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The aim of this study is to obtain information about the mouse tongue muscle rendered using micro-computed tomography (μCT) at low, middle, and high magnifications. Three-dimensional (3D) μCT is used in various fields. Most μCT observations are restricted to hard tissue in biomaterial samples. Recently, with the use of osmium tetroxide, μCT has been effectively employed to observe soft tissue; it is now believed that μCT observation of soft tissue is feasible. On the other hand, the structure of the tongue muscle has been well studied, but cross-sectional imaging enhanced by 3D rendering is lacking. We chose the mouse tongue as a soft tissue case study for μCT and generated cross-sectional images of the tongue enhanced by 3-D image rendering with histological resolution. During this observation, we developed new methods of low-magnification observation to show the relation between the tongue muscles and surrounding tissues. We also applied high-resolution μCT in high-magnification observation of muscle fiber fascicles. Our methodological techniques give the following results: (1) For low-magnification observation (field of view: 12,000 μm), pretreatment with decalcification and freeze drying is suitable for observing the area between the muscle of the tongue and the bone around the tongue using μCT. (2) For middle-magnification observation (Field of view: 3,500 μm), the use of osmium tetroxide to observe the muscle arrangement of the tongue by μCT is suitable. (3) For high-magnification observation (Field of view: 450 μm), high-resolution μCT is suitable for observation of the transversus muscle fiber fascicles.

As a reflection of different life styles and environment, the tongue of vertebrates, which plays a major role in the intake and swallowing of food, displays significant morphological differences. The gross form and microscopic structure of the avian tongue differ greatly according to lifestyle. The avian tongue plays a fundamental role in many functions such as capturing, filtering, sucking and manipulating food in order to compensate absence of subsidiary organs like teeth in the oropharyngeal cavity. Variations in lingual papillae play an important role in feeding of birds, as they represent a structure similar to teeth in the upper and lower beaks and can be used to hold and direct food in the oropharyngeal cavity. Tongues of birds exhibit common as well as varying anatomical characteristics in terms of surface morphology, structure and topographical distribution of lingual papillae as well as distinct specialized structures, epithelial layers, taste buds and lingual glands. This review evaluates the important morphological peculiarities of the tongue in birds, focusing on the relationship between anatomical features and feeding functions., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

Three-dimensional observation during embryogenesis is possible with micro-computed tomography, but there are no observations of organ size. In this paper, three examples of three-dimensional observation of organs by micro-CT are tried. At 13.0 days post-coitum, mouse embryos were fixed in 4% paraformaldehyde for 24 h and stained enbloc by osmium tetroxide overnight. The embryos were then embedded in paraffin using standard methods for 24 h. Specimens were analyzed by micro-computed tomography and image processing was performed. The entire Meckel's cartilage and its relation in the mandible, as well as the complex structure of the otocyst, are easily visualized. Although it is difficult to extract detailed structures of the tongue muscles, it is possible to identify the inner and external tongue muscles. Relation among the organs and other are easily visualized. Three-dimensional observation by micro-computed tomography is an important technology for visualization of embryogenesis and could be used in organ culture.

In an effort to identify a possible role for type III collagen in the morphogenesis of circumvallate papillae on the surface of the rat tongue, we examined its appearance by fluorescent immunostaining, in conjunction with differential interference contrast images and images obtained, after staining with toluidine blue, in the transmission mode by laser-scanning microscopy. We analyzed semi-ultrathin sections of epoxy resin-embedded samples of the lingual mucosa of embryonic and juvenile rats, 13 days after conception (E13) to day 21 after birth (P21). Immunoreactivity specific for type III collagen was recognized first in the mesenchymal connective tissue just beneath the circumvallate papilla placode in fetuses on E13. At this stage, most of the lingual epithelium with the exception of the circumvallate papilla placode was pseudostratified epithelium composed of one or two layers of cuboidal cells. However, the epithelium of the circumvallate papilla placode was composed of several layers of cuboidal cells. Immunoreactivity specific for type III collagen was detected mainly on the lamina propria just beneath the lingual epithelium of the rudiment of the circumvallate papilla and the developing circumvallate papilla in fetuses on E15 and E17, and slight immunostaining was detected on the lamina propria around the rudiment. In fetuses on E19, immunoreactivity specific for type III collagen was widely and densely distributed on the connective tissue around the developing circumvallate papillae and, also, on the connective tissue that surrounded the lingual muscle. However, the immunoreactivity specific for type III collagen was sparsely distributed on the lamina propria of each central papillar structure. After birth, from P0 to P14, morphogenesis of the circumvallate papillae advanced gradually with the increase in the total volume of the tongue. At these postnatal stages, the intensity of the fluorescence due to immunoreactivity specific for type III collagen was distinctively distributed on the lamina propria around each circumvallate papilla, on each central bulge and on the connective tissue that surrounded the lingual muscle. However, immunofluorescence was less distinct on the connective tissue that surrounded the lingual muscle. Thus, type III collagen appeared in conjunction with the morphogenesis of the circumvallate papillae, as well as in the connective tissue that surrounded the lingual muscle during myogenesis of the rat tongue.

Using LM and SEM methods, the study describes microstructures in particular areas of the tongue of the goose. A thick multilayered keratinized epithelium forms the "lingual nail" and covers small and giant conical papillae, whereby the first functions as an exoskeleton of the tongue apex, and the latter are arranged along the lingual and well-developed connective tissue cores, and together with the bill lamellae are involved in cutting. The row of conical papillae on the lingual prominence prevents regurgitation of transported food. In the area of the "lingual nail" and in the anterior part of the lingual prominence, Herbst corpuscles are accumulated, which allow to recognize food position. Filiform papillae, as widely distributed between the conical papillae of the body, are responsible for filtering. They can be explained as long keratinized processes of the epithelium and are devoid of connective tissue cores. During food transport, the flattened areas of the lingual body and the lingual prominence are protected by a parakeratinized epithelium, but the root is covered by a nonkeratinized epithelium. The presence of adipose tissue in the tongue probably reduces pressure during food passage, but also promotes mucus evacuation from the lingual glands, thus facilitating food transport. An entoglossal bone with a continuation as cartilage is the stable structural basis of the tongue system., (Copyright © 2011 Wiley-Liss, Inc.)

We used fluorescence immunohistochemistry, analysis of differential interference contrast (DIC) images and confocal laser-scanning microscopy in the transmission mode, after staining specimens with toluidine blue, to examine the localization of keratin 13 (K13) and keratin 14 (K14) in the lingual epithelium of fetal and juvenile Sprague-Dawley rats during the prenatal and postnatal morphogenesis of circumvallate papillae. No immunoreactivity specific for K13 and K14 was detected in the lingual epithelium of fetuses on day 15 after conception (E15), at which time the primitive rudiment of the circumvallate papillae was detectable by the thickening of several layers of cuboidal epithelial cells. On E17 and E19, the developing circumvallate papillae were clearly recognizable, consisting of a central papilla and the surrounding sulcus. No immunoreactivity specific for K13 and K14 was evident in the lingual epithelium around these structures at this time. K14-specific immunoreactivity was first detected in the basal layer of the epithelium of the circumvallate papillae on postnatal day 0 (P0) and K13-specific immunoreactivity was detected on P7. Morphogenesis of the circumvallate papillae progressed significantly from P0 to P14, and immunoreactivity specific for K13 and K14 was clearly recognizable after P7. The respective patterns of K13-specific and K14-specific immunoreactivity differed during the development of the circumvallate papillae: K13-specific immunoreactivity was generally evident in cells of the intermediate layer of the epithelium, while K14-specific immunoreactivity was detected in cells of the basal and suprabasal layers. The present results are discussed in the context of the previously determined localization of K13 and K14 in the dorsal epithelium of the anterior part of the rat tongue during its morphogenesis., (Copyright © 2010 Elsevier GmbH. All rights reserved.)

