Trova il tuo prossimo libro preferito

Abbonati oggi e leggi gratis per 30 giorni
Optimising Sweet Taste in Foods

Optimising Sweet Taste in Foods

Leggi anteprima

Optimising Sweet Taste in Foods

Lunghezza:
854 pagine
6 ore
Pubblicato:
Jul 17, 2006
ISBN:
9781845691646
Formato:
Libro

Descrizione

A sweet taste is often a critical component in a consumer’s sensory evaluation of a food product. This important book summarises key research on what determines consumer perceptions of sweet taste, the range of sweet-tasting compounds and the ways their use in foods can be optimised.

The first part of the book reviews factors affecting sweet taste perception. It includes chapters on how taste cells respond to sweet taste compounds, genetic differences in sweet taste perception, the influence of taste-odour and taste-ingredient interactions and ways of measuring consumer perceptions of sweet taste. Part two discusses the main types of sweet-tasting compounds: sucrose, polyols, low-calorie and reduced-calorie sweeteners. The final part of the book looks at ways of improving the use of sweet-tasting compounds, including the range of strategies for developing new natural sweeteners, improving sweetener taste, optimising synergies in sweetener blends and improving the use of bulk sweeteners.

With its distinguished editor and international team of contributors, Optimising sweet taste in foods is a standard reference for the food industry in improving low-fat and other foods.
  • Investigates what determines consumer perceptions of sweet taste
  • Looks at improving the use of sweet-tasting compounds
  • Explores strategies for delivering new natural sweeteners
Pubblicato:
Jul 17, 2006
ISBN:
9781845691646
Formato:
Libro

Correlato a Optimising Sweet Taste in Foods

Titoli di questa serie (295)
Libri correlati
Articoli correlati

Anteprima del libro

Optimising Sweet Taste in Foods - Elsevier Science

19.

Part I

Factors affecting sweet taste perception

1

Stimulation of taste cells by sweet taste compounds

M. Naim    The Hebrew University of Jerusalem, Israel

Liquan Huang    Monell Chemical Senses Center, USA

A.I. Spielman    College of Dentistry, New York University, USA

M.E. Shaul; A. Aliluiko    The Hebrew University of Jerusalem, Israel

1.1 Introduction

Perception of sweet sensation is known to be associated in humans and many mammals with a unique pleasure. Studies examining facial expressions of neonates following taste stimulation indicate appealing responses to sweet taste and aversion to bitter taste (Steiner, 1973), and newborns prefer sugar solutions to water and their preference increases with sugar concentration (Desor et al., 1973). Furthermore, their preference for different sugars corresponds to adult perception (Maller and Desor, 1974). These studies suggest that the response to sweet taste is innate. The high threshold for the appealing taste of sugar and actual preference (Cagan and Maller, 1974) lead mammals to select high-caloric foods. In contrast, as a counter defense and screening mechanism, a strong rejection of bitter stimuli is due to the fact that traditionally most bitter compounds were toxic and therefore are perceived at much lower concentrations than sweet compounds. These innate behavioral mechanisms have apparently had a significant effect on survival during phylogenetic development. Moreover, it has long been recognized that the preference for sugars may change with physiological state. Human subjects prefer a 5% sucrose solution to 30% sucrose solution, but can be induced to prefer the more concentrated solution once blood glucose level is decreased to 50 mg% following insulin injection (Mayer-Gross and Walker, 1946). Similar results are found in rats (Jacobs, 1958). A strong preference for sweet high-fat foods and fat is associated with opioids (Marks-Kaufman and Kanarek, 1981; Marks-Kaufman and Lipeles, 1982; Drewnowski et al., 1992; Drewnowski, 1991) which appear to be released under mild stress. Administration of naloxone, an opioid antagonist, reduces the hedonic preference for sugar (Drewnowski et al., 1995). Other studies have shown that oral stimulation by sweet taste substances may, via the cephalic phase of endocrine secretion, release insulin, which in turn may participate in stimulating eating (Brand et al., 1982).

