Ancient vertebrate dermal armor evolved from trunk neural crest.

Bone is an evolutionary novelty of vertebrates, likely to have first emerged as part of ancestral dermal armor that consisted of osteogenic and odontogenic components. Whether these early vertebrate structures arose from mesoderm or neural crest cells has been a matter of considerable debate. To examine the developmental origin of the bony part of the dermal armor, we have performed in vivo lineage tracing in the sterlet sturgeon, a representative of nonteleost ray-finned fish that has retained an extensive postcranial dermal skeleton. The results definitively show that sterlet trunk neural crest cells give rise to osteoblasts of the scutes. Transcriptional profiling further reveals neural crest gene signature in sterlet scutes as well as bichir scales. Finally, histological and microCT analyses of ray-finned fish dermal armor show that their scales and scutes are formed by bone, dentin, and hypermineralized covering tissues, in various combinations, that resemble those of the first armored vertebrates. Taken together, our results support a primitive skeletogenic role for the neural crest along the entire body axis, that was later progressively restricted to the cranial region during vertebrate evolution. Thus, the neural crest was a crucial evolutionary innovation driving the origin and diversification of dermal armor along the entire body axis.

Pre-mandibular pharyngeal pouches in early non-teleost fish embryos.

The vertebrate pharynx is a key embryonic structure with crucial importance for the metameric organization of the head and face. The pharynx is primarily built upon progressive formation of paired pharyngeal pouches that typically develop in post-oral (mandibular, hyoid and branchial) domains. However, in the early embryos of non-teleost fishes, we have previously identified pharyngeal pouch-like outpocketings also in the pre-oral domain of the cranial endoderm. This pre-oral gut (POG) forms by early pouching of the primitive gut cavity, followed by the sequential formation of typical (post-oral) pharyngeal pouches. Here, we tested the pharyngeal nature of the POG by analysing expression patterns of selected core pharyngeal regulatory network genes in bichir and sturgeon embryos. Our comparison revealed generally shared expression patterns, including Shh, Pax9, Tbx1, Eya1, Six1, Ripply3 or Fgf8, between early POG and post-oral pharyngeal pouches. POG thus shares pharyngeal pouch-like morphogenesis and a gene expression profile with pharyngeal pouches and can be regarded as a pre-mandibular pharyngeal pouch. We further suggest that pre-mandibular pharyngeal pouches represent a plesiomorphic vertebrate trait inherited from our ancestor's pharyngeal metameric organization, which is incorporated in the early formation of the pre-chordal plate of vertebrate embryos.

Pre-oral gut contributes to facial structures in non-teleost fishes.

Despite the wide variety of adaptive modifications in the oral and facial regions of vertebrates, their early oropharyngeal development is considered strictly uniform. It involves sequential formation of the mouth and pharyngeal pouches, with ectoderm outlining the outer surface and endoderm the inner surface, as a rule. At the extreme anterior domain of vertebrate embryos, the ectoderm and endoderm directly juxtapose and initial development of this earliest ecto-endoderm interface, the primary mouth, typically involves ectodermal stomodeal invagination that limits the anterior expansion of the foregut endoderm. Here we present evidence that in embryos of extant non-teleost fishes, oral (stomodeal) formation is preceded by the development of prominent pre-oral gut diverticula (POGD) between the forebrain and roof of the forming mouth. Micro-computed tomography (micro-CT) imaging of bichir, sturgeon and gar embryos revealed that foregut outpocketing at the pre-oral domain begins even before the sequential formation of pharyngeal pouches. The presence of foregut-derived cells in the front of the mouth was further confirmed by in vivo experiments that allowed specific tracing of the early endodermal lining. We show that POGD in sturgeons contribute to the orofacial surface of their larvae, comprising oral teeth, lips, and sensory barbels. To our knowledge, this is the first thorough evidence for endodermal origin of external craniofacial structures in any vertebrate. In bichir and gar embryos, POGD form prominent cranial adhesive organs that are characteristic of the ancient bauplan of free-living chordate larvae. POGD hence seem arguably to be ancestral for all ray-finned fishes, and their topology, pharyngeal-like morphogenesis and gene expression suggest that they are evolutionarily related to the foregut-derived diverticula of early chordate and hemichordate embryos. The formation of POGD might thus represent an ancestral developmental module with deep deuterostome origins.

