Dual Embryonic Origin Research Articles

Lineage-specific mutation of Lmx1b provides new insights into distinct regulation of suture development in different areas of the calvaria.

The calvaria (top part of the skull) is made of pieces of bone as well as multiple soft tissue joints called sutures. The latter is crucial to the growth and morphogenesis of the skull, and thus a loss of calvarial sutures can lead to severe congenital defects in humans. During embryogenesis, the calvaria develops from the cranial mesenchyme covering the brain, which contains cells originating from the neural crest and the mesoderm. While the mechanism that patterns the cranial mesenchyme into bone and sutures is not well understood, function of Lmx1b, a gene encoding a LIM-domain homeodomain transcription factor, plays a key role in this process. In the current study, we investigated a difference in the function of Lmx1b in different parts of the calvaria using neural crest-specific and mesoderm-specific Lmx1b mutants. We found that Lmx1b was obligatory for development of the interfrontal suture and the anterior fontanel along the dorsal midline of the skull, but not for the posterior fontanel over the midbrain. Also, Lmx1b mutation in the neural crest-derived mesenchyme, but not the mesoderm-derived mesenchyme, had a non-cell autonomous effect on coronal suture development. Furthermore, overexpression of Lmx1b in the neural crest lineage had different effects on the position of the coronal suture on the apical part and the basal part. Other unexpected phenotypes of Lmx1b mutants led to an additional finding that the coronal suture and the sagittal suture are of dual embryonic origin. Together, our data reveal a remarkable level of regional specificity in regulation of calvarial development.

Intermediate Cells of Dual Embryonic Origin Follow a Basal to Apical Gradient of Ingression Into the Lateral Wall of the Cochlea.

Intermediate cells of the stria vascularis are neural crest derived melanocytes. They are essential for the establishment of the endocochlear potential in the inner ear, which allows mechanosensory hair cells to transduce sound into nerve impulses. Despite their importance for normal hearing, how these cells develop and migrate to their position in the lateral wall of the cochlea has not been studied. We find that as early as E10.5 some Schwann cell precursors in the VIIIth ganglion begin to express melanocyte specific markers while neural crest derived melanoblasts migrate into the otic vesicle. Intermediate cells of both melanoblast and Schwann cell precursor origin ingress into the lateral wall of the cochlea starting at around E15.5 following a basal to apical gradient during embryonic development, and continue to proliferate postnatally.

Prenatal and Postnatal Development and also Morphological and Morphometric Indicators of Thymus in Newborns

The thymus is a lymphoepithelial organ located in the mediastinum and reaches its maximum development in youth. While other lymphoid organs develop exclusively from the mesenchyme (mesoderm), the thymus is of dual embryonic origin. Its lymphocytes develop in the bone marrow from cells of mesenchymal origin; they invade the epithelial bud, which has developed from the endoderm of the third and fourth pharyngeal pockets. The thymus is covered with a connective tissue capsule, which is introduced into the parenchyma and divides it into incomplete lobules, so the cortex and medulla of the adjacent lobules are connected to each other.

Supernumerary teeth in patients with cleft lip and palate: the tooth germs do not separate.

ABSTRACTIntroduction: Supernumerary teeth in cases of cleft lip and palate do not result from the division of normal germs before the formation of hard tissue. Deciduous and permanent teeth odontogenesis begins after the face has formed, either with or without the cleft. Discussion: The most acceptable hypothesis to enable understanding of the presence of supernumerary teeth on one or both sides of the cleft palate is hyperactivity of the dental lamina in its walls. This hyperactivity, with the formation of more tooth germs, must be attributed to mediators and genes related to tooth formation, under strong influence of local epigenetic factors, whose developmental environment was affected by the presence of the cleft. Conclusion: The current concepts of embryology no longer support the fusion of embryonic processes for the formation of the face, but rather the leveling of the grooves between them. All human teeth have a dual embryonic origin, as they are composed of ectoderm and mesenchyme/ectomesenchyme, but this does not make it easy for them to be duplicated to form supernumerary teeth.

