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Distribution of taste buds on fungiform and circumvallate papillae of bovine tongue.

The distribution of taste buds on the fungiform and circumvallate papillae of the cow tongue has been determined. The two tongues studied were from Holstein-Friesian cows four to six years of age; they contained 14,765 and 21,691 taste buds, respectively. The tip of the tongue is well supplied with fungiform papillae, and the posterior portion contains the circumvallate papillae. The midportion of the tongue contains relatively few taste papillae. The fungiform papillae contained 1,580 and 1,838 taste buds on the two tongues, respectively, and the circumvallate papillae were estimated to contain 13,185 and 19,853 taste buds. The highest concentration of taste buds therefore occurs in the circumvallate papillae; these relatively few papillae contain approximately 90% of the taste buds. On a circumvallate papilla, taste buds are found only on the papillary sidewall, with none either on the apical surface of the papilla or on the outer wall of the moat.

Animals

The fine structural effect of sialectomy on the taste bud cells in the rat.

Taste buds in the rat and other mammals share a secretory activity with their transduction function as taste receptor. The present work shows the effect of bilateral removal of the main salivary glands on taste bud cells' components related to secretion in the vallate papilla of the rat. In the sialectomized rats remarkable changes were evidence in the dark and intermediate types of taste bud cells, which are known to be the secretory components. Such changes involve hypertrophy of either the protein synthetizing machinery, the smooth endoplasmic reticulum or the Golgi complex. Lucent and coated vesicles associated to Golgi cisternae increased in number but the amount of dense-core vesicles (secretory vesicles) at the apical cytoplasm of cells decreased. Images of exocytosis of secretory products were observed. The hypertrophy of Golgi complex components was clearly detected with the OsO4 impregnation method for light and electron microscopy. Alteration in the acid phosphatase activity of taste bud cells was not observed in the sialectomized rats. These findings suggest that sialectomy stimulates the entire secretory cycle of dark and intermediate taste bud cells. The light taste bud cells, which are not engaged in secretion, are hardly affected by the treatment. Although taste buds in mammals are neuro-dependent structures, present evidence indicates that they are also sensitive to non-neural influences.

Animals

Liquid-tissue behavior and differential cohesiveness during chick limb budding.

Emerging chick limb-buds at first grow only in length, not width. The growth parameters of limb mesoderm--cell shapes, distributions, division patterns and cleavage orientations--are incompatible with representations of this tissue as an elongating solid composed of proliferating but immobile cells. We observe that samples of both early limb mesoderm and also surrounding flank mesoderm round up like liquid droplets in organ culture. Therefore, liquid-like tissue rearrangments, including cell shuffling movements and neighbor exchanges, may occur in limb and flank mesoderm during in vivo limb budding. If so, differences in limb-flank surface tension properties would have to be present to keep these two fluid cell populations segregated into distinct tissues and properly positioned underneath limb and flank ectoderm. Previous studies have shown that tissue surface tensions are reflected in the spreading behavior of fused pairs of cell aggregates. To determine whether or not they possess differing surface tension properties, we pair excised pieces of early leg-bud, wing-bud or intervening flank mesoderm with pieces of 5 3/4-day heart or liver in hanging drop cultures. For more rapid determinations of relative liquid-tissue cohesiveness than can be obtained in conventional, long-term experiments, aggregate pairs are fixed shortly after fusion. Since partial-envelopment configurations depend upon relative aggregate sizes as well as their tissue surface tensions, new procedures are used to deduce relative aggregate cohesiveness from cross-sections of these briefly fused aggregate pairs. The envelopment tendencies of aggregates fixed 6--9 h after fusion are similar to those fixed 15--19 h after fusion: heart tends to surround leg; heart and wing surround each other with similar frequencies, but flank tends to surround heart. Also, liver tends to surround leg and wing, but flank tends to surround liver. When the effects of relative aggregate size are taken into account, these non-random, tissue-specific patterns of aggregate envelopment indicate that the relative cohesiveness of these tissues falls into the sequence: leg greater than heart congruent to wing greater than liver greater than flank. The in vitro behavior of early limb-bud and neighboring flank mesoderm in these studies suggests that they are not simply mechanically identical portions of a single liquid tissue. We have previously proposed that early limb-bud mesoderm may act like a non-dispersing, cohesive liquid droplet which is embedded within a less cohesive fluid layer of flank tissue (and which is molded distally into paddle-shaped conformations by solid-like limb ectoderm and/or subjacent extracellular matrix). This proposal is not only compatible with the growth parameters of limb-bud mesoderm in vivo, but is also consistent with our observation that flank mesoderm surrounds tissues which surround limb mesoderm in these aggregate-fusion experiments. Our model suggests that differences in the surface tension properties of limb vs...

