Species generalization and differences in Hedgehog pathway regulation of fungiform and circumvallate papilla taste function and somatosensation demonstrated with sonidegib.

Species generalization in the profound, modality-specific effects of Hedgehog pathway inhibition (HPI) in taste organ homeostasis and sensation is shown. With the HPI, cancer drug sonidegib, we demonstrate that the rat taste system, in addition to mouse, is regulated by Hedgehog signaling. After sonidegib treatment for 16-36 days in rat, there is loss of taste buds (TB) in soft palate, in fungiform (FP) and circumvallate papillae (CV), and elimination of taste responses from chorda tympani and glossopharyngeal nerves. The retained innervation in FP and CV during HPI cannot sustain TB. Responses to tactile stimuli are not altered, and temperature responses are reduced only after 28 days treatment, demonstrating modality-specific effects. Rat FP and neural effects are similar to those in mouse whereas TB and neural response effects from the rat CV are much more severe. When recovery is introduced in mouse after prolonged, 48 days HPI, the TB in CV are restored whereas those in FP are not. Overall, Hedgehog signaling regulation is shown to generalize to the rat taste system, and the modality-specific controls in taste organ sensation are affirmed. The reported, debilitating taste disturbances in patients who use HPI drugs can be better understood based on these data.

Embryonic geniculate ganglion neurons in culture have neurotrophin-specific electrophysiological properties.

Geniculate ganglion neurons provide a major source of innervation to mammalian taste organs, including taste buds in the soft palate and in fungiform papillae on the anterior two thirds of the tongue. In and around the fungiform papillae, before taste buds form, neurotrophin mRNAs are expressed in selective spatial and temporal patterns. We hypothesized that neurotrophins would affect electrophysiological properties in embryonic geniculate neurons. Ganglia were explanted from rats at gestational day 16, when growing neurites have entered the papilla core, and maintained in culture with added brain-derived neurotrophic factor (BDNF), neurotrophin 4 (NT4), nerve growth factor (NGF) or neurotrophin 3 (NT3). Neuron survival with BDNF or NT4 was about 80%, whereas with NGF or NT3 less than 15% of neurons survived over 6 days in culture. Whole cell recordings from neurons in ganglion explants with each neurotrophin condition demonstrated distinctive neurophysiological properties related to specific neurotrophins. Geniculate neurons cultured with either BDNF or NT4 had similar passive-membrane and action potential properties, but these characteristics were significantly different from those of neurons cultured with NGF or NT3. NGF-maintained neurons had features of increased excitability including a higher resting membrane potential and a lower current threshold for the action potential. About 70% of neurons produced repetitive action potentials at threshold. Furthermore, compared with neurons cultured with other neurotrophins, a decreased proportion had an inflection on the falling phase of the action potential. NT3-maintained neurons had action potentials that were of relatively large amplitude and short duration, with steep rising and falling slopes. In addition, about 20% responded with a repetitive train of action potentials at threshold. In contrast, with BDNF or NT4 repetitive action potential trains were not observed. The data demonstrate different neurophysiological properties in developing geniculate ganglion neurons maintained with specific neurotrophins. Therefore, we suggest that neurotrophins might influence acquisition of distinctive neurophysiological properties in embryonic geniculate neurons that are fundamental to the formation of peripheral taste circuits and a functioning taste system.

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.

Alterations in size, number, and morphology of gustatory papillae and taste buds in BDNF null mutant mice demonstrate neural dependence of developing taste organs.

