Morphological adaptation of the tongue of okapi (Okapia johnstoni Artiodactyla, Giraffidae)-Anatomy, histology, and ultrastructure.

The aim of this study was to describe the morphology of the tongue of the okapi, and to compare the results with other ruminants including browsers, intermediates and grazers. The material was collected post-mortem from two animals from a Zoological Garden. The structure of the okapi tongue, focusing of the shape of the tongue, lingual surface, its papillae and lingual glands, was examined using gross morphology, light and polarized microscopy, and by scanning electron microscopy. The okapi tongue was characterized by dark pigmentation on the lingual dorsum (except lingual torus) and on the whole ventral surface. Two types of filiform papillae were observed, with additional, even 6-8 projections at their base. The round fungiform papillae were present at a higher density, up to 16/cm2, on the ventro-lateral area of the lingual apex. Round and elongate vallate papillae were arranged in two parallel lines between the body and root of the tongue. Numerous taste buds were detected within the epithelium of their vallum, while fungiform papillae had sparse taste buds. A lack of foliate papillae was noted. Very small conical papillae, some lenticular in shape, were present on the lingual torus. Thick collagen type I fibers were dominant over collagen type III fibers in the connective tissue of the lingual papillae. The mucous acini units were dominant among lingual glands, indicating that the secretion of okapi lingual glands was mostly mucous. In many aspects, the tongue of okapi resembles the tongue of other ruminants. The specific lingual shape and lingual surface, together with the lingual glands, support the processing of plant food, such as young and soft leaves. Although okapi tongue is characterized by smaller conical papillae compared to other ruminants, its high number of vallate papillae is similar that found in other browsers, intermediate and grazers. Thus the number of gustatory papillae rather indicates that this feature is not related to the type of feeding.

Ultrastructural localization of calcium homeostasis modulator 1 in mouse taste buds.

Taste receptor cells are morphologically classified as types II and III. Type II cells form a unique type of synapses referred to as channel synapses where calcium homeostasis modulator 1 (CALHM1) together with CALHM3 forms voltage-gated channels that release the neurotransmitter, adenosine triphosphate (ATP). To validate the proposed structural model of channel synapses, the ultrastructural localization of CALHM1 in type II cells of both fungiform and circumvallate taste buds was examined. A monoclonal antibody against CALHM1 was developed and its localization was evaluated via immunofluorescence and immunoelectron microscopy using the immunogold-silver labeling technique. CALHM1 was detected as puncta using immunofluorescence and along the presynaptic membrane of channel synapses facing atypical mitochondria, which provide ATP, by immunoelectron microscopy. In addition, it was detected along the plasma membrane lined by subsurface cisternae at sites apposed to afferent nerve fibers. Our results support the validity of a previously proposed structural model for channel synapses and provide insights into the function of subsurface cisternae whose function in taste receptor cells is unknown. We also examined the localization of CALHM1 in hybrid synapses of type III cells, which are conventional chemical synapses accompanied by mitochondria similar to atypical mitochondria of channel synapses. CALHM1 was not detected in the six hybrid synapses examined using immunoelectron microscopy. We further performed double immunolabeling for CALHM1 and Bassoon, which is detected as puncta corresponding to conventional vesicular synapses in type III cells. Our observations suggest that at least some, and probably most, hybrid synapses are not accompanied by CALHM1.

Lingual adaptations of the Tarentola annularis with new insights into its papillary system adaptations: Ultrastructure, histochemistry, and immunohistochemical observations.