We have developed a technique, using a combination of immunofluorescent staining of semi-ultrathin sections of epoxy resin-embedded samples and the corresponding differential interference contrast (DIC) images obtained by light microscopy that provides detailed information about the immuno-localization of histological and cellular structures. To demonstrate the effectiveness of our method, we examined the immunofluorescence of immuno-stained keratin 13 (K13) and type III collagen (CIII) and the corresponding DIC images during the morphogenesis of filiform papillae on the rat tongue. Immunoreactivity specific for K13 and CIII was detected on the lingual epithelium of juveniles on postnatal days 7 and 14 (P7 and P14). The immunoreactivity specific for K13 was clearly located in the intermediate-layer cells of the interpapillary cell columns, while that specific for CIII was also distinct in the connective-tissue fibers between the lingual epithelium and the lingual muscle. The DIC images revealed the keratinization of the stratified squamous cells of the lingual epithelium and, also, myogenesis beneath the connective tissue. In addition, immunoreactivity specific for CIII was also recognizable in the endomysium and perimysium around the lingual muscle. Thus, our method demonstrated changes in patterns of immunoreactivity of K13 and of CIII during the morphogenesis of the rat tongue.

We examined the distribution of immunofluorescence due to immunostaining of type III collagen, differential interference contrast (DIC) images and images obtained in the transmission mode after toluidine blue staining by laser-scanning microscopy of semi-ultrathin sections of epoxy resin-embedded samples, during morphogenesis of the filiform papillae, keratinization of the lingual epithelium, and myogenesis of the rat tongue. Immunoreactivity specific for type III collagen was distributed widely in the mesenchymal connective tissue in fetuses on day 15 after conception (E15), at which time the lingual epithelium was composed of one or two layers of cuboidal cells and the lingual muscle was barely recognizable. Immunoreactivity specific for type III collagen was clearly detected on the lamina propria in fetuses on E17 and E19, and it was relatively distinct just beneath the lingual epithelium. Immunoreactivity specific for type III collagen was sparsely distributed on the connective tissue around the developing lingual muscle. In fetuses on E19, the epithelium became clearly stratified and squamous. At postnatal stages from newborn (P0) to postnatal day 14 (P14), keratinization of the lingual epithelium advanced gradually with the development of filiform papillae. On P0, myogenesis of the tongue was almost completed. The intensity of the fluorescence immunoreactivity specific for type III collagen at postnatal stages was almost same as that on E19. The immunoreactivity around the fully mature muscle was relatively distinct between P0 and P14. Thus, type III collagen appeared in conjunction with the morphogenesis of filiform papillae and the keratinization of the lingual epithelium as well as in the connective tissue that surrounded the lingual muscle during myogenesis of the rat tongue.

Objectives: We examined the timing of the appearance and distribution of type II collagen as a possible component of the extracellular matrix that is involved in the morphogenesis of the rat tongue., Methods: We examined the immunofluorescence of type II collagen, differential interference contrast (DIC) images, and images recorded in transmission mode after toluidine blue staining by laser-scanning microscopy (LSM) during the morphogenesis of filiform papillae and the keratinization of the lingual epithelium of rats on semi-ultrathin sections of epoxy resin-embedded samples., Results: Immunoreactivity specific for type II collagen was scattered on cells over a wide area of the mesenchymal connective tissue of the fetal tongue on day 15 after conception (E15), when the lingual epithelium was composed of one or two layers of cuboidal cells. Immunoreactivity specific for type II collagen was recognisable on cells of the lamina propria of the lingual mucosa and around the developing lingual muscle of fetuses at E17 and E19. On E19, the epithelium was clearly of the stratified squamous type. At postnatal stages after birth (P0), immunoreactivity became more and more significant in the connective tissue of the lamina propria with the advancing of morphogenesis of the filiform papillae. In addition, immunoreactivity was widely distributed in the connective tissue around the lingual muscle, as myogenesis in the tongue advanced. The lingual epithelium was composed of stratified squamous cells, and keratinization of the lingual epithelium proceeded gradually as morphogenesis of filiform papillae continued during postnatal development., Conclusion: Type II collagen appeared not only in the connective tissue of the lamina propria as the morphogenesis of filiform papillae occurred and the lingual epithelium became keratinized but also in the endomysium and perimysium around the lingual muscle after myogenesis of the tongue is complete at P0.

We examined the expression of keratin 14 (K14) on the lingual epithelium by immunofluorescent staining while monitoring morphological changes in the filiform papillae of mice by confocal laser-scanning microscopy in the transmission mode of the same sections to define both the histology and the morphology of cells. It is difficult to visualize histological details of the fetal lingual epithelium of the mouse on semi-ultrathin sections by light microscopy after immunohistochemical staining because the histological structures in such sections cannot be distinguished by standard counterstaining. To solve this problem and to visualize the immunoreactivity specific for K14, we analyzed the results of immunofluorescent staining of semi-ultrathin sections in combination with an examination of the corresponding images by laser-scanning microscopy in the transmission mode after staining of specimens with toluidine blue. No immunoreactivity specific for K14 was detected on the lingual epithelium of fetuses on embryonic day 15 (E15), but immunoreactivity was distinct at all postnatal stages from postnatal day 0 (P0) to P21.

We examined the immunofluorescence labelling epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR), as well as differential interference contrast (DIC) images, during the morphogenesis of filiform papillae and the keratinization of the lingual epithelium of rats on semi-ultrathin sections of epoxy resin-embedded samples using laser-scanning microscopy. We also examined semi-ultrathin sections of epoxy resin-embedded, toluidine blue-stained samples by light microscopy to obtain details of cell histology and morphology. No immunoreactivity specific for EGF and EGFR was detected on the lingual epithelium of fetuses on days 12 and 16 after conception (E12 and E16), during which time the number of layers of cuboidal cells in the lingual epithelium increased from one to several. Immunoreactivity specific for EGF and EGFR was first detected on the lingual epithelium of fetuses at birth or on postnatal day 0 (P0). Immunoreactivity specific both for EGF and EGFR appeared in the connective tissue and the basal cells of the papillary and interpapillary cell columns. The lingual epithelium was composed of stratified squamous cells. The rudiments of filiform papillae were compactly arranged and interpapillary cell columns were very narrow. Immunoreactivity specific for EGF and EGFR was distinct on the cell membrane of basal cells of the papillary cell column and weakly positive on the cell membrane of basal cells of the interpapillary cell column on postnatal day 21 (P21). Thus, the patterns of immunoreactivity of EGF and EGFR differed as the filiform papillae developed. Filiform papillae developed gradually from P0 to P21. The width of interpapillary spaces also increased during this period. These observations indicate a possibility that EGF might affect the expression of keratins in the lingual epithelium via epithelium-mesenchymal interactions.

