Taste signaling elements expressed in gut enteroendocrine cells
regulate nutrient-responsive secretion of gut hormones1–4
Zaza Kokrashvili, Bedrich Mosinger, and Robert F Margolskee
ABSTRACT
Many of the receptors and downstream signaling elements involved
in taste detection and transduction are also expressed in enteroendo-
crine cells where they underlie the chemosensory functions of the
gut. In one well-known example of gastrointestinal chemosensation
(the “incretin effect”), it is known that glucose that is given orally, but
not systemically, induces secretion of glucagon-like peptide 1 and
glucose-dependent insulinotropic peptide (the incretin hormones),
which in turn regulate appetite, insulin secretion, and gut motility.
Duodenal L cells express sweet taste receptors, the taste G protein
gustducin, and several other taste transduction elements. Knockout
mice that lack gustducin or the sweet taste receptor subunit T1r3 have
deficiencies in secretion of glucagon-like peptide 1 and glucose-
dependent insulinotropic peptide and in the regulation of plasma
concentrations of insulin and glucose in response to orally ingested
carbohydrate—ie, their incretin effect is dysfunctional. Isolated
small intestine and intestinal villi from gustducin null mice displayed
markedly defective glucagon-like peptide 1 secretion in response to
glucose, indicating that this is a local circuit of sugar detection by
intestinal cells followed by hormone secretion from these same cells.
Modulating hormone secretion from gut “taste cells” may provide
novel treatments for obesity, diabetes, and malabsorption syn-
dromes. Am J Clin Nutr 2009;90(suppl):822S–5S.
MOLECULAR BASIS OF TASTE SIGNALING
The vertebrate sense of taste depends on specialized epithelial
receptor cells contained in taste buds located in the surface
papillae of the tongue. Taste is initiated by the interaction of
tastants with receptors and ion channels in the apical microvilli of
taste receptor cells. Some taste transduction pathways convert
chemical information into a cellular second messenger code (eg,
cyclic nucleotides and inositol trisphosphate) (reviewed in ref-
erence 1). These messengers are components of signaling cas-
cades that typically lead to taste receptor cell depolarization and
Ca2+ release. In other cases, the tastant itself may constitute all
or part of the initial cellular signal (eg, Na+, K+, H+).
The sense of sweet taste is initiated by the binding of sugars or
sweeteners to the sweet taste receptor: a heterodimer of 2 type 1
taste G protein–coupled receptors (T1R2 + T1R3) (2–6). The
sweet taste receptor couples to heterotrimeric gustducin (a-gustducin,
Gb3, and Gc13) (7, 8). Gustducin’s a-subunit (a-gustducin)
activates taste cell phosphodiesterase to decrease cyclic nucle-
otide concentrations (1), whereas gustducin’s bc-unit (Gb3-Gc13)
activates phospholipase Cb2 to generate inositol trisphosphate
and diacylglycerol (9). Ca2+ influx and release from internal
822S Am J Clin Nutr 2009;90(suppl):822S–5S. Printed in USA. Ó 2009 American Society for Nutrition
stores activates taste-cell-expressed transient receptor potential
channel type M5 (TRPM5) (10), a Ca2+-activated cation channel
(11–13), which leads to taste cell depolarization and signaling to
other taste cells in the bud and to gustatory afferent nerves.
TASTE MOLECULES ARE EXPRESSED IN THE GUT
The taste G protein gustducin, originally identified in taste
receptor cells (7), is expressed also in the cells of the stomach and
small intestine (14, 15). In recent work from our group (16, 17)
gustducin’s 3 subunits (a-gustducin, Gb3, Gc13), along with
many other taste-signaling elements (eg, T1r1, T1r2, T1r3,
TRPM5, PLCb2, and others) were also found to be expressed in
L-type enteroendocrine cells of the small intestine. Indeed, en-
tire taste-signaling pathways are present in human duodenal L
cells (16). The roles of gustducin, taste receptors, TRPM5, and
other taste-signaling elements expressed in gut endocrine cells
are now being made clear from physiologic studies in knockout
mice. The expression of taste-signaling elements in gut endo-
crine cells and the characterization of their function as the gut’s
luminal glucose sensor that initiates the incretin response to
elicit the release of glucagon-like peptide 1 (GLP-1) from L
cells are described here and elsewhere (16, 17).
