Spilanthol Enhances Sensitivity to Sodium in Mouse Taste Bud Cells.

Overconsumption of NaCl has been linked to increased hypertension-related morbidity. Compounds that can enhance NaCl responses in taste cells could help reduce human NaCl consumption without sacrificing perceived saltiness. Spilanthol is an unsaturated alkylamide isolated from the Jambu plant (Acmella oleracea) that can induce tingling, pungency, and numbing in the mouth. Structurally similar fatty acid amides, such as sanshool, elicit numbing and tingling sensations by inhibiting 2-pore-domain potassium leak channels on trigeminal sensory neurons. Even when insufficient to induce action potential firing, leak current inhibition causes depolarization and increased membrane resistance, which combine to make cells more sensitive to subsequent depolarizing stimuli, such as NaCl. Using calcium imaging, we tested whether spilanthol alters sensitivity to NaCl in isolated circumvallate taste bud cells and trigeminal sensory neurons of mice (Mus musculus). Micromolar spilanthol elicited little to no response in taste bud cells or trigeminal neurons. These same perithreshold concentrations of spilanthol significantly enhanced responses to NaCl (140 and 200 mM) in taste bud cells. Trigeminal neurons, however, exhibited response enhancement only at the highest concentrations of NaCl and spilanthol tested. Using a combination of potassium depolarization, immunohistochemistry, and Trpm5-GFP and Tas1r3-GFP mice to characterize taste bud cells by type, we found spilanthol enhancement of NaCl responses most prevalent in NaCl-responsive type III cells, and commonly observed in NaCl-responsive type II cells. Our results indicate that spilanthol enhances NaCl responses in taste bud cells and point to a family of compounds that may have utility as salty taste enhancers.

THE IMPACT OF THE ACRYLIC MONOMER ON THE MORPHOLOGICAL STRUCTURE OF RAT LINGUAL MUCOSA.

The analysis of publications shows that diverse multiple factors can induce changes in taste sensitivity and the main irritants are the chemicals of different types. However, the study of the effect of the components of dental structural materials on the state of lingual mucosa, in particular, taste sensors, has not been fully elucidated to date. The purpose of the paper was the study of the effect of monomer of the "Ftoraks" base acrylic resin on the state of the rats' lingual mucosa within 2-4 weeks after its impact. The previous paper [5] presents the findings of the study on the impact of the monomer of the "Ftoraks" base acrylic resin on the state of the rats' lingual mucosa in the early period (1 to 7 days) and its subsequent regeneration. The studies have found that the greatest changes in the lingual mucosa occur on day 3 and 7 after the application of monomer, and are of erosive-inflammatory origin. Regeneration of the lingual epithelium is delayed. The studies confirm that the monomer of acrylic resin causes a number of pathological changes in the mucous membrane and muscles of the rat tongue, the nature of which varies depending on the duration of its impact. On day 14 in the lingual mucosa the destructive processes are significantly delayed, substituting for the sclerotic processes in the proper plate and atrophic processes, observed, first of all, in the papillae of the tongue. It is appropriate to assume that such changes in the papillae will lead to violation of the taste reception, first of all, in the areas of lateral surfaces of the body of the tongue and in the root area. At the same time, it should be noted that at the end of the experimental period (on day 28 of the contact of the monomer with the lingual mucosa), in the mucous membrane of the tongue, along with atrophic and sclerotic processes, the destructive changes and inflammatory reaction are evident. We hypothesize that this may indicate about partial recovery of taste sensitivity due to the decrease in the number of gustatory buds, taste papillae of different types and the increase in the period of their regeneration.

Glucose elicits cephalic-phase insulin release in mice by activating KATP channels in taste cells.

