Presence Of Neural Crest Cells Research Articles

Visualizing mesoderm and neural crest cell dynamics during chick head morphogenesis

Vertebrate head morphogenesis involves carefully-orchestrated tissue growth and cell movements of the mesoderm and neural crest to form the distinct craniofacial pattern. To better understand structural birth defects, it is important that we characterize the dynamics of these processes and learn how they rely on each other. Here we examine this question during chick head morphogenesis using time-lapse imaging, computational modeling, and experiments. We find that head mesodermal cells in culture move in random directions as individuals and move faster in the presence of neural crest cells. In vivo, mesodermal cells migrate in a directed manner and maintain neighbor relationships; neural crest cells travel through the mesoderm at a faster speed. The mesoderm grows with a non-uniform spatio-temporal profile determined by BrdU labeling during the period of faster and more-directed neural crest collective migration through this domain. We use computer simulations to probe the robustness of neural crest stream formation by varying the spatio-temporal growth profile of the mesoderm. We follow this with experimental manipulations that either stop mesoderm growth or prevent neural crest migration and observe changes in the non-manipulated cell population, implying a dynamic feedback between tissue growth and neural crest cell signaling to confer robustness to the system. Overall, we present a novel descriptive analysis of mesoderm and neural crest cell dynamics that reveals the coordination and co-dependence of these two cell populations during head morphogenesis.

Neural Crest Does Not Contribute to the Neck and Shoulder in the Axolotl (Ambystoma mexicanum)

BackgroundA major step during the evolution of tetrapods was their transition from water to land. This process involved the reduction or complete loss of the dermal bones that made up connections to the skull and a concomitant enlargement of the endochondral shoulder girdle. In the mouse the latter is derived from three separate embryonic sources: lateral plate mesoderm, somites, and neural crest. The neural crest was suggested to sustain the muscle attachments. How this complex composition of the endochondral shoulder girdle arose during evolution and whether it is shared by all tetrapods is unknown. Salamanders that lack dermal bone within their shoulder girdle were of special interest for a possible contribution of the neural crest to the endochondral elements and muscle attachment sites, and we therefore studied them in this context.ResultsWe grafted neural crest from GFP+ fluorescent transgenic axolotl (Ambystoma mexicanum) donor embryos into white (d/d) axolotl hosts and followed the presence of neural crest cells within the cartilage of the shoulder girdle and the connective tissue of muscle attachment sites of the neck-shoulder region. Strikingly, neural crest cells did not contribute to any part of the endochondral shoulder girdle or to the connective tissue at muscle attachment sites in axolotl.ConclusionsOur results in axolotl suggest that neural crest does not serve a general function in vertebrate shoulder muscle attachment sites as predicted by the “muscle scaffold theory,” and that it is not necessary to maintain connectivity of the endochondral shoulder girdle to the skull. Our data support the possibility that the contribution of the neural crest to the endochondral shoulder girdle, which is observed in the mouse, arose de novo in mammals as a developmental basis for their skeletal synapomorphies. This further supports the hypothesis of an increased neural crest diversification during vertebrate evolution.

Nestin expression in glial and neuronal progenitors of the developing human spinal ganglia

Expression of the neural crest marker nestin was studied in glial and neuronal progenitors of developing human spinal ganglia using immunohistochemistry in 10 conceptuses, 5–10 weeks old. Quantification was performed by counting the ratio of positive cells in the total cell number, expressed as mean ± SD and by the Mann–Whitney test. Strong expression of nestin in the 5th–6th developmental week (56%) decreased to 49% in the foetal period. During the same period, number of PGP9.5-positive cells increased from 38% to 42%. At earliest stages, the number of cells expressing GFAP was two folds higher (21%) than S100 (11%). During further development, their number nearly levelled (23% and 28%, respectively), and finally reached level of 25% for GFAP and 40% for S100. While expression of nestin was constantly higher in the dorsal parts of spinal ganglia, number of PGP9.5-, GFAP- and S100-positive cells was higher in their ventral parts, thus indicating ventral to dorsal direction of spinal ganglia differentiation. Co-localization of nestin and GFAP or nestin and S100 was observed during the whole investigate period, while PGP9.5 did not co-localize with nestin. Some ganglion cells simultaneously co-expressing GFAP and S100 might be satellite cells and immature Schwann cells. We suggest that some nestin-positive cells might be capable to differentiate into neurons during the earliest stages of development. Gangliogenesis seems to be important process during the whole ganglion development. Continuous presence of neural crest cells during development might be important in regenerative processes following damage of the spinal ganglia.