The remarkable dynamics in the establishment, rearrangement, and loss of dentition during the ontogeny of the sterlet sturgeon.

BACKGROUND: Sturgeons belong to an early-branching lineage often used as a proxy of ancestor-like traits of ray-finned fishes. However, many features of this lineage, such as the transitory presence and the eventual loss of dentition, exemplify specializations that, in fact, provide important information on lineage-specific evolutionary dynamics. RESULTS: Here, we introduce a detailed overview of the dentition during the development of the sterlet sturgeon. The dentition is composed of tooth fields at oral, palatal, and anterior pharyngeal regions. Oral fields are single-rowed, non-renewed and are shed early. Palatal and pharyngeal fields are multi-rowed and renewed from the adjacent superficial epithelium without the presence of the successional dental lamina. The early loss of oral fields and subsequent establishment of palatal and pharyngeal fields leads to a translocation of the functional dentition from the front to the rear of the oropharyngeal cavity until the eventual loss of all teeth. CONCLUSIONS: Our survey shows the sterlet dentition as a dynamic organ system displaying differential composition at different time points in the lifetime of this fish. These dynamics represent a conspicuous feature of sturgeons, unparalleled among extant vertebrates, and appropriate to scrutinize developmental and evolutionary underpinnings of vertebrate odontogenesis.

Migratory patterns and evolutionary plasticity of cranial neural crest cells in ray-finned fishes.

The cranial neural crest (CNC) arises within the developing central nervous system, but then migrates away from the neural tube in three consecutive streams termed mandibular, hyoid and branchial, respectively, according to the order along the anteroposterior axis. While the process of neural crest emigration generally follows a conserved anterior to posterior sequence across vertebrates, we find that ray-finned fishes (bichir, sterlet, gar, and pike) exhibit several heterochronies in the timing and order of CNC emergence that influences their subsequent migratory patterns. First, emigration of the cranial neural crest in these fishes occurs prematurely compared to other vertebrates, already initiating during early neurulation and well before neural tube closure. Second, delamination of the hyoid stream occurs prior to the more anterior mandibular stream; this is associated with early morphogenesis of key hyoid structures like external gills (bichir), a large opercular flap (gar) or first forming cartilage (pike). In sterlet, the hyoid and branchial CNC cells form a single hyobranchial sheet, which later segregates in concert with second pharyngeal pouch morphogenesis. Taken together, the results show that despite generally conserved migratory patterns, heterochronic alterations in the timing of emigration and pattern of migration of CNC cells accompanies morphological diversity of ray-finned fishes.

Interactions of superabsorbent polymers based on acrylamide substances with microorganisms occurring in human dwellings.

Superabsorbent polymers (SAPs) are most often used in hygienic goods or in the agricultural sector but the range of their application is much broader, including the utilization in advanced building materials. Although SAPs were studied widely during the last decades, the data related to the interactions between the natural environment and various organisms occurring on their surface are still lacking. In addition, SAPs can create a variable gel-forming matter in the presence of water but standard ecotoxicological bioassays are mostly not suitable for testing such type of materials. In this study, the SAPs potential for reducing/supporting unwanted indoor microorganism settlement was analyzed by biological methods under controlled laboratory conditions. Three commonly used SAPs (Cabloc CT, Creasorb SIS, Hydropam) were exposed to selected organisms representing green algae (Hematococcus pluvialis), cyanobacteria (Nostoc sp.), yeasts (Saccharomyces cerevisiae), wood-destroying fungi (Gleophyllium trabeum), and aerial molds. The obtained results indicated that Hydropam provided favorable conditions for Hematococcus pluvialis, Nostoc sp., and Saccharomyces cerevisiae. All three tested SAPs inhibited, both with and without nutrient addition, the growth of Gleophyllium trabeum and aerial molds.