The taste of sugar elicits cephalic-phase insulin release (CPIR), which limits the rise in blood glucose associated with meals. Little is known, however, about the gustatory mechanisms that trigger CPIR. We asked whether oral stimulation with any of the following taste stimuli elicited CPIR in mice: glucose, sucrose, maltose, fructose, Polycose, saccharin, sucralose, AceK, SC45647, or a nonmetabolizable sugar analog. The only taste stimuli that elicited CPIR were glucose and the glucose-containing saccharides (sucrose, maltose, Polycose). When we mixed an α-glucosidase inhibitor (acarbose) with the latter three saccharides, the mice no longer exhibited CPIR. This revealed that the carbohydrates were hydrolyzed in the mouth, and that the liberated glucose triggered CPIR. We also found that increasing the intensity or duration of oral glucose stimulation caused a corresponding increase in CPIR magnitude. To identify the components of the glucose-specific taste-signaling pathway, we examined the necessity of Calhm1, P2X2+P2X3, SGLT1, and Sur1. Among these proteins, only Sur1 was necessary for CPIR. Sur1 was not necessary, however, for taste-mediated attraction to sugars. Given that Sur1 is a subunit of the ATP-sensitive K+ channel (KATP) channel and that this channel functions as a part of a glucose-sensing pathway in pancreatic ß-cells, we asked whether the KATP channel serves an analogous role in taste cells. We discovered that oral stimulation with drugs known to increase (glyburide) or decrease (diazoxide) KATP signaling produced corresponding changes in glucose-stimulated CPIR. We propose that the KATP channel is part of a novel signaling pathway in taste cells that mediates glucose-induced CPIR.

Oxaliplatin Alters Expression of T1R2 Receptor and Sensitivity to Sweet Taste in Rats.

As one of the adverse effects of oxaliplatin, a key agent in colon cancer chemotherapy, a taste disorder is a severe issue in a clinical situation because it decreases the quality of life of patients. However, there is little information on the mechanism underlying the oxaliplatin-induced taste disorder. Here, we examined the molecular and behavioral characteristics of the oxaliplatin-induced taste disorder in rats. Oxaliplatin (4-16 mg/kg) was administered to Sprague-Dawley (SD) rats intraperitoneally for 2 d. Expression levels of mRNA and protein of taste receptors in circumvallate papillae (CP) were measured by real-time quantitative polymerase chain reaction (PCR) and immunohistochemistry, respectively. Taste sensitivity was assessed by their behavioral change using a brief-access test. Morphological change of the taste buds in CP was evaluated by hematoxyline-eosin (HE) staining, and the number of taste cells in taste buds was counted by immunohistochemical analysis. Among taste receptors, the expression levels of mRNA and protein of T1R2, a sweet taste receptor subunit, were increased transiently in CP of oxaliplatin-administered rats on day 7. In a brief-access test, the lick ratio was decreased in oxaliplatin-administered rats on day 7 and the alteration was recovered to the control level on day 14. There was no detectable alteration in the morphology of taste buds, number of taste cells or plasma zinc level in oxaliplatin-administered rats. These results suggest that decreased sensitivity to sweet taste in oxaliplatin-administered rats is due, at least in part, to increased expression of T1R2, while these alterations are reversible.

The role of G-protein-coupled receptor 120 in fatty acids sensing in chicken oral tissues.

Clarification of the mechanism of chickens' taste sense will provide meaningful information for creating and improving new feedstuff for chickens, because the character of taste receptors in oral tissues affects feeding behavior in animals. Although fatty acids are partly recognized via G-protein coupled receptor 120 (GPR120) for fat taste in mammalian oral tissues, the fat taste receptor of chickens has not been elucidated. Here we cloned chicken GPR120 (cGPR120) from the chicken palate, which contains taste buds. By using Ca(2+) imaging methods, we identified oleic acid and linoleic acid as cGPR120 agonists. Interestingly, in a behavioral study the chickens preferred corn oil-rich feed over mineral oil (control oil)-rich feed. Because corn oil contains high amounts of oleic acid and linoleic acid, this result was thought to be reasonable. Taken together, the present results suggest that cGPR120 is one of the functional fat taste receptors in chickens.

