Simplification of the enameloid microstructure of large stingrays (Chondrichthyes: Myliobatiformes): a functional approach

The microstructure of neoselachian teeth is characterized by a triple‐layered enameloid, despite its purported absence in batoids since their appearance during the early Jurassic. This feature is thought to have been secondarily lost, as an adaptation to durophagy in batoids. The monophyletic Myliobatiformes are an ecologically diverse clade of batoids that includes very specialized taxa such as the filter‐feeding mobulids and the durophagous myliobatids. Their diversity and recent evolutionary history (Palaeogene) make them an ideal model for assessing adaptive constraints that influence enameloid microstructure. This first study focusing only on batoid enameloid shows a very reduced single‐layered enameloid in the derived filter feeders. However, most other taxa show a double‐layered enameloid, which is probably plesiomorphic for batoids. It is concluded that the triple‐layered enameloid is not a synapomorphy of the neoselachii as a whole, but of Selachimorpha only, and that diet specialization in derived myliobatoids indeeds impact their enameloid microctructure, which is confirmed by their fossil diversity. © 2013 The Linnean Society of London     

The homology and phylogeny of chondrichthyan tooth enameloid

A systematic SEM survey of tooth microstructure in (primarily) fossil taxa spanning chondrichthyan phylogeny demonstrates the presence of a superficial cap of single crystallite enameloid (SCE) on the teeth of several basal elasmobranchs, as well as on the tooth plates of Helodus (a basal holocephalan). This suggests that the epithelial-mesenchymal interactions required for the development of enameloid during odontogenesis are plesiomorphic in chondrichthyans, and most likely in toothed gnathostomes, and provides phylogenetic support for the homology of chondrichthyan and actinopterygian enameloid. Along the neoselachian stem, we see a crownward progression, possibly modulated by heterochrony, from a monolayer of SCE lacking microstructural differentiation to the complex triple-layered tooth enameloid fabric of neoselachians. Finally, the occurrence of fully-differentiated neoselachian enameloid microstructure (including compression-resistant tangle fibered enameloid and bending-resistant parallel fibered enameloid) in Chlamydoselachus anguineus, a basal Squalean with teeth that are functionally "cladodont," is evidence that triple-layered enameloid microstructure was a preadaption to the cutting and gouging function of many neoselachian teeth, and thus may have played an integral role in the Mesozoic radiation of the neoselachian crown group.     

Chondrichthyan tooth enameloid: past,present, and future

Enameloid is a hard mineralized tissue covering chondrichthyan and actinopterygian teeth. Over the past 40 years, it has been extensively studied in various extinct and extant sharks, leading to the broad use of microstructural characters to differentiate between hybodont and neoselachian teeth. However, the chondrichthyan taxic diversity is disproportionately high compared to the number of taxa explored for enameloid microstructure, and the generalization of these few observations to the whole group is problematic. Indeed, many other groups, in particular modern rays and skates, have been completely overlooked, and almost nothing is known about their tooth histology. Furthermore, the recent discovery of typical neoselachian character in cladodontomorph sharks teeth clearly indicates that we have had an over‐simplified perception of the chondrichthyan enameloid distribution, which put into question the previously proposed evolutive scenarios dealing whith this tissue. We propose a brief historical overview of the study and understanding of chondrichthyan enameloid diversity and briefly discuss preparation issues encountered when dealing with the study of chondrichthyan hypermineralized tissues. Then, the variation of enameloid microstructures encountered in ctenacanthiforms, hybodonts, selachimorphs, and batomorphs is explored, summarized, and discussed. Although the full extent of the diversity and variability of the enameloid microstructure in many of these groups and others remains to be fully determined, we are able to show that most possess a much more complex enameloid microstructure than expected, and propose a revised and more fitting chondrichthyan enameloid terminology, based on the recognition of two main units: an external Single Crystallite Enameloid (SCE) and an internal Bundled Crystallite Enameloid (BCE). Our study reveals new insights in the understanding of character distribution among batomorphs and sets a framework for tackling global chondrichthyan tooth enameloid evolution. © 2015 The Linnean Society of London     

The shark fauna from the Middle Triassic (Anisian) of North-Western Nevada

The shark fauna from the Anisian of Nevada is dominated by durophagous hybodontiforms but also shows an important neoselachian component. Two new species of hybodontiform sharks, Acrodus cuneocostatus and Polyacrodus bucheri , are described in addition to a new neoselachian taxon: Mucrovenator minimus. The enameloid of the teeth of Acrodus and Polyacrodus comprises two layers, an outer compact layer and an inner bundled layer. For the typical three-layered enameloid of neoselachian teeth, we propose to replace the terms parallel-fibred enameloid and tangled-fibred enameloid by the more appropriate parallel-bundled and tangled-bundled enameloid.     