Dual embryonic origin of the mammalian enteric nervous system

The enteric nervous system is thought to originate solely from the neural crest. Transgenic lineage tracing revealed a novel population of clonal pancreatic duodenal homeobox-1 (Pdx1)-Cre lineage progenitor cells in the tunica muscularis of the gut that produced pancreatic descendants as well as neurons upon differentiation in vitro. Additionally, an in vivo subpopulation of endoderm lineage enteric neurons, but not glial cells, was seen especially in the proximal gut. Analysis of early transgenic embryos revealed Pdx1-Cre progeny (as well as Sox-17-Cre and Foxa2-Cre progeny) migrating from the developing pancreas and duodenum at E11.5 and contributing to the enteric nervous system. These results show that the mammalian enteric nervous system arises from both the neural crest and the endoderm. Moreover, in adult mice there are separate Wnt1-Cre neural crest stem cells and Pdx1-Cre pancreatic progenitors within the muscle layer of the gut.

A tale of two cities: The genetic mechanisms governing calvarial bone development.

The skull bones must grow in a coordinated, three-dimensional manner to coalesce and form the head and face. Mammalian skull bones have a dual embryonic origin from cranial neural crest cells (CNCC) and paraxial mesoderm (PM) and ossify through intramembranous ossification. The calvarial bones, the bones of the cranium which cover the brain, are derived from the supraorbital arch (SOA) region mesenchyme. The SOA is the site of frontal and parietal bone morphogenesis and primary center of ossification. The objective of this review is to frame our current in vivo understanding of the morphogenesis of the calvarial bones and the gene networks regulating calvarial bone initiation in the SOA mesenchyme.

Embryonic origin of the gnathostome vertebral skeleton.

The vertebral column is a key component of the jawed vertebrate (gnathostome) body plan, but the primitive embryonic origin of this skeleton remains unclear. In tetrapods, all vertebral components (neural arches, haemal arches and centra) derive from paraxial mesoderm (somites). However, in teleost fishes, vertebrae have a dual embryonic origin, with arches derived from somites, but centra formed, in part, by secretion of bone matrix from the notochord. Here, we test the embryonic origin of the vertebral skeleton in a cartilaginous fish (the skate, Leucoraja erinacea) which serves as an outgroup to tetrapods and teleosts. We demonstrate, by cell lineage tracing, that both arches and centra are somite-derived. We find no evidence of cellular or matrix contribution from the notochord to the skate vertebral skeleton. These findings indicate that the earliest gnathostome vertebral skeleton was exclusively of somitic origin, with a notochord contribution arising secondarily in teleosts.

Dual embryonic origin of maxillary lateral incisors: clinical implications in patients with cleft lip and palate.

Introduction: Cleft lip and palate are craniofacial anomalies highly prevalent in the overall population. In oral clefts involving the alveolar ridge, variations of number, shape, size and position are observed in maxillary lateral incisors. The objective of this manuscript is to elucidate the embryonic origin of maxillary lateral incisors in order to understand the etiology of these variations. Contextualization: The hypothesis that orofacial clefts would split maxillary lateral incisor buds has been previously reported. However, recent studies showed that maxillary lateral incisors have dual embryonic origin, being partially formed by both the medial nasal process and the maxillary process. In other words, the mesial half of the lateral incisor seems to come from the medial nasal process while the distal half of the lateral incisor originates from the maxillary process. In cleft patients, these processes do not fuse, which results in different numerical and positional patterns for lateral incisors relating to the alveolar cleft. In addition to these considerations, this study proposes a nomenclature for maxillary lateral incisors in patients with cleft lip and palate, based on embryology and lateral incisors position in relation to the alveolar cleft. Conclusion: Embryological knowledge on the dual origin of maxillary lateral incisors and the use of a proper nomenclature for their numerical and positional variations renders appropriate communication among professionals and treatment planning easier, in addition to standardizing research analysis.

The development of the mammalian outer and middle ear.