We examined the expression of keratin 18 (K18), by immunofluorescence staining, while monitoring morphological changes in the periderm on the lingual epithelium of rats by laser-scanning microscopy of epoxy resin-embedded, semi-ultrathin sections. We also examined differential interference contrast (DIC) images of the same sections to define the histology and morphology of the cells. It is difficult to visualize histological details of the fetal lingual epithelium of the rat on semi-ultrathin sections by light microscopy after immunohistochemical staining, because the histological structures in such sections cannot be distinguished by standard counterstaining. To solve this problem and to visualize keratin 18 (K18), we used a combination of immunofluorescence staining of semi-ultrathin sections and corresponding differential contrast (DIC) images, obtained by laser-scanning microscopy.

We examined the immunofluorescence of keratins 13 (K13) and 14 (K14) and differential interference contrast (DIC) images during the morphogenesis of filiform papillae and the keratinization of the lingual epithelium of rats on semi-ultrathin sections of epoxy resin-embedded samples by laser-scanning microscopy. We also examined semi-ultrathin sections of epoxy resin embedded, toluidine blue stained samples by light microscopy to obtain details of cell histology and morphology. No immunoreactivity specific for K13 and K14 was detected on the lingual epithelium of foetuses on days 13, 15 and 17 after conception (E13, E15 and E17), during which time the number of layers of cuboidal cells in the lingual epithelium increased from one to several. Immunoreactivity specific for K13 and K14 was first detected on the lingual epithelium of foetuses on E19. The immunoreactivity specific for K13 appeared in the suprabasal cells of the papillary and interpapillary cell columns and immunoreactivity specific for K14 was detected in the basal and suprabasal cells of the papillary and interpapillary cell columns. The lingual epithelium was composed of stratified squamous cells. The rudiments of filiform papillae were compactly arranged and interpapillary cell columns were very narrow. Filiform papillae developed gradually from postnatal day 0 (PO) to 21 (P21). The width of interpapillary spaces also increased during this period. Immunoreactivity specific for K13 and K14 was distinct at all postnatal stages examined. Thus, the patterns of immunoreactivity of K13 and K14 differed as the filiform papillae developed.

An immunofluorescence study of the expression of keratin 14 (K14) during the formation of filiform papillae was performed and the progress of keratinization of the epithelium of the rat tongue was monitored on semi-ultrathin sections by laser-scanning microscopy. Differential interference contrast (DIC) images were also examined to provide details of histology and cell morphology. No cells with immunoreactivity specific for K14 were detected on the lingual epithelium of foetuses on embryonic days 12 and 16 (E12 and E16), when the lingual epithelium was composed of a single layer or several layers of cuboidal cells. Immunoreactivity specific for K14 was detected first on basal and suprabasal keratinocytes of the dorsal epithelium of the tongue of new-borns on postnatal day 0 (P0) and was conspicuous in juveniles on P14. The immunoreactivity was particularly strong on the basal and suprabasal keratinocytes along the connective tissue papillae. The immunoreactivity extended over the entire cytoplasm but was not detected in the nucleus. The lingual epithelium was composed of stratified squamous cells and the rounded rudiments of filiform papillae were compactly arranged at equal intervals, for the most part, and the spaces between them were narrow and indistinct. Immunostaining of K14 was distinct on basal and suprabasal keratinocytes of the filiform papillar area of tongues of juveniles on P21, when the filiform papillae were conical. The spaces between them were relatively wide and, as a result, interpapillar cell columns were clearly visible. Immunoreactivity specific for K14 in the basal and suprabasal keratinocytes of the interpapillar cell columns was recognizable but was weaker than that in cells of papillar cell columns. The thickness of the epithelium in papillar and interpapillar areas increased gradually with the development of filiform papillae. However, sizes of basal and suprabasal keratinocytes remained almost unchanged during this process. These results suggest that the basal and suprabasal keratinocytes of the filiform papillar area proliferate with the initiation of the morphogenesis of filiform papillae and the keratinization of the epithelium. In addition, it appears that, after P14, the basal and suprabasal keratinocytes of the interpapillar area proliferate to supply the keratinocytes of the expanding interpapillar regions.

Studies of the comparative morphology of the tongues of living vertebrates have revealed how variations in the morphology and function of the organ might be related to evolutional events. The tongue, which plays a very important role in food intake by vertebrates, exhibits significant morphological variations that appear to represent adaptation to the current environmental conditions of each respective habitat. This review examines the fundamental importance of morphology in the evolution of the vertebrate tongue, focusing on the origin of the tongue and on the relationship between morphology and environmental conditions. Tongues of various extant vertebrates, including those of amphibians, reptiles, birds and mammals, were analysed in terms of gross anatomy and microanatomy by light microscopy and by scanning and transmission electron microscopy. Comparisons of tongue morphology revealed a relationship between changes in the appearance of the tongue and changes in habitat, from a freshwater environment to a terrestrial environment, as well as a relationship between the extent of keratinization of the lingual epithelium and the transition from a moist or wet environment to a dry environment. The lingual epithelium of amphibians is devoid of keratinization while that of reptilians is keratinized to different extents. Reptiles live in a variety of habitats, from seawater to regions of high temperature and very high or very low humidity. Keratinization of the lingual epithelium is considered to have been acquired concomitantly with the evolution of amniotes. The variations in the extent of keratinization of the lingual epithelium, which is observed between various amniotes, appear to be secondary, reflecting the environmental conditions of different species.

Tongues were removed from rat fetuses on d 16 of gestation (E16) and from newborn (P0) and juvenile rats on d 7 (P7) and d 21 (P21) postnatally for examination by light and transmission electron microscopy. In the fetuses at E16, no rudiments of filiform papillae were visible on the dorsal surface of the tongue. No evidence of keratinisation could be recognised over the entire dorsal lingual epithelium. At P0, rudiments of filiform papillae showed a similar distribution to that seen in the adult, but had a more rounded appearance. The columnar structure of cells in the epithelium, with the different degrees of keratinisation as observed in the mature adult, was indistinct, but a keratinised layer was clearly located at the tip of each filiform papilla. In juveniles at P7, the filiform papillae on the anterior part of the tongue were long and slender, and the anterior and posterior cell columns of the filiform papillae and the interpapillary cell columns were clearly distinguishable. In juveniles at P21, the structure of filiform papillae was identical to that in the adult. These results indicate that, in rats, the morphogenesis of filiform papillae advances in parallel with keratinisation of the lingual epithelium from just before birth to a few weeks after birth.