Animals

Intraperitoneal transplants of taste buds in the newt.

Autografts of tongue onto the liver contained taste buds without nerves for up to 30 days. However, these denervated taste buds were generally smaller than normal and distorted in structure. Nerve fibers invaded the graft from the liver and after 30 days normal appearing taste buds were found only in implants of tongue which contained extensive nerve fibers. Thus it appears that certain visceral nerves can maintain and possibly induce taste bud formation in implants of tongue. This is interpreted as additional evidence for the trophic influence of nerves upon taste buds. Moreover, this ability to maintain taste buds is not restricted to gustatory nerves in the newt.

Animals

Surface morphology of taste buds in catfish barbels.

External taste buds abound on barbels of the adult catfish Corydoras arcuatus. When examined by scanning electron microscopy, they are visualized as a series of punctate, conical elevations projecting from the general surface epithelium. All taste buds were found to be of one type. Both their external and internal surface features could be clearly elucidated on intact barbels and in barbels fractured transversely at various positions along their length. An extensive nerve terminal network penetrates the base of each taste bud. Two populations of elongated cells bearing prominent microvilli project through the central pore at the tip of each bud. One set of microvilli is thicker, longer and more club-shaped than its counterpart. While both are randomly distributed within each central pore, the small, short microvilli appear to outnumber the larger ones. A third population of cells, devoid of any apical microvilli, was also seen in some of the taste buds examined internally. These cells do not project to the external surface and are interpreted as "basal" cells described in previous light and transmission electron microscope studies of taste buds in other vertebrate species. The functional significance of some of these morphological findings is discussed.

Animals

Location of taste buds in intact taste papillae by a selective staining method.

Taste buds were found to stain strongly and selectively in intact papillae with highly acidic dyes such as ponceau S. In intact tongues the taste buds in the fungiform, circumvallate and foliate papillae of the cynomolgus monkey and in the fungiform papillae of the rat as well as the taste discs in the fungiform papillae of the frog could be visualized. This method enables a rapid location and counting of taste buds in taste papillae without preparing histological sections. In cynomolgus tongue material fixed in formalin, the dyes penetrate into the buds. In fresh tongues only the taste pore region of the buds stains, which suggests that in vivo taste buds are impenetrable underneath the pore.

Animals

The effect of temperature on the turnover of taste bud cells in catfish.

Renewal of taste bud cells on the barbels of channel catfish was studied. Groups of catfish, held in and acclimitized to 14 degrees C, 18 degrees C, 22 degrees C and 30 degrees C dechlorinated tap water were injected with [3H]thymidine (3.0 muCi/g body weight intraperitoneally). Barbels were sampled at various times after injection and prepared for light microscope autoradiography. Results show that epithelial cells surrounding the taste buds divide and some of their daughter cells migrate into the taste buds. The time at which 50% of the labelled cells have degenerated is taken as the average turnover time or average life span of the taste bud cells. The average life span as well as the time spent inside the taste buds is highly temperature-dependent. At 14 degrees C, 18 degrees C, 22 degrees C and 30 degrees C the average life span is on the order of 40, 30, 15 and 12 days respectively. Further studies indicate that both light and dark staining cells of the taste bud were labelled.

Animals

Ecto-calcium-dependent ATPase activity of mammalian taste bud cells.