The intake of food items containing refined sugars and high caloric density constituents has increased significantly in the last century (Grand, 1974), and has been linked to metabolic disorders such as diabetes and obesity, termed ‘diabesity’ (Shafrir, 1997), considered to be the main threats to human health in the 21st century (Zimmet et al., 2001). In response to such health hazards, during the last four decades many chemical studies were initiated to explore low-calorie alternative sweeteners with high sweet potency (DuBois et al., 1981; DuBois et al., 1993; Nofre and Tinti, 1987; Nofre et al., 1988; Tinti and Nofre, 1996). The resulting synthetic and some natural non-sugar sweeteners include a large collection of diverse compounds such as sulfamates, flavonoids, oximes, amino acids, peptides, proteins, guanidines and terpenoids. These discoveries were based on structure–function relationship studies which have essentially proposed a single sweet taste receptor for all sweeteners (Shallenberger and Acree, 1967; Kier, 1972; van der Heijden et al., 1985a, 1985b). Tinti and Nofre have prepared extremely potent sweeteners and proposed the multipoint attachment (MPA) model of sweetness (Tinti and Nofre, 1996; Nofre and Tinti, 1996). This model was in line with the notion of a single sweet taste receptor but with multiple binding sites between the high potency sweeteners and the receptor molecule. The nutritional justification of alternative sweeteners to sugars has led to increased consumption of low-calorie soft-drinks, creating a multi-billion dollar economic target for the food industry. However, the sweet taste of sugars, especially that of sucrose, is regarded as pure with optimal sensation in humans, whereas many non-sugar sweeteners possess inferior sweet quality. Psychophysical sensory studies in humans using the multi-dimensional similarity (MDS) analysis (Schiffman et al., 1979) clearly showed that the sweetness of a variety of non-sugar sweeteners may be located at a different site in the MDS sweet similarity map from that of sugar sweeteners. Cross-adaptation studies of sweeteners (Schiffman et al., 1981; Lawless and Stevens, 1983) have suggested for a long time that either different sweet taste receptors or different mechanisms are involved for sweet sensation. A further difference between non-sugar sweeteners and sugars is a phenomenon known as synergism. Sweet taste synergism occurs mainly between pairs of non-sugar sweeteners but not between pairs of sugars (Ayya and Lawless, 1992; DuBois, 2004; Schiffman et al., 1995). Moreover, in most cases, stimulation by non-sugar sweeteners does not lead to the maximal sweet taste intensity level achieved with sugars during psychophysical experiments, even though these sweeteners are more potent when intensity is evaluated on a weight basis (Moskowitz, 1977; DuBois et al., 1991). Sugars and sugar alcohols show linear concentration–response relationships while high-potency sweeteners yield hyperbolic dependence. According to Moskowitz (1977), it may be related to the fact that as one increases the concentration of the non-sugar sweeteners, the additional taste qualities that such sweeteners possess become predominant and do not permit a further increase in sweetness intensity as occurs for sugars. In fact, it was suggested that the concentration-intensity slope be used as an additional parameter for evaluation of the sweet taste quality of a sweetener (Moskowitz, 1977).

It is apparent that there are multiple reasons that may be related for the above dissimilarity in sweet taste between sugars and non-sugar sweeteners. It is well known that many (though not all) non-sugar sweeteners possess additional taste sensations such as bitter, metallic and licorice. Although the molecular basis for such sensations is not clear, the sweeteners saccharin and acesulfam K were recently found to stimulate both bitter and sweet taste receptors (Kuhn et al., 2004) (see more discussion on sweet taste receptors below).

A further distinguishing factor that makes clearly non-sugar sweeteners inferior to sugars is their temporal properties. Time–intensity relationship studies indicated that, compared to sucrose, it takes a longer time for the sensation of a non-sugar sweetener to reach a maximal sweet taste intensity (slow taste onset), and more time (sometimes minutes) for the sweetness to be extinguished (lingering aftertaste) (Birch et al., 1980; Larson-Powers and Pangborn, 1978; Naim et al., 1986; DuBois et al., 1977). This lingering phenomenon is also known to occur for a variety of bitter stimuli. Interestingly, sweeteners which produce the sweet aftertaste may also induce ‘sweet water aftertaste’, a phenomenon where water tastes sweet for several minutes after tasting a sweetener (van der Wel, 1972; Bartoshuk, 1987; Naim et al., 1986). This is also true for stimulation by some sweetener receptor antagonists (D’Angelo and DuBois, 1999). The delay in sweet taste termination following the tasting of non-sugar sweeteners is not unique to humans. It was also shown to occur in old world monkeys during behavioral studies (e.g., baboons, (Rogatka et al., 1985)). The mechanism of lingering taste is unknown. One question is whether such a delay is due to peripheral or central nervous system events. For instance, in olfaction, the memory of odors can persist, e.g., in the neuronal tissue of insects, even as cells go through metamorphosis (Ray, 1999). In the brain, taste sensation which is associated with malaise may form long-term conditioned taste aversion through memory which depends on identified biochemical mechanisms (Berman and Dudai, 2001). However, electrophysiological recordings of taste nerves at the periphery (Hellekant, 1994) have indicated ‘lingering aftertaste’ in response to stimulation by non-sugar sweeteners. Furthermore, time-course measurements have shown that the transient onset and termination of IP3 release in taste cells following stimulation by some bitter tastants known to possess lingering aftertaste is delayed beyond 500 ms (Spielman et al., 2002). These results suggest that the delay in taste-signal termination induced by some non-sugar sweeteners and bitter tastants is at the periphery, at the taste-cell level. To date, the molecular basis for the ‘slow taste onset’ and the ‘lingering aftertaste’ phenomena is not known, even though it has significant implications regarding the acceptance of a variety of food products.