The mammalian ear is a complex structure divided into three main parts: the outer; middle; and inner ear. These parts are formed from all three germ layers and neural crest cells, which have to integrate successfully in order to form a fully functioning organ of hearing. Any defect in development of the outer and middle ear leads to conductive hearing loss, while defects in the inner ear can lead to sensorineural hearing loss. This review focuses on the development of the parts of the ear involved with sound transduction into the inner ear, and the parts largely ignored in the world of hearing research: the outer and middle ear. The published data on the embryonic origin, signalling, genetic control, development and timing of the mammalian middle and outer ear are reviewed here along with new data showing the Eustachian tube cartilage is of dual embryonic origin. The embryonic origin of some of these structures has only recently been uncovered (Science, 339, 2013, 1453; Development, 140, 2013, 4386), while the molecular mechanisms controlling the growth, structure and integration of many outer and middle ear components are hardly known. The genetic analysis of outer and middle ear development is rather limited, with a small number of genes often affecting either more than one part of the ear or having only very small effects on development. This review therefore highlights the necessity for further research into the development of outer and middle ear structures, which will be important for the understanding and treatment of conductive hearing loss.

Dual embryonic origin and patterning of the pharyngeal skeleton in the axolotl (Ambystoma mexicanum).

The impressive morphological diversification of vertebrates was achieved in part by innovation and modification of the pharyngeal skeleton. Extensive fate mapping in amniote models has revealed a primarily cranial neural crest derivation of the pharyngeal skeleton. Although comparable fate maps of amphibians produced over several decades have failed to document a neural crest derivation of ventromedial elements in these vertebrates, a recent report provides evidence of a mesodermal origin of one of these elements, basibranchial 2, in the axolotl. We used a transgenic labeling protocol and grafts of labeled cells between GFP+ and white embryos to derive a fate map that describes contributions of both cranial neural crest and mesoderm to the axolotl pharyngeal skeleton, and we conducted additional experiments that probe the mechanisms that underlie mesodermal patterning. Our fate map confirms a dual embryonic origin of the pharyngeal skeleton in urodeles, including derivation of basibranchial 2 from mesoderm closely associated with the second heart field. Additionally, heterotopic transplantation experiments reveal lineage restriction of mesodermal cells that contribute to pharyngeal cartilage. The mesoderm-derived component of the pharyngeal skeleton appears to be particularly sensitive to retinoic acid (RA): administration of exogenous RA leads to loss of the second basibranchial, but not the first. Neural crest was undoubtedly critical in the evolution of the vertebrate pharyngeal skeleton, but mesoderm may have played a central role in forming ventromedial elements, in particular. When and how many times during vertebrate phylogeny a mesodermal contribution to the pharyngeal skeleton evolved remain to be resolved.

Reevaluation of the homology of bones of the cranial vault in tetrapod vertebrates

Homology assessments of bones are made using one or more criteria (e.g., topology, development, phylogeny). In tetrapods, the bones of the cranial vault appear highly conserved topologically. However, recent fate maps have revealed key differences in the embryonic origin of bones of the cranial vault in representatives of two amniote lineages. In mouse, the frontal is derived from cranial neural crest (CNC) and the parietal of mesoderm, placing the CNC-mesoderm boundary at the suture between these bones. In chicken, by contrast, this boundary is located within the frontal, which thus has a dual embryonic origin. This difference is seemingly inconsistent with the traditional assessment of homology of the avian frontal with that of mammals and other tetrapods. To elucidate this apparent conflict, we fate-mapped CNC and mesoderm using GFP-transgenic axolotls to reveal specific contributions to the cranial vault. The CNC-mesoderm boundary in axolotl is located between frontal and parietal, as in mouse, but differs from that in chicken. If, however, the avian frontal is instead regarded as a fused frontal and parietal (i.e., frontoparietal) and the parietal a postparietal, then the avian cranial vault remains topologically congruent, and becomes developmentally congruent, with that of both urodeles and mammals. This hypothesis is supported by data from the fossil record: a separate frontal, parietal and postparietal is present in all stem lineages of extant taxa, including birds. Moreover, it implies that a postparietal is/was present in non-avian archosaurs but likely fuses early in ontogeny to the parietal (or supraoccipital), as it does in many extant mammals.

