Abigail Tucker | King's College London (original) (raw)
Papers by Abigail Tucker
Proceedings of the National Academy of Sciences, 2010
Changes in tooth shape have played a major role in vertebrate evolution with modification of dent... more Changes in tooth shape have played a major role in vertebrate evolution with modification of dentition allowing an organism to adapt to new feeding strategies. The current view is that molar teeth evolved from simple conical teeth, similar to canines, by progressive addition of extra "cones" to form progressively complex multicuspid crowns. Mammalian incisors, however, are neither conical nor multicuspid, and their evolution is unclear. We show that hypomorphic mutation of a cell surface receptor, Lrp4, which modulates multiple signaling pathways, produces incisors with grooved enamel surfaces that exhibit the same molecular characteristics as the tips of molar cusps. Mice with a null mutation of Lrp4 develop extra cusps on molars and have incisors that exhibit clear molar-like cusp and root morphologies. Molecular analysis identifies misregulation of Shh and Bmp signaling in the mutant incisors and suggests an uncoupling of the processes of tooth shape determination and morphogenesis. Incisors thus possess a developmentally suppressed, cuspid crown-like morphogenesis program similar to that in molars that is revealed by loss of Lrp4 activity. Several mammalian species naturally possess multicuspid incisors, suggesting that mammals have the capacity to form multicuspid teeth regardless of location in the oral jaw. Localized loss of enamel may thus have been an intermediary step in the evolution of cusps, both of which use Lrp4-mediated signaling. cusp | Lrp4 | tooth development | evo/devo | multicuspid crown V ertebrates exhibit remarkable diversity in their dentitions, which is a feature of the importance of tooth shape in adaptation to new feeding strategies in evolution. Even quite closely related species of mammals can have different shapes of teeth and thus tooth development provides an excellent model for molecularly based evolutionary developmental biological studies (evo/devo). These tooth evolutional changes took place by the activation or inactivation of gene function, and thus evolutionary lost structures or gene activation/inactivation during evolution are occasionally retained as vestigial structures or latent gene activation/inactivation at embryonic stages.
The air-filled cavity and ossicles of the mammalian middle ear conduct sound to the cochlea. Usin... more The air-filled cavity and ossicles of the mammalian middle ear conduct sound to the cochlea. Using transgenic mice, we show that the mammalian middle ear develops through cavitation of a neural crest mass. These cells, which previously underwent an epithelial-to-mesenchymal transformation upon leaving the neural tube, undergo a mesenchymal-to-epithelial transformation to form a lining continuous with the endodermally derived auditory tube. The epithelium derived from endodermal cells, which surrounds the auditory tube and eardrum, develops cilia, whereas the neural crest–derived epithelium does not. Thus, the cilia critical to clearing pathogenic infections from the middle ear are distributed according to developmental derivations. A different process of cavitation appears evident in birds and reptiles, indicating that this dual epithelium may be unique to mammals.
Background: The contribution of the endoderm to the oral tissues of the head has been debated for... more Background: The contribution of the endoderm to the oral tissues of the head has been debated for many
years. With the arrival of Cre/LoxP technology endoderm progenitor cells can now be genetically labeled and tissues derived from the endoderm traced. Using Sox17-2A-iCre/Rosa26 reporter mice we have followed the fate of the endoderm in the teeth, glands, and taste papillae of the oral cavity. Results: No
contribution of the endoderm was observed at any stage of tooth development, or in development of the major salivary glands, in the reporter mouse during development. In contrast, the minor mucous glands of the tongue were found to be of endodermal origin, along with the circumvallate papilla and foliate pap- illae. The mucous minor salivary glands of the palate, however, were of mixed ectodermal and endodermal origin. Conclusions: In contrast to urodele studies, the epithelium of murine teeth is derived solely from
the ectoderm. The border between the ectoderm- and endoderm-derived epithelium may play a role in determining the position of the lingual glands and taste buds, and may explain differences observed between taste buds in the anterior and posterior part of the tongue.
Background: The stapes, an ossicle found within the middle ear, is involved in transmitting sound... more Background: The stapes, an ossicle found within the middle ear, is involved in transmitting sound waves to
the inner ear by means of the oval window. There are several developmental problems associated with this ossicle and the oval window, which cause hearing loss. The developmental origin of these tissues has not been fully elucidated. Results: Using transgenic reporter mice, we have shown that the stapes is of dual ori-
gin with the stapedial footplate being composed of cells of both neural crest and mesodermal origin. Wnt1cre/Dicer mice fail to develop neural crest-derived cartilages, therefore, have no middle ear ossicles. We have shown in these mice the mesodermal stapedial footplate fails to form and the oval window is induced but underdeveloped. Conclusions: If the neural crest part of the stapes fails to form the mesoder-
mal part does not develop, indicating that the two parts are interdependent. The stapes develops tightly associated with the otic capsule, however, it is not essential for the positioning of the oval window, suggest- ing that other tissues, perhaps within the inner ear are needed for oval window placement.