Development and Evolution of Dentition Pattern and Tooth Order in the Skates And Rays (Batoidea; Chondrichthyes)

Shark and ray (elasmobranch) dentitions are well known for their multiple generations of teeth, with isolated teeth being common in the fossil record. However, how the diverse dentitions characteristic of elasmobranchs form is still poorly understood. Data on the development and maintenance of the dental patterning in this major vertebrate group will allow comparisons to other morphologically diverse taxa, including the bony fishes, in order to identify shared pattern characters for the vertebrate dentition as a whole. Data is especially lacking from the Batoidea (skates and rays), hence our objective is to compile data on embryonic and adult batoid tooth development contributing to ordering of the dentition, from cleared and stained specimens and micro-CT scans, with 3D rendered models. We selected species (adult and embryonic) spanning phylogenetically significant batoid clades, such that our observations may raise questions about relationships within the batoids, particularly with respect to current molecular-based analyses. We include developmental data from embryos of recent model organisms Leucoraja erinacea and Raja clavata to evaluate the earliest establishment of the dentition. Characters of the batoid dentition investigated include alternate addition of teeth as offset successional tooth rows (versus single separate files), presence of a symphyseal initiator region (symphyseal tooth present, or absent, but with two parasymphyseal teeth) and a restriction to tooth addition along each jaw reducing the number of tooth families, relative to addition of successor teeth within each family. Our ultimate aim is to understand the shared characters of the batoids, and whether or not these dental characters are shared more broadly within elasmobranchs, by comparing these to dentitions in shark outgroups. These developmental morphological analyses will provide a solid basis to better understand dental evolution in these important vertebrate groups as well as the general plesiomorphic vertebrate dental condition.     

New Studies of Enamel Microstructure in Mesozoic Mammals: A Review of Enamel Prisms as a Mammalian Synapomorphy

Characters from enamel microstructure have not been used in recent phylogenetic analyses of Mesozoic Mammalia. Reasons are that enamel characters have been perceived as (A) variable without regard to systematic position of taxa, (B) inconsistently reported within the literature, and (C) simply scored as either prismatic or not prismatic in earlier mammals. Our work on Mesozoic mammals such as Sinoconodon, Gobiconodon, Triconodontidae, Docodon, Laolestes, and others suggests that synapsid columnar enamel (SCE) structure was easily transformed into plesiomorphic prismatic enamel (PPE) and that PPE may be described with at least five independent character states. Two PPE characters—a flat, open prism sheath and a planar prism seam—were present in the cynodont Pachygenelus and in several Jurassic and Cretaceous mammals. We propose that appearance of a prism sheath transforms SCE into PPE and that reduction and loss of a prism sheath reverse PPE into SCE, in both phylogeny and ontogeny. We further propose that no amniote vertebrates other than the trithelodontid cynodont, Pachygenelus, plus Mammalia have ever evolved an ameloblastic Tomes process capable of secreting PPE and that the genetic potential to secrete PPE is a synapomorphy of Pachygenelus plus Mammalia, whether or not all lineages of the clade have expressed that potential.     

Enameloid/enamel transition through successive tooth replacements in <Emphasis Type="Italic">Pleurodeles waltl</Emphasis> (Lissamphibia,Caudata)

Study of the evolutionary enameloid/enamel transition suffers from discontinuous data in the fossil record, although a developmental enameloid/enamel transition exists in living caudates, salamanders and newts. The timing and manner in which the enameloid/enamel transition is achieved during caudate ontogeny is of great interest, because the caudate situation could reflect events that have occurred during evolution. Using light and transmission electron microscopy, we have monitored the formation of the upper tooth region in six successive teeth of a tooth family (position I) in Pleurodeles waltl from late embryos to young adult. Enameloid has only been identified in embryonic tooth I₁ and in larval teeth I₂ and I₃. A thin layer of enamel is deposited later by ameloblasts on the enameloid surface of these teeth. From post-metamorphic juvenile onwards, teeth are covered with enamel only. The collagen-rich enameloid matrix is deposited by odontoblasts, which subsequently form dentin. Enameloid, like enamel, mineralizes and then matures but ameloblast participation in enameloid matrix deposition has not been established. From tooth I₁ to tooth I₃, the enameloid matrix becomes ever more dense and increasingly comes to resemble the dentin matrix, although it is still subjected to maturation. Our data suggest the absence of an enameloid/enamel transition and, instead, the occurrence of an enameloid/dentin transition, which seems to result from a progressive slowing down of odontoblast activity. As a consequence, the ameloblasts in post-metamorphic teeth appear to synthesize the enamel matrix earlier than in larval teeth.     