In the current chapter we will discuss the complex signaling which leads to sweet taste sensation events occurring at the level of the taste bud cells. Furthermore, we will analyze current experimental data assumed to provide better understanding of biochemical events related to sweet taste quality.

1.2 Peripheral organization of taste

The peripheral gustatory system is exposed to a variety of physical, chemical and biological insults. Extreme hot, cold, irritating, acidic and non-sterile stimuli may have damaging effects on peripheral taste receptors. Therefore, unlike all other sensory systems that have peripheral neuronal receptors, the peripheral gustatory system has evolved to have receptor cells that are rapidly-renewing, specialized epithelial cells. This adaptation ensures rapid regeneration of taste receptor cells that become damaged. The peripheral organ in the gustatory system is comprised, in decreasing hierarchical order of: taste papilla, taste bud and taste cell. The largest is the taste papilla (Spielman and Brand, 1997). Visible to the naked eye, taste papillae in humans come in at least four different shapes and are located on the tongue, soft and hard palate, pharynx, epiglottis and larynx. The largest of the four types, the circumvallate (CV) papillae, are located on the dorsal surface of the tongue between the anterior 2/3 and posterior 1/3 along a V shaped line. The number of CV papillae varies from species to species. There are between three and 13 papillae in humans. In other animals, their number varies from just one in rodents to 25 in cows.

On the postero-lateral border of the tongue are the foliate papillae, each of which is a pocket-shaped invagination lined with taste buds. The invaginations protect the taste buds from direct physical damage. The third type of papillae, the mushroom shaped fungiform papillae, cover the anterior dorsal surface of the tongue. Compared with other gustatory papillae, they are the largest in number with 50–200 in humans.

Finally, the extralingual papillae are located on the soft and hard palate, the pharynx, epiglottis and larynx, with the most prominent being the taste stripes (or Geschmackstreifen from the original German term), located on both sides of the palatal midline at the transition of the soft and hard palate. Interestingly, the most abundant and recognizable structures, the filiform papillae are non-gustatory. When overgrown and stained by beverages and food dye these thread-like papillae tend to provide a characteristic colour to the tongue. Similarly, excessive shedding of the papillae provides a ‘white coat’ on the tongue. On the other hand, loss of the filiform papillae creates a dry and smooth appearance which is associated with a variety of oral and systemic conditions such as dry mouth, anaemia and Scarlet fever. The filiform papillae act like ‘Velcro®’ to help secure food to the tongue so that the bolus can be moved around the oral cavity (Spielman, 1998).

Within each taste papilla there are varying numbers of taste buds visible under light and electron microscopes. The number of taste buds varies from one papilla to the next. In humans, fungiform papillae contain 1–10, CV papillae contain 100–200, while foliate papillae may have from a few hundred to a few thousand buds.

The taste bud is the functional unit of the peripheral taste organ. It is onion-shaped and contains 50–100 continuously-maturing taste receptor and supporting cells. Most of the taste cells in a bud are shielded from the oral cavity. Only the apical tip of a few taste cells contains taste receptor proteins that is exposed to the oral cavity through a 3–5 micron wide opening, referred to as the taste pore. Unlike any other sensory system, taste cells have a rapid turnover rate of 10.5 days, which is about twice as fast as the surrounding epithelial cells. The progenitor cells, also known as basal cells, are located in an epithelial layer at the base of the taste bud that corresponds to the germinal layer of the epithelium. As taste receptor cells continuously grow and mature, they migrate from the basal area of the bud toward the taste pore over a period of about 10 days, its turnover time. Exposure of an individual receptor cell to taste stimulants is limited usually to a single meal. Within a few hours of a chemosensory experience, the exposed taste receptor cells are shed into the oral cavity and the dead cells are washed away by saliva. The fungiform- and taste-stripe-containing taste buds are bathed in saliva from the bilateral major salivary glands (parotid, submandibular, sublingual). In contrast, taste buds in the circumvallate and foliate taste papillae are washed primarily by the saliva of the von Ebner’s salivary glands located in the body of the tongue with openings into the trenches that surround these papillae.

