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Distribution of taste buds on the epiglottis of the rat and house shrew, with special reference to air and food pathways.

We investigated the positioning of the epiglottis in the pharyngo-laryngeal region and the distribution of taste buds on the epiglottis in the rat and house shrew, animals which have different feeding habits. In the fixed samples of both species, when the mouth was closed or slightly opened, the epiglottis was found to protrude into the nasopharyngeal hiatus above the soft palate. But it retracted from its position when the mouth was widely opened. In omnivorous rats (n = 6), the mean number (mean density +/- s.d.) of taste buds was 52 (12.6 +/- 2.2/mm2) on the laryngeal surface but only 4 (1.3 +/- 1.0/mm2) on the oral surface. The three-dimensional view was reconstructed from serial sections. The taste buds were distributed most densely close to the caudal base and became fewer toward the more rostral tip. In insectivorous house shrews (n = 2), 4 taste buds on average were found only on the laryngeal surface of the epiglottis. Epiglottal taste buds may work as chemosensory detectors to initiate the reflex reaction to protect the airway from oral substances during swallowing and drinking.

Animals↗

Taste buds and neuronal markers in patients with chronic renal failure.

OBJECTIVE: To study the number of taste buds and, with the use of specific markers for peripheral nervous tissue, to study the neuronal pattern in taste buds from 36 patients with chronic renal failure (CRF), 19 renal transplant recipients, and 40 healthy subjects. Of the patients with CRF, 17 patients had not started dialysis, 12 patients were on peritoneal dialysis, and 7 patients were on hemodialysis. DESIGN: From all subjects, two or three fungiform papillae were collected from the anterior part of the tongue. Cryostat sections were cut and inspected under light microscopy to determine the presence of taste buds. The sections were subsequently incubated with primary rabbit antibodies against protein gene product 9.5, substance P, and nerve growth factor receptor. RESULTS: Using these antibodies, no differences between the groups were observed. However, patients with CRF had fewer taste buds than control subjects. CONCLUSION: No immunohistochemical differences were observed between patients with CRF and healthy controls. However, patients with CRF had significantly fewer fungiform taste buds, suggesting an important factor contributing to the well-known impairment of taste acuity in this patient group.

Adult↗

Erb and c-Kit receptors have distinctive patterns of expression in adult and developing taste papillae and taste buds.

Twenty four different protein tyrosine kinases (PTKs) were amplified from a taste-enriched cDNA library using PCR. The expression of four protein tyrosine kinase receptors (EGFR, ErbB2, ErbB3, and c-kit) was examined in adult and developing rat taste papillae. All four of these receptors were expressed in overlapping populations of differentiated taste cells within adult taste buds. Taste bud basal cells were ErbB2(+) but did not express the other Erb receptors. During prenatal development, the Erb receptors were expressed extensively in the basal cells around developing papillae, and ErbB2 and c-kit immunoreactive neuronal fibers were seen in close association with taste papillae. In early postnatal stages, ErbB2(+) and c-kit(+) neuronal fibers were often seen entering the taste papillae epithelium, where new taste buds form, and by postnatal day 2 (P2), individual ErbB2(+) and c-kit(+) cells were seen in this region as well. Between P3 and P8, c-kit was highly expressed at the bottom of foliate papillae trenches. The extensive expression of the Erb and c-kit receptors in adult taste buds and in and around developing papillae suggests that these receptors may play a role in the prenatal and postnatal development of gustatory papillae and taste buds.

Age Factors↗

The mouse polydactylous mutation, luxate (lx), causes anterior shift of the anteroposterior border in the developing hindlimb bud.