Developmental origins of complex radular characters in the Muricidae: the bifid rachidian edge

Muricid gastropod radulae are more complex than those of most other neogastropods, especially in the number and variety of cusps, denticles, and interlocking mechanisms. How this complexity evolved, however, is unknown. Morphological gaps between higher taxa within the Muricidae are substantial, and there are few unambiguous intermediates. Here, we use developmental data from the Patagonian trophonine muricid Trophon geversianus to investigate the evolution of an unusual condition in which there are two marginal cusps at each end of each central rachidian tooth, rather than one or none as in most muricids. Trophon geversianus begins ontogeny with one marginal cusp (the inner marginal cusp), but a second (the outer marginal cusp) appears later, arising from separation of the rachidian base edge from the radular membrane rather than through bifurcation or lateral migration of pre‐existing cusps. Truncation of development (i.e., paedomorphosis) at this second developmental phase in a trophonine ancestor provides an explanation for the lack of transitional forms between most adult trophonine muricids, which have the plesiomorphic condition of one marginal cusp, and sister group ocenebrine muricids, which have the derived condition of two marginal cusps.     

Phylogeny and biogeography of tetralophodont rodents of the tribe Oryzomyini (Cricetidae: Sigmodontinae)

Oryzomyini is the richest tribe among the Sigmodontine rodents, encompassing 32 living and extinct genera and including an increasing number of recently described species and genera. Some Oryzomyini are tetralophodont showing a reduction in the number of molar folds to four, while most taxa in this tribe retain the plesiomorphic pentalophodont state. We applied phylogenetic methods, molecular dating techniques and ancestral area analyses to members of an oryzomyini clade informally named ‘D’ in former studies and included related fossil tetralophodont forms. Based on 98 morphological characters and sequences of five gene fragments, we found that the tetralophodont condition is paraphyletic. Among living taxa, Pseudoryzomys is sister to Holochilus, and Lundomys is derived from a basal divergence. A clade formed by living Holochilus and the fossils Noronhomys and Carletonomys is sister to Holochilus primigenus, making Holochilus paraphyletic. Therefore, we describe a new genus that accommodates the fossil H. primigenus. Because trans‐Andean taxa currently share a common ancestor with taxa of cis‐Adean distribution, the northern Andes uplift may have worked as a postdispersal barrier. The tetralophodont lineages diverged during the Pliocene from a cis‐Andean ancestor, and the Great Plains in South America may have favoured the diversification of tetralophodont forms adapted to open habitats during the Pliocene.     

Larval Planktotrophy—A Primitive Trait in the Bilateria?

The concept of Gösta Jägersten of a primary biphasic metazoan life-cycle, consisting of a planktotrophic larva and a benthic adult, forms the basis for several theories on metazoan phylogeny. In this paper the assumed planktotrophic life-style of the larva is critically analyzed and reconsidered. It is shown, in particular for the Mollusca, that a biphasic life-cycle with a lecithotrophic larva is probably the plesiomorphic condition. Character distribution and structural data suggest a parallel evolution of the downstream collecting system used in planktotrophic larvae or filter-feeding adults of gastropods, bivalves and other spiralian or aschelminth taxa. In the basic metazoans (Parazoa, Placozoa, coelenterates) direct or lecithotrophic development dominates by far. For the acoelomate (Platyhelminthes, Gnathostomulida) and pseudocoelomate taxa direct development is probably the plesiomorphic condition. The structural similarities of the upstream collecting system in tentaculate and deuterostome phyla may also be explained by parallel events of heterochrony out of an ancestor with adult filter-feeding. The main conclusion of this survey is that larval planktotrophy is likely to be secondary and not a plesiomorphic condition among the Bilateria. Accordingly, theories which are based on the assumed plesiomorphy of larval planktotrophy of the Bilateria, need careful reevaluation.     