Our research aims to conduct a comprehensive ultrastructural, histochemical, and immunohistochemical examination of Tarentola annularis' tongue, utilizing various techniques such as light, scanning electron microscopy, and morphometric analysis. The complex papillary system consisted of four conical subtypes and one filiform type. The apex carried three conical subtypes (elongated, quadrilateral, and round); the midtongue carried two papillary types (quadrilateral conical and rectangular pointed filiform); and the hindtongue carried two conical subtypes (quadrilateral and elongated serrated). The dorsal papillary surface carried little taste pores on the foretongue and taste buds on the midtongue. The foretongue had a slightly stratum corneum that spread to coat the papillae, while the mid- and hindtongue did not. The glands are absent from the foretongue but are found in the interpapillary spaces of the mid- and hindtongue. Histochemical analysis reveals the presence of collagen fibers in the muscle bundles and the papillary core. The midtongue glands exhibited a strong reaction to AB and PAS, while the hindtongue showed moderate AB positivity and strong positive PAS. The cytokeratin expression in the foretongue papilla was positive, whereas the papillae in other regions were negative. The Tarentola annularis exhibits distinctive lingual structural characteristics due to its varied feeding habits influenced by available food particles.

Comparative histological and ultrastructural features of the tongue of the mallard domestic duck, Anas platyrhynchos f. domestica, Anatidae (Linnaeus, 1758) in different two age stages (post-hatching [P2] and adult female) captured from Egypt.

The domestic duck is classified as a specialist filter-feeder bird living in the water. These birds also use grazing and pecking as terrestrial feeding methods. The tongues of domestic ducks, similar to those of other Anseriformes, exhibit numerous types and shapes of mechanical papillae that serve a number of purposes when collecting food. The current study attempts to describe the morphological characteristics of the tongue as well as the mechanical papillae's development. In addition, the study aims to determine whether the papillae observed post-hatching (P2) exhibit similar morphology to those found in adult female avian species, as well as to investigate the readiness of the tongue to fulfill its feeding function following hatching. The comprehensive examination of lingual mucosa is examined about the structural modifications necessary for this variety of feeding activities. In this study, the tongues of nine young (P2) and adult female were used. The tongue had three distinct parts: the apex, which had a lingual nail on its ventral surface; the body, which exhibits numerous small and large conical papillae on its lateral sides and a lingual prominence in the caudal region; and the root, which is covered with numerous conical papillae of varying sizes. Conical, filiform, and hair-like mechanical papillae, the three types of food filtration apparatus, are present in both stages. The intraoral transfer involves several structures, including the median groove, lingual combs, and the rostral border of the lingual prominence. The rostral border of the lingual prominence is characterized by distinct rows of conical papillae. The histological analysis demonstrated the presence of both keratinized and nonkeratinized epithelium on different tongue regions. The lingual salivary glands in the rostral and caudal lingual salivary glands exhibit a pronounced periodic acid-Schiff-positive reaction. Additionally, the yellow adipose tissue and sensory receptors, namely the Grandry and Herbst corpuscles, which collectively form the bill-tongue organ that monitors the movement of food. These results conclude the presence of microstructural species-specific alterations in specific tongue areas of domestic ducks' lingual mucosa. These modifications are formed by the filtering mechanism and terrestrial feeding mechanisms, such as grazing or pecking. Following hatching, the tongue of the domestic duck undergoes significant development, primarily in preparation for grazing activities. The anatomical and histological structure of the young (P2) tongue exhibited similarities to that of the adult female domestic duck while also displaying certain variations that could potentially be attributed to the bird's habitat and mode of feeding. RESEARCH HIGHLIGHTS: The results of this study concluded that the domestic duck exhibit a complex tongue structure characterized by the arrangement and morphology of its mechanical papillae, the presence of the lingual prominence with distinctive shape and the lingual comb. These features are believed to be adaptations that enable the duck to actively and efficiently filter food particles from water, serving as its primary feeding mechanism. Additionally, the tongue of domestic ducks is specifically adapted to facilitate various terrestrial activities, such as grazing and pecking. This adaptation is achieved through the presence of conical papillae and a lingual nail. These investigations facilitate our comprehension of both the anatomical and histological characteristics of the domestic duck tongue, as well as enhance our understanding of bird adaptations to various feeding mechanisms.

Tongue microarchitecture and functional characterization of the lingual papillae in the desert hedgehog (Paraechinus aethiopicus).

