Annals of Anatomy 195 (2013) 128136

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Annals of Anatomy
journal homepage: www.elsevier.de/aanat

Research article

Fetal development and variations in the cartilages surrounding the human

external acoustic meatus
Yasutoyo Ikari a , Yukio Katori b, , Aiji Ohtsuka c , Jose Francisco Rodrguez-Vzquez d , Hiroshi Abe e ,
Tetsuaki Kawase f , Gen Murakami g , Shin-ichi Abe a
a
Department of Anatomy, Tokyo Dental College, 1-2-2 Masago, Mihama, Chiba 261-8502, Japan
b
Division of Otorhinolaryngology, Sendai Municipal Hospital, 3-1 Shimizukouji, Wakabayashi, Sendai 984-8501, Japan
c
Department of Anatomy, Okayama University School of Medicine, 2-5-1 Shikadamachi, Kita-ku, Okayama 700-8558, Japan
d
Department of Anatomy and Embryology II, Faculty of Medicine, Complutense University, Madrid 28040, Spain
e
Department of Anatomy, Akita University School of Medicine, 1-1-1 Hondouri, Akita 010-8543, Japan
f
Department of Otorhinolaryngology, Tohoku University Graduate School of Medicine, 1-1 Seiryo-machi, Aoba, Sendai 980-8575, Japan
g
Division of Internal Medicine, Iwamizawa Kojin-kai Hospital, 297 Shibunchou, Iwamizawa 068-0833, Japan

a r t i c l e i n f o s u m m a r y

Article history: In contrast to the osseus part that develops from the tympanic ring of the squamous part of the temporal
Received 24 February 2012 bone after birth, there is little information on fetal development of the cartilages surrounding the human
Received in revised form 9 June 2012 external acoustic meatus. Using routine histology and immunohistochemistry, we examine sections of
Accepted 18 July 2012
22 fetuses (CRL 100270 mm) to study the development of these cartilages. Early external ear cartilages
Available online 2 October 2012
are composed of three groups: (1) a ring-like cartilage at the putative tragus on the anterior side of
the meatus, (2) two or three bar-like cartilages along the inferior wall of the meatus, and (3) a plate-
Keywords:
like cartilage in a skin fold for the putative helix on the posterior side. In contrast to the rst and second
External acoustic meatus
Cartilage
pharyngeal arch cartilages, all the external ear cartilages express glial brillary acidic protein. Notably, the
Tragus bar-like cartilages along the meatus are connected with a fascia-like structure to the second pharyngeal
Tympanic ring arch cartilage. Later, with considerable individual variation, new cartilage bars extend from the inferior
Temporal bone cartilages to the superior side of the meatus. Thus, via an intermediate stage showing a chain of triangular
Human fetuses elastic cartilages, a chain of bar-like cartilages on the inferior side appears to change into a complex of
H-shaped cartilages. Numerous ceruminous glands are seen in the thick subcutaneous tissue overlying
the cartilaginous part of the meatus. However, they do not insert into the cartilage. The external ear
cartilages develop much earlier than, and independently of, the osseus part.
2012 Elsevier GmbH. All rights reserved.