Relationship between Neural Crest Cells and Cranial Mesoderm during Head Muscle Development

BackgroundIn vertebrates, the skeletal elements of the jaw, together with the connective tissues and tendons, originate from neural crest cells, while the associated muscles derive mainly from cranial mesoderm. Previous studies have shown that neural crest cells migrate in close association with cranial mesoderm and then circumscribe but do not penetrate the core of muscle precursor cells of the branchial arches at early stages of development, thus defining a sharp boundary between neural crest cells and mesodermal muscle progenitor cells. Tendons constitute one of the neural crest derivatives likely to interact with muscle formation. However, head tendon formation has not been studied, nor have tendon and muscle interactions in the head.Methodology/Principal FindingsReinvestigation of the relationship between cranial neural crest cells and muscle precursor cells during development of the first branchial arch, using quail/chick chimeras and molecular markers revealed several novel features concerning the interface between neural crest cells and mesoderm. We observed that neural crest cells migrate into the cephalic mesoderm containing myogenic precursor cells, leading to the presence of neural crest cells inside the mesodermal core of the first branchial arch. We have also established that all the forming tendons associated with branchiomeric and eye muscles are of neural crest origin and express the Scleraxis marker in chick and mouse embryos. Moreover, analysis of Scleraxis expression in the absence of branchiomeric muscles in Tbx1−/− mutant mice, showed that muscles are not necessary for the initiation of tendon formation but are required for further tendon development.Conclusions/SignificanceThis results show that neural crest cells and muscle progenitor cells are more extensively mixed than previously believed during arch development. In addition, our results show that interactions between muscles and tendons during craniofacial development are similar to those observed in the limb, despite the distinct embryological origin of these cell types in the head.

Normal distribution of melanocytes in the mouse heart

We report the consistent distribution of a population of pigmented trp-1-positive cells in several important septal and valvular structures of the normal mouse (C57BL/6) heart. The pigmented cell population was first apparent by E16.5 p.c. in the right atrial wall and extended into the atrium along the interatrial septum. By E17.5, these cells were found along the apical membranous interventricular septum near or below the surface of the endocardium. The most striking distribution of dark pigmented cells was found in the tricuspid and mitral valvular leaflets and chordae tendineae. The normal distribution of pigmented cells in the valvuloseptal apparatus of C57BL/6 adult heart suggests that a premelanocytic lineage may participate in the earlier morphogenesis of the valve leaflets and chordae tendineae. The origin of the premelanocyte lineage is currently unknown. The most likely candidate populations include the neural crest and the epicardially derived cells. The only cell type in the heart previously shown to form melanocytes is the neural crest. The presence of neural crest cells, but not melanocytes, in some of the regions we describe has been reported by others. However, previous reports have not shown a contribution of melanocytes or neural crest derivatives to the atrioventricular valve leaflets or chordae tendineae in mouse hearts. If these cells are of neural crest origin, it would suggest a possibly greater contribution and persistence of neural crest cells to the valvuloseptal apparatus than has been previously understood.

Neural tube-ectoderm interactions are required for trigeminal placode formation.