The present work attempted to provide a comprehensive description of the morphoanatomical, histological, and ultrastructural characteristics of the tongue in the desert hedgehog (Paraechinus aethiopicus), and to correlate lingual modifications to the feeding lifestyle. Five adult male hedgehogs were utilized in our investigation. The macroscopic observations revealed elongated, with a moderately pointed apex, tongue and the tongue dorsum lacks both lingual prominence and median sulcus. The main subdivisions of the tongue are radix linguae (root), corpus linguae (body), and apex linguae (apex). The tongue dorsum carries two types of mechanical (conical and filiform) and gustatory (fungiform and circumvallate) papillae. The lingual apex is characterized by the existence of a unique encapsulated muscular structure. Additionally, the lingual glands were interposed between the muscular strands and no lingual glands were detected on the lingual apex. The dorsal surface of the lingual apex exhibited the highest level of keratinization as revealed by histochemical staining while the root showed moderate staining. The topography of the tongue was investigated by scanning electron microscopy (SEM). The obtained results are important to provide basic knowledge that can contribute to better understanding of the nourishment, feeding habits and behavior in this species. Furthermore, the addition of the newly investigated species may help us to determine the evolutionary relationships among species.

Histological and ultrastructural characterization of the dorso-ventral skin of the juvenile and the adult starry puffer fish (Arothron stellatus, Anonymous 1798).

BACKGROUND: The starry puffer fish (Arothron stellatus, Anonymous, 1798) is a poisonous tetradontidae fish inhabiting the Red sea. The skin constitutes an important defense against any external effects. The study aims to characterize the dorso-ventral skin of the juvenile and the adult starry puffer fish using light and scanning electron microscopies. Twenty specimens of juvenile and adult fresh fishes were used. RESULTS: The scanning electron microarchitecture of the skin of the juvenile and adult fish showed delicate irregular-shaped protrusions, and well-defined bricks-like elevations on the dorsal side and interrupted folds as well as irregular-shaped protrusions on the ventral side. In adult fish, the patterned microridges of the superficial and deep epithelial cells (keratinocytes) were larger and well-defined in the dorsal skin than in the ventral side, the contrary was seen in the juvenile fish. The microridges were arranged in a fingerprint or honeycomb patterns. The openings of the mucous cells were more numerous in the dorsal skin in both age stages but more noticeable in adult. Furthermore, the sensory cells were more dominant in the juveniles than the adults. The odontic spines were only seen in adult. Histologically, few taste buds were observed in the epidermis of the dorsal skin surface of the adult fish. Both mucous and club cells were embedded in the epidermis of the juvenile and adult fish with different shapes and sizes. Melanophores were observed at the dorsal skin of both juvenile and adult fishes while fewer numbers were noticed at the ventral surfaces. Several dermal bony plates with different shapes and sizes were demonstrated in the skin of both adult and juvenile fishes. CONCLUSION: The structural variations of skin of the juvenile and adult fishes may reflect the various environmental difficulties that they confront.

A morphological investigation of the tongue of roe deer (Capreolus capreolus Linnaeus, 1758).

The morphological structure of the tongue and papillae that occur on it vary according to an animal's lifestyle, nutrition, and adaptation to various environmental conditions. This study aimed to reveal in detail the morphological, histological, and electron microscopic structure of the tongue of roe deer (Capreolus capreolus Linnaeus, 1758). In this study, nine roe tongues were used. The tongue consists of three parts: the apex, body, and root. When the dorsal surface of the tongue was examined in detail, five different papillae were observed: filiform, lenticular, conical, fungiform, and vallate. Filiform papillae differed in having secondary papillae according to their localization. The opening holes of taste buds were observed on the surface of the round and flat fungiform papillae. The free ends of the filiform papillae were more pointed and thinner than those of the other papillae, while the width of the lenticular papillae was thicker, the surface was flat, and the free ends were blunt. Triangular-shaped conical papillae were observed differently regarding the presence or absence of secondary papillae. The vallate papillae were caudolateral to the lingual torus. On the surface of the vallate papillae, circumferenced by a deep groove, were the opening holes of the taste buds and microridges. From this analysis, it appears to be characteristic of roe deer that mechanical function, filiform, and conical papillae contain secondary papillae; lenticular papillae, absent in many deer species, are found; and a prominent papillary groove surrounds all mechanical and gustatory papillae. RESEARCH HIGHLIGHTS: The lingual papillae of roe deer (Capreolus capreolus Linnaeus, 1758) were examined with this study in detail for the first time. Similarities and differences with ruminant species were determined.