Within the taste bud, several taste receptor cells, certainly not all, will synapse with first order neurons belonging to either the facial (chorda tympani), glossopharyngeal or vagus nerves. In a recent study synapses were observed in a few taste receptor cells (Yang et al., 2000). In another study, the specific receptor cells, called Type II, associated with bitter, sweet and umami taste transduction was devoid of conventional synapse (Clapp et al., 2004). These cells either communicate first with adjacent taste cells in the bud before synapsing, or they have subsurface cysternae of smooth endoplasmic reticulum instead of traditional synapses (Clapp et al., 2004).

The sensory pathway for taste involves a chain of neurons in series. First order neurons connect the periphery to the nucleus of the solitary tract in the central nervous system where they synapse second order neurons, cross the midline and connect with the thalamus on the opposite side, while the third order neurons connect the thalamus to the cortex. The sensory pathway for taste involves neural connections with the salivary nuclei to activate a reflex pathway and the perception of taste depends on the signals reaching the sensory cortex.

1.3 The sweet taste receptor(s)

1.3.1 Sweet taste is mediated by G protein coupled receptors

Earlier biochemical studies indicated that sweet taste is detected by proteinaceous membrane receptors that are located on the tip of taste receptor cells. Proteolytic (pronase) treatment of the surface of human tongue could temporarily remove sweet perception, indicating that the sweet receptors are the membrane-bound surface proteins (Hiji, 1975). Radiolabeled sugars and sweeteners such as sucrose, fructose, glucose, synthetic sweeteners (cyclamate and saccharin) and sweet-tasting proteins monellin and thaumatin could bind to the purified homogenate fractions from human and bovine papillae. This binding activity was taste papillae-dependent, saturable and reversible, could be competitively inhibited by unlabeled sugars and sweeteners, indicating that there were specific receptors on taste bud cells that could reversibly bind to sweet compounds (Cagan, 1971; Lum and Henkin, 1976; Cagan and Morris, 1979).

Additionally, photoaffinity labeling demonstrated that the intensely sweet-tasting protein thaumatin I could bind to a protein of approximately Mr 50,000 from monkey circumvallate papillae (Shimazaki et al., 1986). This finding was further corroborated in other studies. Colloidal gold-labeled thaumatin was shown to bind to the microvilli in the taste pore of monkey taste buds, as expected, where the sweet receptors are located (Farbman et al., 1987; Menco and Hellekant, 1993). In an attempt to purify the sweet receptor proteins, affinity columns of covalently attached thaumatin and monellin were used to isolate thaumatin/monellin-bound proteins from cow, pig and rat gustatory membranes. These bindings could be displaced by the sweet peptide aspartame or the sweet taste modifier gymnemic acid, indicating that there is a common receptor for some sweet proteins and the peptide aspartame or gymnemic acid (Persaud et al., 1988). However, the isolation of sweet receptor proteins has been hampered largely due to three problems: the scarcity of taste buds, the lack of a tight binding between taste membranes and their presumed receptors (Bruch et al., 1988), and lack of cultured taste cell lines.

Further biochemical studies indicated that sweet receptors are G-protein coupled receptors (GPCRs). Activation of these receptors in the membranes derived from taste bud-containing lingual epithelium by sucrose and other sugars significantly enhanced the activity of adenylyl cyclase (AC), one of the effector enzymes in G-protein-mediated signaling cascades. This response was concentration-dependent, and was mediated by guanine-nucleotide-binding proteins (G-proteins) found in the taste cell membranes (Striem et al., 1989; Bruch and Kalinoski, 1987).