Journal of Anatomy, 2009
Since their recruitment in the oral cavity, approximately 450 million years ago, teeth have been ... more Since their recruitment in the oral cavity, approximately 450 million years ago, teeth have been subjected to strong selective constraints due to the crucial role that they play in species survival. It is therefore quite surprising that the ability to develop functional teeth has subsequently been lost several times, independently, in various lineages. In this review, we concentrate our attention on tetrapods, the only vertebrate lineage in which several clades lack functional teeth from birth to adulthood. Indeed, in other lineages, teeth can be absent in adults but be functionally present in larvae and juveniles, can be absent in the oral cavity but exist in the pharyngeal region, or can develop on the upper jaw but be absent on the lower jaw. Here, we analyse the current data on toothless (edentate) tetrapod taxa, including information available on enamel-less species. Firstly, we provide an analysis of the dispersed and fragmentary morphological data published on the various living taxa concerned (and their extinct relatives) with the aim of tracing the origin of tooth or enamel loss, i.e. toads in Lissamphibia, turtles and birds in Sauropsida, and baleen whales, pangolins, anteaters, sloths, armadillos and aardvark in Mammalia. Secondly, we present current hypotheses on the genetic basis of tooth loss in the chicken and thirdly, we try to answer the question of how these taxa have survived tooth loss given the crucial importance of this tool. The loss of teeth (or only enamel) in all of these taxa was not lethal because it was always preceded in evolution by the pre-adaptation of a secondary tool (beak, baleens, elongated adhesive tongues or hypselodonty) useful for improving efficiency in food uptake. The positive selection of such secondary tools would have led to relaxed functional constraints on teeth and would have later compensated for the loss of teeth. These hypotheses raise numerous questions that will hopefully be answered in the near future.
Developmental Dynamics, 2002
AbstractThe neural crest is a migratory population of cells that originates from the dorsal neura... more AbstractThe neural crest is a migratory population of cells that originates from the dorsal neural tube in vertebrates. Recently, the existence of a group of ventrally emigrating neural tube (VENT) cells has been proposed, based upon cell labelling studies in the hindbrain of avian embryos. Like crest cells, these VENT cells have been reported to give rise to numerous cell types. VENT cell emigration is thought to occur after embryonic day (E) 3, when neural crest cell production has ceased. Migration of cells from the ventral neural tube into the periphery was inferred retrospectively after examining numerous embryos harvested at different stages. We have attempted to label VENT cells in vivo by using a green fluorescent protein (GFP) expression vector, electroporated into the ventral neural tube after crest cell migration and before the putative migration of the ventrally localised cells. Because GFP can be visualised strongly in living tissue a few hours after electroporation, the migration of labelled cells within the same embryo can be followed. Fluorescent cells labelled in the mid-hindbrain region were examined in ovo and in explant culture. No GFP-expressing cells were detected emigrating from the ventral neural tube from E3 to E5. Our findings are, thus, in disagreement with those of previous studies, which have indicated the existence of VENT cells in the cranial region. © 2001 Wiley-Liss, Inc.
Evolution & Development, 2004
The skeletal elements of the branchial region are made by neural crest cells, following tissue in... more The skeletal elements of the branchial region are made by neural crest cells, following tissue interactions with the pharyngeal endoderm. Previous transplantation experiments have claimed that the cranial neural crest is morphogenetically prespecified in respect of its branchial skeletal derivatives, that is, that information for the number, size, shape, and position of its individual elements is already determined in these cells when they are still in the neural folds. This positional information would somehow be preserved during delamination from the neural tube and migration into the branchial arches, before being read out as a spatial pattern of chondrogenesis and osteogenesis. However, it now appears that signals from the endoderm are able to specify not only the histogenic differentiation state of neural crest cells but also the identity and orientation of the branchial skeletal elements. It is therefore important to ask whether fine details of branchial skeletal pattern such as those that exist between different species are also governed by extrinsic factors, such as the endoderm, or by the neural crest itself. We have grafted neural crest between duck and quail embryos and show that the shape and size of the resulting skeletal elements is donor derived. The ability to form species-specific patterns of craniofacial skeletal tissue thus appears to be an inherent property of the neural crest, expressed as species-specific responses to endodermal signals.
Developmental Biology, 2002
In the avian hindbrain, premigratory neural crest cells undergo programmed cell death (apoptosis)... more In the avian hindbrain, premigratory neural crest cells undergo programmed cell death (apoptosis) in rhombomeres 3 and 5 (r3, r5). Here, we have attempted to analyze the significance of the loss of neural crest cells from these odd-numbered rhombomeres. When apoptosis is prevented in r3 and r5, r3 crest migrate into the first arch and r5 into the third arch. Interestingly, these extra neural crest cells contributed to the formation of ectopic muscle attachment sites that are also found in those species in which r3 and r5 neural crest cells do not undergo apoptosis. Thus, apoptosis in the odd-numbered rhombomeres appears to be an evolutionarily derived mechanism that is required to eliminate r3 and r5 crest migration into first and third arches and thereby remove these muscle attachment sites. © 2002 Elsevier Science (USA)
BMC Developmental Biology, 2007
The middle ear of mammals is composed of three endochondrial ossicles, the stapes, incus and mall... more The middle ear of mammals is composed of three endochondrial ossicles, the stapes, incus and malleus. Joints link the malleus to the incus and the incus to the stapes. In the mouse the first arch derived malleus and incus are formed from a single Sox9 and Type II collagen expressing condensation that later subdivides to give rise to two separate ossicles. In contrast the stapes forms from a separate condensation derived from the second branchial arch. Fusion of the malleus and incus is observed in a number of human syndromes and results in conductive hearing loss. Understanding how this joint forms during normal development is thus an important step in furthering our understanding of such defects.
Developmental Dynamics, 2006
The malleus, incus and stapes form an ossicle chain in the mammalian middle ear. These ossicles a... more The malleus, incus and stapes form an ossicle chain in the mammalian middle ear. These ossicles are articulated by joints that link the chain together. In humans and mice, fusion of the ossicles leads to hearing loss. However, in the adult guinea pig the malleus and incus are normally found as a single complex. In this report, we investigate how the malleus and incus form during mouse and guinea pig development. The murine malleus and incus develop from a single condensation that splits to form the two ossicles. Even before a morphological split, we show that the ossicles have distinct genetic identities and joint markers are expressed. In the guinea pig embryo, joint formation is initiated but no cavitation is observed, resulting in a single complex divided by a thin suture. The malleal-incudo complex in the guinea pig is, therefore, not caused by a defect in joint initiation. Developmental Dynamics 235:1326–1333, 2006. © 2006 Wiley-Liss, Inc.