TASTE MOLECULES UNDERLIE THE INCRETIN
EFFECT
GLP-1, secreted from enteroendocrine L cells of the gut, is an
incretin hormone that augments the release of insulin from the
pancreas. The incretin effect—the observation that orally in-
gested glucose is a much more effective stimulator of insulin
secretion from the pancreas than is intravenously injected glucose
(18)—is primarily mediated by GLP-1. It was recently deter-
mined that sugars in the gut lumen act on the taste-signaling
proteins T1r3 and gustducin that are expressed in L cells to elicit
1
From the Department of Neuroscience, Mount Sinai School of Medi-
cine, New York, NY.
2
Presented at the “100th Anniversary Symposium of Umami Discovery:
The Roles of Glutamate in Taste, Gastrointestinal Function, Metabolism, and
Physiology,” held in Tokyo, Japan, 10–13 September 2008.
3
Supported in part by NIH grant DC03055 to RFM.
4
Address correspondence to RF Margolskee, Department of Neurosci-
ence, Mount Sinai School of Medicine, 1425 Madison Avenue, Box 1065,
New York, NY 10029. E-mail: robert.margolskee@mssm.edu.
First published online July 1, 2009; doi: 10.3945/ajcn.2009.27462T.
 TASTE ELEMENTS IN GUT ENTEROENDOCRINE CELLS
the release of GLP-1 from enteroendocrine L cells (16). Anal-
ogously with their function in oral taste cells, the gut-expressed
taste-signaling elements respond to sugars and artificial sweet-
eners in the gut’s lumen (16, 17). However, instead of trans-
mitting their signals via gustatory afferents, these gut “taste cells”
act via humoral mediators such as GLP-1 and glucose-dependent
insulinotropic peptide (GIP).
TASTE MOLECULES ARE EXPRESSED IN INTESTINAL
ENTEROENDOCRINE CELLS
Intestinal mucosal cells in humans and mice have been ex-
amined for the presence of a-gustducin, T1R taste receptors, and
other known taste-signaling elements (16, 17). In human duo-
denal biopsy sections, a-gustducin was shown by indirect im-
munofluorescence to be present in 4 populations of intestinal
mucosal cells: 1) GLP-1-expressing enteroendocrine L cells, 2)
GIP-expressing enteroendocrine K cells, 3) GIP and GLP-1 co-
expressing entereoendocrine K/L cells, and 4) mucosal cells
(presumed to be brush cells) that expressed neither GLP-1 nor
GIP (16). Most human duodenal L cells and a sizable minority
of K cells expressed a-gustducin (16). Independent confirmation
of the expression of a-gustducin in human duodenal enter-
oendocrine K and L cells came from laser capture followed
by reverse transcriptase–polymerase chain reaction (16). This
same technique was used to show that a-gustducin was absent
from human duodenal enterocytes (16). In addition, multiple
taste-signaling elements (T1R2, T1R3, Gb3, Gc13, PLCb2, and
TRPM5) were found to be co-expressed with a-gustducin and
GLP-1 in human duodenal L cells (16).
TASTE MOLECULES IN ENTEROENDOCRINE L CELLS
UNDERLIE THE INCRETIN EFFECT
The function of taste-signaling elements in intestinal endo-
crine cells (mouse duodenal cells) has been examined by looking
for the co-expression of taste-signaling elements. In mouse duo-
denum, GLP-1-expressing L cells frequently expressed a-gustducin
FIGURE 1. A: Indirect immunofluorescence staining of mouse duodenum showing co-expression of a-gustducin (a-gust) with glucagon-like peptide 1
(GLP-1), and GLP-2. Intrinsic fluorescence of green fluorescent protein (GFP) shows the expression of GFP and GLP-2 in the a-gustducin-expressing cells of
a-gustducin–GFP transgenic mice. B: Indirect immunofluorescence of mouse jejunum showing the co-expression of a-gustin with GLP-1, GLP-2, and peptide
YY (PYY). Bars, 15 lm. Data are modified with permission from reference 16.