[Taste-masking possibilities in solid dosage forms]. / Izfedési lehetoségek szilárd gyógyszerformák esetén.

Increasing number of new, innovative pharmaceutical products are in the generic market. One of the important dosage forms is the group of orally disintegrating products. The feature of these products is that they disintegrate rapidly in the mouth upon contact with the saliva; therefore the dissolution of the water-soluble components begins in the mouth. Since a large part of the drug molecules is characterised by a more or less bitter taste, even a small amount of the dissolved drug can cause an unpleasant taste in the mouth. Manufacturers apply different techniques to mask the bitter taste of these products, depending on the characteristics of the dosage form and the bioavailability requirements. In this study, we reviewed the most widespread taste-masking methods based on the scientific literature and different patents and the characteristics of some important excipients, and outline an instrumental technique used for bitterness measurements.

Epidermal growth factor attenuates lingual papillae lesions in a rat model of sialoadenectomy.

Salivary epidermal growth factor (EGF) plays an important role in the maintenance of the oral and gastro-esophageal mucosa. Sialoadenectomy delays healing of oral wounds and affects lingual papillae. In this work, we aimed to determine the effect of EGF deficiency induced by sialoadenectomy and evaluate the effect of exogenous EGF administration on the lingual papillae and taste buds in rats. Thirty male adult Wistar albino rats were equally divided into 3 groups; sham-operated control group, sialoadenectomy group and group of sialoadenectomy + EGF. EGF was given 8 weeks after sialoadenectomy in a dose of 1 µg /ml/day in drinking water for 2 weeks. The anterior two-thirds of the tongue was dissected and cut longitudinally into two halves; one half for light microscope and the other for electron microscope examinations. Saliva and blood were collected to determine salivary and plasma EGF. Our results revealed that sialoadenectomy significantly reduced plasma and saliva levels of EGF which resulted in severe disruption of the architecture of lingual papillae. These changes were effectively improved by the exogenous EGF administration. In conclusion, EGF supplementation reversed the effects of sialoadenectomy and restored almost normal architecture of lingual papillae and taste buds.

Saliva and other taste stimuli are important for gustatory processing of linoleic acid.

Paradoxically, bilateral transection of the chorda tympani nerve (CTX) raises the taste discrimination threshold for the free fatty acid, linoleic acid (LA), yet the chorda tympani nerve (CT) is unresponsive to lingual application of LA alone. LA may require a background of saliva to activate taste cells, since CTX decreases saliva production through denervation of the submaxillary and sublingual salivary glands. To assess the role of saliva, we measured LA taste discrimination thresholds for animals whose submaxillary and sublingual salivary glands were removed and also recorded CT responses to LA mixed in artificial saliva. Partial desalivation shifted LA discrimination thresholds from between 5.5 and 11 microM to between 11 and 22 microM. However, this effect was not as pronounced as previously seen with CTX animals. Surprisingly, the CT was unresponsive to LA mixed with artificial saliva, suggesting that artificial saliva may lack components necessary for LA taste. Additionally, fats may primarily enhance other tastes. We previously reported that LA increases CT responses to monosodium glutamate (MSG). Thus we also recorded CT whole nerve responses to taste mixtures of LA and sodium chloride (NaCl), sucrose (SUC), citric acid (CA), or quinine hydrochloride (QHCl) in anesthetized rats. We found that LA increased CT responses to NaCl but did not alter CT responses to SUC, CA, and QHCl. Thus CT recordings either lack the sensitivity to detect small changes to SUC, CA, and QHCl or LA may affect CT responses to MSG and NaCl only, perhaps by specifically modulating gustatory processing of Na(+).

Anatomical stability of human fungiform papillae and relationship with oral perception measured by salivary response and intensity rating.

