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G Hellekant

Publications and source records attributed to G Hellekant.

At least 19 recordsLinked to original sources
PubMed2001

A new gene (rmSTG) specific for taste buds is found by laser capture microdissection.

Getting pure populations of taste buds suitable for molecular analysis has hampered the characterization of genes specifically expressed in taste cells. To solve this problem, we prepared specific cDNA libraries from small numbers of taste cells and surrounding epithelium isolated by laser capture microdissection (LCM) and report the discovery of a rhesus monkey novel gene (rmSTG) expressed specifically in taste cells, as found by differential screening of the cDNA libraries and RNA in situ hybridization. RNA in situ hybridization shows the preferential expression of this gene in taste buds from circumvallate, foliate, and fungiform papillae of the tongue. RT-PCR and Northern analysis of RNA from different non-taste organs showed no expression, pointing to a very specialized function of the protein in taste cells. Analysis of extended cDNAs and genomic DNA showed two exons and one intron. Northern analysis of circumvallate papillae showed a transcript of 1.3 kb as established in the gene model. BLAST search analysis showed that the human homolog is localized in the recently completely sequenced HLA class I region of Chromosome 6p21 and is sublocalized to the main susceptibility region for psoriasis vulgaris. The predicted gene encodes a protein of 314 amino acids with an N-terminal signal peptide and cleavage site, suggesting a membrane-bound or secreted protein with an extracellular role in taste cell physiology. The monkey, human, and mouse STG proteins contain potential O-glycosylation sites and tandem repeats inside a region showing approximately 50% similarity with prion proteins.

Amino Acid Sequence↗
PubMed2000

The taste of ethanol in a primate model. II. Glossopharyngeal nerve response in Macaca mulatta.

The glossopharyngeal nerve (NG) mediates taste from the posterior part of the tongue. Here, we studied the effects of ethanol on the tongue in recordings from both the whole NG and individual taste fibers of the rhesus monkey, Macaca mulatta. The results show that the nerve activity increased at 0.7 M ethanol, reaching half maximum at around 4 M alcohol. Previously, we identified three types of taste fibers in the rhesus monkey NG: S fibers predominantly responding to sweeteners, Q fibers responding to bitter, such as quinine hydrochloride (QHCl), and M fibers responding best to monosodium glutamate, NaCl and acids [Hellekant, G., Danilova, V., & Ninomiya, Y. (1997). Primate sense of taste: behavioral and single chorda tympani and glossopharyngeal nerve fiber recordings in the rhesus monkey, Macaca mulatta. J Neurophysiol 77, 978-993]. Here, this fiber classification was used to elucidate the oral effects of ethanol and ethanol mixtures with NaCl, sucrose, citric acid and QHCl. One and three molar concentrations of ethanol stimulated all fiber types. Mixtures of ethanol with QHCl elicited a smaller response in Q fibers than did QHCl alone. In S fibers, mixtures of ethanol with sucrose gave a larger response than did sucrose alone. The variability of M fibers was too large to allow a conclusion about the effect of ethanol. These results suggest that ethanol suppresses the taste of QHCl. Similarly, the taste of sucrose might be enhanced by adding ethanol to sucrose. These effects and conclusions corroborate an earlier ethanol study of the chorda tympani (CT) nerve [Hellekant, G., Danilova, V., Roberts, T., & Ninomiya, Y. (1997). The taste of ethanol in a primate model: I. Chorda tympani nerve response in Macaca mulatta. Alcohol 14, 473-484].

Analgesics, Non-Narcotic↗
PubMed2000

Responses to umami substances in taste bud cells innervated by the chorda tympani and glossopharyngeal nerves.

