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PubMed2000

Prox 1 in eye degeneration and sensory organ compensation during development and evolution of the cavefish Astyanax.

We have investigated expression of the homeobox gene Prox 1 during eye degeneration and sensory organ compensation in cavefish embryos. The teleost Astyanax mexicanus consists of sighted surface-dwelling forms (surface fish) and several populations of blind cave-dwelling forms (cavefish), which have evolved independently. Eye formation is initiated during cavefish development, but the lens vesicle undergoes apoptosis, and the eye subsequently arrests and degenerates. The requirement of Prox 1 for lens fiber differentiation and gamma-crystallin expression in the mouse suggests that changes in the expression of this gene could be involved in cavefish eye degeneration. Surface fish and cavefish embryos stained with a Prox 1 antibody showed Prox 1 expression in the lens, neuroretina, myotomes, heart, hindbrain, and gut, as reported in other vertebrates. We found that Prox 1 expression is not altered during cavefish lens development. Prox 1 protein was detected in the lens vesicle as soon as it formed and persisted until the time of lens degeneration in each cavefish population. The cavefish lens vesicle was also shown to express a gamma-crystallin gene, suggesting that Prox 1 is functional in cavefish lens development. In addition to the tissues described above, Prox 1 is expressed in developing taste buds and neuromasts in cavefish, which are enhanced to compensate for blindness. It is concluded that the Prox 1 gene is not involved in lens degeneration, but that expansion of the Prox 1 expression domain occurs during taste bud and neuromast development in cavefish.

Animals↗
PubMed1979

Postnatal development of rat soft palate.

The stratified squamous epithelium lining the oral surface of the soft palate is keratinized and serves as a masticatory organ. Taste buds develop from the cells of the stratified squamous epithelium lining the oral surface during the first 3 weeks of life, and after this no new buds are formed. They are present in the median and paramedian regions of the soft palate: 30% are in the epithelium of fungiform papillae, the remainder being in the surface epithelium. Only 6% of the taste buds are in the vicinity of glandular pits. Glands begin to develop from the deeper portion of tubular invaginations of the stratified squamous epithelium lining the soft palate and grow by pouching, resulting in the development of glands with large irregular pouches. The glands are lined by tall columnar cells and secrete mucus; they are absent from the lateral regions of the posterior third of the soft palate.

Aging↗
PubMed2001

Neural networks distinguish between taste qualities based on receptor cell population responses.

Response features of taste receptor cell action potentials were examined using an artificial neural network to determine whether they contain information about taste quality. Using the loose patch technique to record from hamster taste buds in vivo we recorded population responses of single fungiform papillae to NaCl (100 mM), sucrose (200 mM) and the synthetic sweetener NC-00274-01 (NC-01) (200 microM). Features of each response describing both burst and inter-burst characteristics were then presented to an artificial neural network for pairwise classification of taste stimuli. Responses to NaCl could be distinguished from those to both NC-01 and sucrose with accuracies of up to 86%. In contrast, pairwise comparisons between sucrose and NC-01 were not successful, scoring at chance (50%). Also, comparisons between two different concentrations of NaCl, 0.01 and 0.005 M, scored at chance. Pairwise comparisons using only those features that relate to the inter-burst behavior of the response (i.e. bursting rate) did not hinder the performance of the neural network as both sweeteners versus NaCl received scores of 75--85%. Comparisons using features corresponding to each individual burst scored poorly, receiving scores only slightly above chance. We then compared the sweeteners with varying concentrations of NaCl (0.1, 0.01, 0.005 and 0.001 M) using only those features corresponding to bursting rate within a 1 s time window. The neural network was capable of distinguishing between NaCl and NC-01 at all concentrations tested; while comparisons between NaCl and sucrose received high scores at all concentrations except 0.001 M. These results show that two different taste qualities can be distinguished from each other based solely on the bursting rates of action potentials in single taste buds and that this distinction is independent of stimulation intensity down to 0.001 M NaCl. These data suggest that action potentials in taste receptor cells may play a role in taste quality coding.