Fungiform papillae house taste buds on the anterior dorsal tongue. Literature is inconclusive as to whether taste perception correlates with fungiform papillae density (FPD). Gustatory reflexes modulate the amount and composition of saliva subsequently produced, and thus may be a more physiologically objective measure of tastant-receptor interactions. Taste perception fluctuates with time but the stability of individual fungiform papillae is unclear. This study followed ten healthy volunteers longitudinally at baseline, one and six months. FPD, diameter and position were measured and participants rated intensity perception of sucrose, caffeine, menthol and capsaicin solutions. Salivary flow rate, protein concentration and relative changes in protein composition were measured following each tastant. FPD, diameter and position were unchanged at six months. FPD did not correlate with intensity rating for any taste. FPD did correlate with changes in salivary protein output following sucrose (ρ = 0.72, p = 0.02) and changes in levels of proline-rich protein and mucin 7 following capsaicin (ρ = 0.71, p = 0.02, ρ = 0.68, p = 0.04, respectively). These results suggest that over six months fungiform papillae are anatomically stable, playing a greater role in mediating the physiological salivary response to stimuli rather than determining the perceived intensity of taste.

Regional specificity of chlorhexidine effects on taste perception.

Chlorhexidine (CHX) gluconate, a bitter bis-biguanide antiseptic, reduces the intensity of the salty taste of NaCl and bitter taste of quinine in humans. This study addresses regional specificity of CHX's effects on taste. Perceptual intensity and quality were measured for separate taste bud containing oral loci innervated either by afferent fibers of cranial nerve (CN) VII or CN IX. Measurements were obtained following three 1-min oral rinses with either 1.34 mM CHX or water, the control rinse. CHX rinse reduced the intensity of NaCl more at the tongue tip and palate than at posterior oral sites. Thus, fungiform and palatal salt-taste receptors may differ from salt-taste receptors of the foliate and circumvallate taste papillae. The intensity of quinine.HCl was reduced equally by CHX at all sites tested but was frequently tasteless on the less sensitive anterior sites, suggesting quinine receptor diversity. In rodents, a portion of NaCl-taste receptors in the receptive field of CN VII is sensitive to the epithelial Na+ channel blocker amiloride and a portion is amiloride insensitive; all CN IX receptors are amiloride insensitive. The current results are the first to suggest that there may also be distinct, regionally specific populations of NaCl-taste receptors in humans.

Salivary proteins alter taste-guided behaviors and taste nerve signaling in rat.

Taste stimuli are normally dissolved in saliva prior to interacting with their respective receptor targets. There are hundreds of proteins in saliva, and it has been hypothesized that these proteins could interact with either taste stimuli or taste receptors to alter taste signaling and diet acceptance. However, the impact of these proteins on feeding has been relatively unexplored using rodent models. We have developed a novel technique for saliva collection that allows us to link salivary protein expression with feeding behavior. First, we monitored the microstructure of rats' feeding patterns on a 0.375% quinine diet (Q-diet) while tracking changes in salivary protein expression. We found 5 protein bands were upregulated by diet exposure to Q-diet and upregulation of a subset of these bands were statistically related to increased diet acceptance, including changes in behavioral measures that are thought to represent both orosensory and postingestive signaling. In a second experiment, we measured the licking to a range of quinine solutions (0.01-1.0mM) before and after the animals were exposed to a tannic acid diet that altered salivary protein expression. Rats found the quinine solutions less aversive after salivary protein altering diets. In a third experiment we recorded the response of the chorda tympani (CT) nerve while delivering quinine solutions (0.3-30mM) to the front of the tongue dissolved in either "donor saliva" containing salivary proteins or donor saliva which has had the salivary proteins removed. Donor saliva was collected from a separate group of animals using isoproterenol and pilocarpine. The samples containing salivary proteins resulted in lower nerve responses than those without salivary proteins. Together these data suggest that salivary proteins are capable of altering taste-guided behaviors and taste nerve signaling.

[Taste dysfunction (dysgeusia) and radioiodine therapy of thyroid cancer - be aware of side effects by antidepressants and sedatives. Vorschädigung durch Antidepressiva und Sedativa beachten]. / Geschmacksstörung (Dysgeusie) und Radioiodtherapie des Schilddrüsenkarzinoms. Vorschädigung durch Antidepressiva und Sedativa beachten.