Sensory ganglia that innervate taste buds and gustatory papillae (geniculate and petrosal) are reduced in volume by about 40% in mice with a targeted deletion of the gene for brain-derived neurotrophic factor (BDNF). In contrast, the trigeminal ganglion, which innervates papillae but not taste buds on the anterior tongue, is reduced by only about 18%. These specific alterations in ganglia that innervate taste organs make possible a test for roles of lingual innervation in the development of appropriate number, morphology, and spatial pattern of fungiform and circumvallate papillae and associated taste buds. We studied tongues of BDNF null mutant and wild-type littermates and made quantitative analyses of all fungiform papillae on the anterior tongue, the single circumvallate papilla on the posterior tongue, and all taste buds in both papilla types. Fungiform papillae and taste buds were reduced in number by about 60% and were substantially smaller in diameter in mutant mice 15-25 days postnatal. Remaining fungiform papillae were selectively concentrated in the tongue tip region. The circumvallate papilla was reduced in diameter and length by about 40%, and papilla morphology was disrupted. Taste bud number in the circumvallate was reduced by about 70% in mutant tongues, and the remaining taste buds were smaller than those on wild-type tongues. Our results demonstrate a selective dependence of taste organs on a full complement of appropriate innervation for normal growth and morphogenesis. Effects on papillae are not random but are more pronounced in specific lingual regions. Although the geniculate and petrosal ganglia sustain at least half of their normal complement of cell number in BDNF -/- mice, remaining ganglion cells do not substitute for lost neurons to rescue taste organs at control numbers. Whereas gustatory ganglia and the taste papillae initially form independently, our results suggest interdependence in later development because ganglia derive BDNF support from target organs and papillae require sensory innervation for morphogenesis.

The role of innervation in induction and differentiation of taste organs: introduction and background.

To establish lingual receptive fields that are the basic unit of taste function, ganglion cells must extend neurites of peripheral and central targets and form connections. This symposium concerns developmental interactions between the geniculate, trigeminal and petrosal ganglia and peripheral taste organs, the gustatory papillae and resident taste buds. Investigators present data from organ and tissue culture, from mice with targeted gene deletions and from grafting experiments, in pursuit of principles that direct early innervation of the taste system. The lingual ganglia and the taste papillae initially develop independently, but then become reciprocally dependent as ganglia drive neurotrophin support from gustatory papillae and the papillae require sensory innervation for growth and morphogenesis. The issue of subsequent taste bud induction is discussed with results from amphibian and mammalian models, yielding conclusions that are not yet totally convergent. However, an essential role for sensory innervation in mammalian taste bud differentiation and acquisition of appropriate quantitative relations between ganglion cells and target organs is clearly demonstrated. A working outline is presented for periods of ganglion cell/target organ independence and interdependence during early innervation of the peripheral taste system.

Measurement of coronary blood flow and flow reserve using magnetic resonance imaging.

PURPOSE: It was the purpose of this study to demonstrate the feasibility of performing coronary artery flow and coronary flow reserve (CFR) measurements in normal human volunteers using a magnetic resonance (MR) phase contrast technique. MATERIALS AND METHODS: Coronary flow rate, flow velocity, peak flow and CFR were determined at rest and during pharmacologically induced hyperemia in 10 healthy volunteers. The flow measurements were obtained during a single breath-hold by using a fast, prospectively gated, segmented k-space gradient-echo phase contrast acquisition with view sharing (FASTCARD PC) that was modified to improve sampling of the diastolic flow. Data were processed using the standard phase difference (PD) processing techniques as well as a new complex difference (CD) flow measurement method intended to improve the accuracy of flow measurements in small vessels. RESULTS: Mean hyperemic flow velocity (40 +/- 16 cm/s) and blood flow (3.9 +/- 1.5 ml/s) rates differed significantly from resting velocity (13 +/- 6.6 cm/s) and flow (1.1 +/- 0.4 ml/s) measurements (p < 0.0001). PD methods consistently measured larger flow rates at rest (24% larger, p < 0.0005) and stress (29% larger, p < 0.0001). CFR, calculated as the ratio of the mean PD flows (4.7 +/- 2.8), was higher than CFR calculated as the ratio of mean CD flows (4.2 +/- 1.8); however, the differences did not reach statistical significance (p = 0.07). Flow measurements performed in adjacent slices of the same vessel correlated well (r = 0.88). CONCLUSIONS: Coronary flow and CFR measurements using the MR techniques are feasible and are similar to those reported in the literature for healthy volunteers.

Organ cultures of embryonic rat tongue support tongue and gustatory papilla morphogenesis in vitro without intact sensory ganglia.