Pattern formation along the anterior-posterior axis of the vertebrate limb is established upon activation of Sonic Hedgehog (SHH) in the zone of polarizing activity (ZPA). Since many mouse mutants with preaxial polydactyly show ectopic expression of Shh at the anterior margin of the limb buds, it has been thought to be a primary defect caused by these mutations. We show here that the mouse mutation luxate (lx) exhibits dose-dependent reduction in the size of the Fgf8 expression domain in the ectoderm from the initial stage of limb development. This aberration was independent of Fgf10 expression in the limb mesenchyme. Shh was induced in the mesenchyme underlying the posterior end of the Fgf8 expression domain, indicating an anterior shift of Shh expression in lx hindlimb buds. Prior to the ectopic induction of Shh, the expression domains of genes downstream from Shh, namely dHAND, Gli1, Ptc and Gre, which are normally expressed in posterior mesenchyme of limb buds, expanded anteriorly on the lx hindlimb buds. Conversely, the expression domains of anterior mesenchymal markers such as Gli3and Alx4 decreased in size. Thus, ectopic Shh is not a primary defect of the lx mutation. Rather, our results indicate that the lx mutation affects the positioning of the anteroposterior border in developing hindlimb buds.

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↗

Histochemical observations on the taste buds of the marmosets (Callithrix jacchus and Callithrix penicillata).

The epithelial cells in the taste buds of C. jacchus and C. penicillata show a moderate amount of ribonucleic acid an a concentration of a PAS-positive diastase-resistant material at their apical part. These cells are devoid of UDPG-GT, phosphorylases, G-6-PA, alanyl aminopeptidase, leucine aminopeptidase, cholinesterase and MAO; they present a weak reaction of F-1, 6-P Ald, LDH, SDH, MDH, cytochrome oxidase, beta-OHBDH, nonspecific esterase and acid phosphatase and a stronger reaction to ADH, NADPH2-TR, ATPases, alpha-GPDH, alkaline phosphatase, 5-nucleotidase and GDH. Although some enzymes (alkaline phosphatase, 5-nucleotidase and ATPases) have an almost uniform reactivity by the several taste buds, the other ones react with a lesser intensity in the smaller uniform reactivity by the several taste buds, the other ones react with a lesser intensity in the smaller taste buds of the fungiform papillae. As a rule the apical part of the cells shows a stronger enzymatic reactivity. The taste buds of the marmosets are penetrated by acetylcholinesterase positive nerve fibers whereas the autonomic ganglia in the connective tissue contain both-acetyl and butyrylcholinesterase.

Adenosine Triphosphatases↗

Taste bud density in circumvallate and fungiform papillae of the bovine tongue.

Taste bud quantitation may provide useful parameters for interspecies comparisons of the gustatory system. The present study is a morphometric analysis of bovine taste papillae. Circumvallate and fungiform papillae from six bovine tongues were serially sectioned and, following staining, analyzed. Circumvallate papillae were found to have a mean volume of 3.66 +/- 2.82 mm3, a mean number of taste buds per papilla of 445 +/- 279, and a mean taste bud density of 155 +/- 112 buds/mm3. Values for lateral fungiform papillae for the same three parameters were 0.384 +/- 0.184 mm3, 13.2 +/- 13.4, and 40.8 +/- 46.6 buds/mm3, respectively. Values for dorsal fungiform papillae were 0.438 +/- 0.246 mm3, 4.39 +/- 4.78, and 14.0 +/- 17.1 buds/mm3, respectively. Circumvallate papillae were found to have a significantly greater volume, number of taste buds per papilla, and taste bud density than either type of fungiform papilla. These data should serve as background for biochemical, endocrinological, or neurological studies involving the bovine tongue.

Animals↗

Evidence for a novel mechanism of binding and release of stimuli in the primate taste bud.