1.3.2 Discovery of the sweet taste receptors

Over the last three decades, animal behavioral assays demonstrated that inbred mouse strains such as C57BL/6 and DBA/2 displayed a marked difference in detection thresholds for synthetic sweet compounds such as saccharin and other sweeteners (Fuller, 1974). This trait exhibited a simple mendelian inheritance (Lush, 1989; Phillips et al., 1994; Lush et al., 1995). Genetic mapping of recombinant strains identified a Sac locus, named after saccharin, on the distal end of mouse chromosome 4 as the genetic determinant for this trait (Phillips et al., 1994; Lush et al., 1995; Blizard et al., 1999). In addition, the peripheral nerve responses to sucrose were affected by the Sac locus as well (Bachmanov et al., 1997), suggesting that this locus encodes a sweet taste receptor or some other key sweet taste transduction element. In 1999, by random sequencing of subtracted rat taste papilla-enriched cDNA libraries, Hoon and colleagues isolated cDNAs that encoded two putative GPCRs: TR1 and TR2 (now referred to as T1R1 and T1R2) (Hoon et al., 1999). These two genes are expressed in a subset of taste bud cells and expression levels of each gene showed distinct topographical distributions across the tongue and palate that corresponds to the taste sensitivity map of the tongue. Based on its topographical expression pattern, T1R1 was suggested to be putative sweet receptor. However, high resolution genetic mapping eliminated T1R1 as a Sac-encoded sweet receptor (Li et al., 2001).

With the availability of mouse and human genome sequences and bioinformatics tools, data mining of DNA sequences of the Sac locus on mouse chromosome 4 and a syntenous region on human chromosome 1 identified a novel putative 7-transmembrane domain receptor, T1R3 (Bachmanov et al., 2001; Kitagawa et al., 2001; Max et al., 2001; Montmayeur et al., 2001; Sainz et al., 2001). Subsequent analyses revealed that the allelic variations between sweet-sensitive and -insensitive mouse strains resulted in amino acid substitutions in T1R3 receptor that corresponded to the Sac phenotypes, suggesting that it is the sweet receptor that is encoded by this Sac locus. Introgression by serial backcrossing of a small T1R3 gene-containing chromosomal fragment from a sweet taster mouse strain (C57BL/6ByJ) to a sweet non-taster strain (129P3/J) conferred the latter high sensitivity to sweeteners (Bachmanov et al., 2001). Further confirmation of T1R3 gene as Sac is from a transgenic experiment that demonstrated the rescue of the sweet taste deficit by introduction of the T1R3 gene from a sweet taster strain into a sweet non-taster strain (Nelson et al., 2001).

Based on the sequence similarity of the heptahelical domain, T1R3, as well as T1R1 and T1R2, is classified as a member of Class C GPCR family (Pin et al., 2003). Many members of this class including calcium sensing receptor (CaSRs), putative pheromone receptor V2Rs, neurotransmitter receptors mGluRs and GABABRs, as well as T1R1, T1R2 and T1R3 have a large N-terminal extracellular domain. Some of the receptors in this class form homo- or heterodimers. Double labeling in situ hybridization indicated some taste bud cells co-expressed T1R1 and T1R3, or T1R2 and T1R3 while a few cells express T1R3 alone. Thus T1R-expressing cells can be categorized into three types: cells co-expressing T1R1+T1R3; cells co-expressing T1R2+T1R3; cells expressing T1R3 alone (Montmayeur et al., 2001; Nelson et al., 2001). These expression patterns suggested that T1R3, like other members of Class C GPCRs, may form heterodimers with T1R1 and T1R2. Co-expression of T1Rs in a heterologous system in fact confirmed that the dimeric receptor of T1R2 and T1R3 functions as a sweet receptor and human T1R2/T1R3 can be activated by all sweet compounds tested, including sugars: fructose, galactose, glucose, lactose, sucrose and maltose; amino acids: glycine and D-tryptophan; sweet proteins: monellin and thaumatin; synthetic sweeteners: acesulfame K, aspartame, cyclamate, dulcin, neotame and saccharin. The order of the sensitivity of the receptors to these substances approximates that from animal behavioral tests. Interestingly, T1R3 can also form a heterodimeric receptor with T1R1 that can be activated by umami stimuli (Nelson et al., 2001, 2002; Li et al., 2002). Therefore, T1R3 appears to be a common monomer of sweet and umami receptors.