Taste buds on the mammalian tongue are confined to the epithelium of three types of gustatory papillae: the fungiform, circumvallate, and foliate. The gustatory papillae are composed of an epithelium that covers a broad connective tissue core, with extensive innervation to taste bud and nongustatory epithelial locations. Although the temporal sequence of gustatory papilla development is known for several species, factors that regulate initiation, growth, and maintenance of the papillae are not understood. We tested the hypothesis that sensory innervation is required for the initial formation and early morphogenesis of fungiform papillae in a patterned array. An organ culture of the embryonic rat tongue was developed to provide an in vitro system for studying mechanisms involved in fungiform papilla morphogenesis in patterns on the anterior tongue. Tongues were dissected from embryos at 13 days of gestation (E13), a time when the tongue has not yet fully formed and gustatory papillae have not yet appeared, and at 14 days of gestation (E14), when the tongue is well formed and papillae make their initial morphological appearance. Dissected tongues were maintained at the gas/liquid interface in standard organ culture dishes, fed with DMEM/F12 plus 2% B-27 supplement and 1% fetal bovine serum. After 1, 2, 3, or 6 days in culture, tongues were processed for scanning electron or light microscopy, or immunocytochemistry. Tongues cultured from E13 or E14 underwent extensive morphogenesis and growth in vitro. Furthermore, fungiform papillae developed on these tongues on a culture day equivalent to E15 in vivo; that is, after 2 days for cultures begun at E13 and 1 day for those begun at E14. Because E15 is the characteristic time for gustatory papilla formation in the intact embryo, results demonstrate that the cultured tongues retain important temporal information related to papilla development. In addition, fungiform papillae formed in the tongue cultures in the stereotypic pattern of rows. The papillae were large structures with epithelial and mesenchymal cell integrity, and an intact epithelial basement membrane was indicated with laminin immunoreactivity. The cultures demonstrate that gustatory papilla morphogenesis can progress in the absence of an intact sensory innervation. To exclude a potential developmental role for autonomic ganglion cells that are located in the posterior rat tongue, cultures consisting of only the anterior half of E14 tongues were established. Fungiform papilla development progressed in half tongues in a manner directly comparable to whole tongue cultures. Therefore, robust, reproducible development of fungiform papillae in patterns is supported in rat tongue cultures from E13 or E14, without inclusion of intact sensory or major, posterior tongue autonomic ganglia. This is direct evidence that papillae will form and develop further in vitro without sensory ganglion support. The data also provide the first detailed account of in vitro development of the entire embryonic tongue.

Initial innervation of embryonic rat tongue and developing taste papillae: nerves follow distinctive and spatially restricted pathways.

The rat tongue has an extensive, complex innervation from four cranial nerves. However, the precise developmental time course and spatial routes of these nerves into the embryonic tongue are not known, although this knowledge is crucial for studying mechanisms that regulate development and innervation of the lingual taste organs, gustatory papillae and resident taste buds. We determined the initial spatial course of nerves in the developing tongue and papillae, and tested the hypothesis that sensory nerves first innervate the tongue homogeneously and then retract to more densely innervate papillae and taste buds. Antibodies to GAP-43 and neurofilaments were used to label nerve fibers in rat embryo heads from gestational day 11 through 16 (E11-E16). Serial sagittal sections were traced and reconstructed to follow paths of each nerve. In E11 rat, geniculate, trigeminal and petrosal ganglia were labeled and fibers left the ganglia and extended toward respective branchial arches. At E13 when the developing tongue is still a set of tissue swellings, the combined chorda/lingual, hypoglossal and petrosal nerves approached the lingual swellings from separate positions. Only the chorda/lingual entered the tongue base at this stage. At E14 and E15, the well-developed tongue was innervated by all four cranial nerves. However, the nerves maintained distinctive entry points and relatively restricted mesenchymal territories within the tongue, and did not follow one another in common early pathways. Furthermore, the chorda/lingual and glossopharyngeal nerves did not set up an obvious prepattern for gustatory papilla development, but rather seemed attracted to developing papillae which became very densely innervated compared to surrounding epithelium at E15. To effect this dense papilla innervation, sensory nerves did not first innervate the tongue in a homogeneous manner with subsequent retraction and/or extensive redirection of fibers into the taste organs. Results contribute to a set of working principles for development of tongue innervation. Points of entry and initial neural pathways are restricted from time of tongue formation through morphogenesis, suggesting distinctive lingual territories for each nerve. Thus, sensory and motor nerves distribute independently of each other, and sensory innervation to anterior and posterior tongue remains discrete. For taste organ innervation, gustatory papillae are not induced by a prepatterned nerve distribution. In fact, papillae might attract dense sensory innervation because neither chorda/lingual nor glossopharyngeal nerve grows homogeneously to the lingual epithelium and then redistributes to individual papillae.