The origin and diversification of the developmental mechanisms that pattern the vertebrate head skeleton.

The apparent evolvability of the vertebrate head skeleton has allowed a diverse array of shapes, sizes, and compositions of the head in order to better adapt species to their environments. This encompasses feeding, breathing, sensing, and communicating: the head skeleton somehow participated in the evolution of all these critical processes for the last 500 million years. Through evolution, present head diversity was made possible via developmental modifications to the first head skeletal genetic program. Understanding the development of the vertebrate common ancestor's head skeleton is thus an important step in identifying how different lineages have respectively achieved their many innovations in the head. To this end, cyclostomes (jawless vertebrates) are extremely useful, having diverged from jawed vertebrates approximately 400 million years ago, at the deepest node within living vertebrates. From this ancestral vantage point (that is, the node connecting cyclostomes and gnathostomes) we can best identify the earliest major differences in development between vertebrate classes, and start to address how these might translate onto morphology. In this review we survey what is currently known about the cell biology and gene expression during head development in modern vertebrates, allowing us to better characterize the developmental genetics driving head skeleton formation in the most recent common ancestor of all living vertebrates. By pairing this vertebrate composite with information from fossil chordates, we can also deduce how gene regulatory modules might have been arranged in the ancestral vertebrate head. Together, we can immediately begin to understand which aspects of head skeletal development are the most conserved, and which are divergent, informing us as to when the first differences appear during development, and thus which pathways or cell types might be involved in generating lineage specific shape and structure.

Roles for FGF in lamprey pharyngeal pouch formation and skeletogenesis highlight ancestral functions in the vertebrate head.

A defining feature of vertebrates (craniates) is a pronounced head supported and protected by a cellularized endoskeleton. In jawed vertebrates (gnathostomes), the head skeleton is made of rigid three-dimensional elements connected by joints. By contrast, the head skeleton of modern jawless vertebrates (agnathans) consists of thin rods of flexible cellular cartilage, a condition thought to reflect the ancestral vertebrate state. To better understand the origin and evolution of the gnathostome head skeleton, we have been analyzing head skeleton development in the agnathan, lamprey. The fibroblast growth factors FGF3 and FGF8 have various roles during head development in jawed vertebrates, including pharyngeal pouch morphogenesis, patterning of the oral skeleton and chondrogenesis. We isolated lamprey homologs of FGF3, FGF8 and FGF receptors and asked whether these functions are ancestral features of vertebrate development or gnathostome novelties. Using gene expression and pharmacological agents, we found that proper formation of the lamprey head skeleton requires two phases of FGF signaling: an early phase during which FGFs drive pharyngeal pouch formation, and a later phase when they directly regulate skeletal differentiation and patterning. In the context of gene expression and functional studies in gnathostomes, our results suggest that these roles for FGFs arose in the first vertebrates and that the evolution of the jaw and gnathostome cellular cartilage was driven by changes developmentally downstream from pharyngeal FGF signaling.

Dual epithelial origin of vertebrate oral teeth.

The oral cavity of vertebrates is generally thought to arise as an ectodermal invagination. Consistent with this, oral teeth are proposed to arise exclusively from ectoderm, contributing to tooth enamel epithelium, and from neural crest derived mesenchyme, contributing to dentin and pulp. Yet in many vertebrate groups, teeth are not restricted only to the oral cavity, but extend posteriorly as pharyngeal teeth that could be derived either directly from the endodermal epithelium, or from the ectodermal epithelium that reached this location through the mouth or through the pharyngeal slits. However, when the oropharyngeal membrane, which forms a sharp ecto/endodermal border, is broken, the fate of these cells is poorly known. Here, using transgenic axolotls with a combination of fate-mapping approaches, we present reliable evidence of oral teeth derived from both the ectoderm and endoderm and, moreover, demonstrate teeth with a mixed ecto/endodermal origin. Despite the enamel epithelia having a different embryonic source, oral teeth in the axolotl display striking developmental uniformities and are otherwise identical. This suggests a dominant role for the neural crest mesenchyme over epithelia in tooth initiation and, from an evolutionary point of view, that an essential factor in teeth evolution was the odontogenic capacity of neural crest cells, regardless of possible 'outside-in' or 'inside-out' influx of the epithelium.

