Progress in Neurobiology 75 (2005) 295–307

www.elsevier.com/locate/pneurobio

The taste cell-related diffuse chemosensory system

A. Sbarbati *, F. Osculati
Department of Morphological-Biomedical Sciences, Section of Anatomy and Histology, University of Verona,
Strada Le Grazie, 8, 37134 Verona, Italy
Received 7 October 2004; accepted 8 March 2005

Abstract

Elements expressing the molecular mechanisms of gustatory transduction have been described in several organs in the digestive and
respiratory apparatuses. These taste cell-related elements are isolated cells, which are not grouped in buds, and they have been interpreted as
chemoreceptors. Their presence in epithelia of endodermal origin suggests the existence of a diffuse chemosensory system (DCS) sharing
common signaling mechanisms with the ‘‘classic’’ taste organs. The elements of this taste cell-related DCS display a site-related morphologic
polymorphism, and in the past they have been indicated with various names (e.g., brush, tuft, caveolated, fibrillo-vesicular or solitary
chemosensory cells). It may be that the taste cell-related DCS is like an iceberg: the taste buds are probably only the most visible portion, with
most of the iceberg more caudally located in the form of solitary chemosensory cells or chemosensory clusters. Comparative anatomical
studies in lower vertebrates suggest that this ‘submerged’ portion may represent the most phylogenetically ancient component of the system,
which is probably involved in defensive or digestive mechanisms. In the taste buds, the presence of several cell subtypes and of a wide range of
molecular mechanisms permits precise food analysis. The larger, ‘submerged’ portion of the iceberg is composed of a polymorphic population
of isolated elements or cell clusters in which the molecular cascade of cell signaling needs to be explored in detail. The little data we have
strongly suggests a close relationship with taste cells. Morphological and biochemical considerations suggest that the DCS is a potential new
drug target. Modulation of the respiratory and digestive apparatuses through substances, which act on the molecular receptors of this
chemoreceptive system, could be a new frontier in drug discovery.
# 2005 Elsevier Ltd. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 296
2. Anatomy of the peripheral taste system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 296
2.1. Taste transduction mechanisms. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 296
2.2. Taste-related markers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 297
2.3. Gustducin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 297
2.4. Expression of elements of bitter taste transduction mechanisms outside taste buds . . . . . . . . . . . . . . . . . . . . . . . . . 298
2.5. Expression of bitter taste receptors of the T2R family in the gastrointestinal tract . . . . . . . . . . . . . . . . . . . . . . . . . 298
3. A ‘‘classic’’ DCS: brush cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 298
3.1. BCs in different organs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 299
3.2. Phenotypic characteristics of BCs. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 299
3.3. Considerations about BCs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 299
4. Solitary chemosensory cells (SCCs) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 299

Abbreviations: a-gustducin, gustducin alpha subunit; BCs, brush cells; cAMP, adenosine 30 ,50 -cyclic monophosphate; Ca2+, calcium; CCs, chemosensory
clusters; DCS, diffuse chemosensory system; G-protein, guanine nucleotide-binding regulatory protein; CPCR, G-protein-coupled receptors; IP3R3, inositol
1,4,5-triphosphate type III receptor; mGluR4, metabotropic glutamate receptor R4; PLCb2, phospholipase C of the b2 subtype; SCCs, solitary chemosensory
cells; SLSE, specific laryngeal sensory epithelium; T1R/Tas1R, taste receptor family 1; T2R, taste receptor family 2; TRP, transient receptor potential
* Corresponding author. Tel.: +39 045 8027155; fax: +39 045 8027163.
E-mail address: andrea.sbarbati@univr.it (A. Sbarbati).

0301-0082/$ – see front matter # 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.pneurobio.2005.03.001
296 A. Sbarbati, F. Osculati / Progress in Neurobiology 75 (2005) 295–307

4.1. Comparative anatomical considerations . . . . . . . . . . . . . . . . .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 299

4.2. SCCs in the oral cavity . . . . . . . . . . . . . . . . . . . . . . . . . . . .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 301
4.3. SCCs in the nasal cavities . . . . . . . . . . . . . . . . . . . . . . . . . .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 301
4.4. SCCs in the larynx . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 302
4.5. SCCs in the trachea and bronchi . . . . . . . . . . . . . . . . . . . . .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 302
4.6. Structures possibly associated with SCCs . . . . . . . . . . . . . . .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 302
4.7. General consideration about SCCs . . . . . . . . . . . . . . . . . . . .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 302
5. Chemosensory clusters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 303
6. Conclusions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 303
6.1. Similarities and differences between the primary taste system and the DCS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 303
6.2. The role of the DCS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 304
6.3. The DCS and the quorum sensing based strategy . . . . . . . . . .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 304
6.4. General conclusions: uncovering the iceberg . . . . . . . . . . . . .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 304
6.5. Unanswered questions and perspectives. . . . . . . . . . . . . . . . .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 305
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 305
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 305