Developmental Dynamics, 2002
Hox genes have a critical role in controlling the patterning processes of many tissues by imparti... more Hox genes have a critical role in controlling the patterning processes of many tissues by imparting positional information in embryogenesis. Patterning of the pharyngeal component of the skull (the visceroskeleton) has been proposed to be influenced by this “Hox code.” Recently, it has been shown that Hox genes are associated with the evolution of jaws, loss of Hox gene expression in the first branchial arch being necessary for the transition from the agnathan condition to the gnathostome condition. Teeth develop on the first branchial arch in mammals and, therefore, might be expected to be under the control of Hox genes in a manner similar to that of the cranial skeletal elements. However, we show that, unlike cartilage and bone, the development of teeth is not affected by alterations in Hoxa2 expression. Tooth development in the first arch was unaffected by overexpression of Hoxa2, whereas recombinations of second arch mesenchyme with first arch epithelium led to tooth development within a Hoxa2-positive environment. These data demonstrate that teeth develop from local interactions and that tooth formation is not under the axial patterning program specified by the Hox genes. We propose that the evolutionary development of teeth in the first branchial arch is independent of the loss of Hox expression necessary for the development of the jaw. © 2002 Wiley-Liss, Inc.
Journal of Experimental Zoology Part B-molecular and Developmental Evolution, 2009
The amelogenins comprise 90% of the developing extracellular enamel matrix proteins and play a ma... more The amelogenins comprise 90% of the developing extracellular enamel matrix proteins and play a major role in the biomineralization and structural organization of enamel. Amelogenins were also detected, in smaller amounts, in postnatal calcifying mesenchymal tissues, and in several nonmineralizing tissues including brain. Low molecular mass amelogenin isoforms were suggested to have signaling activity; to produce ectopically chondrogenic and osteogenic-like tissue and to affect mouse tooth germ differentiation in vitro. Recently, some amelogenin isoforms were found to bind to the cell surface receptors; LAMP-1, LAMP-2 and CD63, and subsequently localize to the perinuclear region of the cell. The recombinant amelogenin protein (rHAM+) alone brought about regeneration of the tooth supporting tissues: cementum, periodontal ligament and alveolar bone, in the dog model, through recruitment of progenitor cells and mesenchymal stem cells.We show that amelogenin is expressed in various tissues of the developing mouse embryonic cranio-facial complex such as brain, eye, ganglia, peripheral nerve trunks, cartilage and bone, and is already expressed at E10.5 in the brain and eye, long before the initiation of tooth formation. Amelogenin protein expression was detected in the tooth germ (dental lamina) already at E13.5, much earlier than previously reported (E19). Application of amelogenin (rHAM+) beads together with DiI, on E13.5 and E14.5 embryonic mandibular mesenchyme and on embryonic tooth germ, revealed recruitment of mesenchymal cells. The present results indicate that amelogenin has an important role in many tissues of the cranio-facial complex during mouse embryonic development and differentiation, and might be a multifunctional protein. J. Exp. Zool. (Mol. Dev. Evol.) 312B:445–457, 2009. © 2008 Wiley-Liss, Inc.
Developmental Biology, 2011
The parathyroid glands originate from the endoderm of the caudal pharyngeal pouches. How these pa... more The parathyroid glands originate from the endoderm of the caudal pharyngeal pouches. How these parathyroids are restricted to developing in the caudal pouches is unclear. In this paper we investigate the role of Shh signalling in patterning the vertebrate pharyngeal pouches, and show that Hh signalling may be involved in restricting the expression of the parathyroid marker Gcm2 in the pharyngeal epithelium. In the chick and mouse, Shh signalling is excluded or highly reduced in the posterior/caudal pouches, where the parathyroid marker Gcm2 is expressed, while remaining at high levels in the more anterior pouches. Moreover, though the block of Shh signalling at early developmental stages results in the loss of chick Gcm2 expression, at later stages, it induces ectopic Gcm2 expression domains in the second and first pharyngeal epithelium, suggesting that HH signalling prevents Gcm2 in those tissues. These ectopic domains go on to express other parathyroid markers but do not migrate and develop into ectopic parathyroids. Differences in the expression of Gcm2 in the chick, mouse and zebrafish, correlate with changing patterns of Shh signalling, indicating a conserved regulatory mechanism that acts to define pouch derivatives.
Journal of Dental Research, 1999
Development of the mammalian tooth has for many years served as a useful model system for the stu... more Development of the mammalian tooth has for many years served as a useful model system for the study of cell-cell interactions in organogenesis. Early development of teeth (tooth buds) shows many morphological and molecular similarities with early development of other organs such as the lung, hair, kidney, etc. There has been much progress toward understanding epithelial/mesenchymal cell signaling in tooth germ formation. Advances in understanding the formation of different shapes of teeth (morphogenesis) at their correct positions in the jaws (patterning) has, until recently, been less forthcoming. We review here the latest ideas on the control of odontogenic patterning and morphogenesis. The stages of early tooth development are welldefined histologically and have been described in numerous textbooks. The progression from localized thickenings of oral epithelium to bud, cap, and bell stages provides an adequate description of the gross morphological changes seen in the epithelial cells of early developing tooth germs. Less obvious are the concomitant changes taking place in the dental (ecto)mesenchymal cells which originate from the cranial neural crest and which condense around the tooth bud epithelium. However, it is very clear that these mesenchymal cells are equal partners with epithelium during the early stages of tooth germ formation and undergo complex changes which, although not obvious histologically, are revealed with molecular (gene) probes. Genes identified as being important for the early communication between the epithelial and ectomesenchymal cells mainly comprise those which code for proteins which act as secreted signals between the cells (ligands) and those that code for nuclear proteins that act to control gene expression in response to the signals. Little is presently known about the changes in structural proteins such as cell adhesion molecules which are involved in mediating the physical interactions between cells and generating the morphological changes.