In previous work, we showed that thaumatin, an intensely sweet protein, binds to certain formed elements in taste pores of Rhesus monkey foliate papillae, namely, microvilli and small vesicles shed from microvilli, in addition to amorphous secretions (Farbman et al., 1987). We suggested that the taste bud responds to a thaumatin stimulus by shedding the small vesicles containing fragments of microvillar membrane bearing the stimulus-binding site complex. To examine this hypothesis further, we used electron microscopy to examine taste pores of both vallate and foliate papillae from Rhesus monkeys before or after stimulation with thaumatin or sucrose. We also recorded the neural activity from the glossopharyngeal nerve during stimulation with thaumatin, sucrose, citric acid, and NaCl. The results indicate (1) with no stimulation, vesicles are found in pores of foliate papilla taste buds much more frequently than in pores of vallate papilla buds, (2) in both types of papillae, stimulation with sucrose has no apparent effect on the number of pores containing vesicles, (3) stimulation with thaumatin elicits release of vesicles into pores of both foliate and vallate buds, (4) repeated stimulation of taste buds with thaumatin results in a declining neural response, not seen after repeated stimulation with sucrose, citric acid, or NaCl, and (5) stimulation with thaumatin suppresses the neural response to sucrose, but the reverse does not occur. The combined morphological and physiological data support our original hypothesis that, in response to thaumatin stimulation, binding sites on taste microvillar membranes may be shed as a stimulus-receptor complex into the pore. Alternatively, the binding sites may in some way be altered by the shedding of part of the membrane. The data suggest further that the binding site for sucrose may be close to that for thaumatin because it too is lost or altered after thaumatin stimulation. The reduced neural response after repeated thaumatin stimulation indicates that cells may have suffered a net loss of functional receptors or at least a loss in functional capacity of the receptors. The data indicate that the cellular mechanism for handling the protein sweetener, thaumatin, is strikingly different from the adsorption-desorption response to saccharide sweeteners.

Adaptation, Physiological↗

[Histogenesis of the taste buds of the vallate papilla in the in the rat in the postnatal stages of development].

The developing taste buds of vallate papillae were studied with electron microscope in rats during the first 7 days after birth. Two types of cells--light and dark--are identified in the taste buds of a one day old animal. The apical parts of dark cells are characterized by numerous dark granules. A distinguishing feature of light cells is the presence of synaptic contacts with afferent intragemmal nerves. On the 4th day of development on the top of the apical parts of the cell, a microvillar apparatus is seen to form, which does not yet communicate with the oral cavity. On the 7th day, basal cells appear in the taste buds. Some of these cells are seen mitotically dividing. The differentiated microvillar apparatus now communicates with oral cavity. The structure of the taste buds is getting similar to that in the adults. The structural and functional peculiarities of the developing taste buds are discussed in association with the period of ontogenesis under consideration.

Animals↗

[Development of the taste buds in rat embryogenesis].

Electron microscopic studies have been made on the developing taste buds in fungiform and vallate papillae of prenatal rats. Three stages of differentiation of these buds are described. The first stage is characterized by presence of the nervous fibers in the connective tissue of the papillae and dense granules of various size, as well as dense-cored vesicles (500-700 A in diameter) in the basal parts of some epithelial cells at the top of the papillae (16-17th days of gestation). The second stage is characterized by nerve processes entering the epithelium and by formation of afferent synaptic contacts between the differentiating epithelial cells and the nervous fibers (19th day of gestation). At the third stage, the cluster of differentiating epithelial cells attains a form which is similar to mature taste buds (21-22nd days of gestation). Thus, to the birthday of rats, differentiation of the basal parts of the taste buds takes place, whereas the apical parts of the taste buds remain undeveloped and do not communicate with the oral cavity. Peculiarities of fine structure of differentiating epithelial cells at the three stages are discussed.

Animals↗

Taste buds of the fungiform papillae in Cynomolgus monkey.

A study of the number of taste buds borne on 145 fungiform papillae from 20 Cynomolgus monkeys showed that, in contrast to the situation in man, most papillae are bud-bearing. The fine structure of taste buds on fungiform papillae of these monkeys was also examined. The cells contained clear apical vesicles, mitochondria, filament bundles and stacks of Golgi cisternae. Nerve fibres containing mitochondria, as well as clear and dense-cored vesicles, were scattered throughout the whole bud, but were more numerous near the base. It was, however, not possible to distinguish different cell types in these taste buds as a function of cell shape, electron density of the ground cytoplasm, presence of different organelles or relation to nerve processes.

Animals↗

Bovine taste bud cyclic adenosine 3', 5' monophosphate phosphodiesterase is inhibited by divalent metal ions.