Action Potentials↗
PubMed1997

Initial innervation of embryonic rat tongue and developing taste papillae: nerves follow distinctive and spatially restricted pathways.

The rat tongue has an extensive, complex innervation from four cranial nerves. However, the precise developmental time course and spatial routes of these nerves into the embryonic tongue are not known, although this knowledge is crucial for studying mechanisms that regulate development and innervation of the lingual taste organs, gustatory papillae and resident taste buds. We determined the initial spatial course of nerves in the developing tongue and papillae, and tested the hypothesis that sensory nerves first innervate the tongue homogeneously and then retract to more densely innervate papillae and taste buds. Antibodies to GAP-43 and neurofilaments were used to label nerve fibers in rat embryo heads from gestational day 11 through 16 (E11-E16). Serial sagittal sections were traced and reconstructed to follow paths of each nerve. In E11 rat, geniculate, trigeminal and petrosal ganglia were labeled and fibers left the ganglia and extended toward respective branchial arches. At E13 when the developing tongue is still a set of tissue swellings, the combined chorda/lingual, hypoglossal and petrosal nerves approached the lingual swellings from separate positions. Only the chorda/lingual entered the tongue base at this stage. At E14 and E15, the well-developed tongue was innervated by all four cranial nerves. However, the nerves maintained distinctive entry points and relatively restricted mesenchymal territories within the tongue, and did not follow one another in common early pathways. Furthermore, the chorda/lingual and glossopharyngeal nerves did not set up an obvious prepattern for gustatory papilla development, but rather seemed attracted to developing papillae which became very densely innervated compared to surrounding epithelium at E15. To effect this dense papilla innervation, sensory nerves did not first innervate the tongue in a homogeneous manner with subsequent retraction and/or extensive redirection of fibers into the taste organs. Results contribute to a set of working principles for development of tongue innervation. Points of entry and initial neural pathways are restricted from time of tongue formation through morphogenesis, suggesting distinctive lingual territories for each nerve. Thus, sensory and motor nerves distribute independently of each other, and sensory innervation to anterior and posterior tongue remains discrete. For taste organ innervation, gustatory papillae are not induced by a prepatterned nerve distribution. In fact, papillae might attract dense sensory innervation because neither chorda/lingual nor glossopharyngeal nerve grows homogeneously to the lingual epithelium and then redistributes to individual papillae.

Animals↗
PubMed1992

Stereo architecture of the interface of the epithelial cell layer and connective tissue core of the foliate papilla in the rabbit tongue.

The three-dimensional relationship between the epithelial cell layer and the underlying connective tissue core (CTC) of the foliate papilla of the rabbit tongue was studied by scanning electron microscopy after removal of the epithelial cell layer. The foliate papillae were fixed in Karnovsky's fixative, and the epithelial cell layers were exposed to long-term hydrochloric acid treatment (3.5 N HCl for 2-3 weeks at room temperature). The foliate papillae consisted of ridges and grooves located on the posterolateral margin of the tongue. They appeared as linear projections or ridges of lingual mucosa roughly perpendicular to the longitudinal axis of the tongue. These projections or ridges were parallel to one another and separated by grooves. After removal of the epithelium, two kinds of CTC folds appeared: one was the septal fold of CTC which runs in the central portion under each linear projection or ridge, and the other consisted of two sheets of groove side folds of CTC which run along both sides of the former and face the groove side epithelium. It was revealed that there are two sheets of septal epithelial processes, and each of them fits in between each septal fold and groove side fold of CTC. Numerous taste buds were located in the groove side epithelia, and their pores faced the surface of the groove. On the hollow surfaces that appeared on the surface of the groove side fold of CTC after removal of the epithelial cells with taste buds, nerve-terminal-like structures were encountered. Some openings of the ducts of small lingual glands were arranged linearly on the underside of the basal portion of each groove side epithelium.

Animals↗
PubMed2005

The survival of vagal and glossopharyngeal sensory neurons is dependent upon dystonin.

