Morphology of the buccopharyngeal portion of the gill in the fathead minnow Pimephales promelas (Rafinesque)

Buccopharyngeal epithelium covering gill arches and gill rakers of the fathead minnow was studied by light microscopic, scanning, and transmission electron microscopic techniques. Mature mucous cells in goblet pattern and nonmucus containing cells were in the apical one-third of the tissue. The latter cells contributed to a surface microridge system which overlapped apices of goblet cells. The bottom of the epithelium was comprised of a continuous row of darkly stained basal epithelial cells. In this region, two to three epithelial cells of similar staining characteristics were piled up forming apical columns which partially encircled nests of lightly stained cells. A basal lamina and thick basement lamella of about 20 plies of orthogonally arranged collagen supported the epithelium. Numerous taste buds were seen in gill arches and rakers. Taste bud cellular components included marginal cells, light receptor cells, dark receptor cells, and basal cells. These were identical in all taste buds. Taste bud surface morphology differed between gill arch and raker. Pores of the former were depressed, while those of the latter were raised. Thick microvilli of taste pores were apical extensions of light cells, while smaller, more numerous microvilli were projections from dark cells.     

Morphology of taste buds on the gill arches of the mullet Mugil cephalus,and the killifish Fundulus heteroclitus

The morphology of taste buds on the gill arches of two euryhaline teleosts, the mullet Mugil cephalus, and the killifish Fundulus heteroclitus, were investigated using light microscopic and scanning and transmission electron microscopic techniques. On the mullet gill arches, taste buds were limited to the pharyngeal surfaces of the smooth-surfaced gill rakers. On the killifish gill arches, taste buds were located on the pharyngeal surfaces of all gill rakers and on the gill arch itself at the bases of the gill rakers. Despite dramatic differences in gill-raker structure between these two species, the taste buds themselves were similar ultrastructurally and closely resembled those described in other fishes. Cells within the taste buds included spindle-shaped dark and light cells and basal cells. Ultrastructural features of both the light and dark cells could support either receptor or transport functions. Tufts of microvilli, including one thick microvillus per light cell and numerous thin microvilli per dark cell, protruded at the apex of each taste bud between the ridged surface epithelial cells. Light cells contained numerous tubular membrane elements some of which appeared to open onto the apical surface of the taste bud. Dark cells contained numerous microtubules and apical, electron-lucent vesicles possibly involved in transport.     

Confocal imaging of Merkel-like basal cells in the taste buds of zebrafish

The oropharyngeal cavity in fish supports a range of sensory modalities, including detection of chemical and mechanical stimuli. Taste buds are found throughout this tissue and may participate in both processes. We used confocal microscopy and immunohistochemistry to characterize the morphology of Merkel-like cells and their association with other cell types and nerve fibers of the taste bud in the vertebrate model, the zebrafish. In addition, we document procedures for the observation of these structures in whole-tissue preparations from larvae and adults using zebrafish-specific and monoclonal antibodies. A single microvillus Merkel-like cell was found in each taste bud regardless of age or location. Merkel-like cells were neurosecretory, as indicated by labelling with the styryl dye, FM1-43, and the synaptic vesicle marker, SV2. Merkel-like cells were associated with SV2- and calretinin-positive taste receptor cells, received innervation from discoid aggregations of nerve fibers, and retained serotonin-filled synaptic vesicles oriented within the cytoplasm toward adjacent innervation. Moreover, a ring-like formation of nerve endings was identified with the neuronal marker, zn-12 that circumscribed the taste receptor area, surrounding calretinin-immunoreactive taste cell microvilli, and appeared to associate with the nerve plexus adjacent to Merkel-like cells. We suggest that these nerve fibers are somatosensory, perhaps associated with mechanoreception or the common chemical sense.     