Comparative observations of the effects of cholinergic and adrenergic agonists (such as pilocarpine, phenylephrine and isoproterenol) on the secretion of salivary fluid and of secretory granules from the lingual epithelium and the lingual gland in the Tokyo Daruma pond frog, Rana porosa porosa, were made by light and transmission electron microscopy. The effect of pilocarpine on the loss of cytoplasm in both the lingual epithelium and the lingual gland was the strongest, and that of isoproterenol was the weakest. Transmission electron microscopy revealed that electron-dense granules, located in cells in both the lingual epithelium and the lingual gland, were discharged by exocytosis after stimulation by phenylephrine and isoproterenol. Immediately before secretion of these granules, they became somewhat larger and round or distorted in shape. However, after administration of pilocarpine, no obvious discharge of electron-dense granules was apparent and, instead, granules in some cells began deteriorating within the cells and those in other cells developed electron-dense and electron-lucent areas. A dot-like pattern was recognized in the electron-dense areas of these granules. These phenomena were assumed to be secondary effects accompanying the secretion of salivary fluid from cells of the lingual epithelium and the lingual gland. By contrast, mucous granules in the lingual gland were secreted by a holocrine process.

Background: Many mammals have four different types of lingual papilla, namely filiform, fungiform, circumvallate, and foliate papillae, on the dorsal or lateral surface of the tongue. However, details of the morphogenesis of these lingual papillae have not been reported. We have investigated the changes in the three-dimensional ultrastructure that occur during the morphogenesis of filiform, fungiform, and circumvallate papillae in rats during fetal and postnatal development., Methods: Tongues were removed from rat fetuses on days 12 (E12) and 16 (E16) of gestation from newborns (P0) and from juveniles on days 7 (P7) and 14 (P14), and 21 (P21) after birth. Scanning electron microscopy was used for all observations., Results: In fetuses at E12, the rudiments of fungiform papillae could be observed as two rows of bulges that extended bilaterally and parallel to the median sulcus on the anterior half of the dorsal surface of the tongue. In fetuses at E16, the arrangement of the rudiments of fungiform papillae was relatively regular, with a latticelike pattern. At this stage, the outline of the rudiment of the circumvallate papilla could be recognized on the median line between the lingual body and the lingual radix. No rudiments of filiform papillae were visible. At P0, rudiments of filiform papillae were compactly distributed over the dorsal surface in the same way as in the adult. The width of these rudiments was about one-fourth that of fungiform papillae, and their tips were round as compared with those of filiform papillae in the adult. The fungiform and circumvallate papillae were large, and their outlines were somewhat irregular, as in the adult. In juveniles at P7, filiform papillae were long and slender. On the intermolar eminence, filiform papillar structures were quite large. A taste pore was clearly visible at the center of each fungiform papilla at this stage. The shape of the circumvallate papilla was similar to that in the adult. In juveniles at P14 and P21, the shapes of all three types of papilla were almost same as those in the adult., Conclusions: The rudiments of each of the three different kinds of lingual papilla appeared at a different respective stage of development in rats. The rudiments of the fungiform and circumvallate papillae, which are related to the sense of taste, were visible earlier than those of the filiform papillae, which are not involved in this sense.

Background: Comparative studies of ultrastructural features of tongues allow deductions to be made about relationships between structure and function, as reflected by an animal's feeding habits. The present study was performed to serve as a basis for further studies of avian feeding mechanisms and of relationships between the fine structure of the lingual epithelium and the development of the expression of keratins., Methods: The light microscope, scanning electron microscope, and transmission electron microscope were used., Results: The dorsal surface of the tongue of Middendorff's bean goose, Anser fabalis middendorffii, has a distinctive anterior region that extends for five-sixths of its length and has a clear posterior region. The anterior region, when observed macroscopically and by scanning electron microscopy, is distinguished along its forward half by a clear median line. The back half of the anterior region has an indistinct median sulcus in some parts. There are no lingual papillae on the entire dorsal surface of the anterior and posterior regions. Giant conical papillae are located in a transverse row between the anterior and posterior regions. On both lateral sides of the anterior region for five-sixths of the length of the tongue, lingual hairs are compactly distributed, and small numbers of large cylindrical papillae are arranged at almost regular intervals between these lingual hairs. Examination of the dorsal lingual epithelium by light and transmission electron microscopy provided histological and cytological criteria for distinguishing the anterior and posterior regions, both of which were composed of stratified squamous epithelium. Basal cells were similar throughout the dorsal epithelium. The intermediate layer of cells in the anterior region contained numerous tonofibrils in electron-dense bundles composed of tonofilaments of 10 nm in diameter. The outer layer was composed of electron-dense, well-keratinized cells, with layers of electron-lucent cells on the outermost surface. The cells in the intermediate layer in the posterior region of the tongue were almost completely filled with unbundled tonofilaments. The surface layer exhibited features of parakeratinization. In all of the giant conical papillae, the large cylindrical papillae, and the lingual hairs, the epithelium was strongly keratinized., Conclusions: The three-dimensional microanatomy and cytological features of the dorsal lingual epithelium of avians seem to be related to the functional role and shape of the tongue of each species in feeding.

Background: The soft-shell turtle, Trionyx cartilagineus, is classified phylogenetically to the family Trionychidae, whose members live in small rivers or ponds. The purpose of the present study was to examine the ultrastructure of the dorsal epithelium of the tongue of the soft-shell turtle and to compare the results of the observations with those reported for the tongue of other freshwater turtles., Methods: Light microscopy, transmission electron microscopy, and scanning electron microscopy were used to examine the dorsal epithelium of the tongue of the soft-shell turtle., Results: The tongue is triangular with a slightly round apex when viewed dorsally but it appears flattened when viewed laterally. Lingual papillae were visible on the dorsal surface of the tongue with some localized variations. Irregular, dome-shaped or ridge-like papillae were observed on the anterior part of the dorsal lingual surface. Large, cylindrical papillae were located along the midline of the posterior part of the tongue. Low, disk-like papillae were located on both sides of the dorsal surface of the posterior part of the tongue. Taste pores were recognizable in the center of the disk-like papillae. At higher magnification, scanning electron microscopy revealed microridges on the surface of cells located on the outermost side of the anterior part of the tongue, and the thickenings of cell margins were clearly seen. Microvilli were distributed compactly over the entire posterior part of the tongue. Light microscopy revealed that the mucosal epithelium of the anterior part of the tongue was of the keratinized, stratified squamous type, while the mucosal epithelium of the posterior part of the tongue was of the nonkeratinized, stratified cuboidal type. In the latero-posterior part of the tongue, taste buds were recognized. Transmission electron microscopy revealed that the epithelium of the anterior part of the tongue was of a typical keratinized type. Small numbers of keratohyalin granules and membrane-coating granules appeared in the cytoplasm of the shallow intermediate layer. On the apical side of the lingual papillae located on the posterior side of the tongue, cells from the intermediate layer to the surface layer of the non-keratinized epithelium contained many fine, discoidal granules. A large part of the epithelium consisted of mucous cells in the concave area on the posterior side., Conclusions: The dorsal surface and epithelium of the tongue of the soft-shell turtle differed significantly from those of other freshwater turtles, in spite of the similarity in terms of gross morphology among the tongues of such turtles.