Service life assessment of historical building envelopes constructed using different types of sandstone: a computational analysis based on experimental input data.

Service life assessment of three historical building envelopes constructed using different types of sandstone is presented. At first, experimental measurements of material parameters of sandstones are performed to provide the necessary input data for a subsequent computational analysis. In the second step, the moisture and temperature fields across the studied envelopes are calculated for a representative period of time. The computations are performed using dynamic climatic data as the boundary conditions on the exterior side of building envelope. The climatic data for three characteristic localities are experimentally determined by the Czech Hydrometeorological Institute and contain hourly values of temperature, relative humidity, rainfalls, wind velocity and direction, and sun radiation. Using the measured durability properties of the analyzed sandstones and the calculated numbers of freeze/thaw cycles under different climatic conditions, the service life of the investigated building envelopes is assessed. The obtained results show that the climatic conditions can play a very significant role in the service life assessment of historical buildings, even in the conditions of such a small country as the Czech Republic. In addition, the investigations reveal the importance of the material characteristics of sandstones, in particular the hygric properties, on their service life in a structure.

Consequences of omitting some important factors in the environmental analyses of commercial sodium silicate/sodium hydroxide use for alkaline activation in the light of comparison with cement-based composites.

Alkali-activated materials (AAMs) based on various waste precursors were considered mostly as a sustainable alternative to Portland cement-based composites to date. However, a narrow focus on carbon dioxide savings in the environmental assessment of AAMs may not be sufficient to achieve a truly sustainable solution. Therefore, this paper provides a detailed insight into midpoint impact categories related to the production of AAMs based on waste precursors and conventional activators, as compared with common cement-based materials. The obtained results point to a higher environmental load of AAMs in several categories, such as ozone layer depletion, primary resource consumption, and terrestrial and aquatic ecotoxicity. In a hypothetical scenario, it is demonstrated that 10 % replacement of global concrete production by AAMs may result in notably increased emissions of ozone depletion substances (+35 %) and damage to the aquatic environment (+ 40 %). The risk for human health can then be higher. As for the aquatic environment, eutrophication can also lead to a significant increase in indirect emissions of CH4 and N2O having a high impact on the greenhouse effect. Hence, the importance of robust interdisciplinary research in the environmental assessment of AAMs should be emphasized, together with the need to use alternative alkaline substances, which would be more environment-friendly than conventional activators.

Assessment of Clayey Freshwater Sediments as Suitable Precursors for Alkaline Activation.

One of the biggest challenges in the construction industry in recent times is the mitigation of the environmental impact of this sector, the reduction in dependence on primary raw materials, and the reduction in CO2 production while maintaining functional properties. Alkaline activation of a number of waste products represents a promising way to achieve the above-mentioned goals, but the availability of a number of waste products changes over time, especially in Europe. While freshwater sediments were in the past widely utilized as an agricultural fertilizer, recent precautions have significantly decreased such application, and thus new destinations must be delivered. To explore the potential of freshwater sediments, select samples from various locations were subjected to detailed characterization to verify the applicability of the material for alkali activation. As recognized, the selected sediments contain a substantial volume of desired mineralogical compounds that can serve, after 900 °C curing, as suitable precursors. Such samples have consequently activated the mixture of alkaline activators to obtain dense structures and were subjected to detailed investigation aimed at understanding the mechanical parameters. The obtained mechanical results ranging between 14.9 MPa and 36.8 MPa reveal the engineering potential of sediments for valorization through alkali activation and outline new research challenges in this area.