Espin cytoskeletal proteins in the sensory cells of rodent taste buds

Espins are multifunctional actin-bundling proteins that are highly enriched in the microvilli of certain chemosensory and mechanosensory cells, where they are believed to regulate the integrity and/or dimensions of the parallel-actin-bundle cytoskeletal scaffold. We have determined that, in rats and mice, affinity purified espin antibody intensely labels the lingual and palatal taste buds of the oral cavity and taste buds in the pharyngo-laryngeal region. Intense immunolabeling was observed in the apical, microvillar region of taste buds, while the level of cytoplasmic labeling in taste bud cells was considerably lower. Taste buds contain tightly packed collections of sensory cells (light, or type II plus type III) and supporting cells (dark, or type I), which can be distinguished by microscopic features and cell type-specific markers. On the basis of results obtained using an antigen-retrieval method in conjunction with double immunofluorescence for espin and sensory taste cell-specific markers, we propose that espins are expressed predominantly in the sensory cells of taste buds. In confocal images of rat circumvallate taste buds, we counted 21.5 ± 0.3 espin-positive cells/taste bud, in agreement with a previous report showing 20.7 ± 1.3 light cells/taste bud when counted at the ultrastructural level. The espin antibody labeled spindle-shaped cells with round nuclei and showed 100% colocalization with cell-specific markers recognizing all type II [inositol 1,4,5-trisphosphate receptor type III (IP₃R₃)_, α-gustducin, protein-specific gene product 9.5 (PGP9.5)] and a subpopulation of type III (IP₃R₃, PGP9.5) taste cells. On average, 72%, 50%, and 32% of the espin-positive taste cells were labeled with antibodies to IP₃R₃, α-gustducin, and PGP9.5, respectively. Upon sectional analysis, the taste buds of rat circumvallate papillae commonly revealed a multi-tiered, espin-positive apical cytoskeletal apparatus. One espin-positive zone, a collection of ∼3 μm-long microvilli occupying the taste pore, was separated by an espin-depleted zone from a second espin-positive zone situated lower within the taste pit. This latter zone included espin-positive rod-like structures that occasionally extended basally to a depth of 10–12 μm into the cytoplasm of taste cells. We propose that the espin-positive zone in the taste pit coincides with actin bundles in association with the microvilli of type II taste cells, whereas the espin-positive microvilli in the taste pore are the single microvilli of type III taste cells.     

Distribution of taste buds on the lips and inside the mouth in the minnow, Pseudorasbora parva

The distribution and abundance of taste buds were quantitatively examined by observing silver impregnated serial sections. The taste buds were widely dispersed on the skin, the lips, the mucosa in the oro-pharyngeal cavity, the esophagus, and the branchial apparatus. The great majority of them was found on the lips and inside the mouth. The external buds were concentrated especially on the outer lips and the adjacent skin, while their number diminished in a caudal direction. Very few were found on the scaled skin. The total number of external buds in a specimen of 6 cm in length was 1,486. The number of taste buds inside the mouth was 6,600. On the inner lips and the palatal organ densities were found to reach over 140 per mm². High concentrations of taste buds were also found on the gill arches and rakers. These taste buds varied to some extent in size and shape, depending on the thickness of the epithelial layer. It is suggested that the minnow may use the lips, gills and palatal organ as its main taste organs.     