Two fractions from bovine taste buds, the soluble S4 fraction and the membrane P4B fraction, were used to evaluate the effects of divalent metal ions on cyclic adenosine 3', 5' monophosphate (cAMP) phosphodiesterase (PDE) activity. Zn++, Ni++, Cu++, Fe++, Sn++ and Hg++, in the presence of 5mM Mg++, inhibited cAMP PDE activity whereas these divalent metal ions alone did not affect enzyme activity if Mg++ were absent. Zn++ inhibited cAMP PDE activity in S4 and P4B taste bud fractions with Ki values of 100 microM and 90 microM, respectively; this inhibition was noncompetitive with substrate activity but competitive with Mg++. In the presence of Mg++, Zn++ inhibited taste bud cAMP PDE more effectively than any other metal ion studied. Inhibition of taste bud cAMP PDE by divalent metal ions, particularly Zn++, suggests a role for these substances in the taste process through regulation of intracellular concentration of taste bud cAMP.

3',5'-Cyclic-AMP Phosphodiesterases↗

Thyroid hormone inhibits purified taste bud membrane adenosine 3',5'-monophosphate phosphodiesterase activity.

Thyroid hormone inhibited purified taste bud membrane adenosine 3'5'-monophosphate (cAMP) phosphodiesterase (PDE) activity in a dose-dependent manner. Taste bud membrane cAMP PDE was inhibited most effectively by thyroxine (T4) followed in order by triiodothyionine (T3), diiodotyrosine (DIT) and monoiodotyrosine (MIT). Concentrations required for 50% inhibition (IC50) of enzyme activity were about 1 x 10(-6), 1 x 10(-5), 6 x 10(-4) M and greater than 1 x 10(-3) M for T4, T3, DIT and MIT, respectively. Addition of zinc at physiological concentrations found in serum greatly augmented the inhibitory effects of T4 and T3 at lower concentrations (10(-7) and 10(-6) M, respectively) resulting in further inhibition of cAMP PDE by 40-50%. Inhibition of cAMP PDE by T4 appears to be relatively tissue selective as indicated by the IC50 of 1 x 10(-6) M for the taste bud but only 7 x 10(-6) M, 3 x 10(-5) M, and 4 x 10(-5) M, for heart, kidney and brain cAMP PDE, respectively. Inhibition of taste bud membrane cAMP PDE by T4 was competitive with substrate cAMP with a Ki of 4 microM. These results suggest that inhibition of cAMP PDE, which increases taste bud membrane intracellular cAMP, may participate in the action of thyroid hormone in the taste process.

3',5'-Cyclic-AMP Phosphodiesterases↗

A quantitative study of cat epiglottal taste buds during development.

To quantify the development of upper airway chemoreceptors, epiglottal taste buds were counted in cat fetuses, kittens and adults. Taste buds with structural characteristics similar to those in the adult were observed in all fetuses close to term and in kittens from birth. During the first postnatal week a mean of 76 taste buds was present on the kitten epiglottis and by adulthood 800 were observed. The number of taste buds increased as a logarithmic function of both age and weight, but the correlation with weight is better than that with age. The presence of epiglottal taste buds from very early developmental stages suggests that these receptors may mediate the reflex apnoea and swallowing that occur in response to chemical stimulation of the larynx in newborns.

Aging↗

Particular features of the innervation of taste buds of the epiglottis in monkeys.

The work is devoted to the study of the structure of the innervation apparatus of taste buds in the epiglottis of monkeys (Macacus rhesus). The Campos inpregnation method was used. It is established that several afferent myelinated fibers participate in the innervation of each taste bud of the epiglottis. The peculiarity of structure of their preterminal and terminal parts having the appearance of complex windings and spirals is noted. The polyaxonic principle of the innervation of taste buds of the epiglottis in monkeys is considered as a possible mechanism of generalization of the afferent impulses. It is suggested that the innervation apparatus of the taste buds of the epiglottis constitutes part of unique afferent system of this organ, ensuring its defensive function. The incongruity (from the histophysiological standpoint) of the term 'taste bud' in relation to the epiglottis is noted. It is proposed to call these formations special structures of the chemo-receptors.