Development and evolution of the vertebrate primary mouth.

The vertebrate oral region represents a key interface between outer and inner environments, and its structural and functional design is among the limiting factors for survival of its owners. Both formation of the respective oral opening (primary mouth) and establishment of the food-processing apparatus (secondary mouth) require interplay between several embryonic tissues and complex embryonic rearrangements. Although many aspects of the secondary mouth formation, including development of the jaws, teeth or taste buds, are known in considerable detail, general knowledge about primary mouth formation is regrettably low. In this paper, primary mouth formation is reviewed from a comparative point of view in order to reveal its underestimated morphogenetic diversity among, and also within, particular vertebrate clades. In general, three main developmental modes were identified. The most common is characterized by primary mouth formation via a deeply invaginated ectodermal stomodeum and subsequent rupture of the bilaminar oral membrane. However, in salamander, lungfish and also in some frog species, the mouth develops alternatively via stomodeal collar formation contributed both by the ecto- and endoderm. In ray-finned fishes, on the other hand, the mouth forms via an ectoderm wedge and later horizontal detachment of the initially compressed oral epithelia with probably a mixed germ-layer derivation. A very intriguing situation can be seen in agnathan fishes: whereas lampreys develop their primary mouth in a manner similar to the most common gnathostome pattern, hagfishes seem to undergo a unique oropharyngeal morphogenesis when compared with other vertebrates. In discussing the early formative embryonic correlates of primary mouth formation likely to be responsible for evolutionary-developmental modifications of this area, we stress an essential role of four factors: first, positioning and amount of yolk tissue; closely related to, second, endoderm formation during gastrulation, which initiates the process and constrains possible evolutionary changes within this area; third, incipient structure of the stomodeal primordium at the anterior neural plate border, where the ectoderm component of the prospective primary mouth is formed; and fourth, the prime role of Pitx genes for establishment and later morphogenesis of oral region both in vertebrates and non-vertebrate chordates.

Evidence for the prepattern/cooption model of vertebrate jaw evolution.

The appearance of jaws was a turning point in vertebrate evolution because it allowed primitive vertebrates to capture and process large, motile prey. The vertebrate jaw consists of separate dorsal and ventral skeletal elements connected by a joint. How this structure evolved from the unjointed gill bar of a jawless ancestor is an unresolved question in vertebrate evolution. To understand the developmental bases of this evolutionary transition, we examined the expression of 12 genes involved in vertebrate pharyngeal patterning in the modern jawless fish lamprey. We find nested expression of Dlx genes, as well as combinatorial expression of Msx, Hand and Gsc genes along the dorso-ventral (DV) axis of the lamprey pharynx, indicating gnathostome-type pharyngeal patterning evolved before the appearance of the jaw. In addition, we find that Bapx and Gdf5/6/7, key regulators of joint formation in gnathostomes, are not expressed in the lamprey first arch, whereas Barx, which is absent from the intermediate first arch in gnathostomes, marks this domain in lamprey. Taken together, these data support a new scenario for jaw evolution in which incorporation of Bapx and Gdf5/6/7 into a preexisting DV patterning program drove the evolution of the jaw by altering the identity of intermediate first-arch chondrocytes. We present this "Pre-pattern/Cooption" model as an alternative to current models linking the evolution of the jaw to the de novo appearance of sophisticated pharyngeal DV patterning.

Experimental and Environmental Analysis of High-Strength Geopolymer Based on Waste Bricks and Blast Furnace Slag.

