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Noradrenergic innervation of the lingual papillae in certain rodents pre-treated with adriblastin. I. Comparative study on the filiform and fungiform papillae.

A comparative study was carried out on the behaviour of the noradrenergic fibres of the filiform and fungiform papillae of four different species of laboratory animals. The adrenergic network proved to be less important in rats and mice than in guinea-pigs and rabbits; moreover, the extreme branchings proved to be more or less closely connected with the taste buds of the mice, guinea-pigs and rabbits. Hypotheses are advanced concerning the possible significance of adrenergic innervation of these lingual papillae.

Adrenergic Fibers↗

Action potentials in epithelial taste receptor cells induced by mucosal calcium.

Chemosensory cells in the taste bud of the tongue of Necturus generate action potentials in response to electrical stimulation through a microelectrode, as recently described by Roper (Science, 220: 1311-1312, 1983). We report that the epithelial receptor cells also respond to 10 mM CaCl2, applied to the mucosal surface, with a depolarization which elicits action potentials when a threshold of -50 mV is reached. Since CaCl2 is one of the taste stimuli in amphibia, the firing of action potentials by chemoreceptor cells may be part of the signal chain in gustatory reception of Ca ions.

Action Potentials↗

Microvasculature of the dorsum of the rat tongue: a scanning electron microscopic study using corrosion casts.

The purpose of this study was to examine the microvascular system of the dorsum of the rat tongue in relation to the subepithelial connective tissue relief. Methylmethacrylate corrosion casts of the vasculature as well as specimens in which the epithelium had been removed were prepared and examined by scanning electron microscopy (SEM). The SEM images revealed four distinct layers in the vasculature of the lamina propria. Superficially, Layer 1 comprised the irregularly shaped ends of capillary loops. Layer 2 consisted of parallel ascending and descending capillaries. These two layers were drained by the subjacent Layer 3, a fish-net-like venous plexus. More deeply, Layer 4 contained arterioles ascending from the muscular layers of the tongue. The vessels of larger papillary structures formed cylindric networks supplied by several arterioles and drained through a single venule to the venous plexus in Layer 3. At their base, the capillary loops often showed a constriction, suggestive of a sphincter. Arteriovenous anastomoses were rare. The epithelium-connective tissue interface showed short conical papillae as well as larger papillary structures which corresponded in both distribution and size to the capillary loops found in the vascular casts. The results confirm that the tongue is highly vascularized and demonstrate that the vascular supply to the papillae, taste buds, and other surface structures forms a highly organized and specific system which is adapted to the specific functions of this organ in the rat.

Animals↗

Effect of ciguatoxin 3C on voltage-gated Na+ and K+ currents in mouse taste cells.

The marine dinoflagellate Gambierdiscus toxicus produces highly lipophilic, polycyclic ether toxins that cause a seafood poisoning called ciguatera. Ciguatoxins (CTXs) and gambierol represent the two major causative agents of ciguatera intoxication, which include taste alterations (dysgeusiae). However, information on the mode of action of ciguatera toxins in taste cells is scarce. Here, we have studied the effect of synthetic CTX3C (a CTX congener) on mouse taste cells. By using the patch-clamp technique to monitor membrane ion currents, we found that CTX3C markedly affected the operation of voltage-gated Na(+) channels but was ineffective on voltage-gated K(+) channels. This result was the exact opposite of what we obtained earlier with gambierol, which inhibits K(+) channels but not Na(+) channels. Thus, CTXs and gambierol affect with high potency the operation of separate classes of voltage-gated ion channels in taste cells. Our data suggest that taste disturbances reported in ciguatera poisoning might be due to the ability of ciguatera toxins to interfere with ion channels in taste buds.

Animals↗

Characterization of bitter taste responses of intestinal STC-1 cells.