Scanning electron microscopic characterizations of the tongue of the Nubian goat (Capra aegagrus hircus): A specialized focus on its papillary system adaptation to Egyptian environmental conditions.

The current investigation was focused on describing the gross and scanning electron features of the Nubian goat (Capra aegagrus hircus) tongue, with new insights into its papillary adaptation to the Egyptian environment. The elongated tongue had a rostral free and the caudal fixed. The ventral apical surface is classified into the smaller rostral papillary region on the tip and the larger non-papillary region by the U-line of filiform papillae. Functionally, there are two papillary types: mechanical (filiform, conical and lentiform in addition to the longitudinal row of large conical papilla on the lateral of the body) and gustatory (fungiform and circumvallate). Filiform papillae were densely distributed on the dorsal surface of the apex and body, and on the lateral apical border and lateral surface of the body and root, in addition to the ventral surface of the tip. This filiform papillary system gives a raspy appearance to the dorsal surface. The conical and lentiform papillae were limited to the torus linguae. Circumvallate papillae are surrounded by an annular groove and slightly vallum. The lingual root was devoid of any papillae. Lingual papillary subtypes are filiform papillae (elongated and triangular), conical papillae (elongated and oval) and fungiform papillae (round and ovoid). The investigated Nubian goat may have developed a specialized papillary system due to regional differences in the distribution, structure and subtypes of the system, allowing it to adapt to the dried grasses and leaves of trees and bushes that are available in Upper Egypt's dry, hot climate.

Investigations on the light and scanning electron microscopic structure of the lingual papillae in the angora goat (Capra hircus): II. Mechanical papillae.

This study was conducted to examine the light microscopy (LM) and scanning electron microscopic (SEM) structures of mechanical papillae on the tongue in the Angora goat (Capra hircus). As study materials, the tongues of four adult female Angora goats were used. The samples were collected from the dorsal surfaces of the apex, body, root and torus of the tongue and the ventral surfaces of the lingual apex for light and scanning electron microscopic examinations. Three types of mechanical papillae were seen in Angora goat tongue: papilla filiformes, papilla lentiformes and papilla conicae.The filiform papillae were detected in the dorsal surface of the tongue from lingual apex to lingual torus, and in the ventro-lateral of the lingual apex and on both sides of the lingual torus. The morphological differences were observed in filiform papillae according to their location in the tongue. The lenticular papillae settled on the centre of the lingual torus. Two types of these papillae, irregular-round and pyramid-shaped were identified. The conical papillae were scattered all over the lingual torus, except for the central part and were also seen on the root of the tongue. In the light microscopic examination, it was found that mechanical papillae had a stratified squamous epithelium and a varying degree of keratin layer on epithelial surfaces. In this study, by examining the light and scanning electron microscopic structure of the mechanical papillae in the Angora goat tongue, their similarities and differences with other domestic and wild ruminant species were determined.

Morphological features of the Egyptian Ossimi sheep tongue: New scanning electron microscopic insights into its papillary system adaptations to Egyptian ecological conditions.