The rationalization of material flows, together with the utilization of waste raw materials for the production of alternative binders, became a very attractive topic during the last decades. However, the majority of designed materials can be used as a replacement for low-performance products. In this work, the waste materials (brick powder and blast furnace slag) are valorized through geopolymerization to design high-performance material as an alternative to high-performance concrete. Designed mixtures activated by sodium silicate and waste-originated alkali solution are characterized by the meaning of the chemical and mineralogical composition, evolution of hydration heat, and mechanical strength test. To contribute to the understanding of the environmental consequences and potential benefits, the carbon footprint and embodied energy analysis are provided. Obtained results highlight the potential of end-of-life bricks for the design of high-performance composites if mixed together with more reactive precursors. Here, even values over 60 MPa in compressive strength can be achieved with the dominant share of low-amorphous brick powder. The higher crystalline portion of brick powder may lead to the reduction of drying shrinkage and preservation of flexural strength to a greater extent compared to used slag. Performed environmental analysis confirmed the CO2 emission savings; however, the embodied energy analysis revealed a huge impact of using alkaline activators.

Ecotoxicity of Caffeine as a Bio-Protective Component of Flax-Fiber-Reinforced Epoxy-Composite Building Material.

Caffeine is a verified bio-protective substance in the fight against the biodegradation of cellulose materials, but its ecotoxicity in this context has not yet been studied. For this reason, the ecotoxicity of flax-fiber-reinforced epoxy composite with or without caffeine was tested in the present study. Prepared samples of the composite material were tested on freshwater green algal species (Hematococcus pluvialis), yeasts (Saccharomyces cerevisae), and crustacean species (Daphnia magna). Aqueous eluates were prepared from the studied material (with caffeine addition (12%) and without caffeine and pure flax fibers), which were subjected to chemical analysis for the residues of caffeine or metals. The results indicate the presence of caffeine up to 0.001 mg/L. The eluate of the studied material was fully toxic for daphnids and partially for algae and yeasts, but the presence of caffeine did not increase its toxicity statistically significantly, in all cases. The final negative biological effects were probably caused by the mix of heavy metal residues and organic substances based on epoxy resins released directly from the tested composite material.

The odontode explosion: the origin of tooth-like structures in vertebrates.

Essentially we show recent data to shed new light on the thorny controversy of how teeth arose in evolution. Essentially we show (a) how teeth can form equally from any epithelium, be it endoderm, ectoderm or a combination of the two and (b) that the gene expression programs of oral versus pharyngeal teeth are remarkably similar. Classic theories suggest that (i) skin denticles evolved first and odontode-inductive surface ectoderm merged inside the oral cavity to form teeth (the 'outside-in' hypothesis) or that (ii) patterned odontodes evolved first from endoderm deep inside the pharyngeal cavity (the 'inside-out' hypothesis). We propose a new perspective that views odontodes as structures sharing a deep molecular homology, united by sets of co-expressed genes defining a competent thickened epithelium and a collaborative neural crest-derived ectomesenchyme. Simply put, odontodes develop 'inside and out', wherever and whenever these co-expressed gene sets signal to one another. Our perspective complements the classic theories and highlights an agenda for specific experimental manipulations in model and non-model organisms.

Properties of municipal solid waste incineration ashes with respect to their separation temperature.

Fly ashes generated by power and heating plants are commonly used in the production of building materials in some countries, mainly as partial replacement of cement or aggregates in concrete. The ashes from municipal solid waste incinerators can be applied in a similar way. However, their chemical and mineralogical composition, granulometry and toxic constituents have to be taken into account. In this paper, four types of municipal solid waste incinerator (MSWI) ashes produced by the modern MSWI facility in Liberec, Czech Republic, were investigated. The relationship between the particular ash properties (morphology, chemical composition) and its separation temperature in the incinerator line is discussed. A coal fly ash (class F) is characterized as well, for a comparison because its utilization in building industry is more developed. The studied MSWI ashes exhibit high concentration of chlorides and sulfates which is an unfavourable feature for a potential concrete admixture. On the other hand, three of four ashes are found to be pozzolanic active and certain hydration reactions are indicated.

Waste brick dust as a prospective eco-friendly alternative component of artificial soils for ecotoxicological studies.