Paleontological and developmental evidence resolve the homology and dual embryonic origin of a mammalian skull bone, the interparietal

The homologies of mammalian skull elements are now fairly well established, except for the controversial interparietal bone. A previous experimental study reported an intriguing mixed origin of the interparietal: the medial portion being derived from the neural crest cells, whereas the lateral portion from the mesoderm. The evolutionary history of such mixed origin remains unresolved, and contradictory reports on the presence or absence and developmental patterns of the interparietal among mammals have complicated the question of its homology. Here we provide an alternative perspective on the evolutionary identity of the interparietal, based on a comprehensive study across more than 300 extinct and extant taxa, integrating embryological and paleontological data. Although the interparietal has been regarded as being lost in various lineages, our investigation on embryos demonstrates its presence in all extant mammalian "orders." The generally accepted paradigm has regarded the interparietal as consisting of two elements that are homologized to the postparietals of basal amniotes. The tabular bones have been postulated as being lost during the rise of modern mammals. However, our results demonstrate that the interparietal consists not of two but of four elements. We propose that the tabulars of basal amniotes are conserved as the lateral interparietal elements, which quickly fuse to the medial elements at the embryonic stage, and that the postparietals are homologous to the medial elements. Hence, the dual developmental origin of the mammalian interparietal can be explained as the evolutionary consequence of the fusion between the crest-derived "postparietals" and the mesoderm-derived "tabulars."

Dual embryonic origin of the mammalian otic vesicle forming the inner ear

The inner ear and cochleovestibular ganglion (CVG) derive from a specialized region of head ectoderm termed the otic placode. During embryogenesis, the otic placode invaginates into the head to form the otic vesicle (OV), the primordium of the inner ear and CVG. Non-autonomous cell signaling from the hindbrain to the OV is required for inner ear morphogenesis and neurogenesis. In this study, we show that neuroepithelial cells (NECs), including neural crest cells (NCCs), can contribute directly to the OV from the neural tube. Using Wnt1-Cre, Pax3(Cre/+) and Hoxb1(Cre/+) mice to label and fate map cranial NEC lineages, we have demonstrated that cells from the neural tube incorporate into the otic epithelium after otic placode induction has occurred. Pax3(Cre/+) labeled a more extensive population of NEC derivatives in the OV than did Wnt1-Cre. NEC derivatives constitute a significant population of the OV and, moreover, are regionalized specifically to proneurosensory domains. Descendents of Pax3(Cre/+) and Wnt1-Cre labeled cells are localized within sensory epithelia of the saccule, utricle and cochlea throughout development and into adulthood, where they differentiate into hair cells and supporting cells. Some NEC derivatives give rise to neuroblasts in the OV and CVG, in addition to their known contribution to glial cells. This study defines a dual cellular origin of the inner ear from sensory placode ectoderm and NECs, and changes the current paradigm of inner ear neurosensory development.

Comparative morphology of normal and cleft minipigs demonstrates dual origin of incisors

ObjectiveThe incisors of the mammalian dental arch develop from tissues arising from separated facial prominences. These primordial craniofacial structures undergo complex morphogenetic processes as they merge and fuse in a time and space dependent fashion. However, local contributions of precursor facial prominences to the incisors that develop subsequently remain unknown. The purpose of this study was to characterize the development of all three deciduous upper rostral teeth in the pig (Sus scrofa f. domestica) for the identification of the likely facial prominence contributions to the incisors based on normal and pathological developmental relationships. DesignEmbryonic minipigs were collected between gestational days 20–36 (E20–36), processed for histological analysis and subjected to computerized 3D modelling. The location and morphology of the incisors (i) in these specimens were characterized and compared between developmental stages. A second set of neonatal minipigs displaying cleft lip and/or cleft palate defects were also obtained and incisor locations and eruption patterns were morphologically examined. ResultsPalate formation begins during the third week of gestation (E20) in the minipig with ossification of the premaxilla initiating soon afterwards (E24). The third incisor (i3) develops caudally to the contact seam formed by the fusion of the primary and secondary palates in normal embryos. All cleft animals displayed normal i3 and canine, on other hand, development of i1 and i2 was often disrupted similar to human. ConclusionsOur observations suggest a dual embryonic origin of the incisors in minipigs with the first and second incisors originating from the frontonasal prominence whilst the third incisor forms from tissues derived from the maxillary prominence.