The current study aims to illustrate the gross and scanning electron microscopic characterizations and ultrastructural adaptation of the lingual papillary system of Egyptian Ossimi sheep to Egyptian ecological conditions. The tongue had three regions: the apex (with a slightly bifurcated tip), the body (subdivided into rostral and caudal parts) and the root (subdivided into rostral papillary and caudal non-papillary parts). Torus linguae had two parts: the triangular rostral part (the caudal part of the body) and the quadrilateral wide part (the rostral part of the root). The lingual papillary system had mechanical (filiform, conical and lentiform) and gustatory (fungiform and circumvallate) types. Filiform papillae were heavily scattered on the dorsal surface of the apex, the rostral part of the body and the ventral surface of the papillary region of the tip. Filiform papillae had five subtypes (ventral and dorsal processed, triangular, leaf-like and triangular-processed papillae), while the conical papillae had three subtypes: two lingual (small, large) and one paralingual (elongated pointed), and the fungiform had two subtypes: the high-density ovoid (on the ventral surface of the tip) and round papillae (on the dorsal surface) that possessed a high number of taste pores, not previously described. They ranged from 5 to 10 for ovoid and 25 to 25 for round papillae. Each circumvallate papilla had an ovoid bulb (with 2-5 taste pores) encircled by an annular groove and two pads (i.e. not described previously). The papillary system's regional divergence was specialized for their harsh and semi-harsh diet.

Ultrastructural focus on the oral cavity roof of the juvenile and adult white grouper (Epinephelus aeneus): Novel view of its development with carnivorous adaptation.

The present study aims to characterize the ultrastructural roof of the oral cavity roof in juvenile and adult Epinephelus aeneus using SEM. The investigation used 10 roofs of the oral cavity of juvenile and adult stages of age. The results in both age groups showed that the roof had five tooth bands, an upper valve, and a palatine region. The upper lip was divided into the anteriorly median and two lateral parts. The posterior margin of the anterior labial part is not attached to the incisive or canine teeth. The anterior surface of the canine teeth was attached to the transverse part of the upper incisive ridge that was separated from the posterior labial part by a deep groove. The semilunar upper velum showed two surfaces, two borders, and two parts. The median part of the oral surface of the velum carried the extension of the longitudinal part of the upper incisive ridge that had some variation among the two examined age stages. In addition, the median part of the oral surface of the upper velum had two rough areas located laterally on each side of the longitudinal part of the upper incisive ridge. The oral surface had several slightly elevated fungiform papillae with taste buds. The palatine region was divided into the peripheral semilunar part and the wide median part, by palatine teeth and the palatine groove laterally and by vomer teeth anteriorly. The results provided important data for the aquaculture in Egypt in relation to their diet.

Morphology of the tongue dorsal surface of turbot (Scophthalmus maximus).

This study aimed to analyze the morphology of the tongue, which varies among fish species and has not been studied in turbot (Scophthalmus maximus), using macro-anatomical, light and scanning electron microscopy (SEM). As research materials, the tongues of eight adult turbot, which were used for consumption, were examined. The roughly triangular-shaped tongue of the turbot consisted of three parts: apex, body, and root. The thickness of the tongue continued to increase from the apex to the root. Although papillae were not observed in the macroscopic examination of the tongue, microscopic examination revealed the presence of cone-like shaped papillae on the submucosa. The tongue was composed of mucosa, submucosa, and hyaline cartilage. By SEM the presence of the taste buds (Types I and III), taste pores and papillae on the dorsal surface were demonstrated. Taste buds are the first descriptions in S. maximus. Therefore our results could add new data to the fish tongue anatomy. HIGHLIGHTS: Morphological and morphometric data of the turbot (Scophthalmus maximus) tongue were obtained in this study and compared with other species. In addition, the dorsal surface of the turbot tongue was described for the first time using SEM.

Lingual papillary system of the Egyptian mouse-tailed bat (Rhinopoma hardwickii): New insights to its feeding strategies using scanning electron microscope, and immunohistochemical analysis of keratin.