1. Introduction defining the boundaries of this newly discovered anatomical

In recent years, new discoveries have been made about
the transduction mechanisms underlying the sensation of
taste (Kinnamon and Margolskee, 1996; Gilbertson et al.,
2000; Lindemann, 2001; Perez et al., 2003). The availability
of new molecular markers allows a detailed exploration of
taste cells in buds of the oro-pharyngeal cavity, and recently
‘‘taste cells’’ have also been described in other organs of
endodermal origin located in the digestive and respiratory
apparatuses. These taste cell-related elements are isolated
cells, which are not grouped in buds, and they have been
interpreted as chemoreceptors. Their presence in epithelia of
endodermal origin suggests the existence of a diffuse
chemosensory system (DCS) sharing common signaling
mechanisms with the ‘‘classic’’ taste organs. The study of
this DCS is of evident importance for extending our
knowledge of the digestive and respiratory apparatuses, and
could also be pertinent to understand the pathogenesis of
diseases, which may affect them.
The elements of the taste cell-related DCS are mainly
localized in specific areas where they may constitute a
significant percentage of the epithelial population and
display a site-related morphologic polymorphism.
Basically, two cell types have been described with taste
cell characteristics, i.e., brush cells (BCs) and solitary
chemosensory cells (SCCs). The two cell types have
similarities, which were noted several years ago, but only
recently has their molecular resemblance to taste cells been
discovered. The relationship between BCs and SCCs is not
clear, and the situation is further confused by the diverse
names used in the literature to indicate the same cytotype
(BCs, tuft cells, caveolated cells, fibrillovesicular cells and
others).
The present review focuses on the specific cytotypes
discovered so far which appear to be related to taste
cells, drawing on the studies that have contributed to
their definition. In addition, we summarize current knowl-
edge about the taste cell-related DCS, contributing to
system.
2. Anatomy of the peripheral taste system
The peripheral taste system includes the gustatory
sensory organs or taste buds, their innervation and the
papillae within which taste buds are assembled (for a recent
review see also Witt et al. (2003)). On the tongue taste buds
are associated with papillae that are fungiform on the
anterior margin, foliate on the side toward the back and
vallate on the back of the tongue. But taste buds are not
confined to the tongue; there are also taste buds on the soft
palate, cheeks, pharynx, esophagus and larynx. Among
vertebrates, taste buds are remarkably similar in size and
shape, ranging from about 20 to 40 mm in diameter and
about 40–60 mm in length. Mammalian taste buds consist of
an elongated cluster of about 50 epithelial cells.
In mammalian taste buds, recent studies recognize four
cell types. Type I cells are the most frequent and ensheath
axons and other cells as glial cells do. These cells are
characterized by large apical granules and are widely
considered to be supporting and secretory elements. Type II
cells are fusiform elements that do not possess granules or
synapses. The function of these elements is still the subject
of debate, but they are generally considered chemosensory
or secretory. Type III cells have apical and basal processes
that make synapses with axons. These elements are
generally considered to be taste sensory cells. Type IV
cells are poorly differentiated elements located at the bases
of taste buds that are generally considered to be stem cells. In
addition to the above-named cell types, perigemmal (type V)
cells are also found around taste buds.
2.1. Taste transduction mechanisms
Several recent reviews have described the molecular
mechanisms of taste transduction (Lindemann, 2001;
 A. Sbarbati, F. Osculati / Progress in Neurobiology 75 (2005) 295–307
Margolskee, 2002; Perez et al., 2003). Briefly, five different
taste qualities exist (i.e., sodium salt, acids, amino acids,
sweet and bitter), but the transduction mechanisms linked to
those qualities are still partially unknown. All taste pathways
are thought to converge on common elements that mediate a
rise in intracellular Ca2+ followed by transmitter release. An
enormous stimulus to studies of taste has been provided by
the recognition that taste responses to bitter/sweet com-
pounds and amino acids are initiated by G-protein-coupled
receptors (GPCRs) and transduced via G-protein signaling
cascades (for reviews, see Chaudhari and Roper (1998),
Gilbertson et al. (2000) and Lindemann (2001)). During the
past few years, several GPCRs have been identified in taste
cells and implicated in taste signal transduction (Adler et al.,
2000; Chandrashekar et al., 2000; Chaudhari et al., 2000;
Max et al., 2001; Nelson et al., 2001, 2002; Li et al., 2002;
Amrein and Bray, 2003).
A series of recent studies (Adler et al., 2000; Nelson et al.,
2001; Zhang et al., 2003) has demonstrated that differences
between taste qualities are linked to different families of
these receptors expressed in sets of taste receptor cells.
Briefly, bitter compounds activate bitter taste T2R/Trb
receptors, which are encoded by a separate gene family
consisting of about 30 members in mice. T2R receptors then
activate gustducin heterotrimers. Activated alpha-gustducin
stimulates phosphodiesterases to hydrolyze cAMP; the
decrease in cAMP levels may modulate cyclic nucleotide-
regulated ion channels and/or kinases. Beta and gamma
subunits of gustducin activate phospholipase C of the b2
subtype to generate IP3, which leads to release of Ca2+ from
internal stores via activation of inositol 1,4,5-trisphosphate
receptor type III (IP3R3).
Detection of amino acid and sweet compounds is mainly
effected by the Tas1R (or T1R) gene family, which encodes
three conserved receptors that function as heterodimers and
form either a sugar receptor (Tas1R2/Tas1R3) or a general
amino acid receptor (Tas1R1/Tas1R3).
More in detail, the candidate receptors for amino acid
taste transduction are ionotropic glutamate receptors,
metabotropic glutamate receptors and in particular taste-
mGluR4, which is a truncated form of the brain mGluR4,
lacking most of the N-terminal extracellular domain as
well as the Tas1R1–Tas1R3 heteromer (Chaudhari et al.,
2000; Li et al., 2002; Nelson et al., 2002; Ruiz et al.,
2003; He et al., 2004). Tas1R1 and Tas1R3 are co-
expressed in taste buds in the anterior part of the tongue
(Nelson et al., 2001), while taste mGluR4 is expressed in
taste buds of the circumvallate and foliate papillae (Yang
et al., 1999).
Tas1R2–Tas1R3 is a GPCR activated by most known
sweeteners (Nelson et al., 2001).
Natural sugars activate G-protein-coupled receptors
linked via adenylate cyclase to cAMP production. In
contrast, artificial sweeteners seem to activate both
ionotropic receptors linked to cation channels, and GPCRs
linked via phospholipase C to IP3 production.
297
2.2. Taste-related markers
On the basis of an impressive amount of recent data, it has
so far been possible to identify a panel of molecules involved
in taste transduction that can be used as taste-related markers
to identify putative chemosensory cells in the epithelial
lining of different apparatuses, thus establishing a link
between the classic ‘‘taste cells’’ and isolated chemosensory
cells (Finger et al., 2003; Sbarbati et al., 2004a,b).
These molecules include taste receptors, gustducin,
PLCb2 (Rossler et al., 1998), IP3R3 (Clapp et al., 2001)
and Trpm5. This latter is a member of the mammalian family
of transient receptor potential (TRP) channels (Perez et al.,
2002). In taste cells, Trpm5 is coexpressed with taste-
signaling molecules such as a-gustducin, Ggamma13,
PLCb2 and IP3R3. Heterologous expression studies of
Trpm5 indicated that it functions as a cationic channel that is
gated when internal calcium stores are depleted. Trpm5 has
been considered to be responsible for capacitative calcium
entry in taste receptor cells that respond to bitter and/or
sweet compounds (Perez et al., 2002).
The definition of taste-related cells can therefore be
obtained on the basis of a chemical code that has been
clarified in a series of studies including molecular biology
and morphology. These elements express cell signaling
mechanisms, which appear to be specific and which have
been the subject of recent reviews (Margolskee, 2002; Perez
et al., 2003).
A recent example of this approach is the demonstration of
the expression, at mRNA and protein levels, of members of
the Tas1R sweet taste receptor family and the a-gustducin,
in the small intestine (Dyer et al., 2005). The use of taste-
related markers also makes possible comparative evaluation
among different species, as an example Asano-Miyoshi et al.
(2000) individuated in barbels of fishes cells expressing taste
receptors and a PLCb2 (Rossler et al., 1998), suggesting that
the tastant-induced second messenger response in taste cells
is common to vertebrate.
2.3. Gustducin
A key molecule in the cascade of events that lead to bitter
transduction is gustducin, a heterotrimeric guanine-binding
protein (G-protein). Its presence was first demonstrated in
rats as a taste specific G-protein (McLaughlin et al., 1992)
and then confirmed in man (Takami et al., 1994). The
gustducin alpha subunit (a-gustducin) is specifically
expressed in taste cells of the circumvallate, foliate, and
fungiform papillae of rat lingual tissue. In rat vallate taste
buds, a-gustducin has been found in cells with character-
istics of Type II (light) cells (Boughter et al., 1997). Other
authors suggested that in rats, a-gustducin is also expressed
in the cytoplasm of type III cells and probably in microvilli
of type I cells of the vallate taste buds (Menco et al., 1997).
Therefore, a-gustducin is considered to be a potent
marker of chemosensitive cells with an important role in
298 A. Sbarbati, F. Osculati / Progress in Neurobiology 75 (2005) 295–307