Cellular responses of STC-1 cells to two bitter tastants (denatonium and caffeine) were investigated using a calcium-imaging technique and compared with the response to bombesin. Caffeine is known to stimulate taste receptor cells, but the properties of its signaling have not been well studied. STC-1 cells responded to all three molecules in a dose-dependent manner, and when a reverse transcriptase-polymerase chain reaction (RT-PCR) for denatonium receptor was performed, the product of predicted size was detected in STC-1 cells. Furthermore, all three signaling pathways were blocked by a phospholipase C (PLC) inhibitor, demonstrating the essential involvement of PLC in cellular responses. To study the regulatory system of G protein signaling in STC-1 cells, we searched G protein-coupled receptor kinases (GRKs) by the degenerate-primer PCR method and found that GRK2 is expressed. We also demonstrated that three GRKs (GRK2, GRK3 and GRK5) are differentially distributed in the circumvallate papilla while only GRK2 is present in taste bud cells. Finally, we overexpressed GRK2 in SCT-1 cells and found that bombesin-induced response was strongly inhibited by GRK2 but denatonium-activated signaling was not affected. In the case of caffeine, response was decreased by expression of GRK2 only when cells were activated by 1 mM caffeine. Thus, we showed that STC-1 cells emerge as a cell model for studying the molecular mechanism of bitter taste signaling, and could indicate properties of caffeine-induced signaling in comparison with other signaling.

Animals↗

Distinctive spatiotemporal expression patterns for neurotrophins develop in gustatory papillae and lingual tissues in embryonic tongue organ cultures.

Brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) mRNAs are expressed in the developing rat tongue and taste organs in specific spatiotemporal patterns. BDNF mRNA is present in the early lingual gustatory papilla epithelium, from which taste buds eventually arise, prior to the arrival of gustatory nerve fibers at the epithelium, whereas NT-3 initially distributes in the mesenchyme. However, a direct test for neural dependence of neurotrophin expression on the presence of innervation in tongue has not been made, nor is it known whether the patterns of neurotrophin expression can be replicated in an in vitro system. Therefore, we used a tongue organ culture model that supports taste papilla formation while eliminating the influence from sensory nerve fibers, to study neurotrophin mRNAs in lingual tissues. Rat tongue cultures were begun at embryonic day 13 or 14 (E13, E14), and BDNF, NT-3, nerve growth factor (NGF) and neurotrophin-4 (NT-4) mRNAs were studied at 0, 2, 3 and 6 days in culture. BDNF transcripts were localized in the gustatory epithelium of both developing fungiform and circumvallate papillae after 2 or 3 days in culture, and NT-3 transcripts were in the subepithelial mesenchyme. The neurotrophin distributions were comparable to those in vivo at E13-E16. In 6-day tongue cultures, however, BDNF transcripts in anterior tongue were not restricted to fungiform papillae but were more widespread in the lingual epithelium, while the circumvallate trench epithelium exhibited restricted BDNF labeling. The NT-3 expression pattern shifted in 6-day organ cultures in a manner comparable to that in the embryo in vivo, and was expressed in the lingual epithelium as well as mesenchyme. NGF mRNA expression was subepithelial throughout 6 days in cultures. NT-4 mRNA was not detected. The neurotrophin mRNA distributions demonstrate that temporospatial localization of neurotrophins observed during development in vivo is retained in the embryonic tongue organ culture system. Furthermore, initial neurotrophin expression in the developing lingual epithelium, mesenchyme, and/or taste papillae is not dependent on intact sensory innervation. We suggest that patterns of lingual neurotrophin mRNA expression are controlled by the influence of local tissue interactions within the tongue at early developmental stages. However, the eventual loss of restricted BDNF mRNA localization from fungiform papillae in anterior tongue suggests that sensory innervation may be important for restricting the localized expression of neurotrophins at later developmental stages, and for maintaining the unique phenotypes of gustatory papillae.

Animals↗

Immunohistochemical investigation of the nitrergic system in the taste organ of the frog, Rana esculenta.

We have studied by immunocytochemistry, the taste discs of the frog, Rana esculenta, with the aim of providing morphological and neurochemical data on the nitrergic system and of assessing the eventual presence of intrinsic neurons associated with the gustatory organs. In taste discs, antibodies against neuronal nitric oxide synthase (nNOS) revealed a positive immunoreaction in the taste receptor cell bodies and processes. The basal cells were also stained. All the fungiform papillae contained intragemmal nerve fibers showing nNOS immunoreactivity; these fiber were mainly located in the basal plexus. Immunoreactive nerve fibers were also visible at the periphery of the papilla-contacting ciliate cells, which form a ring around the taste disc. In conclusion, the findings obtained in this study suggest that the occurrence of nNOS-immunoreactivity in basal cells, taste cells and nerves might reflect a role for nitric oxide in taste mechanisms of Amphibia. The results may also sustain the physiological implication of NO as a molecule involved in the local target function of maintaining the taste bud mucosal integrity and in regulating the blood flow to the epithelium.

Animals↗

Immunohistochemical study of the innervation of the boundary area of the hard and soft palates of the rat.