Phylogenetic analysis showed that T1R3 is most closely similar to T1R1 and T1R2 with ~30% amino acid identity, while T1R1 and T1R2 are ~40% identical to each other (Hoon et al., 1999; Bachmanov et al., 2001; Kitagawa et al., 2001; Max et al., 2001; Montmayeur et al., 2001; Sainz et al., 2001; Nelson et al., 2001). Unlike most other GPCRs, which usually share high similarity across closely related species (e.g. another receptor of the Class C, calcium-sensing receptor has >90% amino acid identity among human, rodent and bovine sequences) (Chattopadhyay et al., 1996), taste receptors including T1R sweet, umami receptors and T2R bitter receptors display only about 70% sequence identity between human and rodents. This suggests that species-specific taste receptors may have different ligand interaction properties, and each may have evolutionarily tuned to their distinct ecological niches. Psychophysical, behavioral and electrophysiological studies showed that humans and old world primates can detect the sweetness of a subset of substances including monellin, brazzein, thaumatin, neotame, aspartame, cyclamate and neohesperidin dihydrochalcone that new world monkeys and rodents cannot. Furthermore, human sweet taste is subject to the suppression by lactisole while rodents are insensitive to this compound (Danilova et al., 1998, 2002; Hellekant et al., 1997; Sclafani and Perez, 1997; Brouwer et al., 1973; Naim et al., 1982; Schiffman et al., 1999; Johnson et al., 1994). In line with these observations, heterologously expressed human T1R2 and T1R3 dimeric receptors can respond to the stimulation of these sweeteners, and the responses are susceptible to lactisole inhibition while rodent receptors cannot be activated by these compounds and the response to other sweeteners are not suppressed by lactisole (Li et al., 2002).

Unlike bitter T2R receptors, which have a short extracellular amino-terminus, T1R receptor proteins consist of several functional domains (Fig. 1.1): a large extracellular amino-terminal domain (ATD), followed by a cysteine rich domain (CRD), a heptahelical transmembrane domain (HD) and an intracellular carboxyl-terminal tail (Hoon et al., 1999; Bachmanov et al., 2001; Kitagawa et al., 2001; Max et al., 2001; Montmayeur et al., 2001; Sainz et al., 2001; Nelson et al., 2001; Adler et al., 2000). Based on the crystal structure of another member of Class C receptors, mGluR1, T1Rs are likely to have a clam shell-like ‘Venus flytrap module’ (VFTM) in the ATD domain that is responsible for ligand binding. Mutagenesis and interspecies domain swapping between human and rodent T1R2 and T1R3 receptors have determined particular domains and specific amino acid residues to which the species-specific sensitivity to ligands and inhibitors are attributable. For example, human T1R2 VFTM but not rat T1R2 VFTM can be activated by aspartame and neotame; human T1R3 CRD domain and extracellular loops 2 and 3 of HD domain can interact with brazzein and cyclamate, respectively; lactisole docks to the human T1R3 binding pocket comprised of the seven transmembrane helices, while rodent counterparts do not interact with these compounds (Xu et al., 2004; Jiang et al., 2004, 2005; Winnig et al., 2005). These data also demonstrate that multiple ligand binding sites exist in the two monomers of the dimeric sweet receptor T1R2/T1R3, which may explain the earlier observations indicative of multiple sweet receptors (Schiffman et al., 1981). Surprisingly but logically, lactisole and cyclamate that suppress/enhance sweet sensation, respectively, also exert the same effect on umami taste by interacting on the T1R3 moiety of the T1R1/T1R3 umami receptor (Xu et al., 2004).

Fig. 1.1 Schematic representation of a dimeric sweet receptor structure. Each monomer consists of a Venus flytrap module (VFTM), a cysteine rich domain (CRD), a heptahelical transmembrane domain (HD) and an intracellular carboxyl-terminal tail (C-tail) ( Pin et al ., 2003 ). Sweeteners including aspartame and neotame bind to the VFTM module of T1R2 while brazzein and cyclamate bind to the CRD domain and extracellular loops on T1R3, respectively ( Jiang et al ., 2004 ; Xu et al ., 2004 ). Sweet inhibitor lactisole binds to the pocket comprised of T1R3 transmembrane helices ( Jiang et al ., 2005 ; Winnig et al ., 2005 ; Xu et al ., 2004 ).