Temporal and spatial patterns of tenascin and laminin immunoreactivity suggest roles for extracellular matrix in development of gustatory papillae and taste buds.

Gustatory papillae are complex organs that are composed of 1) an epithelium, 2) specialized sensory cells within the epithelium (the taste buds), 3) a broad connective core, and 4) sensory innervation. During papilla development, cells in the various tissue compartments must divide, aggregate, detach, migrate, and reaggregate in relation to each other, but factors that regulate such steps are poorly understood and have not been extensively studied. All of these processes potentially require participation of the extracellular matrix. Therefore, we have studied temporal and spatial patterns of immunoreactivity for two extracellular matrix molecules, tenascin and laminin, in the developing fungiform and circumvallate papillae of fetal, perinatal, and adult sheep tongue. To determine relations of tenascin and laminin to sensory innervation, we used an antibody to growth-associated protein (GAP-43) to label growing nerves. Immunocytochemical distributions of tenascin and laminin alter during development in a manner that reflects morphogenesis rather than histologic boundaries of the taste papillae. In early fungiform papillae, tenascin immunoreactivity is very weak within the mesenchyme of the papilla core. However, there is a subsequent shift to an intense, restricted localization in the apical papilla core only--directly under taste bud-bearing regions of the papilla epithelium. In early circumvallate papillae, tenascin immunoreactivity is patchy within the papilla core and within the flanking, nongustatory papillae. Later, immunoreactivity is restricted to the perimeter of the central papilla core, under epithelium that contains developing taste buds. In fungiform and circumvallate papillae, the shift in tenascin immunolocalization is associated with periods of taste bud formation and multiplication within the papilla epithelium and with extensive branching of the sensory innervation in the papilla apex. Laminin immunoreactivity, although it is continuous throughout the basement membrane of general lingual epithelium, is interrupted in the epithelial basement membrane of early fungiform and circumvallate papillae in regions where taste buds are forming. The breaks are large in young fetuses, when taste buds first develop, and are evidenced later as punctate disruptions. Heparan sulfate proteoglycan immunoreactivity confirms that these are basement membrane discontinuities. GAP-43 label coincides with innervation of the papilla core and is most extensive in regions where tenascin immunoreactivity is weak or absent. GAP-43 immunoreactivity is also found in early taste buds: Later, it is extensive within more mature multiple taste buds, presumably in relation to synaptogenesis. We propose that tenascin has a role in promoting deadhesion of cells in the papilla epithelium during periods of taste bud formation and multiplication. Discontinuities in the epithelial basement membrane under developing taste buds, indicated with laminin and heparan sulfate proteoglycan immunoreactivity, may interact to facilitate taste bud morphogenesis and multiplication, to permit access of papilla innervation to the forming taste buds, and/or to allow epithelial/mesenchymal interactions during papilla and taste bud development.

Development of intrinsic electrophysiological properties in neurons from the gustatory region of rat nucleus of solitary tract.

There is no current understanding of the nature or time course of maturation of intrinsic electrophysiological properties for neurons in the gustatory region of the nucleus of the solitary tract (NST). Therefore, we used whole cell recordings in an in vitro slice preparation of the rat brainstem to characterize development of resting membrane, action potential and repetitive discharge properties of cells in gustatory NST at postnatal days 5, 10, 15, 20, and 30, and adult ages. Neurons were filled with Biocytin to verify location and characterize morphology. Membranes from younger neurons demonstrated a steeper current-voltage relation or higher input resistance, and a longer time constant than mature cells. Action potentials in younger cells had a slower rate of rise and were longer in duration. The afterhyperpolarization that typically follows the spike discharge usually had one phase in younger neurons, but was characterized by two or more phases in an increasing proportion of older cells. The repetitive discharge frequency in response to a range of depolarizing current pulses increased during development, and frequency/current plots were steeper in older compared with younger neurons. However, in all age groups there was clear accommodation of the discharge frequency. The greatest changes in resting membrane, action potential, and discharge properties were observed between P5 and P15, and mature values were generally reached by P20. At each postnatal age, neurons could be categorized in four neuron groups, based on the discharge pattern in response to a hyperpolarizing/depolarizing current protocol. Anatomical reconstructions indicated that although cells increased in overall dendritic expanse during development, neurons became less complex as illustrated by decreases in number of dendritic branch points, and in number and density of spines. The timing of major developmental differences in intrinsic electrical characteristics observed here is associated with a period of previously reported maturational changes in extracellular taste responses to number and concentration of chemical stimuli. However, further alterations in extracellular taste responses proceed after apparent maturation of intrinsic neural properties.

