes and within the branchial arches, separate groups of premigratory neural crest cells were labeled with DiI or DiD. Results showed that r6 and r7 neural crest cells migrated to the same spatial location within the fourth branchial arch. The diversity of migration behaviors suggests that no single mechanism guides in ovo hindbrain neural crest cell migration into the branchial arches. The cell-cell contact between migration streams and the co-localization of neural crest cells from adjoining rhombomeres within a single branchial arch support the notion that the pattern of hindbrain neural crest cell migration emerges dynamically with cell-cell communication playing an important guidance role.

Download Full-text