The palatal mucosa plays an important role for patients using full dentures. The posterior ridge of the denture is designed to fit on the border between the hard and soft palates; accordingly, this boundary area is of importance when the outline of the denture is designed. In the present study, a rich supply of nerve fibers was found in the mucosa of the boundary area of the hard and soft palates of the rat. An immunocytochemical examination revealed nerve fibers containing protein gene product 9.5 (PGP 9.5), calcitonin gene-related peptide (CGRP), substance P (SP), pituitary adenylate cyclase-activating peptide, vasoactive intestinal polypeptide (VIP), C-terminal flanking peptide of neuropeptide Y (c-PON), or nitric oxide synthase (NOS). Thin nerve fibers with PGP 9.5, CGRP, or SP penetrated into the epithelium, reaching beneath the cornified layer and terminated as free nerve endings. VIP-, c-PON- and NOS-containing nerve fibers were distributed in the connective tissue. Many of the VIP- and c-PON-containing nerve fibers were associated with blood vessels. In addition, nerve fibers containing PGP 9.5, CGRP, SP and c-PON were observed around, and penetrating into, the taste buds in the boundary area.

Animals↗

Geriatric nutrition: the role of taste and smell in appetite.

Proper nutrition is essential to the health of the oral tissues, and healthy tissues enhance prosthodontic treatment of the elderly. All dentists should be prepared to offer dietary advice to this expanding population. Taste and smell are essential to proper nutrition. In the elderly the peripheral sensory receptors decline, causing the appetite to wane. Taste and aroma are inextricably intertwined in determining the palatability and acceptance of food. For example, during an upper respiratory infection, the olfactory receptors are blocked. Food becomes tasteless; it loses both flavor and aroma and the appetite declines. As a result of aging, the taste buds on the tongue and the olfactory receptors in the roof of the nasal cavity regress. In addition, the gustatory and olfactory nuclei in the brain decline, causing a reduction in appetite and diet.

Aged↗

Trophic capacity of experimentally lengthened gustatory axons.

Long nerve stumps will maintain end-organs for a longer period of time than short nerve stumps. However, longer maintenance might result not from a longer nerve stump per se but from uncontrolled factors such as less interference with the target's blood supply when transecting a nerve far from its target. To control for the site of nerve transection and to evaluate the trophic capacity of lengthened axons, we extended the gerbil's (Meriones unguiculatus) IXth nerve (glossopharyngeal) by splicing in a 5.0 mm segment of the superior laryngeal nerve. Thus, when assessing nerve stump-length dependency 3-5 months later, both the control regenerated nerve and the lengthened experimental nerve could be transected at the same site on the IXth nerve trunk. The peak magnitude of the summated taste discharges of the experimentally lengthened IXth nerve declined at a rate exactly predicted by an earlier study of nerve stump-length dependency. We conclude that maintenance of synaptic transmission depends upon taste axon stump-length and not upon factors uncontrolled in conventional experimental designs of stump-length dependency which transect nerves at different locations. Control regenerated and experimentally lengthened nerves maintained similar numbers of taste buds. Thus, neither the added burden of maintaining longer axons nor the presence of Schwann cells of the superior laryngeal nerve splice degraded the IXth nerve's trophic performance.

Animals↗

Anatomy and histology of the brain and sense organs of the antarctic plunderfish Dolloidraco longedorsalis (Perciformes: Notothenioidei: Artedidraconidae), with comments on the brain morphology of other artedidraconids and closely related harpagiferids.

In the high-latitude shelf waters of Antarctica, fishes in the perciform suborder Notothenioidei dominate the fish fauna and constitute an adaptive radiation and a species flock. The 25 species of notothenioid plunderfishes, comprising four genera of the family Artedidraconidae, contribute substantially to fish species diversity on the high Antarctic shelf. A mental barbel is an autapomorphy for the family. Dolloidraco longedorsalis is the most abundant artedidraconid at depths over 400 m in these waters. In this article we present the anatomy and histology of the brain and special sense organs of Dolloidraco and compare it to the brains of other artedidraconids, closely related harpagiferids, and more generally to other notothenioids. We provide a detailed drawing of the brain and cranial nerves. The brain of Dolloidraco is simple, without external hypertrophy of sensory or motor regions, but contains several unusual features associated with the ventricular system and CSF, including well-developed circumventricular organs, subependymal expansions, and subarachnoid cisterns; and a ventricle in the corpus cerebellum. The brain of Dolloidraco also contains a lobed chief sensory nucleus of the trigeminal nerve that is correlated across species with barbel length. The eyes are large and contain a small choroid rete, a structure previously thought to be absent from members of this family. We document the histology of the duplex retina, olfactory apparatus, cutaneous taste buds, and barbel musculature and innervation. We discuss the role of pedomorphy in producing simplified brain morphologies. We consider the possibility that Dolloidraco is a somatosensory specialist-an unusual feature among vertebrates-and decide that this is unlikely.