Temporal squama shape in fossil hominins: relationships to cranial shape and a determination of character polarity

In 1943, Weidenreich described the squamosal suture of Homo erectus as long, low, and simian in character and suggested that this morphology was dependent upon the correlation between the size of the calvarium and the face. Many researchers now consider this character to be diagnostic of H. erectus. The relationship between cranial size and shape and temporal squama morphology, however, is unclear, and several authors have called for detailed measurements of squamosal variation to be collected before any conclusions are drawn regarding the nature of the morphology observed in H. erectus. Thirteen fossil and extant taxa were examined to address two questions: 1) Are size and shape of the temporal squama correlated with cranial vault morphology? and 2) Is the H. erectus condition plesiomorphic? To answer these questions, measurements were collected and indices were calculated for squamosal suture height, length, and area in relation to metric variables describing cranial size and shape. A two‐dimensional morphometric study was also completed using High Resolution‐Polynomial Curve Fitting (HR‐PCF) to investigate correlations between curvature of the squamosal suture and curvature of the cranial vault. Results of both analyses indicate that squamosal suture form is related to cranial size and shape. Furthermore, the plesiomorphic condition of the squamosal suture for hominins was identified as high and moderately arched; this condition is retained in H. erectus and is distinct from the great ape condition. It is suggested that this similarity is the result of increased cranial length without a corresponding increase in cranial height. Am J Phys Anthropol, 2008. © 2008 Wiley‐Liss, Inc.     

Molecular Phylogeny of Nepenthaceae Based on Cladistic Analysis of Plastid trnK Intron Sequence Data

Abstract: Nepenthaceae are an exceptional family with regard to carnivory and the uniformity of characters. This makes it difficult to resolve phylogenetic relationships due to convergent evolution of morphological features. Using comparative sequencing of the chloroplast trnK intron, the monophyly of this complex family and hypotheses of infrageneric relationships were tested. Sequences from 71 Nepenthes taxa, representing all groups and two taxa of the closely related Ancistrocladaceae and Dioncophyllaceae as outgroup, were determined and analysed using maximum parsimony methods. Results of this analysis show that the isolated taxa N. distillatoria (Sri Lanka) and N. pervillei (Seychelles) are the most basal, clearly separated from the Madagascan taxa N. madagascariensis and N. masoalensis which are placed in a distinct subclade. This corresponds with some plesiomorphic characters shared by these taxa. N. khasiana (North India) has an intermediate position between these relic Western species and the remaining taxa. The species of the Malay Archipelago can be referred to three distinct lineages which indicate a correlation to biogeography. Thus the recent disjunct distribution of Nepenthes is interpreted as a result of an incisive extinction of progenitors, a process of migration and a subsequent diversification on the islands of Borneo, Sumatra, Sulawesi and New Guinea. Based on our molecular data, two interpretations concerning the origin of Nepenthes are possible: i) evolution in the Northern Tethys which is supported by fossil pollen records from the European Focene, or, ii) a Gondwanaland origin at a time when the Indian plate was separated from Madagascar. Molecular data indicate that colonization of SE Asia started from an ancient Indian stock. Subsequently, in the Malay Archipelago a new secondary centre of diversity developed. Madagascar, the Seychelles and New Caledonia were probably reached by migration via land bridges, starting from widespead common ancestors with subsequent extinction leaving the current taxa. There is no evidence for long‐distance dispersal. Current infragenic classification of Nepenthes is only partly in accordance with the phylogeny inferred from trnK intron data.     