Taste receptor signalling - from tongues to lungs

Taste buds are the transducing endorgans of gustation. Each taste bud comprises 50-100 elongated cells, which extend from the basal lamina to the surface of the tongue, where their apical microvilli encounter taste stimuli in the oral cavity. Salts and acids utilize apically located ion channels for transduction, while bitter, sweet and umami (glutamate) stimuli utilize G-protein-coupled receptors (GPCRs) and second-messenger signalling mechanisms. This review will focus on GPCR signalling mechanisms. Two classes of taste GPCRs have been identified, the T1Rs for sweet and umami (glutamate) stimuli and the T2Rs for bitter stimuli. These low affinity GPCRs all couple to the same downstream signalling effectors that include Gβγ activation of phospholipase Cβ2, 1,4,5-inositol trisphosphate mediated release of Ca(2+) from intracellular stores and Ca(2+) -dependent activation of the monovalent selective cation channel, TrpM5. These events lead to membrane depolarization, action potentials and release of ATP as a transmitter to activate gustatory afferents. The Gα subunit, α-gustducin, activates a phosphodiesterase to decrease intracellular cAMP levels, although the precise targets of cAMP have not been identified. With the molecular identification of the taste GPCRs, it has become clear that taste signalling is not limited to taste buds, but occurs in many cell types of the airways. These include solitary chemosensory cells, ciliated epithelial cells and smooth muscle cells. Bitter receptors are most abundantly expressed in the airways, where they respond to irritating chemicals and promote protective airway reflexes, utilizing the same downstream signalling effectors as taste cells.     

Solitary chemosensory cells in mammals?

In fish, solitary chemosensory cells (SCCs) occur in the oropharynx, gills and skin and have often been found in association with taste buds. Among amphibia, a diffuse chemosensory system has been described on the ventral skin of toads, and a structural resemblance of SCCs to taste bud cells has been reported in frogs. Putative solitary chemoreceptors have been described in mammals too, at specific sites in the digestive or respiratory apparatus. In newborn rodents, a specific set of SCCs (composed of elements positive for alpha-gustducin, a marker of chemosensory cells) is associated with the gustatory epithelium. In conclusion, the available data suggest that a SCC system is not restricted to fish but is present in amphibia and mammals as well. At our present level of knowledge, establishing a precise homology between different species is difficult. However, the data from mammals and amphibia fully confirm previous findings in fish, and the use of chemical markers to study the diffuse chemosensory systems of vertebrates seems promising.     

江豚味蕾的分布及超微结构

目的：了解江豚味觉器官味蕾在口腔中的分布方式和味蕾的超微结构。方法：用石蜡切片观察江豚味蕾的分布，用透射电镜观察味蕾的超微结构。结果：江豚味蕾仅分布于舌小窝侧壁上皮层及舌小窝内疣状突起上皮层中；江豚味蕾主要包括明细胞和暗细胞，味孔表面由暗细胞细胞体的上行突起构成，明细胞细胞体上行突起不到达味孔表面。结论：脊椎动物味蕾在体内外的分布特点与动物的分类地位相关，而脊椎动物舌表面的形态学特点与动物的栖息环境相关。     

Fine structure of taste buds located on the lamb epiglottis

Background: Taste buds located on the aryepiglottal folds and laryngeal surface of the epiglottis are the principal receptors responsible for the initiation of the laryngeal chemoreflex. In contrast to the wealth of information available concerning the ultrastructure of oral taste buds, little comparable data exists for taste buds located at the entrance to the larynx. Therefore, the present study was designed to investigate the fine structure of taste buds located on the lamb epiglottis. Materials: Stained thick and semi-serial thin sections from taste buds located on the lamb epiglottis were examined with light and electron microscopy. Results: Based on morphological criteria, three types of cells could be identified in the taste bud: Type I, Type II, and basal cells. Both Type I and Type II cells extended into the apical taste pore, but there were differences between these two cell types with regard to nuclear profiles, electron density, and the relative density of ribosomes, apical mitochondria, and rough and smooth endoplasmic reticulum. Basal cells did not extend a process into the taste pore. Nerve processes were observed throughout the taste bud. Synapses were observed between both Type I and Type II cells and nerve fibers. These synapses exhibited membrane thickenings and accumulations of clear and dense-cored vesicles of varying proportions in the taste cell cytoplasm adjacent to membrane specializations. Conclusions: The taste buds located on the lamb epiglottis share several structural similarities to taste buds located in the oral cavity and other regions of the pharynx and larynx of many mammalian species. The presence of synapses on both Type I and Type II cells of the lamb epiglottal taste bud suggests that both cell types are involved in laryngeal chemorecption. © 1994 Wiley-Liss, Inc.     