The vagal and glossopharyngeal sensory ganglia and their peripheral tissues were examined in wild type and dystonia musculorum mice to assess the effect of dystonin loss of function on chemoreceptive neurons. In the mutant mouse, the number of vagal and glossopharyngeal sensory neurons was severely decreased (70% reduction) when compared with wild type littermates. The mutation also reduced the size of the circumvallate papilla (45% reduction) and the number of taste buds (89% reduction). In addition, immunohistochemical analysis demonstrated that the dystonin mutation reduced the number of PGP 9.5-, calcitonin gene-related peptide-, P2X3 receptor- and tyrosine hydroxylase-containing neurons. Their peripheral endings also decreased in the taste bud and epithelium of circumvallate papillae. These data together suggest that the survival of vagal and glossopharyngeal sensory neurons is dependent upon dystonin.

Animals↗
PubMed1994

Reduction of electrical coupling between Necturus taste receptor cells, a possible role in acid taste.

Cytoplasmic acidification in taste receptor cells is thought to be involved, at least in part, in acid taste transduction. Since in taste buds about 20% of the receptor cells are electrically coupled, we have tested whether reduction in intracellular pH affects these lateral synaptic interactions. By applying the patch clamp technique to a slice preparation of Necturus lingual epithelium, we found that electrical coupling between taste receptor cells was strongly reduced by cytoplasmic acidification. Therefore, electrical coupling in taste buds might be modified during acid stimulation.

Acids↗
PubMed2005

Rapid quantitative assessment of fungiform papillae density in the human tongue.

Fungiform papillae density, which can be used in a variety of circumstances as an indicator of taste function [L.M. Bartoshuk, V.B. Duffy, I.J. Miller, PTC/PROP tasting: anatomy, psychophysics and sex effects, Physiol. Behav. 56 (1994) 1165-1171; I.J. Miller, F.E. Reedy, Variation in human taste bud density and taste intensity perception, Physiol. Behav. 47 (1990) 1213-1219; J.R. Zuniga, N. Chen, C.L. Phillips, Chemosensory and somatosensory regeneration after lingual nerve repair in humans, J. Oral Maxillofac. Surg. 55 (1997) 2-13], was measured on the dorsal surface of the anterior tongue of living humans using a digital camera and a videomicroscope. Both procedures provided similar results, with the camera providing a more rapid, portable and flexible imaging procedure. Subsequently, the camera was successfully used to identify small regions of the anterior tongue which provide reliable measures of fungiform papillae density that correlate highly with the total number of fungiform papillae on the anterior tongue.

Adult↗
PubMed2006

Age-related expression of alpha-gustducin in the rat larynx.

OBJECTIVES: The age-related changes in distribution of alpha-gustducin-immunoreactive structures in the larynx of Sprague-Dawley rats were studied. METHODS: For this purpose, tissues obtained from 12 male rats ranging in age from 5 to 21 weeks were compared with respect to the distribution and morphology of laryngeal taste buds immunoreactive for alpha-gustducin, the alpha-subunit of a taste cell-specific G protein. RESULTS: Three different morphological types of alpha-gustducin-immunoreactive structures were seen: typical gemmal forms, clusters composed of 2 or 3 cells, and isolated immunoreactive cells not associated with taste buds. alpha-Gustducin-immunoreactive structures were most abundant in the epiglottis in all age groups. alpha-Gustducin-immunoreactive cells in rats seem to be located along the lateral food channels, in which they may come in contact with food. The total number of these alpha-gustducin-immunoreactive structures did not show any age-related changes, but the percentage of solitary immunoreactive cells in 5-week-old rats was significantly larger than the percentages in 8-, 14-, and 21-week-old animals. CONCLUSIONS: Solitary alpha-gustducin-immunoreactive cells, which are abundant in 5-week-old rats but are found in fewer numbers along the base of the epiglottis in mature rats, may be nociceptic in function, whereas the chemosensory clusters or buds that contain alpha-gustducin-positive cells and are distributed along the lateral food channels on the pharyngeal side of the larynx may have a role in gustatory reception.

Aging↗
PubMed1988

A study of the dipeptidylpeptidase IV activity in cat fungiform papillae: light and electron microscope histochemistry.