The evolution of dinosaur tooth enamel microstructure

The evolution of tooth enamel microstructure in both extinct and extant mammalian groups has been extensively documented, but is poorly known in reptiles, including dinosaurs. Previous intensive sampling of dinosaur tooth enamel microstructure revealed that: (1) the three‐dimensional arrangement of enamel types and features within a tooth—the schmelzmuster—is most useful in diagnosing dinosaur clades at or around the family level; (2) enamel microstructure complexity is correlated with tooth morphology complexity and not necessarily with phylogenetic position; and (3) there is a large amount of homoplasy within Theropoda but much less within Ornithischia. In this study, the examination of the enamel microstructure of 28 additional dinosaur taxa fills in taxonomic gaps of previous studies and reinforces the aforementioned conclusions. Additionally, these new specimens reveal that within clades such as Sauropodomorpha, Neotheropoda, and Euornithopoda, the more basal taxa have simpler enamel that is a precursor to the more complex enamel of more derived taxa and that schmelzmusters evolve in a stepwise fashion. In the particularly well‐sampled clade of Euornithopoda, correlations between the evolution of dental and enamel characters could be drawn. The ancestral schmelzmuster for Genasauria remains ambiguous due to the dearth of basal ornithischian teeth available for study. These new specimens provide new insights into the evolution of tooth enamel microstructure in dinosaurs, emphasizing the importance of thorough sampling within broadly inclusive clades, especially among their more basal members.     

The geometry of gender: hyper‐diversification of sexual systems in Urtica L. (Urticaceae)

Urtica L. (Urticaceae) is generally reported as a genus of monoecious and dioecious taxa. However, the gender information found in the literature does not at all reflect the actual diversity of gender patterns in Urtica. Dioecy appears to be truly absent from Urtica, but otherwise there has been a major diversification in the geometry of gender and no comparable patterns exist in other plant groups. Thus, we here define technical terms for all unique architectural types of monoecy found in Urtica and closely related genera and reconstruct the ancestral gender states in a Bayesian framework. Our studies are based on a near‐comprehensive sampling, including 61 of the 63 Urtica species recognized. We report polygamy, two types of gynodioecy and five different architectural types of monoecy. A total of 15 switches appear to have taken place within the genus. Although gender characteristics have diversified strongly, they are relatively conserved within clades. Monoecy is the predominant sexual system within Urtica and specifically basiandrous monoecy (i.e. basal inflorescence branches of each individual male only, apical branches female) is the most widespread type, reported for 11 different clades. In particular, it characterizes the basally branching pilulifera‐clade and the sister genus Zhengyia, and may thus represent the plesiomorphic condition for Urtica. Gender distribution and gross morphology appear to evolve largely independently from each other and gender distribution is largely independent of growth habit. However, polygamous taxa are most common amongst rhizomatous perennials (one‐third of the taxa).     

Morphogenesis and histology of large scales of batoids (Elasmobranchii)

Large scales occur only in three families of batoids. Though they have a wide spectrum of different shapes they all serve protective functions in bottom-dwelling species. The crown consists of enameloid, orthodentine and osteodentine. Three different types of basal plates occur: (a) thin basal plate consisting of acellular bone; (b) basal plate which is secondarily thickened; it consists of massive acellular bone and thin denteons which surround the vascular canals; (c) basal plate which is secondarily thickened, consisting of a peculiar type of microspongy bone which has never been found in other elasmobranchs. The scales have either one or several crown elements. None of the scales, however, belongs to a growing type. All large scales were probably replaced regularly. It is the first time that dentine was found within the basal plate of an elasmobranch scale.     

The anatomy of the basal ornithischian dinosaur Eocursor parvus from the lower Elliot Formation (Late Triassic) of South Africa

Ornithischia is a morphologically and taxonomically diverse clade of dinosaurs that originated during the Late Triassic and were the dominant large‐bodied herbivores in many Cretaceous ecosystems. The early evolution of ornithischian dinosaurs is poorly understood, as a result in part of a paucity of fossil specimens, particularly during the Triassic. The most complete Triassic ornithischian dinosaur yet discovered is Eocursor parvus from the lower Elliot Formation (Late Triassic: Norian–Rhaetian) of Free State, South Africa, represented by a partial skull and relatively complete postcranial skeleton. Here, the anatomy of Eocursor is described in detail for the first time, and detailed comparisons are provided to other basal ornithischian taxa. Eocursor is a small‐bodied taxon (approximately 1 m in length) that possesses a plesiomorphic dentition consisting of unworn leaf‐shaped crowns, a proportionally large manus with similarities to heterodontosaurids, a pelvis that contains an intriguing mix of plesiomorphic and derived character states, and elongate distal hindlimbs suggesting well‐developed cursorial ability. The ontogenetic status of the holotype material is uncertain. Eocursor may represent the sister taxon to Genasauria, the clade that includes most of ornithischian diversity, although this phylogenetic position is partially dependent upon the uncertain phylogenetic position of the enigmatic and controversial clade Heterodontosauridae. © 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 160 , 648–684.     