Histochemistry was utilized to characterize Ca-ATPases associated with lingual taste buds in the golden hamster. Taste buds showed elevated staining for magnesium- or calcium-dependent ATPase (Ca-ATPase) relative to the surrounding epithelium. At low calcium concentrations (0.1-0.5 mM), intracellular staining predominated. Most of the studies were conducted at calcium concentrations of > or = 10 mM, in which most of the staining was localized to the external face of plasma membranes of taste bud cells (including receptor and basal cells) located in the core of fungiform taste buds, or the entire vallate or foliate taste buds. The peripheral fungiform taste bud cells stained much less intensely, but the peripheral cells adjacent to the core showed intermediate levels. GTP and ITP were just as effective substrates as ATP. Millimolar concentrations of magnesium were as effective as calcium. Inhibitors of intracellular ATPases, including quercetin, sodium azide, and 2,4-dinitrophenol, had no effect on the staining. Therefore, the Ca-ATPase staining of plasma membranes at mM concentrations of calcium is thought to correspond to one or more ecto-Ca-ATPase activities with unknown functions. Roles related to increased energy requirements or to the possible function of ATP as a neurotransmitter or -modulator are proposed.

2,4-Dinitrophenol

The occurrence of taste buds in the palate of human adults as evidenced by light microscopy.

There is some uncertainty in the literature as to the existence of taste buds in the palate of the human adult. In those histologic studies in which the ages of the individuals have been reported, taste buds have not been found in the palates of adults, but have been found in fetuses or newborn. However, clinical studies have demonstrated taste perception in the palate of the human adult. Thus, the aim of the present study was to attempt to find taste buds in the human palate in subjects of different ages. In serial sections of selected areas of the palatal mucosa from autopsy material from individuals 0--80 years of age no taste buds could be demonstrated. However, in four of seven subjects aged 25--44 years, one or two taste buds were found in biopsies from areas of the soft palate where taste perception had been demonstrated clinically just prior to excision. Thus the present study indicates that scattered taste buds exist also in the soft palate of human adults.

Adolescent

Retinoic acid respecifies limb bud cells in vitro.

Retinoic acid (RA) is known to have dramatic effects on limb pattern formation and has been shown to exert its effects on limbs by converting anterior limb bud cells into cells with posterior positional properties. In this study we find that dissociated posterior limb bud cells from chick and mouse embryos cultured at high density (micromass cultures) are able to stimulate the formation of supernumerary digits when grafted into developing wing buds and that the positional identity of both chick and mouse limb bud cells can be maintained for finite periods of time in vitro. Furthermore, using this assay system we have tested whether anterior cells from mouse and chick limb buds can be converted into cells with posterior identity by exposure to RA in vitro. We find that anterior limb bud cells acquire posterior properties after culture in the presence of RA.

Animals

Further observations on the site of bone prospective areas in the chick embryo wing bud.

The portion of the pre-axial region of the wing bud anterior to the cranial boundary of the skeletogenous territory of the stylo-zeugopod, according to Stark and Searls' recent maps, was surgically isolated in chicken embryos of the stages 18-25, and autoplastically grafted to the dorsal surface of the hind-limb bud or to the trunk. In other embryos of the same stages the cranial half or the cranial two thirds of the pre-axial region mentioned were isolated and heterotopically implanted. The experimental results consistently showed that the isolate contained a variously large portion of the presumptive radius and a significant part of the proximal half of the prospective humerus. In fact, the skeletal parts mentioned did not develop in situ in the donor wing while they differentiated, in general, in the implantation site. In this regard, the present research confirms data from previous experiments showing that Stark and Searls' maps do not offer an accurate representation of the position and the cranio-caudal width of the presumptive areas of the skeletal pieces of the wing. Our findings seem also to indicate that the major (or longitudinal) axis of the future radius is arranged along the proximodistal axis of the wing bud throughout the developmental stages mentioned; instead, the major axis of the future humerus undergoes a gradual shift, being first (stages 18-20) arranged nearly parallel to the bud base and getting more and more parallel to the proximo-distal axis of the wing bud through stages 21-24. Besides, between stage 18 and 25 the cranio-caudal width of the presumptive skeletogenous territory of the zeugopod, and to a lesser extent of the stylopod, seems to undergo a slight relative reduction with respect to the cranio-caudal thickness of the whole wing bud; this might depend on a process of aggregation of the prospective skeletogenous cells into gradually denser and more defined precartilaginous blastemes.