Maturation of neuron types in nucleus of solitary tract associated with functional convergence during development of taste circuits.

Late fetal through postnatal development in sheep is a period of increasing convergence of afferent taste fibers onto second-order neurons in the nucleus of the solitary tract (NST). To learn whether neuron morphology alters in concert with convergence and neurophysiological development in NST, three-dimensional neuron reconstructions were made of cells in a functionally defined region of gustatory NST from Golgi preparations of the brainstem. Elongate, multipolar, and ovoid neurons were studied in fetuses from 85 days of gestation through the perinatal period (term = 147 days of gestation), to postnatal stages. Somal size and form, and dendritic complexity and extent, increased markedly from 85 to about 110 days of gestation in both of the proposed NST projection neurons, elongate and multipolar. From 130 days of gestation to postnatal ages, growth of dendrites of elongate neurons plateaued or declined, whereas dendrites of multipolar neurons apparently continued to increase in size and extent. In addition, spine density decreased on elongate neurons but remained stable on multipolar neurons. Morphological variables of ovoid cells, proposed interneurons in NST, did not alter over this later period. The data suggest that multipolar, not elongate or ovoid, neurons are logical candidates to receive the increasing afferent fiber input onto NST cells during late gestation. Also, neural activity from taste afferent fibers is more likely to have a role in altering NST neuron morphology at later, rather than earlier, developmental periods.

4-Aminopyridine reduces chorda tympani nerve taste responses to potassium and alkali salts in rat.

To study the potential role of potassium channels in the taste response to potassium salts, we applied 4-aminopyridine (4-AP) to the anterior rat tongue and recorded chorda tympani nerve taste responses to chemical stimuli. 4-aminopyridine is a pharmacological blocker that reduces potassium conductance through potassium channels in nerve and muscle. Summated neural responses to stimuli dissolved in water and in 4-AP were compared. Chemical stimuli included concentration ranges of KCl, KBr, KH2PO4, CsCl, RbCl, NH4Cl, NaCl and sucrose. The blocker reduced chorda tympani responses to KCl and other potassium salts, from 0.025 to 0.25 M. Responses to ammonium, rubidium and cesium salts also were reduced, in order of effectiveness that would be predicted from known ion selectivity properties of potassium channels. Responses to NaCl and sucrose were not reduced. Other channel blockers, including tetraethylammonium chloride (TEA), BaCl2 and quinidine, did not reduce the response to KCl. These are the first detailed reports of effects of potassium channel blockers on the peripheral, neural taste response. The results are consistent with a role for potassium channels in apical taste bud cell membranes in transduction for potassium salts.

Convergence in mammalian nucleus of solitary tract during development and functional differentiation of salt taste circuits.

To determine the type and extent of neural rearrangements that are made during functional differentiation of circuits for salt taste processing, we determined receptive field size and salt response characteristics of second-order taste cells in 3 age groups of sheep. Neurophysiological recordings were made from single cells in the nucleus of the solitary tract (NST) in fetal, perinatal, and postnatal sheep. Responses to NH4Cl, NaCl, and KCl were measured, and location and number of fungiform papillae in the receptive field were determined by stimulating individual papillae with anodal electrical current. The data are compared with previous, parallel measures from chorda tympani nerve afferent taste fibers to permit conclusions about convergence or divergence onto second-order cells. Receptive field size of second-order taste neurons increases during development, in contrast to the decrease in field size observed previously for chorda tympani nerve fibers during the same period. Furthermore, receptive fields of second-order cells are significantly larger than those of first-order fibers at perinatal and lamb ages, but not fetal. Thus, there is convergence of first-order taste afferents onto brain-stem neurons, and the convergence increases remarkably between fetal and perinatal periods. Associated with the increase in convergence are increased salt response frequencies relative to afferent fibers for NaCl in perinatal animals and lambs, and for KCl in lambs. The increase in frequencies occurs before NST neurons are functionally mature, as indicated by the rapid response adaptation of many cells in young animals. Convergence in NST during development apparently functions to maximize gain for processing neural responses to NaCl. In the periphery, response frequencies to NaCl are very low in fetuses, and increase progressively during development. In the NST, NaCl response frequencies are high even in fetuses, and remain high. The process of convergence onto second-order cells is accomplished with maintenance of order in afferent projections because receptive fields of NST neurons are composed of fungiform papillae that are clustered together, not dispersed over the tongue. Our quantification of taste receptive field size at 2 neural levels provides strong evidence for increasing convergence in the NST during development. Altering patterns of afferent neural input and geometry of second-order neurons may have a role in establishing convergence. The convergence has an apparently special function: to increase gain for NaCl taste sensation. Therefore, neural rearrangements during differentiation of salt taste pathways result in specific functional outcomes.