823S
and an a-gustducin marker (green fluorescent protein driven
from the a-gustducin promoter) (Figure 1A). Mouse L cells in
duodenum (Figure 1A), jejunum (Figure 1B), and ileum (16)
frequently expressed a-gustducin along with GLP-2 and peptide
YY (well known to be present in L cells).
Knockout mice lacking either a-gustducin (a-gust2/2) or T1r3
(T1r32/2) were examined for their enteroendocrine cell responses
to glucose in the gut lumen. To determine the effects of directly
stimulating the gut’s enteroendocrine cells and to eliminate the
potential effects of the oral taste receptor cells, glucose was ga-
vage administered by inserting feeding needles directly into the
stomachs of a-gustducin knockout mice (a-gust2/2) and their
wild-type (a-gust+/+) littermates (Figure 2). Gavage-administered
glucose led to a significant rise (P , 0.01) in plasma concen-
trations of GLP-1 in wild-type mice with a peak ’10 min after
gavage (Figure 2A). In contrast, there was no significant rise in
plasma concentrations of GLP-1 in the a-gust2/2 mice after
glucose gavage (Figure 2A). Gavage-administered glucose led to
a significant rise (P , 0.001) in plasma concentrations of insulin
in wild-type mice with a peak ’45 min after gavage (Figure 2B).
In contrast, there was a markedly delayed rise in plasma con-
centrations of insulin in a-gust2/2 mice after glucose gavage
(Figure 2B). In the a-gustducin-null mice, insulin reached a peak
’60 min after gavage and remained elevated for 2 h (Figure 2B),
which is a marked difference from wild-type mice whose insulin
concentrations gradually returned to baseline between 60 and 120
min after gavage.
The a-gustducin-null mice also showed disrupted glucose
homeostasis: plasma glucose concentrations were higher in
a-gust2/2 mice compared with wild type after overnight fasting
followed by feeding (Figure 2C). Furthermore, plasma glucose
concentrations in a-gust2/2 mice remained high for .2 h after
feeding (Figure 2C).
a-Gustducin is expressed in brush cells of the stomach (16) as
well as in intestinal enteroendocrine cells (Figure 1). Thus, it is
possible that glucose administered by gavage into the stomach
might be acting directly on the brush cells and indirectly on the
intestinal L cells via brush-cell-released factors. To rule out such
824S KOKRASHVILI ET AL

FIGURE 2. Secretion of glucagon-like peptide 1 (GLP-1) (A) and insulin (B) in response to glucose gavage (5 g/kg body weight) in a-gustducin knockout
(a-gust2/2) mice and wild-type (a-gust+/+) mice. (C) Postfasting plasma glucose concentrations after feeding standard rodent diet in a-gust2/2 mice and
a-gust+/+ mice. (D) Secretion of GLP-1 in response to 10% glucose injected into surgically isolated duodenum in a-gust2/2 mice, T1r3 knockout (T1r32/2)
mice, and a-gust+/+ mice. The duodenum was ligated away from the stomach and the rest of the intestines, and circulatory contact was maintained. (E)
Secretion of GLP-1 ex vivo from minced proximal duodenum from a-gust2/2 and a-gust+/+ mice in response to the addition of 10% glucose to the culture
medium. (F) Secretion of GLP-1 ex vivo from isolated duodenal villi from a-gust2/2 and a-gust+/+ mice in response to the addition of 10% glucose to the
culture medium. For in vivo experiments, n = 5–12 animals/genotype; in vitro experiments were carried out in triplicate and replicated at least twice. All
values are means 6 SEMs. *,**,*** Statistical significance was determined by ANOVA: *P , 0.05, **P , 0.01, ***P , 0.001. Data are modified with
permission from Reference 16.

indirect effects, glucose was injected directly into the duodena
of wild-type, a-gust2/2, and T1r32/2 mice. In each animal, the
duodenum was surgically isolated from the stomach and the
distal portion of the small intestine, but remained in circulatory
contact so that plasma concentrations of hormones could be
monitored. After duodenal injection with glucose, plasma GLP-
1 in wild-type mice peaked at 10 min after injection and then
returned to baseline at 20 min (Figure 2D). In marked contrast,
plasma concentrations of GLP-1 did not increase at all after
duodenal injection with glucose in either a-gust2/2 or T1r32/2
mice (Figure 2D).