The current investigation described the ultrastructural and histological features of the lingual papillary system in 14 adult Rhinopoma hardwickii using scanning electron and light microscopy, and immunohistochemical staining of keratin. Three types of lingual papillae were recognized; one filiform mechanical and two gustatory (fungiform and circumvallate) of three directions: posterior, posteromedian, and median. Five filiform papillae subtypes were observed in four regions. The lingual tip had numerous giant filiform papillae. The lingual root was divided into two regions, an anterior papillary and a posterior non-papillary region that had only three circumvallate papillae in a triangular pattern. Histologically, the papillae exhibited detached thick keratin that spread over the dorsal epithelial surface. The dorsal surface of the root had stratified keratinized circumvallate papillae. Concerning the pan-cytokeratin (CK) antibody staining, moderate CK expression was observed in the superficial corneal layer of the apical lingual mucosa. While the superficial corneal cell layer in both body and root exhibited increasing of CK antibody. Morphometrically, there was a significantly increased, region-specific number of triangular, pointed filiform papillae, which were relatively increased in length and width. However, the round conical blunt filiform papillae and fungiform papillae were sparse. We concluded that our results showed distinguished adaptations in the lingual structure and its different papillae established on the nutritional and energetic demand of this bat species. This lingual adaptation is summarized in number, position, taste buds, direction, ultrastructural features, and histological that concerned with its ability in eating during the night flight. RESEARCH HIGHLIGHTS: Three types of lingual papillae; one mechanical (filiform) and two gustatory (fungiform and circumvallate) in which there were three circumvallate papillae on the posterior part of the lingual root in a triangular arrangement with an irregular surface, but the filiform papillae had five subtypes arranged in four regions. The lingual root was divided into two regions, an anterior papillary region and a posterior non-papillary region that only had three circumvallate papillae. Concerning staining with a pan-cytokeratin (CK) antibody, moderate CK expression was observed in the superficial corneal layer of the lingual mucosa of the tongue apex while the superficial corneal cell layer of the body and root exhibited increased expression of the CK antibody compared to the lingual apex.

Ultrastructural adaptation of the oropharyngeal cavity of the Eurasian common moorhen (Gallinula chloropus chloropus): Specific adaptive dietary implications.

The present investigation represents the first morphological description of the oropharyngeal cavity of Eurasian common moorhen. Nine oropharyngeal cavities were examined grossly and by stereomicroscope and scanning electron microscopic (SEM) observations. The tongue had a rounded apex with multiple acicular processes on its rostral and lateral borders. The dorsal lingual surface of the apex and body had a median sulcus. The papillary crest carried four caudally directed triangular conical papillae on its median part and four triangular conical papillae on each lateral part. The filiform papillary system; small papillae on apex and long papillae on the rostral part of the body while broad papillae on the caudal part of the body. The lingual root had a special appearance by presence of three areas: mucosal fossa, two lateral ridges, and rhomboidal elevated central part. The caudal border of the mound carried heart-shaped pharyngeal papillae that possessed three papillary rows. The palate had a median palatine ridge rostrally that surrounded by two lateral palatine ridges. The choanae had two equal parts: rostral tapering and wide caudal. The rostral tapering choanal part was surrounded by two longitudinal rows of caudally directed conical papillae, one on each side. There was a single transverse row of conical papillae on each side of the caudal part of the rostral tapering choanal part. The caudal wide choanal part did not encircle by any papillae. Our conclusion exhibited unique structural and functional specifications of the oropharyngeal cavity with the tongue that evident with nutritional behavior.

Morphological structure of the tongue of the European badger (Meles meles).

BACKGROUND: This study aimed to reveal the morphological aspects of the tongue and investigate the histologic and scanning electron microscopic (SEM) findings of the papillae on the tongue of the European badger. MATERIALS AND METHODS: Two adult European badgers were used as material. The tongues of the European badgers were removed via the dissection method, and morphometric measurements were taken. The tongues were fixed in 10% buffered formalin solution for histological examination, and the SEM procedure was applied to the tongues. RESULTS: Three types of papillae were observed on the tongues: filiform, fungiform, vallate papillae. The different shapes (hook or conical structures) of filiform papillae were observed on the whole tongue. Fungiform papillae were found scattered in filiform papillae. Six vallate papillae were found on the dorsal face of the middle and root of the tongues. In the root of the tongue, the lingual tonsil was observed in the lentiform shape. CONCLUSIONS: The authors believe that this research will provide contributions to morphological knowledge about the tongue of the badger, which is a poorly known species.

Ultrastructural features on the oral cavity floor (tongue, sublingual caruncle) of the Egyptian water buffalo (Bubalus bubalis): gross, histology and scanning electron microscope.