Journal of Experimental Zoology Part B-molecular and Developmental Evolution, 2009
This study highlights the dynamic nature of the mesenchymal cells during tooth development from t... more This study highlights the dynamic nature of the mesenchymal cells during tooth development from the bud to the bell stage. Condensing mesenchymal cells, labelled on either side of the developing tooth bud, move toward the presumptive roots forming an arc of cells under the dental papilla. These labelled cells take part in formation of the dental follicle, which contributes to both the tooth and its surrounding periodontium, including the supporting alveolar bone. This study, thus, physically links development of the tooth with the tissue into which it develops. The results obtained clearly indicate that the tooth organ is an entity comprising dental and periodontal tissue. J. Exp. Zool. (Mol. Dev. Evol.) 312B:510–517, 2009. © 2009 Wiley-Liss, Inc.
Developmental Biology, 2009
TBX1 is a principal candidate gene for DiGeorge syndrome, a developmental anomaly that affects th... more TBX1 is a principal candidate gene for DiGeorge syndrome, a developmental anomaly that affects the heart, thymus, parathyroid, face, and teeth. A mouse model carrying a deletion in a functional region of the Tbx1 gene has been extensively used to study anomalies related to this syndrome. We have used the Tbx1 null mouse to understand the tooth phenotype reported in patients afflicted by DiGeorge syndrome. Because of the early lethality of the Tbx1−/− mice, we used long-term culture techniques that allow the unharmed growth of incisors until their full maturity. All cultured incisors of Tbx1−/− mice were hypoplastic and lacked enamel, while thorough histological examinations demonstrated the complete absence of ameloblasts. The absence of enamel is preceded by a decrease in proliferation of the ameloblast precursor cells and a reduction in amelogenin gene expression. The cervical loop area of the incisor, which contains the niche for the epithelial stem cells, was either severely reduced or completely missing in mutant incisors. In contrast, ectopic expression of Tbx1 was observed in incisors from mice with upregulated Fibroblast Growth Factor signalling and was closely linked to ectopic enamel formation and deposition in these incisors. These results demonstrate that Tbx1 is essential for the maintenance of ameloblast progenitor cells in rodent incisors and that its deletion results in the absence of enamel formation.
Seminars in Cell & Developmental Biology, 2010
The pharyngeal arches form the face and neck of the developing embryo. The pharyngeal tissue is d... more The pharyngeal arches form the face and neck of the developing embryo. The pharyngeal tissue is divided into distinct arches by the formation of clefts and pouches in between the arches. These clefts and pouches form at the juxtaposition between the ectoderm and endoderm and develop into a variety of essential structures, such as the ear drum, and glands such as the thymus and parathyroids. How these pouches and clefts between the arches form and what structures they develop into is the subject of this review. Differences in pouch derivatives are described in different animals and the evolution of these structures are investigated. The implications of defects in pouch and cleft development on human health are also discussed.
International Journal of Developmental Biology, 2006
Tooth morphogenesis is accompanied by apoptotic events which show restricted temporospatial patte... more Tooth morphogenesis is accompanied by apoptotic events which show restricted temporospatial patterns suggesting multiple roles in odontogenesis. Dental apoptosis seems to be caspase dependent and caspase-3 has been shown to be activated during dental apoptosis. Caspase-3 mutant mice on different genetic backgrounds were used to investigate alterations in dental apoptosis and molar tooth morphogenesis. Mouse embryos at E15.5 were analyzed to reveal any changes in enamel knots, which are transient structures eliminated by apoptosis. In caspase-3 -/mice on the B57BL/6 background, disorganization of the epithelium was found in the original primary enamel knot area and confirmed by altered expression of Shh. Despite this early defect in molar tooth development, these mutants showed correct formation of secondary enamel knots as indicated by Fgf-4 expression. Analyses of adult molar teeth did not reveal any major alterations in tooth shape, enamel structure or pattern when compared to heterozygote littermates. In caspase-3 -/mice on the 129X1/SvJ background, no defects in tooth development were found except the position of the upper molars which developed more posteriorly in the oral cavity. This is likely, however, to be a secondary defect caused by a physical squashing of the face by the malformed brain. The results suggest that although caspase-3 becomes activated and may be essential for dental apoptosis, it does not seem fundamental for formation of normal mineralised molar teeth.
Developmental Dynamics, 2004
Robinow syndrome (RS) is a human dwarfism syndrome characterized by mesomelic limb shortening, ve... more Robinow syndrome (RS) is a human dwarfism syndrome characterized by mesomelic limb shortening, vertebral and craniofacial malformations and small external genitals. We have analyzed Ror2-/- mice as a model for the developmental pathology of RS. Our results demonstrate that vertebral malformations in Ror2-/- mice are due to reductions in the presomitic mesoderm and defects in somitogenesis. Mesomelic limb shortening in Ror2-/- mice is a consequence of perturbed chondrocyte differentiation. Moreover, we show that the craniofacial phenotype is caused by a midline outgrowth defect. Ror2 expression in the genital tubercle and its reduced size in Ror2-/- mice makes it likely that Ror2 is involved in genital development. In conclusion, our findings suggest that Ror2 is essential at multiple sites during development. The Ror2-/- mouse provides a suitable model that may help to explain many of the underlying developmental malformations in individuals with Robinow syndrome. Developmental Dynamics 229:400–410, 2004, © 2004 Wiley-Liss, Inc.