Animals↗

Preservation of the ability of dissociated quail wing bud mesoderm to elicit a position-related differentiative response.

Previous studies showed that grafting wedges of fresh or cultured anterior quail wing mesoderm into posterior slits in chick wing buds resulted in the formation of supernumerary cartilage in a high percentage of cases. When anterior quail mesoderm, which had been dissociated into single cells and pelleted by centrifugation, was grafted into posterior slits of host chick wing buds, supernumerary rods or nodules of cartilage formed in 74.3% of the cases. Few supernumerary skeletal structures formed following control operations in which pelleted dissociated anterior or posterior mesoderm was grafted into homologous locations in host chick wing buds. When pelleted, dissociated anterior mesoderm was cultured in vitro for 1 or 2 days prior to being implanted in posterior locations, the incidence of supernumerary cartilage formation increased to 95.5% and 93.8%, respectively. The incidence of supernumerary cartilage formation following control orthotopic grafts of cultured mesoderm was 11.8% for 1-day and 31% for 2-day cultured anterior mesoderm; for 1- and 2-day cultured posterior mesoderm, the incidence of supernumerary cartilage formation was 20% and 41.7%, respectively. Longer-term culture resulted in a substantial decrease in the percentage of supernumerary cartilage after anterior to posterior grafts and an increase in the incidence of supernumerary cartilage from control grafts. The results demonstrate that quail anterior wing bud mesodermal cells do not need to maintain constant contact with one another in order to retain the ability to form or stimulate the formation of supernumerary cartilage after being grafted into a posterior location in a host wing bud. This ability is retained when the pelleted dissociated mesoderm is cultured in vitro outside the limb field for at least 1 to 2 days.

Animals↗

Stage- and region-dependent chondrogenesis and growth of chick wing-bud mesenchyme in serum-containing and defined tissue culture media.

During development, limb-bud mesenchymal cells carry out complex spatiotemporal-patterns of growth and differentiation. Tissue and organ culture facilitate analysis of environmental influences on these cell behaviors, allowing their partial dissection into exogenous and endogenous components. Two factors that complicate such in vitro analyses are the heterogeneity of the cultured cells and imprecise knowledge of culture medium composition. Limb mesenchyme comprises a heterogenous cell population with important regional differences in cell type. Dividing the limb into subregions helps limit the cellular heterogeneity and using chemically defined, serum-free medium alloys concerns about medium composition. In the present study, mesenchyme from different regions along the anteroposterior and proximodistal axes of stage 21-22 and stage 23-24 chick wing buds was grown in high-density microtiter cultures in chemically defined and in serum-containing medium. Four-day cultures of the various regions were compared in terms of culture morphology and the accumulation of Alcian blue-positive cartilage matrix and DNA. The results demonstrate stage- and region-dependent differences in the in vitro growth, differentiation, and responsiveness of these cells. For example, mesenchyme from the distal anterior region of the wing bud exhibited lower intrinsic chondrogenic capacity and greater responsiveness to serum than other regions. Patterns of in vitro chondrogenesis also suggest that, at the stages examined, distal wing-bud mesenchyme may be less homogeneous than has been believed. A case is made for the suitability of serum-free medium for future in vitro studies of chick limb-bud mesenchyme. The results are considered in relation to the process of limb development and regional expression of pattern-related genes.

Animals↗

Scanning electron microscope study of the developing trout pelvic fin bud.

The growth of the pelvic fin bud has been studied with the SEM along with the characteristics of the pseudoapical epidermal ridge which occupies the free margin of the bud. SEM revealed fluffy protuberances in many of the epidermal cells, distinguishing the fin bud territory from adjacent areas. When the pseudoapical ridge appears, all the cells show this feature but their relative number decreases and these cells, termed the "tassel cells," are finally restricted to the base of the fin bud. This particular surface structure of the superficial cells may be unique to the fish, since it has not been heretofore reported in SEM studies of tetrapod limb bud.

Animals↗