In addition to xerostomia, taste dysfunction (hypo-, dysgeusia) is an independent side effect of radioiodine therapy of thyroid cancer. Hypogeusia results from damage of the small mucous salivary glands in the vicinity of the taste buds. Particularly in those patients, who are treated with drugs such as antidepressants or sedatives, taste dysfunction becomes frequently clinically symptomatic. If feasible, therapy regimens bearing a potential risk for taste dysfunction should be switched. Additional damage to taste function should be minimalized, including cessation of smoking, change of agents for dental hygiene, and change of toothpaste. If the medical indication for ablative radioiodine therapy is based on the patient's decision, the patient should be informed about alternative strategies. Potential clinical consequences of the sialadenitis (xerostomia, alteration in taste, risk of caries and tooth extraction, lacrimal gland dysfunction, tearing and need of dacryocystorhinostomy) should be revealed completely. Adapted to the initial risk of relapse, the 131I-activity for ablation should be reduced to the latest standard of care. Clinical data are currently too heterogeneous to decide whether rhTSH might reduce the rate of dysgeusia. The specialist in nuclear medicine should be aware of the multifactorial causes of taste dysfunction, in particular if the patient seeks medical advice after radioiodine therapy.

Effects of chemotherapy on gene expression of lingual taste receptors in patients with head and neck cancer.

OBJECTIVES/HYPOTHESIS: We aimed to test the hypothesis that chemotherapy changes the gene expression of taste receptors in the tongue to induce dysgeusia in patients with head and neck cancer. STUDY DESIGN: Prospective observation study. METHODS: We enrolled 21 patients who received chemoradiotherapy and five patients who underwent radiotherapy for head and neck cancer. The messenger RNA (mRNA) levels of the taste receptor subunits T1R1, T1R2, T1R3, and T2R5 were measured in lingual mucosa scrapings obtained with a small spatula. The perception thresholds of umami, sweet, and bitter tastes were assessed by the whole mouth gustatory test. RESULTS: In four patients with severe stomatitis induced by chemoradiotherapy, the mRNA levels of T1R1, T1R2, T1R3, and T2R5 in the lingual mucosa were significantly decreased. However, in 17 patients with mild/moderate stomatitis, the mRNA levels of T1R3 were significantly and transiently decreased, whereas those of T1R1 and T1R2 remained unchanged and those of T2R5 mRNA were significantly and transiently increased after chemotherapy. There was a significant negative correlation between the perception thresholds of umami or sweet tastes and lingual mRNA levels of T1R3 in patients with mild/moderate stomatitis after chemotherapy. Although the perception threshold of bitter taste remained unchanged, lingual mRNA levels of T2R5 were significantly increased in patients who complained of phantogeusia after chemotherapy. CONCLUSION: Chemotherapy specifically changed the gene expression of T1R3 and T2R5 in head and neck cancer patients with mild/moderate stomatitis, resulting in both dysgeusia of umami and sweet tastes as well as phantogeusia. LEVEL OF EVIDENCE: 4. Laryngoscope, 126:E103-E109, 2016.

Characteristics of phosphatidic acid-containing lipoproteins which selectively inhibit bitter taste: high affinity to frog tongue surface and hydrophobic model membranes.

In previous studies (Katsuragi and Kurihara (1993) Nature 365,213--214; Katsuragi et al. (1995) Pharm. Res. 12,658--662) we showed that a lipoprotein composed of phosphatidic acid (PA) and beta-lactoglobulin (LG) selectively suppressed the taste responses to bitter substances without affecting those to other taste stimuli in the frog and man, while complexes composed of other lipids except for phosphatidylserine and LG had little inhibitory activity. In the present study, we found that the lipoproteins having inhibitory activity are adsorbed on the frog tongue surface, while those having no inhibitory activity are not adsorbed. We also examined adsorption of the lipoproteins on model lipid membranes coated on a quartz-crystal microbalance by measuring changes in its frequency. The lipoproteins having inhibitory activity were well adsorbed on the hydrophobic lipid membranes, while the lipoproteins having no inhibitory activity were little adsorbed on the membranes. It seems that receptor sites for bitter substances on the taste cell membranes are hydrophobic and those for other taste stimuli such as salts, acids and sugars are hydrophilic. Hence, the binding of PA-LG to hydrophobic sites of the receptor membranes will lead to selective inhibition of bitterness.