Proceedings of the National Academy of Sciences, 2010
Changes in tooth shape have played a major role in vertebrate evolution with modification of dent... more Changes in tooth shape have played a major role in vertebrate evolution with modification of dentition allowing an organism to adapt to new feeding strategies. The current view is that molar teeth evolved from simple conical teeth, similar to canines, by progressive addition of extra "cones" to form progressively complex multicuspid crowns. Mammalian incisors, however, are neither conical nor multicuspid, and their evolution is unclear. We show that hypomorphic mutation of a cell surface receptor, Lrp4, which modulates multiple signaling pathways, produces incisors with grooved enamel surfaces that exhibit the same molecular characteristics as the tips of molar cusps. Mice with a null mutation of Lrp4 develop extra cusps on molars and have incisors that exhibit clear molar-like cusp and root morphologies. Molecular analysis identifies misregulation of Shh and Bmp signaling in the mutant incisors and suggests an uncoupling of the processes of tooth shape determination and morphogenesis. Incisors thus possess a developmentally suppressed, cuspid crown-like morphogenesis program similar to that in molars that is revealed by loss of Lrp4 activity. Several mammalian species naturally possess multicuspid incisors, suggesting that mammals have the capacity to form multicuspid teeth regardless of location in the oral jaw. Localized loss of enamel may thus have been an intermediary step in the evolution of cusps, both of which use Lrp4-mediated signaling. cusp | Lrp4 | tooth development | evo/devo | multicuspid crown V ertebrates exhibit remarkable diversity in their dentitions, which is a feature of the importance of tooth shape in adaptation to new feeding strategies in evolution. Even quite closely related species of mammals can have different shapes of teeth and thus tooth development provides an excellent model for molecularly based evolutionary developmental biological studies (evo/devo). These tooth evolutional changes took place by the activation or inactivation of gene function, and thus evolutionary lost structures or gene activation/inactivation during evolution are occasionally retained as vestigial structures or latent gene activation/inactivation at embryonic stages.
The air-filled cavity and ossicles of the mammalian middle ear conduct sound to the cochlea. Usin... more The air-filled cavity and ossicles of the mammalian middle ear conduct sound to the cochlea. Using transgenic mice, we show that the mammalian middle ear develops through cavitation of a neural crest mass. These cells, which previously underwent an epithelial-to-mesenchymal transformation upon leaving the neural tube, undergo a mesenchymal-to-epithelial transformation to form a lining continuous with the endodermally derived auditory tube. The epithelium derived from endodermal cells, which surrounds the auditory tube and eardrum, develops cilia, whereas the neural crest–derived epithelium does not. Thus, the cilia critical to clearing pathogenic infections from the middle ear are distributed according to developmental derivations. A different process of cavitation appears evident in birds and reptiles, indicating that this dual epithelium may be unique to mammals.
Background: The contribution of the endoderm to the oral tissues of the head has been debated for... more Background: The contribution of the endoderm to the oral tissues of the head has been debated for many
years. With the arrival of Cre/LoxP technology endoderm progenitor cells can now be genetically labeled and tissues derived from the endoderm traced. Using Sox17-2A-iCre/Rosa26 reporter mice we have followed the fate of the endoderm in the teeth, glands, and taste papillae of the oral cavity. Results: No
contribution of the endoderm was observed at any stage of tooth development, or in development of the major salivary glands, in the reporter mouse during development. In contrast, the minor mucous glands of the tongue were found to be of endodermal origin, along with the circumvallate papilla and foliate pap- illae. The mucous minor salivary glands of the palate, however, were of mixed ectodermal and endodermal origin. Conclusions: In contrast to urodele studies, the epithelium of murine teeth is derived solely from
the ectoderm. The border between the ectoderm- and endoderm-derived epithelium may play a role in determining the position of the lingual glands and taste buds, and may explain differences observed between taste buds in the anterior and posterior part of the tongue.
Background: The stapes, an ossicle found within the middle ear, is involved in transmitting sound... more Background: The stapes, an ossicle found within the middle ear, is involved in transmitting sound waves to
the inner ear by means of the oval window. There are several developmental problems associated with this ossicle and the oval window, which cause hearing loss. The developmental origin of these tissues has not been fully elucidated. Results: Using transgenic reporter mice, we have shown that the stapes is of dual ori-
gin with the stapedial footplate being composed of cells of both neural crest and mesodermal origin. Wnt1cre/Dicer mice fail to develop neural crest-derived cartilages, therefore, have no middle ear ossicles. We have shown in these mice the mesodermal stapedial footplate fails to form and the oval window is induced but underdeveloped. Conclusions: If the neural crest part of the stapes fails to form the mesoder-
mal part does not develop, indicating that the two parts are interdependent. The stapes develops tightly associated with the otic capsule, however, it is not essential for the positioning of the oval window, suggest- ing that other tissues, perhaps within the inner ear are needed for oval window placement.