Trophic ecomorphology in eastern Pacific blennioid fishes: character transformation of oral jaws and associated change of their biological roles

Synopsis The ecomorphological relationships between the oral jaws and food spectra were highlighted in 34 species of Gulf of California blennioid fishes (5 Tripterygiidae, 13 Labrisomidae, 11 Chaenopsidae and 5 Blenniidae). Twenty-nine species are microcarnivorous, two are omnivorous browsers, two are algae grazers and one was an ‘ectoparasite’ feeder. The spectrum of oral (as opposed to pharyngeal) jaw (OJA) morphology ranges from plesiomorphic, suction-feeding (relatively large, protrusible jaws, with many coniform-caniniform teeth) to apomorphic, biting (relatively small, non protrusible jaws, with a single row of incisiform teeth). As species with similar morphology may widely differ in food, it is concluded, that morphology is not a reliable predictor for ecology in this case. With the exception of a few specialists, species with apomorphic, biting OJA utilize sessile items in addition to mobile categories and thus show a higher food diversity as compared to species with plesiomorphic OJA. Thus in the present case morphological differentiation goes along with ecological generalization. Only three blenniid species with the most apomorphic OJA may be considered as specialized also with regard to food resource utilization. Transformation of morphological characters and the ecological role of the OJA of blennioids may serve as a model to illustrate the steps required to achieve a biting-browsing and grazing feeding apparatus in many taxa of modern acanthopterygian reef fishes.     

The phylogenetic relationships of sauropod dinosaurs

A data-matrix of 205 osteological characters for 26 sauropod taxa is subjected to cladistic analysis. Two most parsimonious trees are produced, differing only in the relationships between Euhelopus, Omeisaurus and Mamenchisaurus. The monophyly of the Euhelopodidae (including Shunosaurus) is supported by seven synapomorphies. The Cetiosauridae (Patagosaurus, Cetiosaurus and Haplocanthosaurus) is paraphyletic with respect to the Neosauropoda. The latter clade divides into two major radiations–the ‘Brachiosauria’ (Camarasaurus, brachiosaurids and titanosauroids), and the Diplodocoidea (nemegtosaurids, dicraeosaurids, diplodocids and Rebbachisaurus). Further evidence for the inclusion of Opisthocoelwaudia in the Titanosauroidea is presented. Phuwiangosaurus, a problematic sauropod from Thailand, may represent one of the most plesiomorphic titanosauroids. ‘Peg’-like teeth have evolved at least twice within the Sauropoda. The postspinal lamina, on the neural spines of middle and caudal dorsal vertebrae, represents a neomorph rather than a fusion of pre-existing structures. Forked chevrons may have evolved convergently in the Euhelopodidae and the diplodocid-dicraeosaurid clade, or they may have been acquired early in sauropod evolution and subsequently lost in the ‘Brachiosauria’. The strengths and weaknesses of the data-matrix and tree topologies are explored using bootstrapping, decay analysis and randomization tests. Several nodes are only poorly supported, but this seems to reflect the large proportion of missing data in the matrix (～46%), rather than an abnormally high level of homoplasy. The results of the randomization tests indicate that the ‘data-matrix’ probably contains a strong phylogenetic ‘signal’. The relationships of some forms, such as Haplocanthosaurus, are influenced by the inclusion or exclusion of certain taxa with unusual combinations of character states. Such a result suggests that there are dangers inherent in the view that ‘higher’ level sauropod phylogeny can be accurately reconstructed using only a small number of well-known taxa.     

Serially etched shark enameloid observed by incident light microscopy

Longitudinal and transverse tooth sections of Isurus oxyrinchus were serially etched in 2.6% nitric acid. The changing optical properties of the etched surfaces were observed during the serial etchings, and the descent of the enameloid surfaces was measured. Shark enameloid seems to be less effectively etched by acid than human enamel; this difference may be due to differences in solubility between fluorapatite and hydroxyapatite. Most of the information regarding the structure of the enameloid was gained during the first five of ten etchings. The reflection of light from the surface was influenced by the orientation of the crystallites, longitudinally sectioned crystallites reflecting the light better than transversely sectioned crystallites. The dentinal extensions were continuous with and of the same structure as the underlying dentine. The radial fibers originated from the dentinal extensions, and they both contained organic material and were accompanied by crystallites. When the specimens were imbibed with water the distinctness of the dentinal extensions and radial fibers was improved.