Animals

[Contribution of somite cells to the development of posterior limb buds in mice].

1. The structural changes accompanying the early genesis of the mouse hindlimb bud have been studied in serial Epon thick sections at five successive somitic stages collected during the 10th developmental day. 2. As soon as the 14-somite stage, increased proliferative activities occurring in a limited area of the embryonic somatopleure close to the caudal end of the coelomic cavity point out the onset of limb morphogenesis. From its very beginning on, up to its ceasing at the 27-somite stage, the somatopleural proliferation is more active in the caudal part of the prospective limb territory and in the whole area, it appears stronger ventrally than dorsally. 3. Discrete structural changes occur during this period in the mesoderm interposed between the hindlimb bud territory and the overlying unsegmented sheet of somitic mesoderm, in relation with the genesis of the mesonephritic constituents and of the posterior cardinal vein. A detailed analysis of this area in serial sections reveals that the unsegmented somitic mesoderm does not provide any cell to the limb bud mesoderm. 4. As soon as the somitic mesoderm becomes organized into segmented somites, obvious histological changes located on the lateral aspect of the ventro-lateral somitic edge reveal that numerous undifferentiated somitic cells invade the proximal area of the limb bud mesoderm. Starting at the 27-somite stage in the first two metameres adjacent to the precordonal portion of the limb bud, this processus extends to the five somites of the limb territory at the 33-somite stage. In each metamere, the area of cell migration takes place near the caudal border of the somite and, from one somite to the other, the number of migrating cells increases in a cephalo-caudal direction. No sign of a similar cell migration was observed in the somites located cranially or caudally to the hindlimb bud. 5. The results are discussed in relation with the various experimental and descriptive evidences obtained in non-mammalian Vertebrates about the contribution of the somites to limb morphogenesis and about the factors involved in the onset of cell proliferation in the prospective limb mesoderm.

Animals

Biochemical studies of taste sensation. III. Preparation of a suspension of bovine taste bud cells and their labeling with a fluorescent probe.

A method to prepare suspensions of taste bud cells is described. Bovine circumvallate papillae, which contain most of the taste buds in this animal, are incubated in collagenase-containing medium and the epidermal sidewall tissue is then dissected from the inner gelatinous dermis. The sidewall tissue, which contains the taste buds, is gently homogenized by manual operation of an all-glass homogenizer with a loose-fitting pestle. The suspended material is separated on a discontinous Ficoll gradient (2%, 8%, 10%, 12% w/w). The material banding at the 8-2% interface is greatly enriched in spindle-shaped cells that are morphologically similar to taste bud cells as they appear in situ. These cells are not seen when the procedure is done with tissues devoid of taste buds, namely the upper surface of the circumvallate papilla or epithelium from the intermolar eminence. Fluorescence analysis indicates that the hydrophobic probe, 8-anilino-1-naphthalenesulfonate (ANS), binds to relatively nonpolar sites in the suspension. It is postulated that the probe is adsorbing onto the surface membrane of the cell. These preparations may be useful in studying specificity and transduction in taste sensation.

Anilino Naphthalenesulfonates

On the ultrastructure and permeability of taste buds of the marine teleost Ciliata mustela.