Journal of Anatomy, 2009
Since their recruitment in the oral cavity, approximately 450 million years ago, teeth have been ... more Since their recruitment in the oral cavity, approximately 450 million years ago, teeth have been subjected to strong selective constraints due to the crucial role that they play in species survival. It is therefore quite surprising that the ability to develop functional teeth has subsequently been lost several times, independently, in various lineages. In this review, we concentrate our attention on tetrapods, the only vertebrate lineage in which several clades lack functional teeth from birth to adulthood. Indeed, in other lineages, teeth can be absent in adults but be functionally present in larvae and juveniles, can be absent in the oral cavity but exist in the pharyngeal region, or can develop on the upper jaw but be absent on the lower jaw. Here, we analyse the current data on toothless (edentate) tetrapod taxa, including information available on enamel-less species. Firstly, we provide an analysis of the dispersed and fragmentary morphological data published on the various living taxa concerned (and their extinct relatives) with the aim of tracing the origin of tooth or enamel loss, i.e. toads in Lissamphibia, turtles and birds in Sauropsida, and baleen whales, pangolins, anteaters, sloths, armadillos and aardvark in Mammalia. Secondly, we present current hypotheses on the genetic basis of tooth loss in the chicken and thirdly, we try to answer the question of how these taxa have survived tooth loss given the crucial importance of this tool. The loss of teeth (or only enamel) in all of these taxa was not lethal because it was always preceded in evolution by the pre-adaptation of a secondary tool (beak, baleens, elongated adhesive tongues or hypselodonty) useful for improving efficiency in food uptake. The positive selection of such secondary tools would have led to relaxed functional constraints on teeth and would have later compensated for the loss of teeth. These hypotheses raise numerous questions that will hopefully be answered in the near future.
Developmental Dynamics, 2002
AbstractThe neural crest is a migratory population of cells that originates from the dorsal neura... more AbstractThe neural crest is a migratory population of cells that originates from the dorsal neural tube in vertebrates. Recently, the existence of a group of ventrally emigrating neural tube (VENT) cells has been proposed, based upon cell labelling studies in the hindbrain of avian embryos. Like crest cells, these VENT cells have been reported to give rise to numerous cell types. VENT cell emigration is thought to occur after embryonic day (E) 3, when neural crest cell production has ceased. Migration of cells from the ventral neural tube into the periphery was inferred retrospectively after examining numerous embryos harvested at different stages. We have attempted to label VENT cells in vivo by using a green fluorescent protein (GFP) expression vector, electroporated into the ventral neural tube after crest cell migration and before the putative migration of the ventrally localised cells. Because GFP can be visualised strongly in living tissue a few hours after electroporation, the migration of labelled cells within the same embryo can be followed. Fluorescent cells labelled in the mid-hindbrain region were examined in ovo and in explant culture. No GFP-expressing cells were detected emigrating from the ventral neural tube from E3 to E5. Our findings are, thus, in disagreement with those of previous studies, which have indicated the existence of VENT cells in the cranial region. © 2001 Wiley-Liss, Inc.
Evolution & Development, 2004
The skeletal elements of the branchial region are made by neural crest cells, following tissue in... more The skeletal elements of the branchial region are made by neural crest cells, following tissue interactions with the pharyngeal endoderm. Previous transplantation experiments have claimed that the cranial neural crest is morphogenetically prespecified in respect of its branchial skeletal derivatives, that is, that information for the number, size, shape, and position of its individual elements is already determined in these cells when they are still in the neural folds. This positional information would somehow be preserved during delamination from the neural tube and migration into the branchial arches, before being read out as a spatial pattern of chondrogenesis and osteogenesis. However, it now appears that signals from the endoderm are able to specify not only the histogenic differentiation state of neural crest cells but also the identity and orientation of the branchial skeletal elements. It is therefore important to ask whether fine details of branchial skeletal pattern such as those that exist between different species are also governed by extrinsic factors, such as the endoderm, or by the neural crest itself. We have grafted neural crest between duck and quail embryos and show that the shape and size of the resulting skeletal elements is donor derived. The ability to form species-specific patterns of craniofacial skeletal tissue thus appears to be an inherent property of the neural crest, expressed as species-specific responses to endodermal signals.
Developmental Biology, 2002
In the avian hindbrain, premigratory neural crest cells undergo programmed cell death (apoptosis)... more In the avian hindbrain, premigratory neural crest cells undergo programmed cell death (apoptosis) in rhombomeres 3 and 5 (r3, r5). Here, we have attempted to analyze the significance of the loss of neural crest cells from these odd-numbered rhombomeres. When apoptosis is prevented in r3 and r5, r3 crest migrate into the first arch and r5 into the third arch. Interestingly, these extra neural crest cells contributed to the formation of ectopic muscle attachment sites that are also found in those species in which r3 and r5 neural crest cells do not undergo apoptosis. Thus, apoptosis in the odd-numbered rhombomeres appears to be an evolutionarily derived mechanism that is required to eliminate r3 and r5 crest migration into first and third arches and thereby remove these muscle attachment sites. © 2002 Elsevier Science (USA)
BMC Developmental Biology, 2007
The middle ear of mammals is composed of three endochondrial ossicles, the stapes, incus and mall... more The middle ear of mammals is composed of three endochondrial ossicles, the stapes, incus and malleus. Joints link the malleus to the incus and the incus to the stapes. In the mouse the first arch derived malleus and incus are formed from a single Sox9 and Type II collagen expressing condensation that later subdivides to give rise to two separate ossicles. In contrast the stapes forms from a separate condensation derived from the second branchial arch. Fusion of the malleus and incus is observed in a number of human syndromes and results in conductive hearing loss. Understanding how this joint forms during normal development is thus an important step in furthering our understanding of such defects.
Developmental Dynamics, 2006
The malleus, incus and stapes form an ossicle chain in the mammalian middle ear. These ossicles a... more The malleus, incus and stapes form an ossicle chain in the mammalian middle ear. These ossicles are articulated by joints that link the chain together. In humans and mice, fusion of the ossicles leads to hearing loss. However, in the adult guinea pig the malleus and incus are normally found as a single complex. In this report, we investigate how the malleus and incus form during mouse and guinea pig development. The murine malleus and incus develop from a single condensation that splits to form the two ossicles. Even before a morphological split, we show that the ossicles have distinct genetic identities and joint markers are expressed. In the guinea pig embryo, joint formation is initiated but no cavitation is observed, resulting in a single complex divided by a thin suture. The malleal-incudo complex in the guinea pig is, therefore, not caused by a defect in joint initiation. Developmental Dynamics 235:1326–1333, 2006. © 2006 Wiley-Liss, Inc.