Developmental neurobiology of salt taste sensation.

A principal process in the homeostatic control of sodium levels is salt intake, and the sense of taste has a primary role in regulating ingestion. Because ingestion of sodium chloride (NaCl) is essential for life, the taste system for salt sensation might be expected to exhibit mature functional characteristics from very early development. However, major changes in gustatory nerve responses to NaCl take place during development. In sheep and rat, the peripheral nerve responses to NaCl are of low magnitude during early development. Progressively, the taste system acquires an increasing proportion of fibers that respond maximally to NaCl. The sodium responsiveness emerges in the context of shifting peripheral innervation patterns and the apparent addition of functional receptor membrane channels sensitive to the sodium transport blocker, amiloride. These developmental processes can be altered by early manipulation of sodium in the diet.

Anatomy and neurophysiology of the taste system in aged animals.

In summary, the recent data from quantitative studies of taste buds in old humans, rhesus monkeys, and rats complement neurophysiological data on taste responses from aged rats and lead to the general conclusion that the peripheral taste system is maintained structurally and functionally across the life span. Although some statistically significant differences were observed, the magnitude of these differences would not lead one to predict altered taste preferences or feeding behavior in old animals. The robust nature of the peripheral sense of taste in old age appears to be in sharp contrast to other sensory systems. Not only do receptors alter with age, but accessory organs, such as the lens in the visual system, also alter structurally so that sensory function is compromised. One factor contributing to maintenance of the receptor organ of taste bud and papilla is the turnover and replacement of taste bud cells and surrounding epithelial cells. Recent studies in aged mice indicate no differences in turnover time in epithelial mucosa lining the mouth. There are no data on turnover time, however, for taste bud cells or for gustatory papilla epithelium in old animals. Alterations in taste bud cell or membrane receptor turnover could affect function. In other cell types and receptors, age-related differences in membrane lipids, proteins, and fluidity characteristics are reported. Membrane changes such as these are proposed to account for age-related differences in response properties of the system of cardiac muscarinic receptors, a system in which density of receptors does not alter. The small, but statistically significant, differences in neurophysiological responses to taste stimuli from the chorda tympani nerve might well relate to age-related differences in membrane receptors. Not only is there an absence of data on turnover time or membrane characteristics for taste bud cells in old age, but there are no observations on the ultrastructure of taste buds in old animals. The taste bud is a complex structure, composed of many cells and different cell types, with a specific orientation and access to the oral cavity via the taste pore, and with extensive associations with the innervating afferent fibers. Ultrastructural observations could provide information on changes in synapses and other nerve/cell contacts--in microvilli and taste pore structure, and in numbers of various cell types within the bud. Such data will be essential to understanding the nature and extent of age-related alterations in the taste system.(ABSTRACT TRUNCATED AT 400 WORDS)

Developmental decrease in size of peripheral receptive fields of single chorda tympani nerve fibers and relation to increasing NaCl taste sensitivity.