Cranial sensory ganglia in vertebrates develop from the ectodermal placodes, the neural crest, or both. Although much is known about the neural crest contribution to cranial ganglia, relatively little is known about how placode cells form, invaginate and migrate to their targets. Here, we identify Pax-3 as a molecular marker for placode cells that contribute to the ophthalmic branch of the trigeminal ganglion and use it, in conjunction with DiI labeling of the surface ectoderm, to analyze some of the mechanisms underlying placode development. Pax-3 expression in the ophthalmic placode is observed as early as the 4-somite stage in a narrow band of ectoderm contiguous to the midbrain neural folds. Its expression broadens to a patch of ectoderm adjacent to the midbrain and the rostral hindbrain at the 8- to 10-somite stage. Invagination of the first Pax-3-positive cells begins at the 13-somite stage. Placodal invagination continues through the 35-somite stage, by which time condensation of the trigeminal ganglion has begun. To challenge the normal tissue interactions leading to placode formation, we ablated the cranial neural crest cells or implanted barriers between the neural tube and the ectoderm. Our results demonstrate that, although the presence of neural crest cells is not mandatory for Pax-3 expression in the forming placode, a diffusible signal from the neuroectoderm is required for induction and/or maintenance of the ophthalmic placode.

Cytotactin expression in somites after dorsal neural tube and neural crest ablation in chicken embryos.

The spatiotemporal expression of the extracellular matrix protein cytotactin/tenascin during somitogenesis suggests that it plays a role in the morphogenetic events that give rise to the pattern of neural crest (NC) development. In the present study, the spatial distribution and molecular forms of cytotactin in somites were examined using in situ hybridization, Western blotting, and immunohistochemistry during normal development and after injury. In situ hybridization showed that prior to NC cell invasion cytotactin mRNA was restricted to the caudal half of the newly formed epithelial somites. As each epithelial somite matured, giving rise to a sclerotome and dermamyotome, the mRNA was first restricted to the dermamyotome and later restricted to the rostral protion of the sclerotome, consistent with the previously reported protein distribution. Immunocytochemical analysis of the distribution of cytotactin and NC cells in embryos with ablations that removed NC cells, or with simple wounds that left NC cells in place, demonstrated that the presence of NC cells is neither necessary nor sufficient for the correct positioning of cytotactin. Immunoblotting analysis showed that cytotactin synthesized by sclerotomes in the absence of NC cells was of similar molecular mass to that produced in their presence. These findings are in accord with the notion that the abnormalities of cytotactin distribution are related to the wounding process. We conclude that, contrary to the suggestion of Stern et al. [Stern, C. D., Norris, W. E., Bronner-Fraser, M., Carlson, G. J., Faissner, A., Keynes, R. J. & Schachner, M. (1989) Development 107, 309-319], there is no causal link between the presence of NC cells and the distribution and molecular mass of sclerotomal cytotactin.

PUTATIVE MESENCHYMAL RECEPTOR CELLS FOR MIGRATING NEURAL CREST CELLS IN ANEURAL SEGMENTS OF EMBRYONAL CHICK GUT

To clarify the regulation of the neural crest cell migration in the gut, we investigated whether cell adhesion molecules, that are expressed by neural crest cells (NCAM, NgCAM, HNK-1), are also expressed by other cells in aneuronal segments of the gut. We therefore cultured isolated segments of aneuronal gut of four and five day old embryos on the chorioallantoic membrane. Immunostaining, with antibodies directed against cell adhesion molecules, revealed a diffuse band of HNK-1 stained mesenchymal cells, located inside the developing smooth muscle layer. We call this HNK-1 mode I expression. Immunostaining of cultured proximal, innervated segments of the gut of E5 embryos revealed a concentration of the HNK-1 epitope at the sites of the myenteric and submucous plexus. We call this HNK-1 mode II expression. The E/C8 antibody, directed against a neurofilament associated protein, was used to demonstrate presence or absence of neural crest cells in the gut. To determine whether the difference in HNK-1 expression was related to the presence of neural crest cells, we cultured aneuronal gut together with the neural anlage (neural tube and neural crest). Immunostaining revealed that in these co-cultures HNK-1 mode II replaced HNK-1 mode I expression. These results suggest the presence of a subpopulation of mesenchymal cells in the gut, that is involved in neural crest cell migration and aggregation.