molecular mechanisms of bitter and sweet taste transduction

(Margolskee, 2002).

2.4. Expression of elements of bitter taste transduction

mechanisms outside taste buds

Recent findings showed that molecules of the bitter taste

transduction mechanism are expressed outside taste buds.
The presence of gustducin was first reported in the stomach
and intestine of the rat (Höfer et al., 1996), but later findings
demonstrated that several elements of the signaling cascade
colocalize, including taste receptors, gustducin, PLCb2
(Finger et al., 2003) and IP3R3 (Sbarbati et al., 2004a).
These taste cell-related elements have so far been described
as isolated elements in tissues of endodermic origin (i.e., the
digestive and respiratory apparatuses).

2.5. Expression of bitter taste receptors of the T2R family

in the gastrointestinal tract

The newly discovered bitter taste receptor family (T2R)

has also been found in epithelial cells in the rat stomach and
Fig. 1. BC/SCC in the rat larynx (13,000).
duodenum. Wu et al. (2002) identified putative taste receptor
gene transcripts in the gastrointestinal tract. Using reverse
transcriptase-PCR, they demonstrated the presence of
transcripts corresponding to multiple members of the T2R
family of bitter taste receptors in the antral and fundic gastric
mucosa, as well as in the lining of the duodenum. In
addition, cDNA clones of T2R receptors were detected in a
rat gastric endocrine cell cDNA library, suggesting that these
receptors are expressed, at least partly, in enteroendocrine
cells. Expression of multiple T2R receptors was also found
in STC-1 cells, an enteroendocrine cell line. The expression
of alpha subunits of G proteins implicated in intracellular
taste signal transduction, namely Ga(gust) and Ga(t)-(2) (a
member of the transducin family), was shown in the
gastrointestinal mucosa as well as in STC-1 cells, as
revealed by reverse transcriptase-PCR and DNA sequen-
cing, immunohistochemistry, and Western blotting. Further-
more, the addition of compounds widely used in bitter taste
signaling (e.g., denatonium, phenylthiocarbamide, 6-n-
propil-2-thiouracil, and cycloheximide) to STC-1 cells
promoted a rapid increase in intracellular Ca2+ concentra-
tion. These results demonstrate the expression of bitter taste
receptors of the T2R family in the mouse and rat
gastrointestinal tracts.

