The red ruffed lemur, Varecia rubra (É. Geoffroy Saint-Hilaire, 1812): a comparative morphology investigation of lingual papillae and connective tissue cores.

The morphology of the lingual papillae (filiform, foliate, fungiform, and vallate papillae) and the underlying connective tissue core of the red ruffed lemur (Varecia rubra) of a strepsirrhines species were studied using light and scanning electron microscopy. The filiform papillae distributed at the root of the tongue were larger than the structures distributed at the body and apex. Six to eight vallate papillae were arranged in a Y-shape at the border between the lingual body and the lingual root. Foliate papillae were observed at the posterior lateral border of the tongue. Scanning electron microscopy revealed a primary process and numerous auxiliary processes in the epithelial layer of filiform papillae. After epithelial removal, the connective tissue core of the filiform papilla showed several protrusions surrounding an oval-shaped depression that extended slightly posteriorly, and a large, maple-shaped filiform papilla was seen in the posterior portion of the tongue. The connective tissue cores of the fungiform papillae exhibited a longitudinally ridged cylindrical structure. The connective tissue core of the foliate papillae had numerous tubular projections arranged along a groove with a salivary gland conduit at the base. As a Lemuridae species, the appearance of the fungiform and filiform papillae of the red ruffed lemur is similar to that reported in previous studies of the ring-tailed lemur, with some differences, especially in the filiform papillary connective tissue core at the base and tongue body border. These findings suggest the taxonomic and phylogenetic origins of the lemurs as well as the influence of dietary diversity.

Comparative morphology and physiology of the vocal production apparatus and the brain in the extant primates.

Objective data mainly from the comparative anatomy of various organs related to human speech and language is considered to unearth clues about the mechanisms behind language development. The two organs of the larynx and hyoid bone are considered to have evolved towards suitable positions and forms in preparation for the occurrence of the large repertoire of vocalization necessary for human speech. However, some researchers have asserted that there is no significant difference of these organs between humans and non-human primates. Speech production is dependent on the voluntary control of the respiratory, laryngeal, and vocal tract musculature. Such control is fully present in humans but only partially so in non-human primates, which appear to be able to voluntarily control only supralaryngeal articulators. Both humans and non-human primates have direct cortical innervation of motor neurons controlling the supralaryngeal vocal tract but only human appear to have direct cortical innervation of motor neurons controlling the larynx. In this review, we investigate the comparative morphology and function of the wide range of components involved in vocal production, including the larynx, the hyoid bone, the tongue, and the vocal brain. We would like to emphasize the importance of the tongue in the primary development of human speech and language. It is now time to reconsider the possibility of the tongue playing a definitive role in the emergence of human speech.

Non-invasive intravital observation of lingual surface features using sliding oral mucoscopy techniques in clinically healthy subjects.

To investigate intravital morphological features of the broader area of the lingual mucosa in clinically healthy subjects, and to attempt to evaluate subclinical conditions, we evaluated detailed intravital morphological features of the lingual mucosa using our newly developed oral contact mucoscopy techniques. Clinically healthy subjects (female: 19-22 years, average age: 20.27 years, and n = 28) were enrolled. A position indicator stain was placed on the lingual mucosal surface, and sliding images were captured and then reconstructed. In addition, the lingual mucosa was divided into six areas, and morphometry of the fungiform and filiform papillae was performed. The results were statistically analyzed. There were two morphological features among clinically healthy subjects involving the filiform papillae: the length of the papillae and the degree of biofilm (tongue coat) deposition. We defined a modified tongue coat index (mTCI) with scores ranging from 0 (tongue coating not visible) to 0.5, 1, 1.5, and 2 (thick tongue coating) for six sections of the tongue dorsum. No subjects received a score of 2. Significant differences were found in the mTCI between the six sections of the tongue dorsum, especially between the posterior areas and the lingual apex. The fungiform papillae of some subjects exhibited elongated morphological changes. Our findings suggest that magnified lingual dorsum examination of a broader area is especially important in accurate screening for subclinical or transient conditions of potential lingual mucosal diseases. For this purpose, our new oral mucoscopy and non-invasive intravital observational techniques were especially effective.

Patterns of immunoreactivity specific for gustducin and for NCAM differ in developing rat circumvallate papillae and their taste buds.