云豹味蕾的分布与结构

目的：了解云豹味觉器官味蕾在口腔中的分布和结构。方法：用光镜观察云豹舌的形态结构和味蕾的分布，并用透射电镜观察味蕾的结构。结果：云豹味蕾分布于舌尖及轮廓乳突的上皮层中，主要由明细胞和暗细胞组成。结论：云豹味蕾的分布及舌的形态学特点与它的捕食和吞咽习性相适应。     

The taste cell-related diffuse chemosensory system

Elements expressing the molecular mechanisms of gustatory transduction have been described in several organs in the digestive and respiratory apparatuses. These taste cell-related elements are isolated cells, which are not grouped in buds, and they have been interpreted as chemoreceptors. Their presence in epithelia of endodermal origin suggests the existence of a diffuse chemosensory system (DCS) sharing common signaling mechanisms with the "classic" taste organs. The elements of this taste cell-related DCS display a site-related morphologic polymorphism, and in the past they have been indicated with various names (e.g., brush, tuft, caveolated, fibrillo-vesicular or solitary chemosensory cells). It may be that the taste cell-related DCS is like an iceberg: the taste buds are probably only the most visible portion, with most of the iceberg more caudally located in the form of solitary chemosensory cells or chemosensory clusters. Comparative anatomical studies in lower vertebrates suggest that this 'submerged' portion may represent the most phylogenetically ancient component of the system, which is probably involved in defensive or digestive mechanisms. In the taste buds, the presence of several cell subtypes and of a wide range of molecular mechanisms permits precise food analysis. The larger, 'submerged' portion of the iceberg is composed of a polymorphic population of isolated elements or cell clusters in which the molecular cascade of cell signaling needs to be explored in detail. The little data we have strongly suggests a close relationship with taste cells. Morphological and biochemical considerations suggest that the DCS is a potential new drug target. Modulation of the respiratory and digestive apparatuses through substances, which act on the molecular receptors of this chemoreceptive system, could be a new frontier in drug discovery.     

Histochemical analysis of glycoproteins in the gill epithelium of an Indian major carp, Cirrhinus mrigala

Glycoproteins were analyzed by a range of histochemical methods in the epithelium of gills of Cirrhinus mrigala, a valuable food fish of great economic importance cultured extensively in India. The gills consist of gill arches, gill rakers, gill filaments and secondary lamellae. Major components of the epithelium of gill arches and gill rakers are epithelial cells, mucous goblet cells, rodlet cells, lymphocytes, eosinophilic granular cells and taste buds. In contrast, in the gill filament epithelium, rodlet cells and taste buds, and in secondary lamellae epithelium, rodlet cells, lymphocytes, eosinophilic granular cells and taste buds are not discernible. The epithelial cells, the mucous goblet cells and the eosinophilic granular cells elaborate glycoproteins with oxidizable vicinal diols and glycoproteins with sialic acid residues without O-acyl substitution. In addition, glycoproteins with O-sulphate esters are secreted by the mucous goblet cells. The rodlet cells elaborate glycoproteins with oxidizable vicinal diols. Different types of glycoproteins elaborated on the epithelial surface of gills are discussed in relation to physiological significance of glycoprotein classes with special reference to their roles in lubrication, protection and inhibition of invasion and proliferation of pathogenic micro-organisms.     