Current artificial soils for ecotoxicological studies contain non-renewable materials that must be mined, and their production and processing consume a lot of energy and generate a significant amount of carbon dioxide (CO2). In this paper, waste brick dust is proposed as an alternative to calcium carbonate (CaCO3), which is used for pH adjustment of the Organization for Economic Co-operation and Development (OECD) soils. The artificial soils containing brick dust are contaminated with boric acid as a reference substance in the concentration range of 100-500 mg/kg and studied in the tests with enchytraeids (E. crypticus), springtails (F. candida), and plants (L. sativa and B. napus). Experimental results shows the suitability of replacing calcium carbonate with waste brick dust, as neither toxicity nor ability of model organisms to inhabit the analyzed soil is found. A comparison with the standard OECD soil does not reveal any substantial differences between the parameters (survival, reproduction, and root elongation) of the applied ecotoxicological tests. The brick dust as waste material is found to have a lower carbon footprint than CaCO3, while a similar amount of water is necessary for the adjustment of tests with both kinds of artificial soil. The waste brick dust can be considered as a suitable eco-friendly alternative to CaCO3 in artificial soils for ecotoxicological studies.

The conundrum of pharyngeal teeth origin: the role of germ layers, pouches, and gill slits.

There are several competing hypotheses on tooth origins, with discussions eventually settling in favour of an 'outside-in' scenario, in which internal odontodes (teeth) derived from external odontodes (skin denticles) in jawless vertebrates. The evolution of oral teeth from skin denticles can be intuitively understood from their location at the mouth entrance. However, the basal condition for jawed vertebrates is arguably to possess teeth distributed throughout the oropharynx (i.e. oral and pharyngeal teeth). As skin denticle development requires the presence of ectoderm-derived epithelium and of mesenchyme, it remains to be answered how odontode-forming skin epithelium, or its competence, were 'transferred' deep into the endoderm-covered oropharynx. The 'modified outside-in' hypothesis for tooth origins proposed that this transfer was accomplished through displacement of odontogenic epithelium, that is ectoderm, not only through the mouth, but also via any opening (e.g. gill slits) that connects the ectoderm to the epithelial lining of the pharynx (endoderm). This review explores from an evolutionary and from a developmental perspective whether ectoderm plays a role in (pharyngeal) tooth and denticle formation. Historic and recent studies on tooth development show that the odontogenic epithelium (enamel organ) of oral or pharyngeal teeth can be of ectodermal, endodermal, or of mixed ecto-endodermal origin. Comprehensive data are, however, only available for a few taxa. Interestingly, in these taxa, the enamel organ always develops from the basal layer of a stratified epithelium that is at least bilayered. In zebrafish, a miniaturised teleost that only retains pharyngeal teeth, an epithelial surface layer with ectoderm-like characters is required to initiate the formation of an enamel organ from the basal, endodermal epithelium. In urodele amphibians, the bilayered epithelium is endodermal, but the surface layer acquires ectodermal characters, here termed 'epidermalised endoderm'. Furthermore, ectoderm-endoderm contacts at pouch-cleft boundaries (i.e. the prospective gill slits) are important for pharyngeal tooth initiation, even if the influx of ectoderm via these routes is limited. A balance between sonic hedgehog and retinoic acid signalling could operate to assign tooth-initiating competence to the endoderm at the level of any particular pouch. In summary, three characters are identified as being required for pharyngeal tooth formation: (i) pouch-cleft contact, (ii) a stratified epithelium, of which (iii) the apical layer adopts ectodermal features. These characters delimit the area in which teeth can form, yet cannot alone explain the distribution of teeth over the different pharyngeal arches. The review concludes with a hypothetical evolutionary scenario regarding the persisting influence of ectoderm on pharyngeal tooth formation. Studies on basal osteichthyans with less-specialised types of early embryonic development will provide a crucial test for the potential role of ectoderm in pharyngeal tooth formation and for the 'modified outside-in' hypothesis of tooth origins.