Nicotinic acetylcholine receptor gene expression in developing chick autonomic ganglia

The developmental expression patterns of ten genes encoding nicotinic acetylcholine receptor subunits were analyzed using Northern blots and in situ hybridization in chick peripheral ganglia of neural crest, placodal and dual embryonic origin. The superior cervical and ciliary ganglia were investigated in detail because they accumulated relatively abundant transcripts of the α3, β4, α5 and α7 genes. In the superior cervical ganglion, these four mRNA species had similar developmental time-courses. They appeared at embryonic day 8 (E8), increased steadily until E16 and maintained a rather high plateau level until E18. In the ciliary ganglion, α7 transcripts were already abundant at E6, increased until E10, and considerably decreased thereafter. High-resolution in situ hybridization showed that α7 transcripts were present in all cell types of the E6 ciliary ganglion, whereas they were restricted to large neuronal somas at E16. Transfections with a reporter gene under the control of the α7 promoter demonstrated that a sharp developmental divide occurred at E11-12, after which stage the promoter was activatable in neurons exclusively.

Induction of the epibranchial placodes.

The cranial sensory ganglia, in contrast to those of the trunk, have a dual embryonic origin arising from both neurogenic placodes and neural crest. Neurogenic placodes are focal thickenings of ectoderm, found exclusively in the head of vertebrate embryos. These structures can be split into two groups based on the positions that they occupy within the embryo, dorsolateral and epibranchial. The dorsolateral placodes develop alongside the central nervous system, while the epibranchial placodes are located close to the top of the clefts between the branchial arches. Importantly, previous studies have shown that the neurogenic placodes form under the influence of the surrounding cranial tissues. In this paper, we have analysed the nature of the inductive signal underlying the formation of the epibranchial placodes. We find that epibranchial placodes do not require neural crest for their induction, but rather that it is the pharyngeal endoderm that is the source of the inductive signal. We also find that, while cranial ectoderm is competent to respond to this inductive signal, trunk ectoderm is not. We have further identified the signalling molecule Bmp7 as the mediator of this inductive interaction. This molecule is expressed in a manner consistent with it playing such a role and, when added to ectoderm explants, it will promote the formation of epibranchial neuronal cells. Moreover, the Bmp7 antagonist follstatin will block the ability of pharyngeal endoderm to induce placodal neuronal cells, demonstrating that Bmp7 is required for this inductive interaction. This work answers the long standing question regarding the induction of the epibranchial placodes, and represents the first elucidation of an inductive mechanism, and a molecular effector, underlying the formation of any primary sensory neurons in higher vertebrates.

MyoD and Myf-5 define the specification of musculature of distinct embryonic origin

Mounting evidence supports the notion that Myf-5 and MyoD play unique roles in the development of epaxial (originating in the dorso-medial half of the somite, e.g. back muscles) and hypaxial (originating in the ventro-lateral half of the somite, e.g. limb and body wall muscles) musculature. To further understand how Myf-5 and MyoD genes cooperate during skeletal muscle specification, we examined and compared the expression pattern of MyoD-lacZ (258/2.5lacZ and MD6.0-lacZ) transgenes in wild-type, Myf-5, and MyoD mutant embryos. We found that the delayed onset of muscle differentiation in the branchial arches, tongue, limbs, and diaphragm of MyoD-/- embryos was a consequence of a reduced ability of myogenic precursor cells to progress through their normal developmental program and not because of a defect in migration of muscle progenitor cells into these regions. We also found that myogenic precursor cells for back, intercostal, and abdominal wall musculature in Myf-54-/- embryos failed to undergo normal translocation or differentiation. By contrast, the myogenic precursors of intercostal and abdominal wall musculature in MyoD-/- embryos underwent normal translocation but failed to undergo timely differentiation. In conclusion, these observations strongly support the hypothesis that Myf-5 plays a unique role in the development of muscles arising after translocation of epithelial dermamyotome cells along the medial edge of the somite to the subjacent myotome (e.g., back or epaxial muscle) and that MyoD plays a unique role in the development of muscles arising from migratory precursor cells (e.g., limb and branchial arch muscles, tongue, and diaphragm). In addition, the expression pattern of MyoD-lacZ transgenes in the intercostal and abdominal wall muscles of Myf-5-/- and MyoD-/- embryos suggests that appropriate development of these muscles is dependent on both genes and, therefore, these muscles have a dual embryonic origin (epaxial and hypaxial).