The chorda tympani (CT) and glossopharyngeal (GL) nerves of several mammalian species respond differently to umami substances (US) such as monosodium glutamate (MSG), disodium 5'-inosinate (IMP) and disodium 5'-guanylate (GMP). In mice and rhesus monkeys, responses to US are greater in the GL than the CT nerve, with the GL nerve containing larger numbers of MSG-sensitive fibers. Gurmarin, a sweet response inhibitor, suppresses the mouse CT responses to the mixture of MSG and IMP to approximately 65% of control levels but not to the metabotropic and ionotropic glutamate agonists 2-amino-4-phophonobutyrate and N-methyl-D-aspartate. Gurmarin does not inhibit any taste responses in the GL. In mice, CT responses to MSG may be masked by their greater sensitivity to sodium ions. Calcium imaging studies demonstrate that some mouse taste cells isolated from the fungiform papilla innervated by the CT respond selectively (as indicated by a rise in intracellular Ca(2+) concentrations) to MSG and/or IMP or GMP. These MSG responses are not suppressed notably by reducing the Ca(2+) concentration of the stimulus solution, suggesting that the observed Ca(2+) release is from intracellular stores. Measurements of second messengers in the mouse fungiform papilla have revealed consistently that MSG elicits increases in both inositol 1,4,5-trisphosphate and adenosine 3', 5'-cyclic monophosphate levels. Together, these results suggest that US may stimulate two different transduction mechanisms in the fungiform papilla. They also suggest that gurmarin-insensitive components of receptors for US, including metabotropic and ionotropic glutamate receptors, may be commonly involved in transduction for umami taste in taste cells on both anterior and posterior parts of the tongue.

Animals↗
PubMed1999

Solution conformation of brazzein by 1H nuclear magnetic resonance: resonance assignment and secondary structure.

Brazzein is a sweet-tasting protein isolated from the fruit of the West African plant Pentadiplandra brazzeana Baillon. It is the smallest and the most water-soluble sweet protein discovered so far, it is also highly thermostable. The proton NMR study of brazzein at 600 MHz (pH 3.5, 300K) is presented. Complete sequence specific assignment of the individual backbone and sidechain proton resonances were achieved using through-bond and through-space connectivities obtained from standard two-dimensional NMR techniques. The secondary structure of brazzein contains one alpha-helix (residues 21-29), one short 3(10)-helix (residues 14-17), two strands of antiparallel beta-sheet (residues 34-39, 44-50) and probably a third strand (residues 5-7) near the N-terminus.

Amino Acid Sequence↗
PubMed1999

Responses of single taste fibers and whole chorda tympani and glossopharyngeal nerve in the domestic pig, Sus scrofa.

Whole nerve, as well as single fiber, responses in the chorda tympani proper (CT) and glossopharyngeal (NG) nerves of 1- to 7-week-old pigs were recorded during taste stimulation. In the CT acids and in the NG bitter compounds gave the largest responses. Both nerves exhibited large responses to monosodium glutamate (MSG), MSG with guanosine 5'-monophosphate (GMP) and MSG with inositine 5'-monophosphate (IMP) as well as to glycine, xylitol, sucrose, fructose and glucose. Alitame, aspartame, betaine, neohesperedin dihydrochalcone (NHDHC), super-aspartame, saccharin and thaumatin elicited no or little response. Hierarchical cluster analysis of 49 CT fibers separated four major clusters. The M cluster, comprising 28.5% of all fibers, is characterized by strong responses to MSG, KCl, LiCl and NaCl. The responses to NaCl and LiCl were unaffected by amiloride. The H cluster (24.5%) includes units responding principally to acids. The Q cluster (18.5%) responds to quinine hydrochloride (QHCl), sucrose octaacetate (SOA) and salts with amiloride. The S cluster (28.5%) exhibits strong responses to xylitol, glycine and the carbohydrates as well as to MSG alone and to MSG with GMP or IMP. In 31 NG fibers, hierarchical cluster analysis revealed four clusters: the M cluster (10%), responding to MSG and MSG with GMP or IMP; the H cluster (13%), responding to acids; the Q cluster (29%), responding strongly to QHCl, SOA and tilmicosinR; and the S cluster (48%), responding best to xylitol, carbohydrates and glycine but also to the umami compounds. Multidimensional scaling analysis across fiber responses to all stimuli showed the best separation between compounds with different taste qualities when information from both nerves was utilized.

Age Factors↗
PubMed1998

Behavioral and single chorda tympani taste fiber responses in the common marmoset, Callithrix jacchus jacchus.

Gustatory responses of the common marmoset were studied using single fiber recordings from chorda tympani (CT) nerve and two bottle preference (TBP) tests. Hierarchical cluster analysis of 43 fibers' response profiles revealed 3 major clusters of fibers characterized by predominant sensitivity to sweeteners (S cluster), bitter compounds (Q cluster) or acids (H cluster). NaCl as well as LiCl did not stimulate CT taste fibers. The TBP tests showed relationship between a compound's ability to stimulate the taste fibers and the animals' consumption. Activity in the S cluster was associated with preference, while the activity in the Q cluster was associated with rejection. Marmosets neither preferred nor rejected sweeteners which did not stimulate any CT fibers.