Developmental Dynamics, 2002
Hox genes have a critical role in controlling the patterning processes of many tissues by imparti... more Hox genes have a critical role in controlling the patterning processes of many tissues by imparting positional information in embryogenesis. Patterning of the pharyngeal component of the skull (the visceroskeleton) has been proposed to be influenced by this “Hox code.” Recently, it has been shown that Hox genes are associated with the evolution of jaws, loss of Hox gene expression in the first branchial arch being necessary for the transition from the agnathan condition to the gnathostome condition. Teeth develop on the first branchial arch in mammals and, therefore, might be expected to be under the control of Hox genes in a manner similar to that of the cranial skeletal elements. However, we show that, unlike cartilage and bone, the development of teeth is not affected by alterations in Hoxa2 expression. Tooth development in the first arch was unaffected by overexpression of Hoxa2, whereas recombinations of second arch mesenchyme with first arch epithelium led to tooth development within a Hoxa2-positive environment. These data demonstrate that teeth develop from local interactions and that tooth formation is not under the axial patterning program specified by the Hox genes. We propose that the evolutionary development of teeth in the first branchial arch is independent of the loss of Hox expression necessary for the development of the jaw. © 2002 Wiley-Liss, Inc.
Journal of Experimental Zoology Part B-molecular and Developmental Evolution, 2009
The amelogenins comprise 90% of the developing extracellular enamel matrix proteins and play a ma... more The amelogenins comprise 90% of the developing extracellular enamel matrix proteins and play a major role in the biomineralization and structural organization of enamel. Amelogenins were also detected, in smaller amounts, in postnatal calcifying mesenchymal tissues, and in several nonmineralizing tissues including brain. Low molecular mass amelogenin isoforms were suggested to have signaling activity; to produce ectopically chondrogenic and osteogenic-like tissue and to affect mouse tooth germ differentiation in vitro. Recently, some amelogenin isoforms were found to bind to the cell surface receptors; LAMP-1, LAMP-2 and CD63, and subsequently localize to the perinuclear region of the cell. The recombinant amelogenin protein (rHAM+) alone brought about regeneration of the tooth supporting tissues: cementum, periodontal ligament and alveolar bone, in the dog model, through recruitment of progenitor cells and mesenchymal stem cells.We show that amelogenin is expressed in various tissues of the developing mouse embryonic cranio-facial complex such as brain, eye, ganglia, peripheral nerve trunks, cartilage and bone, and is already expressed at E10.5 in the brain and eye, long before the initiation of tooth formation. Amelogenin protein expression was detected in the tooth germ (dental lamina) already at E13.5, much earlier than previously reported (E19). Application of amelogenin (rHAM+) beads together with DiI, on E13.5 and E14.5 embryonic mandibular mesenchyme and on embryonic tooth germ, revealed recruitment of mesenchymal cells. The present results indicate that amelogenin has an important role in many tissues of the cranio-facial complex during mouse embryonic development and differentiation, and might be a multifunctional protein. J. Exp. Zool. (Mol. Dev. Evol.) 312B:445–457, 2009. © 2008 Wiley-Liss, Inc.
Developmental Biology, 2011
The parathyroid glands originate from the endoderm of the caudal pharyngeal pouches. How these pa... more The parathyroid glands originate from the endoderm of the caudal pharyngeal pouches. How these parathyroids are restricted to developing in the caudal pouches is unclear. In this paper we investigate the role of Shh signalling in patterning the vertebrate pharyngeal pouches, and show that Hh signalling may be involved in restricting the expression of the parathyroid marker Gcm2 in the pharyngeal epithelium. In the chick and mouse, Shh signalling is excluded or highly reduced in the posterior/caudal pouches, where the parathyroid marker Gcm2 is expressed, while remaining at high levels in the more anterior pouches. Moreover, though the block of Shh signalling at early developmental stages results in the loss of chick Gcm2 expression, at later stages, it induces ectopic Gcm2 expression domains in the second and first pharyngeal epithelium, suggesting that HH signalling prevents Gcm2 in those tissues. These ectopic domains go on to express other parathyroid markers but do not migrate and develop into ectopic parathyroids. Differences in the expression of Gcm2 in the chick, mouse and zebrafish, correlate with changing patterns of Shh signalling, indicating a conserved regulatory mechanism that acts to define pouch derivatives.
Journal of Dental Research, 1999
Development of the mammalian tooth has for many years served as a useful model system for the stu... more Development of the mammalian tooth has for many years served as a useful model system for the study of cell-cell interactions in organogenesis. Early development of teeth (tooth buds) shows many morphological and molecular similarities with early development of other organs such as the lung, hair, kidney, etc. There has been much progress toward understanding epithelial/mesenchymal cell signaling in tooth germ formation. Advances in understanding the formation of different shapes of teeth (morphogenesis) at their correct positions in the jaws (patterning) has, until recently, been less forthcoming. We review here the latest ideas on the control of odontogenic patterning and morphogenesis. The stages of early tooth development are welldefined histologically and have been described in numerous textbooks. The progression from localized thickenings of oral epithelium to bud, cap, and bell stages provides an adequate description of the gross morphological changes seen in the epithelial cells of early developing tooth germs. Less obvious are the concomitant changes taking place in the dental (ecto)mesenchymal cells which originate from the cranial neural crest and which condense around the tooth bud epithelium. However, it is very clear that these mesenchymal cells are equal partners with epithelium during the early stages of tooth germ formation and undergo complex changes which, although not obvious histologically, are revealed with molecular (gene) probes. Genes identified as being important for the early communication between the epithelial and ectomesenchymal cells mainly comprise those which code for proteins which act as secreted signals between the cells (ligands) and those that code for nuclear proteins that act to control gene expression in response to the signals. Little is presently known about the changes in structural proteins such as cell adhesion molecules which are involved in mediating the physical interactions between cells and generating the morphological changes.