BACKGROUND: The present work was focused on the evaluation of morphological characteristics of the lingual caruncles and tongue with its papillae of Egyptian water buffalo (Bubalus bubalis) using gross examination, light and scanning electron microscopy. MATERIALS AND METHODS: The ventral surface of the sublingual caruncle carried a small opening of the duct of both monostomatic and mandibular salivary gland. The lingual mucosa of dorsal, lateral border and, to some extent, of ventral surface of apex had lingual papillae (filiform, fungiform), while the lingual mucosa of the lingual body especially at torus linguae had conical papillae, but circumvallate papillae observed at the caudal part of body and root. The dorsal surface of the apex and body carried numerous long, thread-like, with blunt apex, caudally directed filiform papillae that covered with keratinised scales without secondary papillae. RESULTS: The degree of keratinisation classified filiform papillae into rostral part of high keratinisation and caudal of less keratinisation. Conical papillary surface carried exfoliated epithelium with longitudinal groove on its rostral surface and carried secondary papillae. Fungiform papillae were scattered among filiform papillae on the dorsal and ventral surface of the apex and its convex surface had exfoliated keratinised epithelium. Circumvallate papillae were surrounded by circular deep groove bordered by vallum that carried small secondary papillae that ended into the primary groove. Taste buds of circumvallate papillae opened in the lateral lining epithelium facing the groove. CONCLUSIONS: Von Ebner's glands were observed in computed tomography under papillae especially toward the groove and their ducts open into the base of the groove.

Taste Bud Connectome: Implications for Taste Information Processing.

Taste buds contain multiple cell types, two of which mediate transduction of specific taste qualities: Type III cells transduce sour while Type II cells transduce either sweet, or bitter or umami. In order to discern the degree of interaction between different cell types and specificity of connectivity with the afferent nerve fibers (NFs), we employed serial blockface scanning electron microscopy (sbfSEM) through five circumvallate mouse taste buds. Points of contact between Type II and Type III cells are rare and lack morphologically identifiable synapses, suggesting that interaction between these cell types does not occur via synapses. Of the 127 NFs that make synaptic contacts with taste cells in the sampling volume, ∼70% (n = 91) synapse with only one taste cell while 32 fibers synapse exclusively with multiple Type II cells or multiple Type III cells. Our data do not rule out multimodal fibers innervating Type II cells of separate taste qualities. Notably, four fibers (∼3%) synapse with both Type II and Type III cells, forming both mitochondrial and vesicular synapses on the different cell types. Since Type II and Type III cells transduce different taste qualities, these dual connected fibers are not consistent with a absolute labeled-line encoding system. Further, our data reveal considerable variation in both the number of synapses per cell/nerve pair and the number of innervating NFs per taste cell, both of which likely have consequences for encoding taste quality and concentration. Finally, we identify a subset of Type II cells which may represent an immature stage.SIGNIFICANCE STATEMENT Taste buds, the sensory end organs for the sense of taste, contain multiple types of sensory cells, with each responding to one of the primary tastes: salt, sweet, sour, bitter, and umami. In order to determine the degree of interaction between cell types and specificity of connectivity to afferent nerves, we employed serial blockface electron microscopy (EM) of mouse circumvallate taste buds. We find no synapses between cell types within the taste bud suggesting that any interactions are indirect. While the majority of nerve fibers (NFs) connect to a single type of taste cell, 3.1% of the fibers branch to receive input from taste cells of different specificities. Thus, taste cannot entirely be carried along NFs dedicated to single taste qualities.

Variable Branching Characteristics of Peripheral Taste Neurons Indicates Differential Convergence.