Animals↗
PubMed1998

Taste in chimpanzees. III: Labeled-line coding in sweet taste.

In peripheral taste the coding mechanism remains an enigma. Among coding theories the "across-fiber pattern" argues that activity across fibers codes for taste, whereas the "labeled line" claims that activity in a particular set of fibers underlies a taste quality. We showed previously that chimpanzee chorda tympani taste fibers grouped according to human taste qualities into an S-cluster, responding predominantly to sweet stimuli, a Q-cluster, sensitive to bitter tastants, and an N-cluster, stimulated by salts. The analysis showed that information in the S-line suffices to distinguish stimuli of one taste quality from the others. However, one condition for the labeled line remained: that blockage of activity in a particular line must cause blockage of one taste quality, but of no other, or its onset give rise to the sensation of a taste quality. Here we studied this requirement with gymnemic acids and miraculin. In humans and chimpanzees, gymnemic acids suppress the sweet taste of all sweeteners whereas miraculin adds a sweet taste quality to sour stimuli. Gymnemic acids also abolish miraculin-induced sweet taste. We found that gymnemic acids practically abolished the response to every sweetener in the chimpanzee S-cluster. Equally important, they had no effect on the responses of the Q- and N-clusters. After miraculin, the S-cluster fibers responded to acids as well as to sweeteners, although they had not responded to acids before miraculin. Gymnemic acids abolished this miraculin-induced response to acids and responses to sweeteners in the S-fibers. These results link the sweet taste quality to activity in fibers of the S-cluster. Thus the S-cluster fibers satisfy the definition of the labeled-line theory: "that activity in a particular fiber type represents a specific taste quality."

Animals↗
PubMed1998

Solution structure of the thermostable sweet-tasting protein brazzein.

The fruit of Pentadiplandra brazzeana Baillon contains a small, sweet-tasting protein named brazzein. The structure of brazzein in solution was determined by proton nuclear magnetic resonance spectroscopy at pH 5.2 and 22 degrees C. The brazzein fold, which contains one alpha-helix and three strands of antiparallel beta-sheet, does not resemble that of either of the other two sweet-tasting proteins with known structures, monellin and thaumatin. Instead, the structure of brazzein resembles those of plant gamma-thionins and defensins and arthropod toxins. Sequence comparisons predict that members of a newly-identified family of serine proteinase inhibitors share the brazzein fold.

Amino Acid Sequence↗
PubMed1998

Gustatory responses of the hamster Mesocricetus auratus to various compounds considered sweet by humans.

The taste of 30 compounds was studied in the golden hamster with three different methods: single-fiber recordings, two-bottle preference (TBP), and conditioned taste aversion (CTA) tests. On the whole, the results showed that the sense of taste in the hamster differs in many respects from that in humans because, of 26 tested compounds known as sweet to humans, 11 had no taste or tasted differently. The results also supported the notion that activity in S-fibers elicits liking and activity in Q- or H-fibers rejection. Specifically hierarchial cluster analysis of 36 single fibers from the chorda tympani proper nerve separated N-, H-, and S-clusters consisting of 11 sucrose-, 14 NaCl-, and 11 citric-best fibers. Ace-K, cyanosuosan, N-4-cyanophenyl-N'-cyanoguanidineacetate (CCGA), -tryptophan, N-3, 5-dichlorophenyl-N'-(S)-alpha-methylbenzylguanidineacetate (DMGA), saccharin, SC-45647, and suosan stimulated only the S-fibers, were significantly preferred in TBP tests, and generalized to sucrose in the CTA tests. Ethylene glycol stimulated the N-fibers in addition to the S-fibers. This explains its generalization to sucrose in CTA. Its toxicity may contribute to its rejection in TBP tests. Sodium cyclamate stimulated a few N- but no S-fibers, which may explain the nondiscriminatory TBP and CTA results. Glycine elicited its largest response in the S-fibers, although it also stimulated other fibers. The resulting mixed taste sensation may explain why it was not preferred in TBP, although it generalized to sucrose in the CTA. Alitame, aspartame, N-4-cyanophenylcarbamoyl--aspartyl-(R)-alpha-methylbenzylamine (CAM), N-4-cyanophenylcarbamoyl-(R, S)-3-amino-3-(3, 4-methylenedioxyphenyl) propionic acid (CAMPA), N-(S)-2-methylhexanoyl--glutamyl-5-amino-2-pyridinecarbonitrile (MAGAP), N-1-naphthoyl--glutamyl-5-amino-2-pyridinecarbonitrile (NAGAP), NHDHC, superaspartame, and thaumatin were among the compounds considered sweet by humans that gave no response, were not discriminated in the TBP test, and gave no generalization in the CTA tests.