The present paper describes histochemical study of the dipeptidylpeptidase IV activity in the nerve structures of cat fungiform papillae at the light and electron microscope levels. The dipeptidylpeptidase IV activity was found in blood vessels and nerve bundles entering the connective tissue stroma of fungiform papillae. The taste buds exhibited a moderate staining for the dipeptidylpeptidase IV activity. Ultracytochemical findings revealed this enzyme as membrane-bound in the endothelium of blood vessels, in plasma membrane of the Schwann cells at the axon-Schwann cell interface as well as in the taste bud cells. A possible function of the dipeptidylpeptidase IV activity in the peripheral nerve structures is discussed in view of the ability of this enzyme to cleave the substance P to the minor fragments with inherent physiological roles.

Animals↗
PubMed1997

Organ cultures of embryonic rat tongue support tongue and gustatory papilla morphogenesis in vitro without intact sensory ganglia.

Taste buds on the mammalian tongue are confined to the epithelium of three types of gustatory papillae: the fungiform, circumvallate, and foliate. The gustatory papillae are composed of an epithelium that covers a broad connective tissue core, with extensive innervation to taste bud and nongustatory epithelial locations. Although the temporal sequence of gustatory papilla development is known for several species, factors that regulate initiation, growth, and maintenance of the papillae are not understood. We tested the hypothesis that sensory innervation is required for the initial formation and early morphogenesis of fungiform papillae in a patterned array. An organ culture of the embryonic rat tongue was developed to provide an in vitro system for studying mechanisms involved in fungiform papilla morphogenesis in patterns on the anterior tongue. Tongues were dissected from embryos at 13 days of gestation (E13), a time when the tongue has not yet fully formed and gustatory papillae have not yet appeared, and at 14 days of gestation (E14), when the tongue is well formed and papillae make their initial morphological appearance. Dissected tongues were maintained at the gas/liquid interface in standard organ culture dishes, fed with DMEM/F12 plus 2% B-27 supplement and 1% fetal bovine serum. After 1, 2, 3, or 6 days in culture, tongues were processed for scanning electron or light microscopy, or immunocytochemistry. Tongues cultured from E13 or E14 underwent extensive morphogenesis and growth in vitro. Furthermore, fungiform papillae developed on these tongues on a culture day equivalent to E15 in vivo; that is, after 2 days for cultures begun at E13 and 1 day for those begun at E14. Because E15 is the characteristic time for gustatory papilla formation in the intact embryo, results demonstrate that the cultured tongues retain important temporal information related to papilla development. In addition, fungiform papillae formed in the tongue cultures in the stereotypic pattern of rows. The papillae were large structures with epithelial and mesenchymal cell integrity, and an intact epithelial basement membrane was indicated with laminin immunoreactivity. The cultures demonstrate that gustatory papilla morphogenesis can progress in the absence of an intact sensory innervation. To exclude a potential developmental role for autonomic ganglion cells that are located in the posterior rat tongue, cultures consisting of only the anterior half of E14 tongues were established. Fungiform papilla development progressed in half tongues in a manner directly comparable to whole tongue cultures. Therefore, robust, reproducible development of fungiform papillae in patterns is supported in rat tongue cultures from E13 or E14, without inclusion of intact sensory or major, posterior tongue autonomic ganglia. This is direct evidence that papillae will form and develop further in vitro without sensory ganglion support. The data also provide the first detailed account of in vitro development of the entire embryonic tongue.

Animals↗
PubMed1995

Distribution of vasoactive intestinal peptide-like immunoreactivity in the taste organs of teleost fish and frog.

Using immunohistochemistry, vasoactive intestinal peptide (VIP) was visualized in taste bud cells of the carp, Cyprinus carpio, and the European catfish, Silurus glanis, by means of light and electron microscopy. Intracellular membrane systems, presumably smooth endoplasmic reticulum, of light (sensory) cells, but not of dark (supporting) cells and basal cells, were densely labelled with antibody. In the frog (four species: Rana temporaria, R. ridibunda, R. arvalis, R. pipiens), taste bud cells did not label. However, the dense basal nerve fibre plexus, some subepithelial ganglionic cells, but no ascending intragemmal fibres, were immunoreactive. In fish, the results support evidence that VIP is involved in the modulation of taste transduction at the level of receptor cells. In the frog, an indirect, possibly vasodilatatory effect on taste perception may be considered.