3. A ‘‘classic’’ DCS: brush cells

BCs are isolated elements with ultrastructural features

suggesting a chemosensory role. In the past, the identifica-
tion of these cells was morphologic and mainly based on the
presence of a brush of rigid apical microvilli (Figs. 1 and 2).
However, in other organs cells with similar characteristics
have been designated with different names. In the Fig. 2. Apical extremity of a BC/SCC in the human duodenum (40,000).
 A. Sbarbati, F. Osculati / Progress in Neurobiology 75 (2005) 295–307
gastrointestinal tract, the term ‘‘tuft cell’’ is the most
commonly used (Jarvi and Keyrilainen, 1955; Sato et al.,
2000). The term ‘‘caveolated cells’’ was used by Nabeyama
and Leblond (1974) for similar elements in the gastric
mucosa. The BCs located in the stomach were also called
fibrillovesicular cells 2 (Hammond and LaDeur, 1968;
Wattel and Geuze, 1978; Wattel et al., 1977a,b). Other terms
used to indicate similar elements are multivesicular cells
(Silva, 1966), undifferentiated cells (Johnson and Young,
1968), s-cells (Ferguson, 1969) or agranular light cells
(Riches, 1972).
3.1. BCs in different organs
BCs are present in the trachea of several mammalian
species (Rhodin and Dalhamn, 1956; Leeson, 1961; Rhodin,
1966; Luciano et al., 1968a,b; Jeffery and Reid, 1975; Ishida,
1977; Taira and Shibasaki, 1978; Christensen et al., 1987).
BCs have also been found in the alveolar epithelium of the
lung (Meyrick and Reid, 1968; Luciano et al., 1969; Hijiya
et al., 1977; Hijiya, 1978; Allan, 1978; Filippenko, 1978;
Foliguet and Grignon, 1980; Chang et al., 1986) and in the
non-sensory epithelium of the vomeronasal organ (Höfer
et al., 2000). Several studies have also confirmed the presence
of BCs in the human respiratory apparatus (Rhodin, 1959,
1966; Watson and Brinkman, 1964; Basset et al., 1971).
The tuft cells of the salivary ducts (Sato and Miyoshi,
1996, 1997; Sato et al., 2000) present strong analogies with
BCs. BCs have been found in the stomach in several species
(Luciano et al., 1980, 1993; Luciano and Reale, 1992). The
BCs of the gallbladder and bile duct were studied in a series
of papers by Luciano and Reale (1969, 1979, 1990), Luciano
et al. (1981) and Iseki (1991). Intestinal BCs were described
in the rat (Luciano et al., 1968a,b). The major pancreatic
excretory ducts have been shown to contain a large number
of BCs (Kugler et al., 1994; Höfer and Drenckhahn, 1996,
1998).
3.2. Phenotypic characteristics of BCs
The most characteristic ultrastructural feature of BCs is
the presence of an apical tuft of stiff microvilli, which often
displays long rootlets that do not appear to integrate into a
terminal web. Usually these cells express villin and fimbrin
(Höfer and Drenckhahn, 1992), cytokeratin 18 (Kasper et al.,
1994), nitric oxide synthase (Kugler et al., 1994) or
neurofilaments (Luciano et al., 2003).
In the rat stomach, intestine and pancreatic ducts, BCs
express gustducin and other taste-related molecules sharing
apical cytoskeletal features of taste receptor cells of the
tongue (Höfer and Drenckhahn, 1996, 1998). The gustducin
is concentrated in the apical tuft of microvilli, and it should
be noted that in specific portions of the digestive apparatus
(i.e., in terminal portions of extralobular ducts and in the
major pancreatic duct), these elements are present in great
density, making up as much as 22% of the epithelium.
299
The presence of gustducin and other molecules of the
taste transductory pathway strongly supports the chemor-
eceptive role of BCs. The presence in these cells of nitric
oxide synthase-I suggests that they are involved in the
chemosensory control of pancreatic secretion.
Taken together, the data reported above prove the
presence of solitary chemoreceptors in the gut, indicating
that BCs are involved in chemoreceptive signaling and
demonstrating a link between BCs and elements of the taste
buds (Höfer et al., 1998).
Although not all BCs display gustducin immunoreactiv-
ity (Höfer et al., 2000), a similar situation seems to exist in
the airway. BCs expressing gustducin were found in the
larynx (Sbarbati et al., 2004a) suggesting that these elements
might have a similar chemosensory role in both the digestive
and the respiratory apparatus. In the latter they are probably
active in driving reflexes against airborne substances.
3.3. Considerations about BCs
An enormous amount of data has demonstrated the
widespread distribution of BCs in large regions of the
respiratory and digestive apparatuses, although (as has been
known for a long time) these cells are often designated by
different names (Luciano and Reale, 1967). In the past, the
vast majority of authors concluded that these cells have a
chemoreceptive role, even if several other roles have been
proposed (mechanoceptive, secretory paracrine, secretory
exocrine, secretory endocrine, absorptive and regenerative).
The recent applications of the markers considered typical of
taste cells has strongly supported the hypothesis that these
elements have a chemoreceptorial role, as suggested by their
structural features. Obviously this does not exclude the
coexistence of other roles in elements that have chemor-
eceptive capacity.
4. Solitary chemosensory cells (SCCs)
SCCs (also called solitary chemoreceptor cells, Finger
et al., 2003) are slender epithelial elements, recently
discovered in mammals, which display cytological char-
acteristics suggesting a chemosensory role and which
possess signaling mechanisms typical of taste cells
(Figs. 3–6) (Sbarbati et al., 1998, 2004a; Finger et al., 2003).
4.1. Comparative anatomical considerations
At their cephalic extremity, vertebrates have different
chemoreceptors for smell and taste. The first chemosense is
effected by olfactory mucosa, the second by taste buds. In
addition, chemosensory free endings of the trigeminal nerve
are present. Most aquatic vertebrates also possess a further
type of chemosensory system based on secondary sensory
elements usually called solitary chemosensory cells (SCCs)
(Kotrschal, 1991, 1996; Whitear, 1992). In fish, SCCs form
300 A. Sbarbati, F. Osculati / Progress in Neurobiology 75 (2005) 295–307

Fig. 3. SCC in the developing gustatory epithelium of the rat tongue.