Hereditary syndactyly in Angus cattle.

Twenty-five syndactylous Angus cattle, characterized pathologically, were reported from 16 herds in 10 states from 1979 to 1994. Twenty-one (84%) had all 4 legs syndactylous, 3 (12%) had 3 legs syndactylous, and 1 (4%) had 2 legs syndactylous. All syndactylous cattle walked with considerable difficulty. Hooves of aged animals became curled and bent laterally or medially. Affected hooves had the appearance of a truncated cone, the base of which was located at the coronary band. Most hooves were fused completely with no indication of dual anlage. An occasional hoof had a distal notch, and other hooves had a dorsally located groove indicating dual embryonic origin. Lateral dewclaws were enlarged in most cases. Radiographs and dissections of limbs of 19 animals revealed a consistent pattern of fusion in most affected calves. Lesions included 1 or more of the following: disappearance of the large metacarpal and metatarsal intertrochlear notches, horizontal fusion of 1 or more carpals and tarsals, fusion of proximal sesamoids, 1 distal sesamoid, and fusion of paired phalanges. Evidence of a genetic cause consisted of 11 syndactylous calves among 70 offspring of 4 3/4 sib families, 8 preterm syndactylous fetuses among 148 preterm fetuses and 13 calves in progenies of 19 animals tested for possible heterozygosity, and 5 syndactylous calves from matings of an Angus syndactylous bull with 1 Angus and 4 Holstein syndactylous cows. Data were consistent with recessive inheritance at a single locus. Angus cattle with sydactytly had a larger number of affected limbs than did syndactylous Holsteins and their Angus crosses, suggesting existence of 2 recessive alleles. The allele of Holsteins (syH) appeared to influence phenotypic expression in a dominant pattern over the Angus allele (syA). Both syA and syH alleles acted as recessives to the normal SY allele. Phenotypic effects on limb development were most dramatic in calves with the syA/syA genotype.

Embryonic origin and fate of chondroid tissue and secondary cartilages in the avian skull.

Chondroid tissue is an intermediate calcified tissue, mainly involved in desmocranial morphogenesis. Often associated with secondary cartilages, it remained of unprecise embryonic origin. The latter was studied by performing isotopic isochronic grafts of quail encephalon onto 30 chick embryos. The so-obtained chimeras were sacrificed at the 9th, 12th, and 14th day of incubation. The contribution of graft- and host-derived cells to the histogenesis of chondroid tissue, bone, and secondary cartilages was analyzed on both microradiographs of thick undecalcified sections and on classical histological sections after several DNA or ECM specific staining procedures. Chondroid tissue is deposited in the primitive anlage of all membranous bones of the avian skull. Also present on their sutural edges, it uniformly arises from the neural crest. In the face, bone and secondary cartilages share this mesectodermal origin. However, secondary cartilages located along the basal chondrocranium and bone formed on the chondroid primordium of the cranial vault, originate from the cephalic mesoderm. These facts provide evidence that chondroid tissue arises from a specific differentiation of neural crest derived cells and that this original skeletogenic program differs from that of secondary chondrogenesis. Moreover, they obviously indicate that in membraneous bone ontogenesis, chondroid tissue replaces functions devoted to mesodermal primary cartilages of the cranial base, and so corroborates at the tissue level, the dual embryonic and phyletic origin of the skull.

Organization of motor pools supplying axial muscles in the chicken

Intramuscular injections of the retrograde tracers 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI) and horseradish peroxidase (HRP) were used to map the motor pools innervating axial muscles in the cervical and thoracic regions of the chicken. We found that motor pool position is well correlated with the muscle's embryonic origin, and not necessarily with its position. Muscles of myotomal (exclusively somitic) origin were innervated by medially positioned motoneurons in the median motor column, and the motor pools supplying these muscles were somatotopically organized. Muscles having a dual embryonic origin, from the somites and lateral plate, were innervated by motoneurons positioned further laterally within the median motor column. The relationship between motor pool position and embryonic origin of the muscle may be a general principle of ventral horn organization.