Taste neurons are functionally and molecularly diverse, but their morphologic diversity remains completely unexplored. Using sparse cell genetic labeling, we provide the first reconstructions of peripheral taste neurons. The branching characteristics across 96 taste neurons show surprising diversity in their complexities. Individual neurons had 1-17 separate arbors entering between one and seven taste buds, 18 of these neurons also innervated non-taste epithelia. Axon branching characteristics are similar in gustatory neurons from male and female mice. Cluster analysis separated the neurons into four groups according to branch complexity. The primary difference between clusters was the amount of the nerve fiber within the taste bud available to contact taste-transducing cells. Consistently, we found that the maximum number of taste-transducing cells capable of providing convergent input onto individual gustatory neurons varied with a range of 1-22 taste-transducing cells. Differences in branching characteristics across neurons indicate that some neurons likely receive input from a larger number of taste-transducing cells than other neurons (differential convergence). By dividing neurons into two groups based on the type of taste-transducing cell most contacted, we found that neurons contacting primarily sour transducing cells were more heavily branched than those contacting primarily sweet/bitter/umami transducing cells. This suggests that neuron morphologies may differ across functional taste quality. However, the considerable remaining variability within each group also suggests differential convergence within each functional taste quality. Each possibility has functional implications for the system.SIGNIFICANCE STATEMENT Taste neurons are considered relay cells, communicating information from taste-transducing cells to the brain, without variation in morphology. By reconstructing peripheral taste neuron morphologies for the first time, we found that some peripheral gustatory neurons are simply branched, and can receive input from only a few taste-transducing cells. Other taste neurons are heavily branched, contacting many more taste-transducing cells than simply branched neurons. Based on the type of taste-transducing cell contacted, branching characteristics are predicted to differ across (and within) quality types (sweet/bitter/umami vs sour). Therefore, functional differences between neurons likely depends on the number of taste-transducing cells providing input and not just the type of cell providing input.

N-cadherin localization in taste buds of mouse circumvallate papillae.

Taste buds, the receptor organs for taste, contain 50-100 taste bud cells. Although these cells undergo continuous turnover, the structural and functional integrity of taste buds is maintained. The molecular mechanisms by which synaptic connectivity between taste buds and afferent fibers is formed and maintained remain ambiguous. In the present study, we examined the localization of N-cadherin in the taste buds of the mouse circumvallate papillae because N-cadherin, one of the classical cadherins, is important for the formation and maintenance of synapses. At the light microscopic level, N-cadherin was predominantly detected in type II cells and nerve fibers in the connective tissues in and around the vallate papillae. At the ultrastructural level, N-cadherin immunoreactivity appears along the cell membrane and in the intracellular vesicles of type II cells. N-cadherin immunoreactivity also is evident in the membranes of afferent terminals at the contact sites to N-cadherin-positive type II cells. At channel type synapses between type II cells and nerve fibers, N-cadherin is present surrounding, but not within, the presumed neurotransmitter release zone, identified by large mitochondria apposed to the taste cells. The present results suggest that N-cadherin is important for the formation or maintenance of type II cell afferent synapses in taste buds.

Anatomical, histological and immunohistochemical study of the tongue in the rainbow trout (Oncorhynchus mykiss).

The rainbow trout (Oncorhynchus mykiss Walbaum, 1792) is a fish commercially farmed all over the world. These fish are usually fed, in aquaculture, with pellets rich in proteins and fat. It is well known that there are close relationships among the adaptation of vertebrates to their environment, the capacity and the modality of feeding and the oral cavity morphology, especially the tongue one. No data are so far available about the morphology of the rainbow trout tongue, and therefore, the aim of this study was to investigate by light, scanning electron and confocal laser microscopy, the morphological characteristics of the tongue. An apex, a body and a root can be distinguished in the tongue, and the presence of teeth, taste buds and fungiform-like papillae was demonstrated. Light microscopy shows the presence of an adipose tissue pad in the deeper layer of the apex and in the most superficial layer of the root. In the deeper layer of the body, a triangular-shaped pad consisting of fusiform cells immersed in abundant extracellular matrix of the mesenchymal tissue was observed. The confocal laser microscopy shows the presence of cells with a fibroblast-like morphology positive for vimentin. In the deepest layer of the tongue root, a large area of osteo-cartilaginous tissue was observed. The results, besides the description of the morphological characteristics of the tongue, related to studies regarding the feeding, could be considered for the eventual applications of the use of mesenchymal cells, observed in adult fish, in cell therapies in different pathologies.