The fungiform papilla is a gustatory organ that provides a specific tissue residence for taste buds on the anterior tongue. Thus, during development there must be a progressive differentiation to acquire papilla epithelium, then taste cell progenitor epithelium, and finally taste cells within the papilla apex. Arranged in rows, the patterned distribution of fungiform papillae requires molecular regulation not only to induce papillae, but also to suppress papilla formation in the between-papilla tissue. Intact sensory innervation is not required to initiate papilla development or pattern. However, members of several molecular families have now been identified with specific localization in developing papillae. These may participate in papilla development and pattern formation, and subsequently in taste progenitor and taste cell differentiation. This review focuses on development of fungiform papillae in embryonic rat and mouse. Basic morphology, cell biology and molecular phenotypes of developing papillae are reviewed. Regulatory roles for molecules in several families are presented, and a broad schema is proposed for progressive epithelial differentiation to form taste cell progenitors in parallel with the temporal course, and participation of lingual sensory innervation.
Mammalian taste buds are maintained through continuous cell renewal so that taste bud cells are constantly generated from progenitor cells throughout life. Taste bud cells are composed of basal cells and elongated cells. Elongated cells are derived from basal cells and contain taste receptor cells (TRC). Morphologically, elongated cells consist of three distinct types of cells: Types I, II and III. In contrast to the remarkable progress in understanding of the molecular basis for taste reception, the mechanisms of taste bud maintenance have remained a major area of inquiry. In this article, we review the expression of regulatory genes in taste buds and their involvement in taste bud cell differentiation. Three major topics include: 1) the Sonic hedgehog (Shh)-expressing cell in the basal cell in taste buds as a transient precursor of elongated cells and as a signal center for the proliferation of progenitor cells; 2) the Mash1-expressing cell as an immature cell state of both Type II and Type III cells and as a mature cell state of Type III cell; and 3) the nerve dependency of gene expression in taste buds. Problems in the application of NCAM for the type III cell marker are also discussed.
Taste receptor cells detect gustatory stimuli using a complex arrangement of ion channels, G protein-coupled receptors, and signaling cascades. Sour and salty tastes are detected by ion channels in the rat. Using a combination of homology screening and functional expression approaches, we screened a rat circumvallate papilla cDNA library and identified acid-sensing ion channel-2a (ASIC2a) and ASIC2b as candidates for the rat sour-sensing channels. In situ hybridization and reverse transcription-polymerase chain reaction experiments revealed that ASIC2a and ASIC2b transcripts were localized in taste bud cells. Immunohistochemistry and immunoprecipitation also revealed that both subunits were expressed in a subset of taste cells and that some of the cells expressed ASIC2a/ASIC2b heteromeric assemblies. Electrophysiological studies demonstrated that stimulation of acetic acid produced larger ASIC2 currents than did hydrochloric acid at the same pH. ASIC2a/ ASIC2b channels generated maximal inward currents at pH ≤ 2.0, which agrees well with the in vivo pH-sensitivity of rat taste cells. The amiloride-sensitivity of ASIC2a/ ASIC2b heteromer lessened with decreasing pH and almost completely disappeared at pH 2.0. These data suggest that ASIC2a and ASIC2b may play roles in sour taste transduction.
Taste signals are first detected by the taste receptor cells, which are located in taste buds existing in the tongue, soft palate, larynx and epiglottis. Taste receptor cells contact with the chemical compounds in oral cavity through the apical processes which protrude into the taste pore. Interaction between chemical compounds and the taste receptor produces activation of taste receptor cells directly or indirectly. Then the signals are transmitted to gustatory nerve fibers and higher order neurons. A recent study demonstrated many similarities between response properties of taste receptor cells with action potentials and those of the gustatory nerve fibers innervating them, suggesting information derived from receptor cells generating action potentials may form a major component of taste information that is transmitted to gustatory nerve fibers. These findings may also indicate that there is no major modification of taste information sampled by taste receptor cells in synaptic transmission from taste cells to nerve fibers although there is indirect evidence. In the peripheral taste system, gustatory nerve fibers may selectively contact with taste receptor cells that have similar response properties and convey constant taste information to the higher order neurons.
Taste is unique among the sensory systems in that, besides its recognition of quality, it is innately associated with hedonic aspects of reward and aversion. This review of the literature will show how taste information is conveyed through the central gustatory pathways to the cortical gustatory area and is processed in terms of qualitative and quantitative aspects. Taste information is also sent to the reward system and feeding center via several brain sites including the prefrontal cortex, insular cortex, and amygdala. The reward system contains the ventral tegmental area, nucleus accumbens, and ventral pallidum; it finally sends information to the lateral hypothalamic area, the feeding center. The dopamine system originating from the ventral tegmental area mediates the motivation to consume palatable food. The actual ingestive behavior is promoted by the orexigenic neuropeptides from the hypothalamus. In the last section, the neural substrate of learning and memory of taste is introduced and the biological mechanisms are elucidated.