1.3.3 Discrimination of taste modalities at taste bud cells

In situ hybridization and immunohistochemistry showed that T1R3 is expressed in about 30% of taste bud cells from all types of taste papillae and palate while T1R1 in turn, is mostly expressed in fungiform and palate, less in foliate, and rarely in circumvallate (Fig. 1.2). In contrast, T1R2 is expressed mostly in circumvallate, less in palate and foliate, and rarely in fungiform (Montmayeur et al., 2001; Max et al., 2001; Nelson et al., 2001). Each of T1R1 and T1R2 is co-expressed with T1R3 in a taste bud cell. And some taste bud cells may express T1R3 alone. The bitter T2R receptors are expressed in a different subset of taste bud cells (Adler et al., 2000; Nelson et al., 2001). This non-overlapping cellular expression profile suggests that sweet, umami and bitter taste modalities are discriminated at the taste bud cell level. Knockout of T1R3 receptors markedly reduced the sensitivity to sweet stimuli in behavioral tests but had no effect on bitter, sour and salty tastes (Damak et al., 2003; Zhao et al., 2003). The residual response to sweeteners in T1R3-null mice was abolished by double knockout of both T1R2 and T1R3 receptors. Transgenic introduction of human T1R2 into mice generated animals with humanized sweet taste preferences. These mice could detect substances that taste sweet to humans, but normally not to rodents. In addition, transgenic expression of a modified opioid receptor in T1R2-expressing cells resulted in the attraction of animals to a synthetic opiate that is normally tasteless to wild type mice. And expression of a human receptor T2R16 for a bitter compound phenyl-B-D-glucopyranoside in mouse T1R2-expressing sweet cells generated animals that exhibit strong attraction to this bitter compound (Mueller et al., 2005). Theses studies demonstrated that taste modalities are detected and segregated by the taste bud cells, rather than receptors themselves.

Fig. 1.2 Expression of taste GPCRs in taste bud cells. Sweet receptor (T1R2/T1R3 or T1R3 alone), umami receptor (T1R1/T1R3) and bitter receptors (T2Rs) are expressed in discrete subsets of taste bud cells ( Adler et al ., 2000 ; Hoon et al ., 1999 ; Nelson et al ., 2001 ) . Sweet receptor cells are most abundant in circumvallate papilla, less in foliate and very few in fungiform. In contrast, umami receptor cells are rare in circumvallate, more in foliate and abundant in fungiform ( Nelson et al ., 2001 ). Bitter receptor cells, which can express multiple different bitter receptors, are more abundant in circumvallate and foliate than in fungiform ( Adler et al ., 2000 ).

1.3.4 Could sweet tastants stimulate non-taste receptors in taste cells?

In a recent study (Zubare-Samuelov et al., 2003) we found that some sweet and bitter tastants, e.g. the non-sugar sweeteners, saccharin, D-tryptophan and neohesperidin dihydrochalcone (NHD), and the bitter tastants cyclo(Leu-Trp) and limonin, stimulated pigment aggregation in a Xenopus laevis melanophore cell line as does the native hormone melatonin. Thus, these tastants were able to generate physiological responses independent of taste sensation. Furthermore, as the native melatonin, these tastants stimulated the pigment aggregation by stimulation of the inhibitory pathway of AC as pre-treatment of melanophores with pertussis toxin almost abolished the tastant-induced cAMP reduction. Importantly, the presence of luzindole (melatonin receptor antagonist) almost abolished the inhibition of cAMP formation induced by saccharin, D-tryptophan, and cyclo(Leu-Trp) and the presence of α2-adrenergic receptor antagonist, yohimbine, almost abolished the inhibition of cAMP formation induced by NHD. These data demonstrated that saccharin and D-tryptophan are agonists of the melatonin receptors and NHD is an agonist of the α2-adrenergic receptors under the experimental conditions. It becomes evident that these tastants may stimulate receptors that are not identified taste receptors. Interestingly, both the α2-adrenergic and the melatonin receptors were reported to be present in taste bud cells (Zubare-Samuelov et al., 2003; Herness et al., 2002). If such sweeteners, in addition to their ability to stimulate the T1R2/T1R3 sweet receptors, can also stimulate melatonin, α2-adrenergic receptors, and perhaps other non-taste receptors in taste cells, they may affect the signaling output in taste cells. Should these events occur within the subsecond time course of sweet taste sensation, they may affect taste sensation which may be related to the lack of pure sweetness when tasting non-sugar sweeteners, or even modify time-taste intensity relationships.

Although the PLCβ2 pathway appears to be essential for the cellular transduction of sweet taste (Zhang et al., 2003), to the best of our knowledge no biochemical data are yet available to show the direct activation of specific signaling pathways in taste cells by either the T1R2/R3 or the T1R3 homodimeric sweet taste receptors. In a recent study (Ozeck et al., 2004), sucrose and non-sugar sweeteners such as aspartame, cyclamate and saccharin stimulated the human T1R2/T1R3 sweet receptor expressed in HEK293 cells to activate the inhibitory G-protein signaling pathways (in particular AC) in a pertussis toxin sensitive manner. These data suggest that either the T1R2/T1R3 GPCRs are coupled to the Gαi subunits in taste cells to induce sweet taste transduction or these receptors can be coupled to more than one transduction pathway, further demonstrating a lack of specificity of the transduction output.