Histological and ultrastructural studies ot the dorsal lingual epithelium of the juvenile Pacific ridley turtle, Lepidochelys olivacea, were performed by light and electron microscopy, and the results were compared to those of freshwater turtles in order to clarify the relationship between the histological and cellular differences of the lingual epithelium and the habitat of the turtles. The tongue of the juvenile Pacific ridley turtle is triangular with a round apex when viewed from above, but it appears flattened in lateral view. Scanning electron microscopy (SEM) revealed no lingual papillae on the dorsal surface of the tongue. Instead, transverse plicae are found on the surface of the body and the radix. The surface of the apex is smooth. Microridge-like structures are present on the surfaces of the cells, and the cell margins are thickened. The mucosal epithelium is keratinized, stratified squamous with a relatively thick layer of desquamating cells. Cells of the basal and deep intermediate layers appear elliptical in shape; and their nuclei are elliptical and centrally located. Numerous desmosomes join the processes of adjacent cells; and hemidesmosomes anchor the basal cells to the basal lamina. The cytoplasm of these cells contains mitochondria, free ribosomes, rough endoplasmic reticulum, vacuoles, and bundles of tonofilaments. Cells and their nuclei in the intermediate layer display gradual flattening. In the shallow intermediate layer, the cells are significantly flattened, with nuclei condensed or absent. The cytoplasm contains many tonofibrils or bundles of tonofilaments, free ribosomes and keratohyalin granules, with numerous ribosomes attached to their surfaces. A few collapsed mitochondria are visible. Cell membranes of the shallow intermediate cells are smooth and attached to those of adjacent cells by desmosomes. The keratinized layer is located on top of the shallow intermediate layer, and consists of significantly flattened cells lacking nuclei and filled with keratin fibers. Very fine cellular processes joined by desmosomes are visible. The desquamating cells located on top of the keratinized layer contain keratin fibers that are somewhat thicker than tonofibrils and tonofilaments, and clearly distinguishable individually. The microridge-like structures visible by SEM could be attributed to the persistence of cells formed in underlying layer. In conclusion, the histology of the lingual epithelium of the juvenile Pacific ridley turtle differs significantly from that of the adult freshwater turtle in spite of the similarity of the gross morphology of their tongues.

Background: Many studies have been performed to clarify the relationship between behavioral performance of the tongue and Jacobson's organ. The purpose of the present study was to examine the ultrastructural features of the surface of the tongue of the rat snake, Elaphe climacophora, and to delineate the functional relationship between the tongue and Jacobson's organ from a morphological perspective., Methods: The three-dimensional ultrastructure of the surface of the tongue of the rat snake Elaphe climacophora was investigated by scanning electron microscopy., Results: Most of the surface of the bifurcated apex of the tongue was relatively smooth. Dome-shaped, hemispherical bulges or microfacets were compactly arranged on the epithelial cell surface over this entire region. Intercellular borders were clearly recognizable as striations. These features were almost the same as those of the dorsal surface of the transitional area between the bifurcated lingual apex and the anterior part of the lingual body. In the posterior half of the lingual body, no microfacets were seen at all. Both microridges and microvilli were compactly distributed on cell surfaces., Conclusions: No evidence was obtained from our ultrastructural analysis for an important role of the lingual apex in the vomeronasal system. By contrast, the epithelial surface of the body of the tongue appeared suitable for retaining stimulating compounds.

Background: Various species of turtles are adapted to different environments, such as freshwater, seawater, and terrestrial habitats. Comparisons of histological and ultrastructural features of the tongue of the juvenile Hawksbill turtle, Eretmochelys imbricata bissa, with those of freshwater turtles should reveal some aspects of the relationship between the structure of the lingual epithelium and the environment., Methods: The light microscope, scanning electron microscope and transmission electron microscope were used., Results: Light microscopy revealed that the mucosal epithelium of the tongue was of the keratinized, stratified squamous type. Under the scanning electron microscope, no lingual papillae were visible on the dorsal surface of the tongue. Micropits and the thickening of cell margins were clearly seen on the surface of cells located on the outermost side. The transmission electron microscope revealed that the cells in the intermediate layer were gradually flattened from the basal side to the surface side, as were their nuclei. In the shallow intermediate layer, the cells were significantly flattened, and their nuclei were condensed or had disappeared. The cytoplasm contained keratohyalin granules, tonofibrils, free ribosomes, mitochondria, and rough endoplasmic reticulum. Numerous free ribosomes were attached to the surface of small keratohyalin granules. The cells of the keratinized layer were significantly flattened, and their nuclei had completely disappeared. Most of cytoplasm was filled with keratin fibers of high electron density. Keratin fibers of the shedding cells, which were located on the outermost side of the keratinized layer, appeared looser, and each fiber, which was somewhat thicker than the tonofibrils and tonofilaments, was clearly distinguishable., Conclusions: The lingual epithelium of the juvenile Hawksbill turtle differs significantly from that of the adult freshwater turtle, in spite of the similarity in gross morphology of the tongues of these species.

The Asian snail-eating turtle, Malayemys subtrijuga, is classified phylogenetically as a member of the family Emydinae. Members of this family usually live in small rivers or ponds. However, this species is relatively well-adapted to terrestrial life. We describe here the light, scanning electron and transmission electron microscopic appearance of the dorsal lingual epithelium of the snail-eating turtle and we compare the results to those obtained from other freshwater turtles in an attempt to clarify the relationship between the histological and ultrastructural differences in the lingual epithelium and the living circumstances of the turtles. The tongue is triangular with a rounded apex when viewed dorsally but it appears flattened when viewed laterally. Under the scanning electron microscope, no lingual papillae were visible on the dorsal surface of the tongue. Instead, plicae were seen all over the dorsal surface. On the surface of the epithelium of the outermost side, dome-shaped bulges, each of which was coincident with an individual cell, were compactly distributed. At higher magnification, scanning electron microscopy revealed numerous microvilli and microridges on the surface of these cells, and the thickening of cell-margins was clearly seen. Light microscopy revealed that the mucosal epithelium of the tongue was of the non-keratinized, stratified squamous type. Under the transmission electron microscope, the cells of the basal and deep intermediate layers of the epithelium appeared irregularly elliptical in shape. The nucleus was large and also irregularly elliptical, lying in the central region of each epithelial cell. The cytoplasm of these cells contained mitochondria, free ribosomes, rough endoplasmic reticulum and bundles of tonofibrils. Cell membranes formed processes around individual cells. Desmosomes were intercalated between the processes of adjacent cells. In the shallow intermediate layer, the cells were also elliptical, and the elliptical nucleus was located in the central area of each cell. A large part of the cytoplasm was occupied by electron-dense, discoid granules. Filamentous structures filled the spaces between these granules. Small numbers of free ribosomes, mitochondria and rough endoplasmic reticulum were scattered in the cytoplasm. Cell membranes still formed processes around cells. Desmosomes were intercalated between the processes of adjacent cells. The cells of the surface layer were still elliptical, as were their nuclei. Most of the cytoplasm was filled with electron-dense, discoid granules. Fine filamentous structures were dispersed between these granules. Cell membranes formed processes around cells which were coincident with microvilli and microridges. Intercalated desmosomes were also seen. In some cells, many of the electron-dense, discoid granules were secreted into the oral cavity. In conclusion, the histology of the lingual epithelium of the snail-eating turtle is very similar to that of the freshwater turtle, reflecting similarities in the gross morphology of the tongues of these species, in spite of the differences in their life styles.