The abundant taste buds of the barbels and free fin rays of the five bearded rockling, Ciliata mustela contain an average of 100-150 cells, falling into two types. Tubule-containing cells ('t-cells'), tentatively identified as receptor cells, and each surrounded by fibril-containing cells ('f-cells') in the central part of the bud. t-Cells also occur in two concentric shells separated by indifferent epithelial cells at the periphery of the bud. f-Cells are characterized by their concentrations of fine fibrils, and by granules or vesicles of 180-190 mmu diameter. The 100 or so receptor cells in a taste bud are innervated by some 250 axons. Lanthanum penetrates more deeply into the extracellular space of taste buds than into the extracellular space of the general epithelium, perhaps indicating that a greater area than the mere protruding tip of receptor cells may be accessible to chemical stimulation. Degenerating cells may provide an important route of entry for such external agents.

Animals

Effect of the length of the distal stump of transected nerve upon the rate of degeneration of taste buds.

The present work was carried out to study the effect of the length of the distal stump of transected nerve upon the rate of degeneration and the time course of disappearance of mammalian taste buds. Twelve adult rabbits were anaesthetized and the glossopharyngeal nerves of both sides were exposed and transected so as to leave a long distal stump on the right and a short one on the left side. The animals were sacrificed at different post-operative periods ranging from 2 to 14 days and the circumvallate and foliate papillae of both sides were examined. The taste buds on the side of the short distal stump always showed a greater decrease in number, size and cell contents than those of the side of the long distal stump. The taste buds on the side of the short distal stump disappeared earlier than those of the side of the long distal stump. Vallate taste buds disappeared earlier than the foliate: possible reasons for this were put forwards. The validity of the neurohumoral theory to the mammalian taste buds was discussed.

Animals

An electron microscopic study on the innervation in the taste buds of the mouse circumvallate papillae.

Taste buds of the mouse circumvallate papillae were studied by electron microscopy to elucidate the innervation involving the adrenergic nerve supply. Typical afferent synaptic contacts, with increased density of the membranes and aggregations of synaptic vesicles in the cytoplasm adjacent to the nerve endings, were demonstrated between the type III cells and the nerve endings. Along the regions of contact between the type II cells and the nerve endings, cisternae of endoplasmic reticulum were often seen beneath the cell membrane, and the nerve endings contained relatively many synaptic-sized vesicles. Such an innervation seems to be efferent in nature. For the detection of the adrenergic nerve supply, 5-hydroxydopamine (5-OH-DA) was injected after pretreatment with L-DOPA and nialamide. The mice showed numerous adrenergic nerve fibers in the connective tissue underlying the taste buds. On very few occasions, the adrenergic nerves penetrated the basal lamina of the taste buds and came into contact with the bud cells. Some adrenergic nerves were distributed among the epithelial cells around the taste buds. The reaction product from acetylcholine esterase activity was found around the adrenergic nerve fibers labeled with 5-OH-DA.

Acetylcholinesterase

Taste buds in the vallate papillae of the rat studied with freeze-fracture preparation.

The taste bud of the vallate papillae of the rat has been examined in the electron microscope by using the freeze-fracture technique. Tight junctions as a junctional complex are located at the taste pore, and form a seal between the oral environment and the taste buds. Tight junctions are not only within the taste buds, but also are demonstrated in the granular cell layers of the surrounding lingual epithelium. The present finding suggests that tight junctions of the taste pore link with those of the lingual epithelium. Desmosomes are observed in the buds, but they are smaller in size than in the surrounding lingual epithelium. Besides these junctions, within the buds, gap junctions containing particle-free zones are demonstrated which are called a subcompartment type.

Animals

Neural cell adhesion molecule of taste buds.

The distribution of the neural cell adhesion molecule (N-CAM) in the taste buds of adult and developing mice was studied by use of an immunocytochemical technique. In adult mice, nerve fibers and some elongated taste bud cells reacted with anti-N-CAM antibody. Immunoelectron microscopic observation showed that only type-III (gustatory) cells, which have afferent synaptic contacts with the nerves, were N-CAM positive in the taste buds. At day 0 after birth, taste buds of the developing mice first appeared in the circumvallate papillae, and N-CAM positive cells, which corresponded to type-III cells having afferent synapses, were found. This result suggests that N-CAM is involved in formation of the synaptic contacts between type-III cells and nerves in the taste buds.

Aging