Proton currents through amiloride-sensitive Na channels in hamster taste cells. Role in acid transduction.

The activity of taste cells maintained in the intact hamster tongue was monitored in response to acid stimulation by recording action currents from taste receptor cells with an extracellular "macro" patch pipette: a glass pipette was pressed over the taste pore of fungiform papillae and perfused with citric acid, hydrochloric acid, or NaCl. Because this technique restricted stimulus application to the small surface area of the apical membranes of the taste cells, many nonspecific, and potentially detrimental, effects of acid stimulation could be avoided. Acid stimulation reliably elicited fast transient currents (action currents of average amplitude, 9 pA) which were consistently smaller than those elicited by NaCl (29 pA). The frequency of action currents elicited by acid stimuli increased in a dose-dependent manner with decreasing pH from a threshold of about pH 5.0. Acid-elicited responses were independent of K+, Na+, Cl-, or Ca2+ at physiological (salivary) concentrations, and were unaffected by anthracene-9-carboxylic acid, tetraethylammonium bromide, diisothiocyanate-stilbene-2,2'-disulfonic acid, vanadate, or Cd2+. In contrast, amiloride (< or = 30 microM) fully and reversibly suppressed acid-evoked action currents. At submaximal amiloride concentrations, the frequency and amplitude of the action currents were reduced, indicating a reduction of the taste cell apical conductance concomitant with a decrease in cell excitation. Exposure to low pH elicited, in addition to transient currents, an amiloride-sensitive sustained d.c. current. This current is apparently carried by protons instead of Na+ through amiloride-sensitive channels. When citric acid was applied while the taste bud was stimulated by NaCl, the action currents became smaller and the response resembled that produced by acid alone. Because of the strong interdependence of the acid and salt (NaCl) responses when both stimuli are applied simultaneously, and because of the similarity in the concentration dependence of amiloride block, we conclude that amiloride-sensitive Na+ channels on hamster taste receptor cells are permeable to protons and may play a role in acid (sour) taste.

Objective assessment of terbinafine-induced taste loss.

OBJECTIVES: Terbinafine (Lamisil), a widely prescribed oral antifungal agent, reportedly induces taste loss in 0.6% to 2.8% of those taking the drug. However, many so-called taste problems reflect olfactory problems, and the sole empirical study published on this topic, based on whole-mouth testing of a single subject, found no terbinafine-related deficit. In this study, we quantitatively assessed, using well-validated taste and smell tests, chemosensory function in six patients complaining of taste disturbance after terbinafine treatment and compared their test scores to those of six age-, race-, and sex-matched normal controls. METHODS: Taste function was measured using a 96-trial regional test that assesses sweet, sour, bitter, and salty taste perception within the anterior (cranial nerve [CN] VII) and posterior (CN IX) lingual taste bud fields. Smell function was bilaterally evaluated using the 40-item University of Pennsylvania Smell Identification Test. RESULTS: Taste function for sweet-, sour-, and bitter-tasting stimuli was significantly depressed in both the anterior and posterior lingual regions. For sodium chloride, the decrements were confined to the posterior region. Olfactory function was within normal limits. CONCLUSION: These findings 1) support anecdotal case reports of taste loss after terbinafine use, 2) demonstrate that all four major taste qualities are affected, and 3) suggest that olfactory dysfunction is not involved. Because self-report markedly underestimates chemosensory deficits, more extensive quantitative testing of patients receiving terbinafine will likely reveal a much higher prevalence of terbinafine-induced taste loss than currently reported. Since being older than 65 years of age and having a low body mass index are reportedly risk factors for terbinafine-induced taste loss, physicians should be particularly on the alert for elderly persons taking this medication who may become depressed or alter their food intake in response to decreased taste sensation.