To determine whether the glucose-stimulated release of GLP-1
was intrinsic to the enteroendocrine L cells of the duodenum, ex
vivo experiments were carried out with dissected proximal du-
odena and isolated duodenal villi from wild-type and a-gustducin
knockout mice. The addition of 10% glucose to the culture
medium increased the release of GLP-1 from duodenal tissue
from both wild-type and a-gust2/2 mice (Figure 2E). However,
the glucose stimulation of GLP-1 release was much greater in
the wild-type mice. Similar results were observed when glucose
stimulated a greater release of GLP-1 from isolated villi of wild-
type mice than from those of a-gust2/2 mice (Figure 2F).
CONCLUSIONS
In 2 recent studies (16, 17), gustducin-coupled sweet taste
receptors were found to be present in enteroendocrine cells in the
proximal intestines of mice and humans. In the human duode-
num, a-gustducin was found in 3 types of enteroendocrine cells:
K (GIP), L (GLP-1), and K/L (GIP and GLP-1) cells (16). In
addition to expressing a-gustducin and the sweet receptor sub-
units T1R2 and T1R3, human duodenal L cells also expressed
several other taste-signaling molecules, including Gb3, Gc13,
PLCb2, and TRPM5 (16).
In mice, a-gustducin was frequently found in L-type enter-
oendocrine cells but rarely in K cells. a-Gustducin knockout mice
were disrupted in their glucose homeostasis and hormonal re-
sponses to glucose within the lumen of the small intestine. The
primary deficit was the failure of enteroendocrine cells to release
GLP-1 in response to glucose within the gut lumen (16). The
 TASTE ELEMENTS IN GUT ENTEROENDOCRINE CELLS
absent GLP-1 response to glucose led to an abnormal insulin re-
sponse and prolonged elevation of postprandial blood glucose.
T1r3 knockout mice also failed to release GLP-1 from their du-
odenal cells after glucose injection into the intestine. Such data
indicate that sweet receptors in intestinal L cells couple to heter-
otrimeric gustducin to detect extracellular glucose and then re-
spond with secretion of GLP-1. In 2 L-cell lines, both a-gustducin
and T1r3 were required for stimulating GLP-1 release in response
to either sugars or noncaloric sweeteners (16, 17). The results
indicate that the sensing of sugars and sweeteners by taste-
signaling elements expressed in intestinal L cells in vivo leads to
GLP-1 release from these same cells. (Other articles in this sup-
plement to the Journal include references 19–47.)
The authors’ responsibilities were as follows—ZK, BM, and RFM: con-
tributed to the design and analysis of the experiments and to writing the man-
uscript; RFM: wrote the final version of the manuscript. The travel expenses
of the presenting author (RFM) associated with participation in the sympo-
sium and an honorarium were paid by the conference sponsor, the Interna-
tional Glutamate Technical Committee, a nongovernmental organization
funded by industrial producers and users of glutamate in food. ZK and
BM had no conflicts of interest. RFM has a personal financial interest in
the form of stock ownership in the Redpoint Bio company, receives consulting
fees from Redpoint Bio, and is an inventor on patents and patent applications
that have been licensed to Redpoint Bio. Redpoint Bio is a biotechnology
company that identifies and develops compounds to improve the taste of phar-
maceutical, food, and beverage products.
REFERENCES
1. Kinnamon SC, Margolskee RF. Taste transduction. In: Basbaum A,
Bushnell M, Smith S, et al, eds. The senses: a comprehensive reference,
Vol 4. Amsterdam, Netherlands; Boston, MA: Elsevier, 2008:219–36.
2. Max M, Shanker YG, Huang L, et al. Tas1r3, encoding a new candidate
taste receptor, is allelic to the sweet responsiveness locus Sac. Nat Genet
2001;28:58–63.