Animals↗

Postnatal development of von Ebner's glands: accumulation of a protein of the lipocalin superfamily in taste papillae of rat tongue.

Antibodies produced against rat von Ebner's gland (VEG) protein, a recently characterized member of a lipophilic ligand carrier protein family, detect this protein immunocytochemically in von Ebner's gland acini and show that it is present at high concentrations in the clefts of circumvallate and foliate papillae. During embryonic development, von Ebner's gland anlagen are innervated (as shown immunocytochemically using neuronal specific antibodies) as early as embryonic day 20, before lateral glandular outgrowth and VEG protein can be observed. Expression of the VEG protein as determined by in situ hybridization and immunocytochemistry begins at postnatal day-2 cells in differentiating and branching off from von Ebner's gland ducts, and sharply increases with further enlargement and maturation of the gland. The close temporal correlation of von Ebner's gland innervation and VEG protein expression with papilla innervation and taste-bud development suggests a functional relationship of both structures. VEG protein might control access of lipophilic sapid molecules, such as bitter substances, to the gustatory receptors.

Amino Acid Sequence↗

Enkephalin immunoreactivity in the paraneurons of the tiger salamander (Ambystoma tigrinum) tongue.

Immunohistochemical tests have demonstrated the presence of leu-5-enkephalin and other bioactive compounds (serotonin and neuron-specific enolase) in the basal cells of lingual taste buds in Ambystoma tigrinum; there was also a weak reaction for met-5-enkephalin. Similar reactions were obtained from particular cells dispersed within the lingual epithelium, which are provisionally identified as Merkel cells.

Ambystoma↗

A comparison of the effects of bilateral sections of the chorda tympani nerve and extirpation of the submaxillary and sublingual salivary glands on the eating and drinking patterns of the rat.

The chorda tympani nerve (CT) innervates the fungiform papillae on the tip of the tongue and has been considered an important nerve for the sense of taste. The CT also contains the parasympathetic supply to the submaxillary and sublingual salivary glands. Therefore, changes in taste or feeding behavior following bilateral sections of CT are caused by both degeneration of fungiform papillae and the inevitable partial desalivation of the rat. In the present experiments we compared the effects of bilateral chorda tympani nerve sections with extirpation of submaxillary and sublingual glands on daily home cage eating and drinking patterns in the rat. Before and after surgery we analyzed the daily eating and drinking patterns, including such measures as intake, bout number, bout length, interbout interval and rate of consumption during bouts. The results of desalivation and bilateral CT sections were indistinguishable. The most profound change was that eating bout duration was increased following surgery. Since food intake did not increase, the results indicate a marked loss in eating efficiency over the daily ingestion periods. Although the eating patterns of desalivated and chorda tympani sectioned rats are quite similar, the evidence is not compelling that they have the same physiological basis. A second experiment was designed to test the hypothesis that the atypical eating patterns observed following bilateral sectioning of CT were the direct result of partial desalivation resulting from the denervation of the salivary glands. In this experiment a unilateral section was made of one CT and it was shown that the eating behavior was not affected. Then the contralateral submaxillary and sublingual salivary glands were removed. This resulted in a six-fold increase in feeding bout length. In all cases a unilateral CT section combined with extirpation of the contralateral salivary glands resulted in rats whose eating behavior was indistinguishable from the earlier data following either the bilateral CT sections or bilateral desalivations. The conclusion is drawn that the eating irregularities noted following bilateral CT sections result from this partial desalivation. CT sections were verified by taste bud counts in the fungiform papillae and histological examinations were made of salivary glands in rats receiving CT sections.

Animals↗

Response of the taste receptor cell to the umami-substance stimulus. An electron-microscopic study.