Tongues were removed from fetuses of mice on the 15th day of gestation (E15), from newborns (P0), from juveniles on the 7th day (P7) and on the 14th day (P14) after birth for examination by scanning electron microscopy. In the fetuses at E15, rudiments of fungiform papillae with a relatively regular, lattice-like pattern were visible on the anterior half of the dorsal surface of the tongue. The outline of the rudiment of a circumvallate papilla could be recognized on the median line between the lingual body and the lingual radix. No rudiments of filiform papillae could be seen. At P0, rudiments of filiform papillae were compactly distributed over the dorsal surface, as are the filiform papillae in the adult, their width was approximately one-third of that of fungiform papillae, and their tips were rounded than those of the filiform papillae in the adult. The fungiform papillae then increased in size and became somewhat irregular in shape. In juveniles at P7, the filiform papillae were long and slender, being relatively large on the intermolar eminence. A taste pore was visible in the center of each fungiform papilla at this stage. The shape of the circumvallate papilla was similar to that in the adult. In juveniles at P14, the shapes of all three types of papilla were almost the same as those in the adult. The rudiments of each of the three kinds of lingual papilla appeared at a different stage of development of mice; rudiments of the fungiform and circumvallate papillae, which are related to the sense of taste, were formed earlier than those of the filiform papillae, which are not involved in taste.

The histological characteristics and ultrastructure of the dorsal lingual epithelium of the rat snake, Elaphe quadrivirgata, were investigated by light microscopy and scanning and transmission electron microscopy. Most of the surface of the bifurcated part of the tongue was relatively smooth. Dome-shaped, hemispherical bulges were compactly arranged on the epithelial cell surface of the basal area of this region. Intercellular borders were clearly recognizable as striations. Microridges were densely distributed on the epithelial cell surface of the lingual body. Intercellular borders were thickened. A keratinized layer was clearly visible in the epithelium of the anterior bifurcated area, namely, at the apex of the tongue. Although keratohyalin granules were not found in any layer of the epithelium in this area, the cells of the surface layer were filled with keratin filaments. The dorsal lingual epithelium of the posterior area, namely, the lingual body, did not show any evidence of keratinization. Each cell on the surface side still had a large, oval nucleus and intact organelles, such as mitochondria, rough endoplasmic reticulum, ribosomes, tonofibrils, and tonofilaments. Cellular interdigitation was evident between adjacent cells and clear microridges or microvilli were observed on the cell membranes on the free-surface side of cells located in the surface layer. The phylogenetic relevance of these findings is discussed.

The amphibian tongue contains two types of papilla which are believed to function in gustation and in the secretion of salivary fluid. Scanning electron microscopy reveals that columnar, filiform papillae are compactly distributed over nearly the entire dorsal surface of the tongue of the frog, Rana cancrivora, and fungiform papillae are scattered among the filiform papillae. Microridges and microvilli are distributed on the epithelial cell surface of the extensive area of the filiform papillae. Light microscopy shows that the apex of each filiform papilla is composed of stratified columnar and/or cuboidal epithelium and its base is composed of simple columnar epithelium. Transmission electron microscopy reveals that most of the epithelium of the filiform papillae is composed of cells that contain numerous round electron-dense granules 1-3 microns in diameter. Cellular interdigitation is well developed between adjacent cells. On the free-surface of epithelial cells, microridges or microvilli are frequently seen. Between these granular cells, a small number of ciliated cells, mitochondria-rich cells and electron-lucent cells are inserted. In some cases, electron-dense granules are present in the ciliated cells. At higher magnification, the electron-dense granules appear to be covered with patterns of spots and tubules. Overall, the morphology and ultrastructure of the lingual epithelium of the three species of Rana that have been studied are quite similar, but they can be easily distinguished from those of Bufo japonicus. Therefore, it appears that lingual morphology is phylogenetically constrained among members of the predominantly freshwater genus Rana to produce uniformity of papillary structure and this morphology persists in Rana cancrivora despite the distinct saline environment in which it lives.

Light and electron microscopic observations of the dorsal lingual epithelium of the crab-eating monkey, Macaca irus, revealed three different regions: the epithelium on the anterior side of the filiform papillae, the epithelium on the posterior side of the filiform papillae, and the interpapillar epithelium. Whereas the basal and suprabasal cells are similar throughout, differences characterize the intermediate and surface layers. Keratohyalin granules appear predominantly in the intermediate layer of the epithelium on the anterior side of filiform papillae. In the epithelium on the posterior side of the filiform papillae, no keratohyalin granules are seen and, instead, tonofibrils are prominent. The cells begin to be significantly flattened. In the interpapillar epithelium, no keratohyalin granules and tonofibrils are seen, and the tonofilaments occupy almost the entire cytoplasm of the cells of the intermediate and surface layers, with the cells having larger volumes in these layers.

The tip and the body of the tongue of the domestic kitten, Felis catus, were examined by light and transmission electron microscopy. On the tip of the tongue, no filiform papillae were observed, but the connective tissue papillae of the lamina propria were recognized. On the lingual body, there were filiform papillae composed of an anterior, a posterior and interpapillar epithelium. Under the transmission electron microscope, the epithelium on the tip of the kitten tongue was found to be of the stratified squamous type. The epithelium contained no cells filled with keratin fibers. In the lingual body, the interpapillar epithelium contained very few keratohyalin granules, and no cells with keratin fibers. In the epithelium on the anterior side of the filiform papillae, numerous keratohyalin granules appeared in the intermediate layer. In the surface layer, a thin layer of typical keratinized cells was visible. In the epithelium on the posterior side of the filiform papillae, a thick layer of keratinized cells was located on the surface layer.

As reptiles are situated phylogenetically between the amphibians and the mammals, they exhibit considerable variation in the structure of their tongues. The present study, one of a series of studies on reptile tongues, aims to demonstrate the three-dimensional structure of the dorsal lingual surface of a turtle, the Japanese terrapin Clemmys japonica, and to clarify the ultrastructural features of the lingual epithelial cells. In the study lingual papillae were observed by scanning electron microscopy to be widely distributed over the dorsal surface of the tongue. Irregularly shaped (conical, columnar or angular) papillae were located in the anterior and central areas, and ridge-like ones, in the latero-posterior area. Histological examination revealed that the connective tissue penetrated into the core of the papillae, and the epithelium was of a stratified squamous and/or cuboidal type. Under the transmission electron microscope, two types of cells were identified in the intermediate layer of the apical epithelium of the lingual papilla: one type was probably an immature mucous cell, whereas the other was elongated in a baso-apical direction, its cytoplasm containing fine granules. In the surface layer of the apical epithelium, typical mucous cells and cells containing numerous, fine, electron-lucent granules were recognized. Both types of cells possessed microvilli on their free-surfaces. In the lateral epithelium of the lingual papillae, the cytological features from the basal layer to the superficial intermediate layer were essentially the same as in the apical epithelium. However, in the surface layer, mucous cells were significantly larger in number than in the apical epithelium.