3. Montmayeur JP, Liberles SD, Matsunami H, et al. A candidate taste
receptor gene near a sweet taste locus. Nat Neurosci 2001;4:492–8.
4. Nelson G, Hoon MA, Chandrashekar J, et al. Mammalian sweet taste
receptors. Cell 2001;106:381–90.
5. Li X, Staszewski L, Xu H, et al. Human receptors for sweet and umami
taste. Proc Natl Acad Sci USA 2002;99:4692–6.
6. Damak S, Rong M, Yasumatsu K, et al. Detection of sweet and umami
taste in the absence of taste receptor T1r3. Science 2003;301:850–3.
7. McLaughlin SK, McKinnon PJ, Margolskee RF. Gustducin is a taste-cell-
specific G protein closely related to the transducins. Nature 1992;357:563–9.
8. Huang L, Shanker YG, Dubauskaite J, et al. Gc13, a new G protein c
subunit expressed in gustducin-positive taste receptor cells, mediates IP3
responses to bitter denatonium. Nat Neurosci 1999;2:1055–62.
9. Rossler P, Kroner C, Freitag J, et al. Identification of a phospholipase C
beta subtype in rat taste cells. Eur J Cell Biol 1998;77:253–61.
10. Perez CA, Huang L, Rong M, et al. A transient receptor potential
channel expressed in taste receptor cells. Nat Neurosci 2002;5:1169–76.
11. Hofmann T, Chubanov V, Gudermann T, et al. TRPM5 is a voltage-
modulated and Ca(2+)-activated monovalent selective cation channel.
Curr Biol 2003;13:1153–8.
12. Liu D, Liman ER. Intracellular Ca2+ and the phospholipid PIP2 regulate
the taste transduction ion channel TRPM5. Proc Natl Acad Sci USA
2003;100:15160–5.
13. Prawitt D, Monteilh-Zoller MK, Brixel L, et al. TRPM5 is a transient
Ca2+-activated cation channel responding to rapid changes in [Ca2+]i.
Proc Natl Acad Sci USA 2003;100:15166–71.
14. Höfer D, Drenckhahn D. Identification of the taste cell G-protein,
alpha-gustducin, in brush cells of the rat pancreatic duct system. His-
tochem Cell Biol 1998;110:303–9.
15. Höfer D, Puschel B, Drenckhahn D. Taste receptor-like cells in the rat
gut identified by expression of alpha-gustducin. Proc Natl Acad Sci USA
1996;93:6631–4.
16. Jang HJ, Kokrashvili Z, Theodorakis MJ, et al. Gut-expressed gustducin
and taste receptors regulate secretion of glucagon-like peptide-1. Proc
Natl Acad Sci USA 2007;104:15069–74.
825S
17. Margolskee RF, Dyer J, Kokrashvili Z, et al. T1R3 and gustducin in gut
sense sugars to regulate expression of Na+-glucose cotransporter 1. Proc
Natl Acad Sci USA 2007;104:15075–80.
18. McIntyre N, Holdsworth CD, Turner DS. New interpretation of oral
glucose tolerance. Lancet 1964;4:20–1.
19. Fernstrom JD. Introduction to the symposium. Am J Clin Nutr 2009;90
(suppl):705S–6S.
20. Krebs JR. The gourmet ape: evolution and human food preferences. Am
J Clin Nutr 2009;90(suppl):707S–11S.
21. Curtis RI. Umami and the foods of classical antiquity. Am J Clin Nutr
2009;90(suppl):712S–8S.
22. Kurihara K. Glutamate: from discovery as a food flavor to role as a basic
taste (umami). Am J Clin Nutr 2009;90(suppl):719S–22S.
23. Beauchamp GK. Sensory and receptor responses to umami: an overview
of pioneering work. Am J Clin Nutr 2009;90(suppl):723S–7S.
24. Sano C. History of glutamate production. Am J Clin Nutr 2009;90
(suppl):728S–32S.
25. Li X. T1R receptors mediate mammalian sweet and umami taste. Am J
Clin Nutr 2009;90(suppl):733S–7S.
26. Chaudhari N, Pereira E, Roper SD. Taste receptors for umami: the case
for multiple receptors. Am J Clin Nutr 2009;90(suppl):738S–42S.