Mouse taste cells with glialike membrane properties

Taste buds are sensory structures made up by tightly packed, specialized epithelial cells called taste cells. Taste cells are functionally heterogeneous, and a large proportion of them fire action potentials during chemotransduction. In view of the narrow intercellular spaces within the taste bud, it is expected that the ionic composition of the extracellular fluid surrounding taste cells may be altered significantly by activity. This consideration has led to postulate the existence of glialike cells that could control the microenvironment in taste buds. However, the functional identification of such cells has been so far elusive. By using the patch-clamp technique in voltage-clamp conditions, I identified a new type of cells in the taste buds of the mouse vallate papilla. These cells represented about 30% of cells patched in taste buds and were characterized by a large leakage current. Accordingly, I named them "Leaky" cells. The leakage current was carried by K(+), and was blocked by Ba(2+) but not by tetraethylammonium (TEA). Other taste cells, such as those possessing voltage-gated Na(+) currents and thought to be chemosensory in function, did not express any sizeable leakage current. Consistent with the presence of a leakage conductance, Leaky cells had a low input resistance (approximately 0.25 G Omega). In addition, their zero-current ("resting") potential was close to the equilibrium potential for potassium ions. The electrophysiological analysis of the membrane currents remaining after pharmacological block by Ba(2+) revealed that Leaky cells also possessed a Cl(-) conductance. However, in resting conditions the membrane of these cells was about 60 times more permeable to K(+) than to Cl(-). The resting potassium conductance in Leaky cells could be involved in dissipating rapidly the increase in extracellular K(+) during action potential discharge in chemosensory cells. Thus Leaky cells might represent glialike elements in taste buds. These findings support a model in which specific cells control the chemical composition of intercellular fluid in taste buds.     

Cell lineage and differentiation in taste buds

Mammalian taste buds are maintained through continuous cell renewal so that taste bud cells are constantly generated from progenitor cells throughout life. Taste bud cells are composed of basal cells and elongated cells. Elongated cells are derived from basal cells and contain taste receptor cells (TRC). Morphologically, elongated cells consist of three distinct types of cells: Types I, II and III. In contrast to the remarkable progress in understanding of the molecular basis for taste reception, the mechanisms of taste bud maintenance have remained a major area of inquiry. In this article, we review the expression of regulatory genes in taste buds and their involvement in taste bud cell differentiation. Three major topics include: 1) the Sonic hedgehog (Shh)-expressing cell in the basal cell in taste buds as a transient precursor of elongated cells and as a signal center for the proliferation of progenitor cells; 2) the Mash1-expressing cell as an immature cell state of both Type II and Type III cells and as a mature cell state of Type III cell; and 3) the nerve dependency of gene expression in taste buds. Problems in the application of NCAM for the type III cell marker are also discussed.     

Intermediate filaments in mouse taste bud cells

The intermediate filaments in mouse taste bud cells were studied by immunocytochemistry using antikeratin antibodies, and by conventional electron microscopy. Taste bud cells (types I, II, and III) possessed less densely aggregated bundles of intermediate filaments than the surrounding epithelial cells. Type III cells, however, contained more densely aggregated bundles than type I or II cells. Basal cells in the taste buds showed aggregations of filaments as dense as those seen in the epithelial cells, although their bundles were more slender than those of the epithelial cells. The antibodies to keratins from the bovine muzzle and human stratum corneum stained all types of the taste bud cells as well as the surrounding epithelial cells. PKK2 antibody reacted with the surrounding epithelial cells, but did not react with the taste bud cells. These results show that keratins are present in both taste bud and surrounding epithelial cells, although the keratin subtype differs between those cells. This finding has led us to the supposition that all cell types comprising the taste buds--including type III (receptor) cells--originate from the epithelial cells surrounding the taste buds. It is also suggested that both keratin subtypes and aggregation patterns of intermediate filament bundles change during differentiation from surrounding epithelial cells to taste bud cells, and from basal cells in the taste buds to types I, II, or III cells.     