Alpha-gustducin immunostaining (300).

a system of differentiated sensory epithelial cells, which are

not organized into discrete end organs and may occur in the
epithelia of the oropharynx, the gills and the skin (Whitear,
1992). The elementary unit of this chemoreceptorial structure Fig. 5. SCC in the rat trachea. Ultrastructural immunocytochemistry for
alpha-gustducin (8000).
differs from the taste buds of the gustatory apparatus,
consisting of a single bipolar epithelial cell contacted by
be recorded electrophysiologically from the moving fin and
nerves and lacking a specialized connective bed. Glial-like
were elicited by a narrow spectrum of stimuli, including
epithelial cells can surround SCCs. In fish, these elements are
dilutions of heterospecific fish body mucus fish bile or
innervated by facial or spinal nerves (Kotrschal et al., 1993).
human sputum. In general, electrophysiological recordings
In some species, the corresponding cells were termed
oligovillous cells due to the ultrastructural appearance of
their apical extremity (Whitear and Lane, 1983).
The most frequently used animal model for studies of
SCC function is the anterior dorsal fin of the rockling
(Gaidropsarus and Ciliata, Teleostei), which is covered by a
large number of these cells (Whitear and Kotrschal, 1988;
Kotrschal and Whitear, 1988). Chemoresponses could only