27. San Gabriel A, Maekawa T, Uneyama H, Torii K. Metabotropic gluta-
mate receptor type 1 in taste tissue. Am J Clin Nutr 2009;90(suppl):
743S–6S.
28. Yasumatsu K, Horio N, Murata Y, et al. Multiple receptors underlie
glutamate taste responses in mice. Am J Clin Nutr 2009;90(suppl):
747S–52S.
29. Kinnamon SC. Umami taste transduction mechanisms. Am J Clin Nutr
2009;90(suppl):753S–5S.
30. Bachmanov AA, Inoue M, Ji H, Murata Y, Tordoff MG, Beauchamp
GK. Glutamate taste and appetite in laboratory mice: physiologic and
genetic analyses. Am J Clin Nutr 2009;90(suppl):756S–63S.
31. Shigemura N, Shirosaki S, Ohkuri T, et al. Variation in umami per-
ception and in candidate genes for the umami receptor in mice and
humans. Am J Clin Nutr 2009;90(suppl):764S–9S.
32. Chen Q-Y, Alarcon S, Tharp A, et al. Perceptual variation in umami
taste and polymorphisms in TAS1R taste receptor genes. Am J Clin Nutr
2009;90(suppl):770S–9S.
33. Mennella JA, Forestell CA, Morgan LK, Beauchamp GK. Early milk
feeding influences taste acceptance and liking during infancy. Am J Clin
Nutr 2009;90(suppl):780S–8S.
34. Raliou M, Wiencis A, Pillias A-M, et al. Nonsynonymous single nucleo-
tide polymorphisms in human tas1r1, tas1r3, and mGluR1 and individual
taste sensitivity to glutamate. Am J Clin Nutr 2009;90(suppl):789S–99S.
35. Donaldson LF, Bennett L, Baic S, Melichar JK. Taste and weight: is
there a link? Am J Clin Nutr 2009;90(suppl):800S–3S.
36. Rolls ET. Functional neuroimaging of umami taste: what makes umami
pleasant? Am J Clin Nutr 2009;90(suppl):804S–13S.
37. Blachier F, Boutry C, Bos C, Tomé D. Metabolism and functions of
L-glutamate in the epithelial cells of the small and large intestines. Am J
Clin Nutr 2009;90(suppl):814S–21S.
38. Akiba Y, Kaunitz JD. Luminal chemosensing and upper gastrointestinal
mucosal defenses. Am J Clin Nutr 2009;90(suppl):826S–31S.
39. Kondoh T, Mallick HN, Torii K. Activation of the gut-brain axis by
dietary glutamate and physiologic significance in energy homeostasis.
Am J Clin Nutr 2009;90(suppl):832S–7S.
40. Tomé D, Schwarz J, Darcel N, Fromentin G. Protein, amino acids, vagus
nerve signaling, and the brain. Am J Clin Nutr 2009;90(suppl):838S–43S.
41. Yamamoto S, Tomoe M, Toyama K, Kawai M, Uneyama H. Can dietary
supplementation of monosodium glutamate improve the health of the
elderly? Am J Clin Nutr 2009;90(suppl):844S–9S.
42. Burrin DG, Stoll B. Metabolic fate and function of dietary glutamate in
the gut. Am J Clin Nutr 2009;90(suppl):850S–6S.
43. Brosnan ME, Brosnan JT. Hepatic glutamate metabolism: a tale of 2
hepatocytes. Am J Clin Nutr 2009;90(suppl):857S–61S.
44. Stanley CA. Regulation of glutamate metabolism and insulin secretion
by glutamate dehydrogenase in hypoglycemic children. Am J Clin Nutr
2009;90(suppl):862S–6S.
45. Hawkins RA. The blood-brain barrier and glutamate. Am J Clin Nutr
2009;90(suppl):867S–74S.
46. Magistretti PJ. Role of glutamate in neuron-glia metabolic coupling. Am
J Clin Nutr 2009;90(suppl):875S–80S.
47. Fernstrom JD. Symposium summary. Am J Clin Nutr 2009;90(suppl):
881S–5S.