Taste bud development in chickens (Gallus gallus domesticus)

Oral epithelium in the anterior mandibular glands region was examined in embryonic, hatchling, and mature chickens to establish the timing of morphologic events during taste bud ontogeny. Hematoxylin-and-eosin-stained sections (10 microns) from 27 Anak (broiler breed) chickens were examined serially, and buds were quantified at 16-20 days of incubation (E) and, posthatch days 1 and 50-60. Taste buds were first recognized at the beginning of E17 as small clusters of cells in the basal epithelium. Only spherical-shaped buds were observed on E17 and E18, and these spherical clusters never penetrated to the surface of the stratified epithelial layer. E19 marked a transitional stage when mature bud features began to emerge: the buds assumed a more elongate shape, several kinds of cells comprising the bud were distinguishable and the first taste pores were observed. During the ensuing embryonic days, buds continued to elongate commensurate with the deepening oral epithelium and by hatching virtually all buds opened to the oral cavity. No marked morphological changes in taste bud structure were observed on the day of hatching and at 50-60 days posthatching. Taste bud numbers increased dramatically during E17 and E18, peaked on E19, and remained relatively constant thereafter. It is concluded that the morphological sequence of taste bud development in chickens is similar to that in mammals. The timing of bud ontogeny, though initiated only during the third trimester in ovo, essentially is completed by hatching, thus providing the precocial hatchling with the sensory apparatus essential for gustatory experience.     

Light and electron microscopical demonstration of methylene blue accumulation sites in taste buds of fish and mouse after supravital dye injection

Electron microscopical data regarding methylene blue staining of taste buds in the epithelia of the goldfish lip and the cirumvallate papilla of the mouse tongue after supravital dye application are presented for the first time. The ultrastructural details were compared with the corresponding light microscopical findings. The dye was applied in different concentrations by injection or in crystalline form directly to the surface of the tissues. Both methylene blue and tissue were simultaneously fixed by immersion in a paraformaldehyde-glutaraldehyde solution with the addition of phosphomolybdic acid. The ensuing dye precipitate was further stabilized by ammonium heptamolybdate. On the light microscopical level, the taste bud's receptive structures, i.e. the receptor area (fish) and the taste pit (mouse), exhibited the highest affinity for the dye. Additionally, the mucous material within the trenches around the circumvallate papillae in mice was intensely stained. On the electron microscopical level, the cationic phenothiazine dye bound to the receptor villi or to the mucus coating the receptive structures. In the case of higher dye concentrations, a staining of single taste bud cells took place starting apically and proceeding down to the base. Dye accumulations within the intercellular clefts between the epithelial cells or within other structures were observed only if the dye concentration was further increased. Since similar results were also obtained with the cationic phenazo dye Janus green, dye accumulation in the mucus covering the receptor villi may be representative of the general binding of organic cations, which are known to induce bitter taste sensations.     

Taste buds and nerve fibers in the rat larynx: an ultrastructural and immunohistochemical study

We investigated the rat laryngeal taste buds and their innervation by electron microscopy and immunohistochemical methods. Taste buds were densely arranged in the surface facing the laryngeal cavity of the epiglottis, the aryepiglottic fold, and the cuneiform process of the arytenoid cartilages. The cells of the buds were classified into types I, II, III, and basal cells, the ultrastucture of which was almost the same as that previously reported in lingual taste buds. The type III cells that had synaptic contacts with nerve fibers were considered to be sensory cells. Immunohistochemical analysis revealed thick calbindin D28k-immunoreactive fibers and thin varicose fibers immunoreactive for calcitonin gene-related peptide or substance P in and around the taste bud. Serotonin-immunoreactive cells were also observed here. The results revealed the innervation pattern of laryngeal taste buds to be the same as that in lingual taste buds. Carbonic anhydrase (CA) is known to catalyze the hydration of CO2 and dehydration of H2CO3, and seems to be essential in CO2 reception. Immunoreactivity for CAI was detected in slender cells and that for CAIII was observed in barrel-like cells in the laryngeal taste buds. The pH-sensitive inward rectifier K+ (Kir) channel in the cell membrane may be involved in CO2 reception as well. CAII-reactive cells were also reactive to Kir4.1, PGP 9.5 and serotonin. Our results indicated that CAII and Kir4.1 are located in type III cells of the laryngeal taste buds, and supported the idea that the buds may be involved in the recognition of CO2.