During development in rats, sheep, and humans, the taste system acquires increasing responsiveness to NaCl, compared with a variety of other salts and chemicals. To better understand the neural basis of changes in salt taste responses, we studied receptive field size and response properties of single chorda tympani nerve fibers in fetal, perinatal, and postnatal sheep. Individual fungiform papillae were stimulated electrically with 5 microA anodal current to determine the location and number of papillae in receptive fields. Response characteristics of NH4Cl, NaCl, and KCl were determined for the entire field. Receptive fields were dissected for later histological reconstruction and taste bud identification. Median receptive field size decreased during development. Field sizes in lambs were smaller than those in younger animals. This decrease was accompanied by an increase in the NaCl/NH4Cl response ratio of single fibers and an increase in the proportion of fibers and associated fields that responded with higher frequency to NaCl, compared with NH4Cl. In addition, for fibers across all age groups, receptive field size correlated negatively with the NaCl/NH4Cl response ratio; that is, fields most responsive to NaCl had fewer papillae than those most responsive to NH4Cl. For all fibers, receptive field size correlated with response frequencies to NH4Cl and KCl but not NaCl. For NaCl-best fibers, receptive field size correlated with the response frequencies to all 3 salts. There was no relation between number of taste buds in a single fungiform papilla and the response frequency elicited during electrical stimulation of the papilla.(ABSTRACT TRUNCATED AT 250 WORDS)

Morphology of chorda tympani fiber receptive fields and proposed neural rearrangements during development.

The average number of fungiform papillae in receptive fields of single chorda tympani nerve fibers decreases during development in sheep, and a greater proportion of small receptive fields that are highly responsive to NaCl, compared with NH4Cl, is acquired. To learn whether there also are developmental differences in the number of taste buds within the papillae in mapped receptive fields, we studied the morphology of receptive fields and fungiform papillae, and also counted fibers in the chorda tympani nerve, in fetal, perinatal, and postnatal sheep. Whether defined as the number of fungiform papillae or as the number of taste buds within papillae, receptive fields of chorda fibers decrease developmentally. Initially, however, there is an increase, and subsequently a decrease, in the number of taste buds per field. The differences in field size cannot be attributed to developmental alterations in numbers of fungiform papillae because the total number of papillae on the tongue remains constant. The average number of taste buds per papilla, however, also increases and then decreases, and the increase in perinatal animals is accompanied by the appearance of large, multipored taste buds. Because there is a significant relation between fungiform papilla size and number of taste buds in the papilla, papilla size could be one regulating factor for taste bud number. Furthermore, the number of chorda tympani nerve fibers apparently increases up to perinatal stages and then decreases postnatally, providing another potential regulating factor for the number of taste buds.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of dietary NaCl deprivation during early development on behavioral and neurophysiological taste responses.

In order to determine whether the gustatory system can be modified by restricting dietary NaCl during early development, neurophysiological taste responses were recorded in rats at various times after deprivation, and behavioral taste preferences were measured in adults. Rats deprived of dietary NaCl from the third day of gestation to 12 days postnatally and then placed on a NaCl-replete diet had chorda tympani nerve responses similar to those of nondeprived rats when recordings were made at 28 days of age and older; however, preferences for NaCl solutions over water were significantly less than those of controls when tested at adulthood. NaCl deprivation in pups from the third day of gestation to approximately 35 days postnatally resulted in altered chorda tympani nerve responses to NaCl but not to other stimuli such as NH4Cl and KCl. Therefore, restriction of dietary NaCl at a period in the rat's development when peripheral and central taste responses are changing results in short-term alterations in peripheral neural responses and in long-term changes in preference behaviors.

Taste responses from the chorda tympani nerve in young and old Fischer rats.

To determine whether neurophysiological taste responses are different in young and old rats, recordings were made from the whole chorda tympani nerve which innervates taste buds on the anterior tongue. Fischer 344 male and female rats in three age groups were studied: 5 to 7 months, 23 to 25 months, and 29 to 32 months. Chemical stimuli included single concentrations of five salts, two acids, sucrose and quinine hydrochloride, and concentration series of the five salts. Neural response magnitudes were expressed as ratios relative to the NaCl(0.1M) response. Substantial neural responses were obtained to all chemicals at all ages. Response ratios for NH4Cl and sucrose, however, increased significantly with age; ratios for MgCl2 and citric acid decreased. Ratios for HCl decreased when data from either sex were analyzed separately. Shapes of response/concentration functions for NaCl, but not other salts, also differed with age. In general, though, the magnitudes of all age-related differences were small and neural recordings demonstrate that the peripheral taste system functions well in old age.