Fig. 6. Schematic IMAGES of a BC and an SCC. The yellow marks axons,

the red marks the apical areas with the highest gustducin expression. M,
microvilli, roots of the microvilli; V, vesicles; F, filaments; G, Golgi
Fig. 4. SCC in the rat trachea. Alpha-gustducin immunostaining (900). complex.
 A. Sbarbati, F. Osculati / Progress in Neurobiology 75 (2005) 295–307
supported the hypothesis that SCCs are chemosensory
(Peters et al., 1991) and that they respond to predator-
avoidance or food-related stimuli, although they do not res-
pond to some typical taste stimuli (Silver and Finger, 1984).
Until recently, SCCs have been considered typical of
fishes. However, several findings suggest that SCCs are also
present in other species. Among amphibians, a good
example is, those described on the ventral skin of the
desert toad, Bufo alvarius. This species discriminates salt
taste through chemosensory function of the ventral skin
(Nagai et al., 1999), and the spinal nerves innervate putative
chemosensory cells in this area (Koyama et al., 2001).
In the oral cavity, some studies have reported the fine
structural resemblance of SCCs to taste bud cells in
amphibians, as has been described in fishes. Cellular
elements with ultrastructural features resembling those of
SCCs (i.e., bipolar cells with microvilli, apical vesicles,
packed cisternae of smooth endoplasmic reticulum and
nerve contact) were found in the frog (Rana esculenta) taste
disk, in which the presence of three different neuroepithelial
systems has been demonstrated (Sbarbati et al., 1990;
Osculati and Sbarbati, 1995).
4.2. SCCs in the oral cavity
In fishes, although SCCs may be located in the skin of the
whole body, a preferential site seems to be the oral cavity,
and a frequent association with taste buds has been reported.
In the past, it was observed that taste buds developed during
the early phylogeny of (agnathan) vertebrates by accumula-
tion of SCCs (Kotrschal, 1991; Whitear, 1971). In some
species structures of intermediate morphology between taste
buds and SCCs have been described. In the oral valves of
Raja clavata, Whitear and Moate (1994) found not taste
buds but groups of bipolar chemosensory cells. Recently, in
the zebrafish, it was confirmed that SCCs can surround taste
buds (Hansen et al., 2002). Finger (1997) suggested that
some cells in the taste buds may be related to the SCC
system on the grounds of their apical pole morphology and
receptor sites for amino acids. According to this hypothesis,
taste buds may be viewed as compound sensory organs
containing several cell types, one of which may be related to
the SCC system (Finger, 1997).
In mammals, a specific set of SCCs associated with
gustatory epithelium has been described. During the first
days of post-natal life, the epithelia of the rat’s circumvallate
papillae contain isolated cells with a bipolar shape, nerve
contacts and neuroendocrine-type granules (Sbarbati et al.,
1998). Ultrastructural features of these cells prove that they
are epithelial elements, suggesting that they could be
homologous to the SCCs described in aquatic vertebrates.
The presence of SCCs in the rat’s circumvallate papilla
(Fig. 3) during the first week of postnatal life was also
studied using a-gustducin immunocytochemistry (Sbarbati
et al., 1999). In newborn rats, isolated a-gustducin-
immunoreactive cells were found within the epithelium.
301
In the following days small taste buds appeared, but isolated,
bipolar shaped a-gustducin-immunoreactive cells were also
found. El-Sharaby et al. (2001a,b) studied the differentiation
of oral epithelium in the newborn rat, demonstrating that
solitary a-gustducin-immunoreactive cells are frequently
present in the circumvallate, foliate and nasoincisor papilla
but rarely in the soft palate. They suggested that solitary a-
gustducin-immunoreactive cells could be related to the early
responsiveness to sweet substances previously described in
newborn rats (Hall and Bryan, 1981).
The difference between taste chemoreception in neonate
and adult mammals is well known, and it was reported that in
newborn mice, receptor cells are characterized by a reduced
speed of repolarization accompanied by a low repetitive
firing capacity (Bigiani et al., 2002). These findings
probably relate to the particular nutrition of neonate
mammals.
In the vallate papilla of the newborn rat, the presence of
SCCs is paralleled by a rapid development of intrinsic
neurones and nerves (Sbarbati et al., 2000, 2002). This
seems to suggest that an anatomical and functional
relationship exists between the chemosensory cells and
the intrinsic nervous system of the developing gustatory
apparatus. Taken together, the data obtained on newborn rats
demonstrate that a phylogenetically primitive system of
SCCs develops and is rapidly replaced by taste buds. This
finding suggests that three different pathways (i.e., gustatory
system, common chemical sense, and solitary chemosensory
cell system) are involved in the oral chemoreception of
suckling rats.
In humans, data about SCCs are still lacking and the only
pertinent finding is the presence, during the early ontogen-
esis of the tongue, of individual slender cells which are
immunopositive for cytokeratin 20, an intermediate filament
protein that is exclusively present in taste bud and epidermal
Merkel cells (Witt et al., 2003).
4.3. SCCs in the nasal cavities
In mammals, solitary a-gustducin-immunoreactive cells
are not restricted to the digestive system but are also found in
another chemoreceptor, the vomeronasal organ (Zancanaro
et al., 1999). The experiments were performed in mice and
immunocytochemical data were confirmed by Northern blot
analysis of the organ. Typically, immunoreactive cells were
only found in the portion of the neuroepithelium close to the
boundary with neighbouring non-receptor epithelium. Here,
cell proliferation took place in adult mice and rats, possibly
to subserve neuroepithelial growth and/or renewal. This
location seems similar to those described in fish, where
SCCs are often located at the boundary between gustatory
epithelium and non-receptor epithelium (Hansen et al.,
2002). However, species-specific differences seem to exist
because Höfer et al. (2000), using both immunocytochem-
istry and immunoblotting, did not find gustducin in the
vomeronasal organ.
302 A. Sbarbati, F. Osculati / Progress in Neurobiology 75 (2005) 295–307
Isolated gustducin-immunoreactive cells of bipolar shape
were found scattered through the respiratory epithelium of
the nasal cavity of the mouse (Zancanaro et al., 1999), which
proves that these elements are not restricted to the digestive
system.
These findings were confirmed by the recent experiments
of Finger et al. (2003) which showed an extensive population
of chemosensory cells in both rats and mice that extend to
the surface of the nasal epithelium and form synaptic
contacts with trigeminal afferent nerve fibers. These
chemosensory cells express T2R ‘‘bitter-taste’’ receptors
and a-gustducin. Functional studies indicate that bitter
substances applied to the nasal epithelium activate the
trigeminal nerve and evoke changes in respiratory rate. By
extending to the surface of the nasal epithelium, these
chemosensory cells serve to expand the repertoire of
compounds that can activate protective trigeminal reflexes.
On the basis of these findings the authors hypothesized that
trigeminal chemoreceptor cells are likely to be remnants of
the phylogenetically ancient population of SCCs found in
the epithelium of all anamniote aquatic vertebrates.
4.4. SCCs in the larynx
A specific laryngeal sensory epithelium (SLSE), which
includes arrays of solitary chemoreceptor cells, has recently
been described in the supraglottic region of the rat (Sbarbati
et al., 2004a). Two plates of SLSE were found, one on each
side of the larynx. The first plate was located in the
ventrolateral wall of the larynx, the second in the inter-
arytenoidal region. Immunoblotting revealed the presence of
a-gustducin in SLSE. At immunocytochemistry, the lar-
yngeal immunoreactivity for a-gustducin was mainly
localized in SCCs. At ultrastructural immunocytochemistry,
these cells showed packed apical microvilli, clear cytoplas-
mic vesicles and cytoneural junctions. Double label immu-
nocytochemistry using confocal microscopy demonstrated
that a-gustducin and a T2R colocalize completely. A double