We studied the earliest stages of the palate in rat embryos using scanning electron microscopy and immunohistochemistry of growth-associated protein-43 (GAP-43) to investigate the role of nerves in the development of the palatal taste buds. Chronological sequences of the palatal gustatory structures revealed characteristic several stages: 1) At embryonic day 13.5 (E13.5), the palatal shelves were widely separated, and no nerves could be observed in the vicinity of their epithelium which was formed of an undifferentiated single cell layer. 2) At E14, intraepithelial GAP-43-immunoreactive fine nerves were first observed along the medial border of the palatal shelves which became several layers thick but still separate along their entire length. 3) At E15, the fusion process resulted in the formation of cranial parts of the soft palate, the epithelium of which was heavily innervated and revealed small fungiformlike papillae devoid of nerves. 4) As the fusion process continued more caudally at E15, there was a substantial increase in palatal innervation and number of fungiformlike papillae. Primordial stages of taste buds were first distinguished in the papillae where they coincided with sparsely distributed GAP-43-immunoreactive nerve fibers. 5) At E16, the whole soft palate was eventually differentiated and attained its definitive morphology. Different stages of taste buds (i.e. pored and non-pored) were recognized, and an extensive subgemmal plexus characteristic for the adult palatal taste buds was observed. 6) Mature taste buds with α-gustducin-immunopositive cells were observed at E18, and their numbers increased gradually with age. The present study reveals that the gustatory nerves preceded the development of taste buds in the palate of rats, and therefore may have some roles in the initial induction of taste buds as proposed in lingual taste buds.
Galanin, a 29-amino-acid neuropeptide, was initially isolated from porcine intestine. It has a wide spread distribution in the central nervous system and is also present in the primary sensory neuron. Galanin has been suggested to be involved in numerous neuronal and endocrine functions as a neurotransmitter and neuromodulator. We examined the expression of galanin and galanin receptors by using a reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry, and in situ hybridization. RT-PCR analysis showed that mRNA of galanin and GalR2 were detected in the taste bud-containing epithelium of the circumvallate papilla of rats. Immunohistochemical analyses detected galanin was detected in a subset of taste bud cells of the circumvallate papillae. Double-label studies showed that galanin colocalized with α-gustducin, NCAM, and PLCβ2. Our results of double staining with galanin and taste cell markers indicate that galanin-expressing taste cells are type II and type III cells. Taken together with previous studies, these findings show that galanin may function as a taste bud neurotransmitter. Furthermore, GalR2 mRNA was expressed in some taste bud cells. This suggests that, galanin release may not only excite the peripheral afferent nerve fiber but also may act on neighboring taste receptor cells via the activation of GalR2.
It has recently become evident that ATP and other extracellular nucleotides could play an important role in signal transductions. ATP mediates excitatory signaling by means of P2X receptors. P2X3, one of its subtypes, a membrane ligand-gated ion channel, is strongly expressed in peripheral sensory neurons. The aim of the present study was to examine the distribution of nerve fibers expressing P2X3 receptors in taste buds in the gustatory papillae and soft palate of rats by immunohistochemistry. We found that the fluorescence ATP marker quinacrine stained subsets of taste bud cells. Numerous nerve fibers innervating taste buds were intensely immunostained with the P2X3 receptor antibody. These nerve fibers ascended among intragemmal cells and terminated just below the taste pores. In order to examine whether P2X3 receptors are involved in signal modulation within taste buds, we used fluorescent double stainings to analyze the distribution of P2X3 receptors and their relationship to α-gustducin immunopositive taste receptor cells. Many varicose nerve fibers expressing P2X3 receptor-immunoreactivities were entangled with α-gustducin-immunopositive taste receptor cells and ended closely below the taste pores. In fungiform papillae, nerve fibers expressing both P2X3 receptors and PGP 9.5 were observed. In contrast, only PGP 9.5 immunoreactive nerve fibers were recognized in filiform papillae. These results suggest that P2X3 receptors might be involved in taste transmission pathways within taste buds. ATP may act as a neurotransmitter, co-transmitter, or neuromodulator at P2X3 receptors to generate activating gustatory nerve fibers.
Double immunohistochemistry of soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor (SNARE) proteins [synaptosomal-associated protein of 25kDa (SNAP-25), syntaxin and vesicle-associated protein-2 (VAMP-2)], and specific cell markers of taste buds cells [α-gustducin and phospholipase Cβ2 (PLCβ2) for type II cells; neural cell adhesion molecule (NCAM) for type III cells] was applied to gustatory epithelia of the rat circumvallate papillae. All three SNARE proteins were present in some elongated taste buds cells as well as intra-, peri- and subgemmal nerve fibers. Double immunohisotochemistry revealed that nearly all α-gustducin and PLCβ2 immunoreactive cells expressed SNAP-25, syntaxin, and VAMP-2. A majority of NCAM immunoreactive cells showed immunoreactivity for these SNARE proteins. These results indicate that these synapse-associated proteins (SNAP-25, syntaxin and VAMP-2) are present in both type II cells and type III cells. Moreover, more than 50% of intragemmal cells containing SNARE proteins showed immunoreactivities for α-gustducin, PLCβ2, and NCAM, suggesting the possible presence of transitional cells having histochemical proterties of both type II and type III cells.