1.4 Downstream signaling components

In general, signaling pathways for GPCRs are characterized by the presence of a GTP binding protein (G-protein) which is a coupling component between GPCRs, effector enzymes and ionic channels (Simon et al., 1991; Kristiansen, 2004). There are two main classes of G-proteins. The first are heterotrimeric G-proteins that associate with GPCRs and are involved in signal transduction (Wong, 2003; Kristiansen, 2004). The second are small cytoplasmic G-proteins. The heterotrimeric G-proteins are composed of α, β and γ subunits, with molecular masses of about 39–45, 35–39 and 6–8 kDa, respectively. About 28 distinct G-protein α subunits, which are products of 16 different genes and splice variants have been identified. β and γ subunits are tightly associated and can be regarded as one functional unit. Five different β and 12 different γ subunits have been identified (Kristiansen, 2004). Although not all interactions are possible, many combinations occur which increased the diversity and specificity of signaling. The heterotrimeric G-proteins are divided into four families based on the degree of primary sequence similarities of their α subunits: Gs (Gs and Golf), Gi/o subfamily which inhibits AC and regulates ion channels (Gtr, Gtc, Ggust, Gi1-3, Go, and Gz), Gq/11 which activates PLCβ (Gq, G11, G14, and G15/16), and G12(G12 and G13) that activates Na+/H+ exchanger pathway (Wong, 2003; Kristiansen, 2004; Cabrera-Vera et al., 2003). The G-protein gustducin (Ggust) was first identified in taste cells along with some other G-proteins (McLaughlin et al., 1992). Upon activation by the GPCR, the α-subunit binds GTP in exchange for GDP but both α- and βγ subunits interact specifically with various effectors in cells including ion channels in cell membranes and enzymes both in the cytosol and in cell membranes. In several signaling systems, specific ‘regulators of G-protein signaling’ (RGS) proteins accelerate GTPase activity of the α-subunits. Interestingly, RGS21, a novel RGS was recently found to be co-expressed with sweet T1R2/T1R3 and bitter T2R taste receptors in subpopulations of taste cells (von Buchholtz et al., 2004).

Even though considerable research efforts have been made during the last 15 years trying to elucidate possible cellular mechanisms, the nature of the cellular signals responsible for sweet taste transduction is complex and to date is far from being fully clarified. Early studies indicated high AC (Kurihara and Koyama, 1972) and PDE (Kurihara, 1972) activities in gustatory epithelium, and the presence of these enzymes in the microvilli of cells in rabbit taste buds was also shown (Asanuma and Nomura, 1982). Furthermore, cAMP content could increase in intact bovine taste papillae in response to sucrose stimulation (Cagan, 1974). When the patch-clamp technique became available to record electrophysiological responses in single taste cells, intracellular administration of the cyclic nucleotides cAMP and cGMP into taste cells of frogs and mice (Avenet et al., 1988; Tonosaki and Funakoshi, 1988) decreased potassium conductance, leading to depolarization. In hamsters, fungiform taste buds responded to stimulation by sucrose and some non-sugar sweeteners by inducing action potentials. These responses were mimicked by stimulation with membrane-permeant analogs of cAMP and cGMP (Cummings et al., 1993). Additional biochemical studies employing crude membrane preparations of taste tissue and isolated circumvallate (CV) taste bud sheets have suggested the involvement of the AC cascade in sugar taste transduction (Naim et al., 1991; Striem et al., 1989, 1991). However, additional biochemical experiments with isolated CV taste buds indicated that the non-sugar sweeteners SC45647 and saccharin did not induce cellular increase in cAMP, but rather an increase in IP3 and parallel elevation of intracellular Ca²+, as observed by confocal microscopy (Bernhardt et al., 1996). This indicates two different signaling pathways for sweet taste in a single cell responsive to sweet sucrose and SC45647 but not to the bitter

Hai raggiunto la fine di questa anteprima. Registrati per continuare a leggere!
Pagina 1 di 1

Recensioni

Cosa pensano gli utenti di Optimising Sweet Taste in Foods

0
0 valutazioni / 0 Recensioni
Cosa ne pensi?
Valutazione: 0 su 5 stelle

Recensioni dei lettori