The histology and the ultrastructure of the dorsal lingual epithelium of the Japanese lizard, Takydromus tachydromoides, were investigated by light and transmission electron microscopy. The epithelium of the anterior and middle portions of the tongue showed "orthokeratinization," being composed of a thick layer of keratinized cells. The epithelium of the posterior portion of the tongue showed "parakeratinization" and "nonkeratinization." Most of the nonkeratinized cells contained large numbers of secretory granules.

The dorsal surface of the tongue of the little tern, Sterna albifrons, has a distinctive anterior region for five-sixths of its length and a terminal posterior region. The anterior region observed by scanning electron microscopy is distinguished along its forward half by a median line from which median papillae protrude. The hind half of the anterior region has a median sulcus without papillae. The deciduous epithelium on both sides of the median line and sulcus bears scattered epithelial protrusions. The posterior lingual region has neither median papillae nor deciduous epithelium. So-called giant conical papillae are located in a transverse row between anterior and posterior regions. Delicate microridges adorn the surfaces of all outer epithelial cells in both regions. Examination of the dorsal lingual epithelium by light and electron microscopy provides histologic and cytologic criteria for distinguishing anterior and posterior regions. Basal cells are nearly alike throughout the dorsal epithelium. Intermediate layer cells of the anterior region contain numerous tonofibrils in electron-dense bundles composed of 10 nm tonofilaments. The outer layer is composed of electron-dense, well-keratinized cells, and electron-lucent epithelial protrusions are present on the exposed surface of the outermost cells. Median papillae are composed of typical keratinized cells, which are nearly filled with keratin filaments. Intermediate layer cells in the posterior region of the tongue are nearly filled with unbundled tonofilaments. There is only a very thin outer keratinized layer in this region.

Filiform papillae, which were densely distributed all over the dorsal surface of the lingual body, were crown-shaped, with a central, circular area that sloped in the anterior direction and several branches that surrounded it in a semicircle from the back of the central area. Dome-shaped, fungiform papillae were scattered among these filiform papillae. At the posterior end of the lingual body, there were four circumvallate papillae. Prominent microridges and elevated intercellular borders were widely distributed in the central area of the filiform papillae and the interpapillar region. On the surface of the branches of the filiform papillae, microridges were rarely seen. On the surface of the fungiform papillae, indistinct microridges were observed. Histologically, the dorsal lingual epithelium revealed three different regions: the epithelium on the anterior side of the filiform papillae, the epithelium on the posterior side of the filiform papillae and the interpapillar epithelium. Whereas the basal and suprabasal cells are similar throughout, differences characterize the intermediate and surface layers. Keratohyalin granules appear predominantly in the intermediate layer in the epithelium on the anterior side of filiform papillae. In the epithelium on the posterior side of the filiform papillae, no keratohyalin granules occur and, instead, tonofibrils are prominent. The cells become significantly flattened. In the interpapillar epithelium, no keratohyalin granules are visible, and the tonofilaments occupy almost the entire cytoplasm of most cells in the intermediate and surface layers. The cells are larger in volume in these layers.

Scanning and transmission electron microscopy was employed to investigate the ultrastructure of the lingual dorsal epithelial cells of the frog, Rana rugosa. The specimens for scanning electron microscopy were prepared by a method that involved osmium postfixation and treatment with acid to remove extracellular material that adhered to the surface of the tongue. Over almost the entire dorsal surface, filiform papillae, consisting of a large number of non-ciliated cells with microridges and a very small number of ciliated cells, were compactly distributed. Fungiform papillae were scattered among these filiform papillae. A round sensory disk was located on the top of each fungiform papilla. Each sensory disk was encircled by a band of ciliated cells. Transmission electron microscopy revealed that a large part of the filiform papillar epithelium was composed of cells that contained numerous electron-dense granules. These cells were coincident with the non-ciliated cells observed by scanning electron microscopy. In these cells, the nucleus was located on the basal side, and the ergastoplasm was well-developed on the basal side of the nucleus.

The three-dimensional structure of the connective tissue core (CTC) of each type of lingual papillae of Mogella wogura wogura was studied by scanning electron microscopy after removal of the epithelial cell layer and compared with the results obtained from light microscopy as well as transmission electron microscopy. Filiform papillae are densely distributed on the dorsal surface of the anterior part of the tongue. They were conical in shape and their connective tissue cores (CTCs) resembled wooden spoons at the tip of the tongue, while they were flower-shaped (Lysichiton camtschatense) at the middle part of the tongue. Fungiform papillae which had a round depression on the top were distributed sporadically among the filiform papillae, and contained columnar CTC with several plane striations running longitudinally along the lateral surface. A pair of vallate papillae was located at the boundary between the anterior and posterior tongue. Their CTC were flower shaped closely resembling a carnation. Giant conical papillae occupied the posterior marginal region of the tongue. These papillae contained much smaller conical CTC similar to the outer form. Light and transmission electron microscopic observations of the dorsal lingual epithelium revealed three different regions: anterior region to the filiform papillae, posterior to the papillae and interpapillary region. In the intermediate layers between the germinal layer and the surface layer of the anterior region to the filiform papillae, a large number of keratohyaline granules was observed, but the cornified layer was obscured. In the posterior region, keratohyaline granules were fewer in number and the cornified layer was clear and thick. In the interpapillary region, keratohyaline granules were few and a thin cornified layer was recognized. At higher magnification, small sized keratohyaline granules contained a large number of free ribosomes, suggesting a close relationship between the two. Odland bodies were found only on the interpapillary region.

The epithelium of the lingual dorsum of the mongoose, Herpestes edwardsi, was composed of two main areas: the interpapillary area, characterised by the absence of keratinisation; and the papillary area, characterised by hard keratinisation. The number of tonofibrils in the cytoplasm gradually increased, and the volume of the individual cells also increased, from the germinal layer to the intermediate layer of the interpapillary epithelium. No keratohyalin granules were recognised in these cells. In the cells of the surface layer of the interpapillary epithelium, most of the cytoplasm was filled with tonofilaments. The germinal layer of the papillary epithelium was almost identical to that of the interpapillary epithelium. In the intermediate layer, three different types of keratohyalin granule were apparent. The first type consisted of loosely aggregated ribosomes and had moderate electron density. The second type consisted of electron-dense granules derived from granules of the first type. The third type consisted of conjugated structures made up of tonofibrils, which were embedded in electron-dense material. Free ribosomes gradually disappeared as the keratinised layer was approached and most of the cytoplasm of cells in this layer was filled with keratin fibres.

Three different types of lingual papilla were observed by scanning electron microscopy on the dorsal lingual epithelium of the lizard Gekko japonicus. Dome-shaped lingual papillae were located at the apex. Flat, fan-shaped lingual papillae were seen in the widest area of the lingual body. Long, scale-like lingual papillae were arranged on the latero-posterior dorsal surface. At higher magnification, microvilli and microridges were seen to be widely distributed over the surface of the papillae. By light microscopy, the epithelium of the dome-shaped papillae was composed of single, columnar epithelial cells filled with secretory granules. The tip of the epithelium of the fan-shaped and scale-like papillae was composed of stratified squamous epithelial cells without granules. The major part of the epithelium of these two types of papilla, except the tip area, was also composed of single, columnar epithelial cells with secretory granules. By transmission electron microscopy, a nucleus without a defined shape was seen to be located in the basal part of each of the single, columnar epithelial cells. Rough-surfaced endoplasmic reticulum and Golgi apparatus were well developed around the nucleus. The other, major part of the cytoplasm was filled with the spherical secretory granules, a large number of which had very electron-dense cores and moderately electron-dense peripheral regions. In the stratified squamous epithelium, a nucleus, which tended to be condensed on the free-surface side, was located in the center of each cell. Mitochondria, endoplasmic reticulum, and vesicles were observed in the cytoplasm.