Animals↗
PubMed2001

Characteristics of dorsal lingual papillae of the Bactrian camel (Camelus bactrianus).

The morphology of dorsal lingual papillae of the Bactrian camel (Camelus bactrianus) was studied by using light and scanning electron microscopy. Filiform and lenticular papillae were considered as mechanical papillae but fungiform and vallate papillae were considered as gustatory papillae. Filiform papillae were distributed mostly in the anterior two-thirds region of the tongue. Each filiform papilla consisted of one primary papilla and a few smaller secondary papillae. Lenticular papillae were distributed on the torus linguae. The larger papillae were arranged in two parallel lines medially whereas the smaller papillae were laterally located. Most of the fungiform papillae were found on the lateral margins of the anterior two-thirds of the tongue. These papillae were small and round. Intra-epithelial taste buds were located on the dorsal surface of each papilla. The vallate papillae were arranged in two rows on each rim of the torus linguae. Each round- and flat-shaped vallate papilla was surrounded by a prominent gustatory groove and an annular pad. A few taste buds were observed in the lateral epithelium of the papillae. The keratinization of the covering stratified squamous epithelium of the mechanical lingual papillae was relatively thicker than those of the gustatory papillae. The lingual papillae of the Bactrian camel exhibited some different characteristics from other domestic ruminants. These morphological characteristics of the tongue of the Bactrian camel might have evolved to assist the camel in prehension and manipulating of the inorganic stiff plants that grow in its environment and therefore might relate to the feed and feeding habits of the animal.

Animals↗
PubMed1986

Effects of gustatory deafferentation on ingestion of taste solutions as seen by licking behavior in rats.

The role of the taste nerves in licking behavior to various taste solutions was examined in rats. The study consisted of two experiments. In the electrophysiological experiment, the whole nerve recording of the anterior palatine nerve (PN) was performed to examine the response properties of this nerve to taste stimulation applied to the taste buds in the nasoincisal duct. When the moderate concentrations of solutions were used, the PN responded best to HCl, followed by sucrose and NaCl. Quinine hydrochloride elicited the smallest response. In the behavioral experiment, the number of licks/20 sec was measured for each of the test solutions such as 0.5 M sucrose, 0.01-1.0 M NaCl, 0.03 M HCl and 0.0005 M-0.01 M quinine in normal control and experimental rats. The experimental animals received bilateral deafferentation of the PN, chorda tympani (CT) and glossopharyngeal nerve (GN) alone or in various combinations. Rats without one of the 3 taste nerves still rejected the aversive HCl and quinine solutions. However, after lesions of both CT and GN, or all the 3 taste nerves, the rats showed a significant increase in the number of licks to these aversive solutions. These results suggest that taste aversion disappears after denervation of more than 80% of the total taste buds. The interlick interval, lick duration and the amount of intake per lick did not change significantly after sections of the taste nerves in any combinations.

Afferent Pathways↗
PubMed1994

Distribution patterns of the paraneuronal endocrine cells in the skin, gills and the airways of fishes as determined by immunohistochemical and histological methods.

The neuro-endocrine cells of fish skin and respiratory surfaces, and their bioactive secretion as far as is known, are reviewed, and compared with similar elements in tetrapods, particularly amphibians. In the skin of teleost fish, immunohistochemistry has shown that Merkel cells react for serotonin, neuron-specific enolase and enkephalins. The pharmacology is not established in dipnoans or lampreys. In some teleosts, neuromasts react for substance P and leu-enkephalins; substance P is also reported from some ampullary organs (electroreceptors). Taste buds of teleosts may react for enkephalin and substance P. Basal cells of taste buds react for serotonin and neuron-specific enolase. Some unicellular skin glands of teleosts express bioactive compounds, including serotonin and some peptides; this ectopic expression is paralleled in amphibian skin glands. The dipnoan Protopterus has innervated pulmonary neuro-endocrine cells in the pneumatic duct region with dense-cored vesicles. In Polypterus and Amia the lungs have serotonin-positive neuro-endocrine cells that are apparently not innervated. In fish gills, a closed type of neuro-endocrine cell reacts for serotonin, an open type for enkephalins and some calcium-binding proteins (calbindin, calmodulin and S-100 protein). The functions of neuro-endocrine cells in fishes await investigation, but it is assumed they are regulatory.