Animals↗
PubMed1997

Taste in chimpanzees II: single chorda tympani fibers.

Data are presented from 48 taste fibers in chorda tympani nerves of 10 chimpanzees during taste stimulation with 29 stimuli. The results demonstrated a higher taste fiber specificity than in any other mammalian species reported; breadth of tuning equals 0.3. Hierarchical cluster analysis separated an S-cluster (50% of all fibers), an N-cluster (31%), and a Q-cluster (19%). The S-cluster showed the highest specificity. Its fibers responded, with few exceptions, to every sweetener tested, including the sweet proteins brazzein and monellin. The response grew with increasing sweetener concentration. A large response to one sweetener was generally accompanied by a large response to all other sweeteners, and vice versa. Except for one broadly tuned fiber, the fibers of the S-cluster never responded to the bitter compounds. The fibers of the Q-cluster were more broadly tuned than any other fibers. Quinine hydrochloride was their best stimulus, but most fibers were also stimulated by KCl and NaCl with amiloride. Acids stimulated some of these fibers. The N-cluster could be divided into 3 subclusters: an Na-subcluster (3 fibers), Na-K subcluster (10 fibers), and M-subcluster (3 fibers). The Na-fibers responded strongly to, and were quite specific to, NaCl and LiCl stimulation but not to KCl, and fibers of the Na-K subcluster responded equally well to NaCl and KCl. The response to NaCl was suppressed by amiloride in the fibers of the Na-subcluster, but not in the fibers of the Na-K subcluster. Umami compounds elicited the strongest responses in the M-subcluster.

Animals↗
PubMed1997

Primate sense of taste: behavioral and single chorda tympani and glossopharyngeal nerve fiber recordings in the rhesus monkey, Macaca mulatta.

The responses of 51 chorda tympani proper (CT) and 33 glossopharyngeal (NG) neural taste units from the rhesus monkey (Macaca mulatta) were recorded during stimulation of either the anterior (CT) or posterior (NG) part of the tongue with 26 stimuli that taste salty, umami, sour, bitter, and sweet to humans. In the CT, hierarchical cluster analysis separated four major clusters. The N and S clusters were most populous, followed by the H cluster and a small Q cluster. NaCl, monosodium glutamate (MSG), and MSG with guanosine 5'-monophosphate were the best stimuli in the N cluster. Amiloride suppressed responses to NaCl. KCl did not stimulate fibers from this cluster. S cluster fibers were characterized by strong responses to all sweeteners. The H cluster responded best to acids but also to some of the sweeteners such as xylitol, fructose, and sucrose. Q fibers responded well to quinine hydrochloride (QHCl) and caffeine, but not to denatonium benzoate. In the NG, hierarchical cluster analysis separated three major clusters. Q fibers formed the largest cluster. QHCl, caffeine, and sucrose octa-acetate but not denatonium benzoate elicited very strong responses in these fibers. S fibers formed a second cluster. Although most of the sweeteners stimulated the S fibers, their responses were not so pronounced as in CT S fibers. The small M cluster was formed by fibers that responded best to MSG. They also responded to NaCl and acids. Two bottle preference tests showed a positive relationship between a sweetener's ability to stimulate the taste fibers and the animals' consumption. Thus the most-liked sweeteners stimulated the S cluster fibers of CT best, whereas less-liked sweeteners such as D-phenylalanine elicited a response in Q fibers and sodium cyclamate stimulated N fibers. The results show that both CT and NG taste fibers of M. mulatta group according to the human concepts of taste qualities.

Animals↗
PubMed1996

Understanding the mechanism of sweet taste: synthesis of ultrapotent guanidinoacetic acid photoaffinity labeling reagents.

Azido-functionalized analogs of potently sweet guanidinoacetic acids have been synthesized for use as sweetener receptor photoaffinity labeling reagents. These compounds have been synthesized using readily available starting materials. One of the azido-labeled guanidinoacetic acids has been evaluated in an electrophysiological model in the Rhesus monkey. We found that the photoaffinity-labeling reagent caused irreversible inhibition in electrophysiological response to sweeteners upon exposure of the monkey tongue to a combination of the reagent and UV light.