The chemosensory gustatory cells in taste buds form chemical synapses with intragemmal neurites. We investigated the ultrastructure of the guinea pig gustatory cells after stimulation with a mixture of monosodium L-glutamate and guanosine 5'-monophosphate. The gustatory cells responded to the stimulus. The dense-cored vesicles localized in the presynaptic regions discharged the contents into the synaptic cleft by means of exocytosis, which resulted in a marked decrease of their population. These findings strongly suggest that the transmitter or transmitters contained in the vesicles are released in response to the taste stimulation at the cell apex, to conduct the excitement of the cell to the nerves.

Animals↗

Peptide 19 in the rat vagal and glossopharyngeal sensory ganglia.

Peptide 19 (PEP 19) is a 7.6-kDa polypeptide which binds to calmodulin and inhibits calcium-calmodulin signaling. In this study, PEP 19-immunoreactivity (PEP 19-IR) was examined in the rat vagal and glossopharyngeal sensory ganglia. Twenty-nine percent, 59%, and 41% of sensory neurons contained PEP 19-IR in the jugular, petrosal, and nodose ganglia, respectively. These neurons were of various sizes (jugular, mean +/- SD = 635.8 +/- 392.6 microm2, range = 105.9-1695.9 microm2; petrosal, mean +/- SD = 370.9 +/- 228.5 microm2, range = 57.7-1662.7 microm2; nodose, mean +/- SD = 380.5 +/- 157 microm2, range = 87.5-950.4 microm2) and scattered throughout these ganglia. Double immunofluorescence method revealed that PEP 19-IR neurons which had parvalbumin-IR were rare in the ganglia (jugular, 4%; petrosal, 10%; nodose, 8%). PEP 19-IR neurons which contained calbindin D-28k were abundant in the petrosal (20%) and nodose (22%) ganglia but not in the jugular ganglion (8%). Retrograde tracing method indicated that many PEP 19-IR neurons projected to the circumvallate papilla and soft palate. In the soft palate, taste buds were innervated by PEP 19-IR nerve fibers. The present study suggests that PEP 19-IR neurons include chemoreceptors in the vagal and glossopharyngeal sensory ganglia.

Animals↗

Co-expression of VRL-1 and calbindin D-28k in the rat sensory ganglia.

The co-expression of vanilloid receptor 1-like receptor (VRL-1), a newly cloned capsaicin-receptor homologue, with calbindin D-28k was examined in the rat sensory ganglia. The co-expression was rare in the dorsal root, trigeminal and jugular ganglia and abundant in the petrosal and nodose ganglia. In the dorsal root ganglion, none of VRL-1-immunoreactive (ir) neuron co-expressed calbindin D-28k-immunoreactivity (ir). Of the VRL-1-ir neurons, 9 and 5% showed calbindin D-28k ir in the trigeminal and jugular ganglia, respectively. On the other hand, 35 and 63% of VRL-1-ir neurons in the petrosal and nodose ganglia, respectively, co-expressed these substances. The retrograde tracing method indicated that petrosal neurons which co-expressed VRL-1-and calbindin D-28k-ir innervated taste buds in the circumvallate papilla. The present findings may suggest that VRL-1 is associated with chemosensory functions in visceral sensory neurons.

Animals↗

Postnatal development of membrane excitability in taste cells of the mouse vallate papilla.

The mammalian peripheral taste system undergoes functional changes during postnatal development. These changes could reflect age-dependent alterations in the membrane properties of taste cells, which use a vast array of ion channels for transduction mechanisms. Yet, scarce information is available on the membrane events in developing taste cells. We have addressed this issue by studying voltage-dependent Na+, K+, and Cl- currents (I(Na), I(K), and I(Cl), respectively) in a subset of taste cells (the so-called "Na/OUT" cells, which are electrically excitable and thought to be sensory) from mouse vallate papilla. Voltage-dependent currents play a key role during taste transduction, especially in the generation of action potentials. Patch-clamp recordings revealed that I(Na), I(K), and I(Cl) were expressed early in postnatal development. However, only I(K) and I(Cl) densities increased significantly in developing Na/OUT cells. Consistent with the rise of I(K) density, we found that action potential waveform changed markedly, with an increased speed of repolarization that was accompanied by an enhanced capability of repetitive firing. In addition to membrane excitability changes in putative sensory cells, we observed a concomitant increase in the occurrence of glia-like taste cells (the so called "leaky" cells) among patched cells. Leaky cells are likely involved in dissipating the increase of extracellular K+ during action potential discharge in chemosensory cells. Thus, developing taste cells of the mouse vallate papilla undergo a significant electrophysiological maturation and diversification. These functional changes may have a profound impact on the transduction capabilities of taste buds during development.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