Animals↗
PubMed1990

Leucine-enkephalin-, neurokinin A- and cholecystokinin-like immunoreactivities in the guinea pig tongue.

The occurrence of these neuropeptides was examined by immunofluorescence. Leu-Enk-like immunoreactivity was seen in nerve fibres associated with the epithelium, blood vessels and lingual salivary glands as well as in ganglionic cells within the tongue. Neuropeptide A-like immunoreactivity was found in nerve fibres associated with the epithelium, taste buds, blood vessels and lingual salivary glands. Cholecystokinin-like immunoreactivity was found in some nerve fibres around blood vessels as well as in ganglionic cells. The coexistence of these neuropeptides and substance P was also demonstrated in some nerve fibres and ganglionic cells within the tongue. Like substance P, these neuropeptides could be involved in blood flow regulation, salivation and as trophic factors for taste buds.

Animals↗
PubMed1998

Responses to monosodium glutamate and guanosine 5'-monophosphate in rat fungiform taste cells.

Monosodium glutamate (MSG) elicits a unique taste sensation called umami. The umami sensation is potentiated by the presence of 5'-ribonucleotides such as guanosine 5'-monophosphate (5'-GMP). We have used giga-seal whole cell recording to examine glutamate transduction in individual cells of isolated rat fungiform taste buds. Approximately 56% of fungiform taste cells responded to bath application of glutamate. Three types of responses occurred: decrease in inward holding current; increase in inward holding current; and a biphasic response, with an increase followed by a decrease in holding current. Similar responses were observed in response to 5'-GMP. Further, responses to 5'-GMP may occur in cells that are glutamate-insensitive, suggesting that different receptors mediate the transduction of glutamate and 5'-GMP. Simultaneous bath application of glutamate and 5'-GMP resulted in a synergistic response in some taste cells.

Animals↗
PubMed2000

An immunohistological study of cytokeratin 20 in human and mammalian oral epithelium.

Cytokeratin (CK) 20 is a low molecular-weight intermediate filament reportedly expressed only by benign and malignant gastrointestinal epithelium, urothelium and Merkel cells. The main aims here were to map its expression in normal oral mucosa of humans and other mammals, and to determine whether it was expressed by abnormal human oral epithelium. Salivary and odontogenic epithelium were also analysed. An immunoperoxidase method was used on wax-embedded and cryostat sections. In addition, double-labelling experiments were undertaken to determine the association between CK 20 expression and that of CK 8/18 or S100 protein. Normal human oral mucosa from four sites, together with abdominal skin, was studied in autopsy samples from 32 individuals. CK 20-positive, basally situated, round or angular cells, consistent with Merkel cells, were recorded in 24/32 (75.0%) samples of mandibular gingiva, 25/32 (78.1%) samples of hard palate, 7/32 (21.9%) samples of buccal mucosa, 0/32 samples of lateral border of tongue, and 2/32 (6.3%) samples of abdominal skin. Double-labelling showed that all CK 20-positive Merkel cells also expressed CK 8/18 and S100. The only other cells to express CK 20 were human taste buds. There was no expression by dysplastic or invasive oral epithelium from biopsy samples. Colonic mucosa showed luminal-cell positivity in man, marmoset, ferret, rabbit and guinea-pig, but oral mucosa was universally negative in non-human species. It is concluded that in oral mucosa CK 20 is a specific marker of Merkel cells and taste buds, that Merkel cells are more frequently present in keratinized than non-keratinized oral mucosa, that CK 20-positive Merkel cells are also S100-positive, that there may be interspecies variations in CK 20 polypeptide composition and that, by contrast to urothelium, CK 20 has no value in the diagnosis of oral epithelial dysplasia.

Adult↗