Affinity Labels↗
PubMed1996

Immunohistochemical, electrophysiological, and electron microscopical study of rat fungiform taste buds after regeneration of chorda tympani through the non-gustatory lingual nerve.

The sensory innervation of fungiform papillae on the rat dorsal tongue is derived from branches of two cranial nerves: the lingual branch of the trigeminal nerve which provides somatosensory innervation and the chorda tympani (CT) branch of the facial nerve, which provides innervation to the taste buds. Removal of the CT results in degeneration of the taste buds. Removal of both nerves results in reduction in size of fungiform papillae and an altered pattern of keratinization in its epithelium. Regeneration of nerves to the epithelium restores the pre-operative condition. Thus, in addition to their sensory functions, both the CT and lingual seem to exert trophic effects on the phenotypic expression of epithelial cells in the fungiform papillae. We severed both the CT and lingual nerves in rats and sutured the proximal stump of the CT to the distal stump of the lingual to promote regeneration of the CT along the lingual nerve pathway. At the same time, we prevented the proximal stump of the lingual from regenerating into the tongue. Our purpose was to determine whether and how the innervation pattern of the regenerated taste bud might be different from normal under these experimental conditions. We found that reinnervation by the CT through the lingual nerve occurs, that this restores the anatomical and functional integrity of the fungiform taste buds and papillae, and that some papillae, but not all, were richly innervated with subgemmal, extragemmal, and perigemmal neuron-specific enolase, calcitonin gene-related peptide, substance P, and neurokinin A-positive fibers. Moreover, responses to taste stimuli were recorded electrophysiologically from the CT.

Animals↗
PubMed1996

Assignment of the disulfide bonds in the sweet protein brazzein.

The thermostable sweet protein brazzein consists of 54 amino acid residues and has four intramolecular disulfide bonds, the location of which is unknown. We found that brazzein resists enzymatic hydrolysis at enzyme/substrate ratios (w/w) of 1:100-1:10 at 35-40 degrees C for 24-48 h. Brazzein was hydrolyzed using thermolysin at an enzyme/substrate ratio of 1:1 (w/w) in water, pH 5.5, for 6 h and at 50 degrees C. The disulfide bonds were determined, by a combination of mass spectrometric analysis and amino acid sequencing of cystine-containing peptides, to be between Cys4-Cys52, Cys16-Cys37, Cys22-Cys47, and Cys26-Cys49. These disulfide bonds contribute to its thermostability.

Amino Acid Sequence↗
PubMed1996

Taste in chimpanzee: I. The summated response to sweeteners and the effect of gymnemic acid.

The purpose of this study was to investigate further with electrophysiological and behavioral techniques the similarity between the sense of taste of humans and chimpanzees, especially with regard to the effects of gymnemic acid. Gymnemic acid (GA) is a powerful suppressor of sweet taste in humans but lacks this ability in nonprimates and lower primates. The summated taste responses from the chorda tympani nerve were recorded in nine adult chimpanzees, Pan troglodytes. The results show that all tested compounds that taste to humans elicited nerve responses in chimpanzees. GA suppressed or abolished the response to all sweeteners, but had no effects on the responses to the nonsweet compounds. The suppression varied from complete abolishment (aspartame, saccharin), to about 50% reduction (xylitol). When the effect of GA was tested on concentration series, 20% remained of the response to sucrose, whereas the responses to aspartame and saccharin were basically abolished. Higher concentrations of GA suppressed more. The effects of GA developed also in the presence of saccharin, but seemed less pronounced. The behavioral results were obtained with a one-bottle preference technique before and after GA. The results demonstrated that after exposure of the tongue to GA, the animals' liking for sweet diminished. These results parallel psychophysical and electrophysiological findings in humans. The way GA suppressed sweet taste in chimpanzees added one more characteristic to those that set chimpanzees apart from monkeys and close to humans.

Animals↗
PubMed1994

Brazzein, a new high-potency thermostable sweet protein from Pentadiplandra brazzeana B.

We have discovered a new high-potency thermostable sweet protein, which we name brazzein, in a wild African plant Pentadiplandra brazzeana Baillon. Brazzein is 2,000 times sweeter than sucrose in comparison to 2% sucrose aqueous solution and 500 times in comparison to 10% of the sugar. Its taste is more similar to sucrose than that of thaumatin. Its sweetness is not destroyed by 80 degrees C for 4 h. Brazzein is comprised of 54 amino acid residues, corresponding to a molecular mass of 6,473 Da.

Amino Acid Sequence↗