Journal of Experimental Zoology Part B-molecular and Developmental Evolution, 2009
This study highlights the dynamic nature of the mesenchymal cells during tooth development from t... more This study highlights the dynamic nature of the mesenchymal cells during tooth development from the bud to the bell stage. Condensing mesenchymal cells, labelled on either side of the developing tooth bud, move toward the presumptive roots forming an arc of cells under the dental papilla. These labelled cells take part in formation of the dental follicle, which contributes to both the tooth and its surrounding periodontium, including the supporting alveolar bone. This study, thus, physically links development of the tooth with the tissue into which it develops. The results obtained clearly indicate that the tooth organ is an entity comprising dental and periodontal tissue. J. Exp. Zool. (Mol. Dev. Evol.) 312B:510–517, 2009. © 2009 Wiley-Liss, Inc.
Developmental Biology, 2009
TBX1 is a principal candidate gene for DiGeorge syndrome, a developmental anomaly that affects th... more TBX1 is a principal candidate gene for DiGeorge syndrome, a developmental anomaly that affects the heart, thymus, parathyroid, face, and teeth. A mouse model carrying a deletion in a functional region of the Tbx1 gene has been extensively used to study anomalies related to this syndrome. We have used the Tbx1 null mouse to understand the tooth phenotype reported in patients afflicted by DiGeorge syndrome. Because of the early lethality of the Tbx1−/− mice, we used long-term culture techniques that allow the unharmed growth of incisors until their full maturity. All cultured incisors of Tbx1−/− mice were hypoplastic and lacked enamel, while thorough histological examinations demonstrated the complete absence of ameloblasts. The absence of enamel is preceded by a decrease in proliferation of the ameloblast precursor cells and a reduction in amelogenin gene expression. The cervical loop area of the incisor, which contains the niche for the epithelial stem cells, was either severely reduced or completely missing in mutant incisors. In contrast, ectopic expression of Tbx1 was observed in incisors from mice with upregulated Fibroblast Growth Factor signalling and was closely linked to ectopic enamel formation and deposition in these incisors. These results demonstrate that Tbx1 is essential for the maintenance of ameloblast progenitor cells in rodent incisors and that its deletion results in the absence of enamel formation.
Seminars in Cell & Developmental Biology, 2010
The pharyngeal arches form the face and neck of the developing embryo. The pharyngeal tissue is d... more The pharyngeal arches form the face and neck of the developing embryo. The pharyngeal tissue is divided into distinct arches by the formation of clefts and pouches in between the arches. These clefts and pouches form at the juxtaposition between the ectoderm and endoderm and develop into a variety of essential structures, such as the ear drum, and glands such as the thymus and parathyroids. How these pouches and clefts between the arches form and what structures they develop into is the subject of this review. Differences in pouch derivatives are described in different animals and the evolution of these structures are investigated. The implications of defects in pouch and cleft development on human health are also discussed.
International Journal of Developmental Biology, 2006
Tooth morphogenesis is accompanied by apoptotic events which show restricted temporospatial patte... more Tooth morphogenesis is accompanied by apoptotic events which show restricted temporospatial patterns suggesting multiple roles in odontogenesis. Dental apoptosis seems to be caspase dependent and caspase-3 has been shown to be activated during dental apoptosis. Caspase-3 mutant mice on different genetic backgrounds were used to investigate alterations in dental apoptosis and molar tooth morphogenesis. Mouse embryos at E15.5 were analyzed to reveal any changes in enamel knots, which are transient structures eliminated by apoptosis. In caspase-3 -/mice on the B57BL/6 background, disorganization of the epithelium was found in the original primary enamel knot area and confirmed by altered expression of Shh. Despite this early defect in molar tooth development, these mutants showed correct formation of secondary enamel knots as indicated by Fgf-4 expression. Analyses of adult molar teeth did not reveal any major alterations in tooth shape, enamel structure or pattern when compared to heterozygote littermates. In caspase-3 -/mice on the 129X1/SvJ background, no defects in tooth development were found except the position of the upper molars which developed more posteriorly in the oral cavity. This is likely, however, to be a secondary defect caused by a physical squashing of the face by the malformed brain. The results suggest that although caspase-3 becomes activated and may be essential for dental apoptosis, it does not seem fundamental for formation of normal mineralised molar teeth.
Developmental Dynamics, 2004
Robinow syndrome (RS) is a human dwarfism syndrome characterized by mesomelic limb shortening, ve... more Robinow syndrome (RS) is a human dwarfism syndrome characterized by mesomelic limb shortening, vertebral and craniofacial malformations and small external genitals. We have analyzed Ror2-/- mice as a model for the developmental pathology of RS. Our results demonstrate that vertebral malformations in Ror2-/- mice are due to reductions in the presomitic mesoderm and defects in somitogenesis. Mesomelic limb shortening in Ror2-/- mice is a consequence of perturbed chondrocyte differentiation. Moreover, we show that the craniofacial phenotype is caused by a midline outgrowth defect. Ror2 expression in the genital tubercle and its reduced size in Ror2-/- mice makes it likely that Ror2 is involved in genital development. In conclusion, our findings suggest that Ror2 is essential at multiple sites during development. The Ror2-/- mouse provides a suitable model that may help to explain many of the underlying developmental malformations in individuals with Robinow syndrome. Developmental Dynamics 229:400–410, 2004, © 2004 Wiley-Liss, Inc.