label approach using immunocytochemistry for a-gustducin
and PGP 9.5, which is a marker of endocrine cells in the
airways, showed that PGP 9.5-immunoreactive elements
were more numerous that a-gustducin-immunoreactive
elements and that a large number of PGP 9.5-immunoreactive
elements were negative for a-gustducin-immunoreactivity.
However, laryngeal a-gustducin-immunoreactive cells gen-
erally colocalize PGP 9.5. The immunocytochemical and
ultrastructural data suggest that SLSE is a chemoreceptor
located in an optimal position to detect substances entering
the larynx from either the pharynx or the trachea.
4.5. SCCs in the trachea and bronchi
In a recent study (Merigo et al., 2005), we examined the
immunohistochemical localization of a-gustducin and
PLCb2 in rat airway epithelium using both light and
electron microscopy. The expression of a-gustducin was
found in solitary cells, which presented ultrastructural
features of chemoreceptor cells (i.e., flask- or pear-shaped,
with an apical process with thin microvilli protruding into
the lumen) (Figs. 4 and 5). Alpha-gustducin immunostaining
was mainly concentrated in the apical process and along the
basolateral cell surface. Immunoblotting for a-gustducin
was positive in all the airway regions tested. This work
showed that a chemosensory system composed of taste cell-
related SCCs exists throughout the whole airway and is not
restricted to the nasal cavity and larynx.
4.6. Structures possibly associated with SCCs
A recent hypothesis posits that SCCs are not always
randomly arranged, but can be anatomically related to
specialized epithelia. For instance, in the larynx they are
embedded in a specific laryngeal sensory epithelium (SLSE)
characterized by high permeability to intraluminal sub-
stances (Sbarbati et al., 2004a). This permeability allows the
penetration of material present in the laryngeal lumen into
the extracellular spaces of the epithelium, where it comes
into widespread contact with cells and nerve fibers. Direct
contact between nerves and exogenous substances is a rather
unusual event and could be a cause of the great sensitivity of
the larynx to chemical substances. The anatomical basis of
this high permeability is a poorly developed system of apical
junctions (Sbarbati et al., 2004a). The complex system of
pits formed by the mucosa causes an expansion of the free
surface, which promotes contact with external substances.
The existence of high permeability is indirectly demon-
strated by the high density of globule leukocytes in the
intercellular spaces. In the areas of high permeability, there
are arrays of cells making specialized contacts with nerve
fibers. The cytoneural junctions appear to be similar to those
present in the basal plexus of gustatory organs.
4.7. General consideration about SCCs
To establish homology among SCCs in fish, amphibians
and mammals is difficult, partly because these cells form
heterogeneous systems. So far, findings in mammals have
fully confirmed previous findings in fish about the general
morphology of SCCs, despite the fact that in mammals the
SCCs seem to be used as internal rather than as external
chemoreceptors.
In the oral cavity, the homology between SCCs described
in the different species is evident, even if the relationship
with the taste system requires further clarification. It is
interesting to note that SCCs develop before taste buds in
mammals, and it has been shown that in fish also they appear
earlier in ontogeny than taste buds (Hansen et al., 2002;
Kotrschal et al., 1997). In mammals, the demonstration of
elements with the morphological characteristics of SCCs at
an early stage of development of the gustatory epithelium
may be an important phenomenon in the ontogeny of the
chemoreceptorial system of the oral cavity. The SCCs of the
 A. Sbarbati, F. Osculati / Progress in Neurobiology 75 (2005) 295–307
oral cavity have never been described in adult mammals;
therefore it is possible that their importance is limited to a
short phase of postnatal life. In the following days, the SCCs
may be removed by apoptosis or be incorporated in the taste
buds.
A more complex question is the parallelism between
SCCs and chemosensory systems found in invertebrates,
because comparisons between vertebrates and invertebrates
are difficult due to differences in terminology. The discovery
of similarities in molecular transductory pathways (see e.g.,
Matsunami and Amrein (2003)) make the research in this
field easier. Several chemosensory systems in invertebrates
utilize seven-transmembrane protein receptors in the same
way as taste cells and SCCs do (Mombaerts, 1999). In
general, the anatomical organization of the DCS strongly
resembles systems, which are widespread in invertebrates.
The number of chemosensors that an invertebrate possesses
can be enormous. These receptors cover large areas of body
surface and their number generally exceeds the number of
central neurons that process sensory information. Derby and
Steullet (2001) discuss these aspects, noting that in
invertebrates, multiple sensors offer many functional
advantages to an animal (i.e., they extend the range of
spatial sampling by increasing the sensory surface area; they
extend the range of stimuli that are discriminated by using a
diversity of sensors; they maintain the functioning of the
system in the event of damage to sensors; they increase the
sensitivity of the system through response summation). It
seems that similar considerations could be applied to the
vertebrate DCS.
5. Chemosensory clusters
Taste buds are present in the most rostral portion of the
larynx (Nishijima and Atoji, 2004), but recently, a new form
of chemosensory structure has been described in this organ,
the chemosensory cluster (Sbarbati et al., 2004b). These
clusters are multicellular organizations, which differ from
taste buds, being generally composed of 2–3 chemoreceptor
cells. Compared with lingual taste buds, chemosensory
clusters show lower height and smaller diameter. In
laryngeal chemosensory clusters, immunocytochemistry
using antibodies against either a-gustducin or PLCb2
identified a similar cytotype. This was rather unlike the a-
gustducin-immunoreactive and PLCb2-immunoreactive
cells visible in lingual taste buds. The laryngeal immunor-
eactive cells were shorter than the lingual ones, with poorly
developed basal processes, and their apical process was
shorter and thicker. Some cells showed a flask-like shape
with a large body and the absence of basal processes.
Chemosensory clusters lacked pores, and delimitation from
the surrounding epithelium was poor. This absence of clear
boundaries between chemosensory clusters and the sur-
rounding structures makes them resemble groups of SCCs
more than true taste buds.
303
The demonstration of the existence of chemosensory
clusters strengthens the hypothesis of a phylogenetic link
between gustatory and solitary chemosensory cells. Lar-
yngeal chemosensory clusters appear to be a transitional
structure between the rostrally located buds and the SCCs,
which are more distally located in specific areas of the
larynx (Sbarbati et al., 2004a). The presence of such clusters
strengthens the analogy between the chemoreceptorial
system of the larynx and that previously reported in the
skin and oral cavity of fish (Sbarbati et al., 2004a). In both
cases, SCCs or clusters of chemosensory cells are located in
areas rich in intraepithelial axons and not particularly
exposed to drying.
6. Conclusions
6.1. Similarities and differences between the primary
taste system and the DCS
Both the primary taste system and the DCS are mainly
composed of bipolar epithelial cells with a morphology
indicating a chemoreceptive role. In addition, the molecular
mechanisms of signal transduction in the two systems
present evident analogies. In view of these similarities, the
taste system could be viewed as a specialized portion of the
DCS located in the oro-pharyngeal cavity and devoted to
food analysis.
The differences between the primary taste system and the
DCS are mainly due to different anatomical organizations. A
first evident difference is the apparent absence of ancillary
structures around SCCs or chemosensory clusters. However,
specific studies have never been made so it is not possible to
definitively affirm the absence of these structures. The DCS
detects substances in very large areas (the respiratory