α-Gustducin and neural cell adhesion molecule (NCAM) are molecules previously found to be expressed in different cell types of mammalian taste buds. We examined the expression of α-gustducin and NCAM during the morphogenesis of circumvallate papillae and the formation of their taste buds by immunofluorescence staining and laser-scanning microscopy of semi-ultrathin sections of fetal and juvenile rat tongues. Images obtained by confocal laser scanning microscopy in transmission mode were also examined to provide outlines of histology and cell morphology. Morphogenesis of circumvallate papillae had already started on embryonic day 13 (E13) and was evident as the formation of placode. By contrast, taste buds in the circumvallate papillae started to appear between postnatal day 0 (P0) and P7. Although no cells with immunoreactivity specific for α-gustducin were detected in fetuses from E13 to E19, cells with NCAM-specific immunoreactivity were clearly apparent in the entire epithelium of the circumvallate papillary placode, the rudiment of each circumvallate papilla and the developing circumvallate papilla itself from E13 to E19. However, postnatally, both α-gustducin and NCAM became concentrated within taste cells as the formation of taste buds advanced. After P14, neither NCAM nor α-gustducin was detectable in the epithelium around the taste buds. In conclusion, α-gustducin appeared in the cytoplasm of taste cells during their formation after birth, while NCAM appeared in the epithelium of the circumvallate papilla-forming area. However, these two markers of taste cells were similarly distributed within mature taste cells.

Immunohistochemical analysis of type III collagen expression in the lingual mucosa of rats during organogenesis of the 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.

Immunohistochemical expression of type II collagen in the lingual mucosa of rats during organogenesis of the 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.

Newly developed technique for dual localization of keratins 13 and 14 by fluorescence immunohistochemistry.

It is difficult to visualize histological details on semi-ultrathin sections by light microscopy after immunohistochemical labeling because the histological structures in such sections cannot be distinguished by standard counterstaining. To solve this problem and to visualize the immunoreactivity of keratins 13 (K13) and 14 (K14), we used a newly developed technique for dual localization of antigens by fluorescence immunohistochemistry and confocal laser-scanning microscopy in transmission mode, after staining specimens with toluidine blue. Using this approach, we examined the immunolocalization of K13 and K14 on the lingual epithelium of fetal and juvenile rats by immunofluorescence while monitoring morphological changes in the filiform papillae by laser-scanning microscopy, in transmission mode, of the same sections. No K13 and K14 immunoreactivity was detected on the lingual epithelium of fetuses on day 15 after conception (E15), at which time the lingual epithelium was composed of a few layers of cuboidal cells. K14 immunoreactivity was first detected on the lingual epithelium of fetuses on E17 and K13 immunoreactivity on E19. The number of layers of cuboidal cells in the lingual epithelium also increased from E17 to E19. K13 and K14 immunoreactivity was distinct at all postnatal stages examined. Although the respective patterns of K13 and K14 immunoreactivity differed as the filiform papillae developed, K13 immunoreactivity was generally evident in the suprabasal cells of the interpapillary cell columns and K14 immunoreactivity was detected in the basal and suprabasal cells of the papillary and interpapillary cell columns. Our newly developed technique for dual localization of antigens should be useful for investigations of very small specimens, such as fetal tissues and organs.

Expression of keratin 18 in the periderm cells of the lingual epithelium of fetal rats: visualization by fluorescence immunohistochemistry and differential interference contrast microscopy.

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.

Morphological study of lingual papillae in the fetal cat at mid-gestation, as revealed by light and scanning electron microscopy.

Tongues were removed from five cat fetuses, after approximately 1 month of gestation, for examination by light and scanning electron microscopy. Rudiments of filiform papillae were visible over the entire dorsal surface of the tongue. The epithelium of the dorsal surface of the tongue was of the stratified cuboidal type. No evidence of keratinization was detected anywhere in the entire dorsal lingual epithelium. Rudiments of fungiform papillae were recognizable only at the lingual apex; none were recognizable on other parts of the dorsal surface of the tongue at this stage. By contrast, rudiments of circumvallate and foliate papillae were already distinguishable from the filiform papillae. Differences between these results and those obtained previously in rats and mice are discussed.