Temporal gustatory and salivary responses to capsaicin upon repeated stimulation.

Repeated oral stimulation with solutions of 2 ppm capsaicin increased time to maximum intensity, but yielded a decrease in sequential mouth-burn intensity due to a rising minimum irritation rating between stimuli. Two response patterns were observed: for 18 subjects mouth-burn increased, while 14 subjects showed depression upon repeated stimulation. Because of across-subject variability, there was little correlation between parotid salivary response and perceived sensory responses. Capsaicin-stimulated salivary flow was significantly higher than flow stimulated by distilled water in 9 high-flow subjects, but not in 14 low-flow subjects. Oral adaptation to capsaicin may attenuate parotid salivary response, although eaters and noneaters of chili peppers did not differ significantly in their perception of mouth-burn or in salivary flow.

Temporal effectiveness of mouth-rinsing on capsaicin mouth-burn.

Oral rinsing with different solutions significantly reduced mouth-burn of capsaicin solutions in both eaters and noneaters of chili peppers. Cold solutions (5 degrees C) were more effective in reduction of mouth-burn than solutions at 20 degrees C. Sucrose solutions (10%) at 20 degrees C and whole milk at 5 degrees C were equally effective while 5% ethanol was no more effective in mouth-burn reduction than water at 20 degrees C. Reduction of mouth-burn by sucrose was not dose dependent. Noneaters of chili peppers experienced a slightly greater reduction of mouth-burn from sucrose solutions than eaters. Oral rinsing with sweetened milk containing 0 and 10% fat, of varying globule size, resulted in similar degree of mouth-burn reduction. The first (control) sample was rated higher in intensity than subsequent ones, suggesting desensitization, which appears to be due to the interaction of stimulation of chemo-, mechano-, thermo- and gustatory receptors.

The gustin (CA6) gene polymorphism, rs2274333 (A/G), as a mechanistic link between PROP tasting and fungiform taste papilla density and maintenance.

Taste sensitivity to PROP varies greatly among individuals and is associated with polymorphisms in the bitter receptor gene TAS2R38, and with differences in fungiform papilla density on the anterior tongue surface. Recently we showed that the PROP non-taster phenotype is strongly associated with the G variant of polymorphism rs2274333 (A/G) of the gene that controls the salivary trophic factor, gustin. The aims of this study were 1) to investigate the role of gustin gene polymorphism rs2274333 (A/G), in PROP sensitivity and fungiform papilla density and morphology, and 2) to investigate the effect of this gustin gene polymorphism on cell proliferation and metabolic activity. Sixty-four subjects were genotyped for both genes by PCR techniques, their PROP sensitivity was assessed by scaling and threshold methods, and their fungiform papilla density, diameter and morphology were determined. In vitro experiments examined cell proliferation and metabolic activity, following treatment with saliva of individuals with and without the gustin gene mutation, and with isolated protein, in the two iso-forms. Gustin and TAS2R38 genotypes were associated with PROP threshold (p=0.0001 and p=0.0042), but bitterness intensity was mostly determined by TAS2R38 genotypes (p<0.000001). Fungiform papillae densities were associated with both genotypes (p<0.014) (with a stronger effect for gustin; p=0.0006), but papilla morphology was a function of gustin alone (p<0.0012). Treatment of isolated cells with saliva from individuals with the AA form of gustin or direct application of the active iso-form of gustin protein increased cell proliferation and metabolic activity (p<0.0135). These novel findings suggest that the rs2274333 polymorphism of the gustin gene affects PROP sensitivity by acting on fungiform papilla development and maintenance, and could provide the first mechanistic explanation for why PROP super-tasters are more responsive to a broad range of oral stimuli.