apparatus has a surface of about 70 mm2 and the
gastrointestinal tube has a length of about 8 m in humans),
and this explains the polymorphic appearance of its
elements.
Basically, the DCS displays a simple organization that
resembles similar systems which have been described in
lower vertebrates or invertebrates (Derby and Steullet,
2001). The cells with chemoreceptorial capacity are isolated
elements or small clusters and are not organized in
differentiated end organs. These elements are sited in
strategic locations. They are densely distributed in excretory
ducts of the pancreas, liver and salivary gland, but not in
their respective parenchyma. The same is true for SCCs in
the developing tongue, which are distributed along the ducts
of the serous von Ebner glands.
Despite their simplicity, the elements of the DCS are
very polymorphic and may have an open or a closed
conformation. The open cell type extends as far as the free
surface while the closed type rests on the basal membrane
without any evident contact with the free surface. Elements
with a closed conformation have been described in the
304 A. Sbarbati, F. Osculati / Progress in Neurobiology 75 (2005) 295–307
developing epithelium of the rat vallate papilla (Sbarbati
et al., 1999) and airway (Merigo et al., 2005). A similar
structural dichotomy is found in endocrine elements of the
digestive apparatus. It is not clear whether the closed
elements are immature, or are mature cells involved in
chemoreception in the intercellular milieu. It has been
clearly demonstrated that a-gustducin is present not only at
the apical but also at the basolateral membrane of BCs.
Therefore, the role of these elements in evaluation of the
extracellular liquid may be important. It has also been
hypothesized that BCs of the respiratory and digestive
apparatuses could also have a mechanoreceptive function as
the specialization (villin-rich microvilli) of their basolateral
membrane suggests.
As for the chemical substances liberated at their basal
pole, the data about gut BCs were reviewed by Höfer et al.
(1998), who suggested NO as a possible mediator. It seems
possible that this hypothesis could be also extended to the
elements of the DCS located in the airways.
6.2. The role of the DCS
If the main taste system plays a role in food sampling, the
DCS seems rather to be an alarm system. The respiratory and
digestive apparatuses have a relatively high autonomy from
the central nervous system due to an intrinsic nervous
system that regulates motor and secretory structures
(Schemann and Neunlist, 2004). In these apparatuses, the
DCS could provide information about the luminal or
intramural microenvironment. The DCS is probably the
afferent branch of intrinsic mechanisms, which might
involve gland secretion and muscle contraction. Local
reflexes could be generated by interaction between the DCS
and surrounding elements. The transmission of information
from the DCS to the central nervous system is possible, but
appears to be limited to the chemosensory cells innervated
by afferent axons. Such cells seem to be a minority and are
mainly localized in the tongue and larynx. For the non-
innervated elements, a paracrine action seems probable.
Areas of the tongue and larynx, which are rich in SCCs are
also rich in intraepithelial innervations. This pattern is rather
unusual, considering that nerve endings usually do not
penetrate into the epithelia, and it may be linked to the
existence of areas with high sensitivity in the tongue and
larynx.
6.3. The DCS and the quorum sensing based strategy
The chemoreceptive capacity of the DCS seems to protect
against exogenous substances. In addition, recently pub-
lished data suggest that the DCS could have an important
role in defense against bacteria.
There is a growing body of evidence that several bacterial
species operate a quorum sensing strategy (Kolter, 2005).
Briefly, these bacteria co-ordinate their activities using
extracellular signals, i.e., auto-inducers or pheromones
(Hardman et al., 1998). When such compounds reach a
sufficient concentration (i.e., when the total population is
large enough), the bacteria activate genetic pathways often
involved in the initiation of aggressive behaviour. Quorum
sensing appears therefore to be a strategy used by bacteria to
co-ordinate their activities, and it is based on the release of
small molecules, generally proteins or acyl-lactones. For its
structural and biochemical characteristics, the DCS appears
to be able to intercept communication among bacteria (it is
interesting to note that lactones are usually bitter taste
compounds) and predict their movements. If messages are
indeed detected in this way, it may be that the organism
mounts a highly localized and efficient response to bacterial
activation. This would be based on defenses like the
quenching of auto-inductors/inducers, the dilution or
removal of bacteria, or secretion of antibiotic agents, and
it might precede and avoid the need for intervention by
immune cells.
6.4. General conclusions: uncovering the iceberg
This work represents a further step in the definition
of a recently identified DCS composed of taste cell-
related elements. In recent years, the identification of
cells with gustatory characteristics located outside the
oro-pharyngeal cavity has been made possible by the
discovery of the molecular mechanisms of taste trans-
duction. This first allowed the chemical coding of the
gustatory cells in taste buds and subsequently permitted
the detection of cells with a similar chemical code in other
organs. Cells with similar characteristics have been
identified in endodermic derivatives, but they always
appear in the form of isolated elements (SCCs/BCs).
New findings suggest that taste cell-related elements may
also be present outside the oro-pharyngeal cavity, in
the airway, in a multicellular form of organization (i.e.,
chemosensory clusters). Due to their structure and
location, chemosensory clusters seem to represent the
missing link between buds and SCCs. Their existence
strengthens the hypothesis of a phylogenetic link between
the gustatory and the SCC systems, demonstrating that the
latter, at specific sites, can reach a multicellular level of
complexity. Investigation of the presence of chemosensory
clusters in other organs seems to be a promising field of
research.
At the present level of knowledge, it seems that an
iceberg-like organization of the taste cell-related DCS could
be hypothesized. Taste buds are probably only the most
visible portion of the iceberg, most of which is more
caudally located in form of SCCs or clusters in which the
molecular cascade of cell signaling needs to be explored in
more detail. Comparative anatomical studies in lower
vertebrates suggest that this submerged part of the iceberg
may be the most phylogenetically ancient component of the
system, which is probably involved in defensive or digestive
mechanisms.
 A. Sbarbati, F. Osculati / Progress in Neurobiology 75 (2005) 295–307
6.5. Unanswered questions and perspectives
Several questions remain about the morphology and
physiology of the DCS. In particular, the links between
the molecular mechanisms of taste and secretory apparatus
have not yet been studied, and the existence of BCs not
containing a-gustducin raises the possibility of alternative
G-proteins.
Such questions could be answered by a detailed chemical
code for the different elements of the DCS. Chemical coding
might also clarify more exactly the organization of the DCS
in different subfamilies of cell types. The rapid expansion of
research in this field seems to promise that the remaining
problems could soon be solved.
The DCS seems to be a potential new drug target
because several elements indicate that information obtained
by this system induces secretory reflexes. Therefore,
modulation of the respiratory and digestive apparatuses
by substances acting on their chemoreceptors could
be important in the treatment of diseases such as cystic
fibrosis and asthma, and might open a new frontier in drug
discovery.
Acknowledgements
This study was supported by the University of Verona,
grant numbers SBAREX6002 and OSCUEX6002, and by
the Italian Cystic Fibrosis Research Foundation. Donatella
Benati, Paolo Bernardi, Caterina Crescimanno, Flavia
Merigo and Marco Tizzano are thanked for providing
excellent technical assistance.
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