The epithelial cell layer was removed from the underlying connective tissue papillae by the method of long term hydrochloric acid treatment and the connective tissue surface was observed by scanning electron microscopy. Tongues of insectivora (Suncus murinus), rodentia (mouse, rat, guinea pig), carnivores (dog, cat) and primates (man) were studied. Each animal species had proper architecture of connective tissue papillae as to four types of lingual papillae. There was a tendency for the stereo structure of connective tissue papillae of these lingual papillae to become more complicated from insectivora to primates.

The three-dimensional structure of the connective tissue core (CTC) of the four types of lingual papillae of the crab-eating macaque (Macaca fascicularis) was studied by scanning electron microscopy after removal of the epithelial cell layer by long-term treatment with hydrochloric acid at room temperature and compared with the results obtained from light microscopy. 1) Filiform papillae are densely distributed on the dorsal surface of the anterior two thirds of the tongue. Fungiform papillae are scattered among these filiform papillae, and are numerous at the anterior margin of the tongue. The connective tissue core of each filiform papilla looked like a lifting human hand with its palm towards the tongue tip. By using light microscopy, AF-positive elastic fibers were accumurated very densely just beneath the basal region of the anterior column epithelium of each filiform papilla. 2) CTC of fungiform papilla showed coralliform structure whose branches were ramified a few times. On the top of each branch there was a small depression where a taste bud was situated. 3) After removal of the epithelial cell layer of the foliate papilla, longitudinal grooves coinciding with the epithelial slits were observed. Some glandular ducts were seen protruding towards the exfoliated epithelium. 4) After removal of the covering epithelium of the vallate papilla, numerous small rod-shaped secondary CT cores appeared on the large primary CT core.

The surface structure of the connective tissue papillae (CP) of Suncus murinus tongue was observed by SEM after fixing with Karnovsky's fixative and removal of the epithelial cell layer with 3N or 8N HCl. On the surface of the slender conical tongue, there are densely distributed filiform papillae among which fungiform papillae are seen sporadically. A pair of vallate papillae are situated in the posterior region of the tongue. Filiform papillae appear somewhat different externally depending on the dorsal surface of the anterior tongue. At the tip of the tongue, filiform papillae are of a slender conical shape and have a slight depression in the anterior basal portion. The CP of these is seen as a spherical protrusion on which a shallow groove runs in the anteroposterior direction. In the middle region, somewhat large filiform papillae contain CP having one or two small round head-like structures on each spherical protrusion. These head-like structures are increased in number in the posterior region. In the most posterior region of the anterior tongue, there are distributed large filiform papillae having several slender protrusions that surround a basal anterior depression. These large branched filiform papillae have a glove finger like CP. Small conical filiform papillae are distributed in the posterior marginal region of the anterior tongue which have CP of a horse-shoe like protrusion that opens in the anterior direction. Spherical fungiform papillae have CP which are thick columnar in shape with many lateral thin folds running vertically and having a round depression on the top of each. CP of the vallate papillae appear as a beehive like structure.(ABSTRACT TRUNCATED AT 250 WORDS)

Light and electron microscopic examination of the dorsal lingual epithelium of beagle dogs (Canis domesticus) revealed three different regions: that anterior to the filiform papillae, that posterior to the papillae, and an interpapillary region. Whereas the basal and suprabasal cells are similar throughout, differences characterize the intermediate and surface layers. Keratohyalin granules are common in the intermediate layers in the anterior and interpapillary regions, tonofibrils are prominent in the posterior region, and no keratohyalin granules occur. The surface layer of the interpapillary region is not keratinized, that of the anterior region shows soft keratinization, and that of the posterior region shows hard keratinization. The perimeter of keratohyalin granules is composed of ribosomes 10-20 nm in diameter.

The morphological relationship between lingual papillae and underlying connective tissue papillae of mouse was studied because it is conceivable that the differentiation of epithelium may be affected by its connective tissue. Tongues of adult male mice were fixed in formol or Karnovsky's fixative. After removal of the epithelium by long-term hydrochloric acid treatment at room temperature, the surface of the connective tissue papillae was observed by scanning electron microscopy. Connective tissue papillae that were fungiform in shape and which were distributed at the anterior part of the tongue showed barnacle-like protrusion after removal of the epithelium. Their surface was covered by numerous long filaments running vertically and there was a round depression on the top of each fungiform papilla that may be found to correspond to a taste bud when the results of light and electron microscopy are compared. Filiform papillae in a narrow sense were closely distributed in the anterior part of the tongue. They had a tapered tip declining posteriorly. Each filiform connective tissue papilla was conical in shape and had a round depression that slightly declined antero-downward, and a long narrow depression ran along the anterior edge of each connective tissue papilla. Large conical papillae which distributed at the anterior margin of the intermolar prominence had shovel-like connective tissue papillae which had a depression at the posterior surface unlike that of the filiform papillae. Branched papillae distributed in the posterior part of the prominence had a depression at the anterior surface. Under the light microscope, numerous keratohyaline granules were seen to be contained only in the posterior epithelial cell line of the large conical papillae distributed in the anterior margin of the prominence, while these granules were found only in the anterior epithelial cell line of both filiform and branched papillae. It became clear that the axes of each connective tissue papilla of large conical papillae distributed radically around a single midpoint. Connective tissue papillae of vallate papillae had a beehive-like shape and in follicate papillae there were several vertical elliptical gaps, seen when the epithelium was peeled from the connective tissue.

The structure of the lingual papillae and the ultrastructure of the surface of the lingual dorsal epithelial cells of squirrel monkeys were observed by scanning electron microscopy. Filiform papillae were distributed over the entire dorsal surface of the tongue, except for the radix zone. Fungiform papillae were scattered among these filiform papillae. In the middle of the posterior end of the lingual body, a single vallate papilla was located. Higher magnification of the lingual dorsal epithelium revealed that prominent microridges and elevated intercellular borders occurred widely in the basofrontal area of the filiform papillae, interpapillar area and lingual radix zone. On the surface of the upper part of the filiform papillae, fine pits and hollows were observed. Indistinct microridges were distributed over the surface of the fungiform papillae.

Comparative features of the dorsal tongue epithelia in musk shrews, mongooses and rats were described. The shapes of the filiform papillae were different in each of the species. The distribution pattern of filiform papillae was similar both in the musk shrews and mongoose, in that the form of filiform papillae changed gradually from the lingual apex to the posterior part of the lingual body. By contrast, the different types of filiform papillae were distributed on definite areas of the dorsal lingual surface in the rat. Microridges on the interpapillar surface in the musk shrew and mongoose presented a clear outline, but those of the rat were not so distinct. In all species, the upper surface of filiform papillae did not show any distinct microridges.