1. Introduction narrower than the cartilaginous part (Williams, 1995). However,

Fetal development of the epithelial lining of the external acous-
tic meatus has been a major focus of interest for embryologists
because of the special topographical relationship with the middle
ear and the cell death that occurs in the epithelial plug (Michaels
and Soucek, 1989; Nishimura and Kumoi, 1992; Mallo et al., 2000;
Masumoto et al., 2010). Likewise, much is known about the osseus
part of the external acoustic meatus. In the adult, the lateral, carti-
laginous part is about 8 mm long and continuous with the auricular
cartilage, while the medial, osseus part is about 16 mm long, and
Corresponding author at: Department of Otorhinolaryngology, Sendai Municipal
Hospital, 3-1 Shimizukouji, Wakabayashi, Sendai 984-8501, Japan.
Tel.: +81 22 266 7111; fax: +81 22 214 7706.
E-mail addresses: yasuikari@nifty.com (Y. Ikari), yukatori@concerto.plala.or.jp
(Y. Katori), aiji@md.okayama-u.ac.jp (A. Ohtsuka), jfrodvaz@med.ucm.es (J.F.
Rodrguez-Vzquez), abe@med.akita-u.ac.jp (H. Abe), kawase@orl.med.tohoku.ac.jp
(T. Kawase), g.murakami@asuka-gp.or.jp (G. Murakami), abeshi@tdc.ac.jp (S.-i. Abe).
at birth, the osseus part is composed of only a small, incom-
plete ring originating from the squamous part of the temporal
bone, i.e., the tympanic ring. After birth, the tympanic ring extends
posterolaterally to become cylindrical, growing into a brocar-
tilaginous tympanic plate, which forms the adjacent part of the
external meatus at this stage (Williams, 1995). With ossication,
this brocartilage develops into the osseus part. Conversely, even
in late-stage fetuses, most or all of the external acoustic meatus is
likely to be surrounded by only cartilages.
The cartilages surrounding the external meatus do not seem to
be brocartilage, but rather elastic cartilage that does not readily
ossify. Do they originate from the auricular cartilages? If so, does
the tube-like cartilage develop from the supercial side and extend
to the deep side of the meatus? In contrast to the osseus part
of the external acoustic meatus, there is little information on the
fetal development of cartilages surrounding the meatus. According
to Kepes and Perentes (1988) and Viale et al. (1988), glial bril-
lary acidic protein (GFAP) is expressed in elastic cartilage for some

0940-9602/$ see front matter 2012 Elsevier GmbH. All rights reserved.
http://dx.doi.org/10.1016/j.aanat.2012.07.009
 Y. Ikari et al. / Annals of Anatomy 195 (2013) 128136 129

Fig. 1. Cartilage development at the tragus and along the inferior wall of the external acoustic meatus: frontal sections of a CRL 113-mm specimen. HE staining. Panels A and
B (and panels E and F) represent the most anterior (and posterior) side in the gure: the distance between panels B and E is 3 mm. Panels A and F display the topographical
anatomy at lower magnication (scale bars in panels A and F). Panels BE are prepared at the same magnication (scale bar in panel B). Panels B and E are higher-magnication
views of the external acustic meatus (EAM) in panel A and F, respectively. A ring-like cartilage is evident at the putative tragus (panels A and B). Along this ring, two notches
are present (arrow and arrowheads in panels B and C). The tragus cartilage issues a bar- or plate-like process, and these cartilages extend along the inferior wall of the
external meatus (panels CE). A fascial structure is evident (arrows in panels A and F) extending from the second pharyngeal arch [Reicherts] cartilage (RC) to the inferior
wall cartilages as well as the tragus cartilage. ICA, internal carotid artery; LC, longus capitis muscle; MC, the rst pharyngeal arch [Meckels] cartilage; OG, os goniale; PG,
parotid gland; RC, the second pharyngeal arch [Reicherts] cartilage; STA, supercial temporal artery; TB, tympanic bone; tegmen, tegmen tympani of the temporal bone;
TM, tympanic membrane; TT, tensor tympani muscle.
130 Y. Ikari et al. / Annals of Anatomy 195 (2013) 128136

unknown reason, and its expression has been identied in long-
preserved parafn-embedded specimens after autopsy. Therefore,
with the aid of immunohistochemistry, the aim of the present study
was to clarify when and how the external ear cartilages develop in
human fetuses before establishment of the tympanic ring.
2. Materials and methods
The study was performed in accordance with the provisions of
the Declaration of Helsinki 1995 (as revised in Edinburgh 2000).
We examined parafn-embedded, frontal, sagittal or horizontal

Fig. 2. Immunohistochemistry for glial brillary acidic protein: tilted frontal sections of a CRL 130-mm specimen. Immunohistochemistry for glial brillary acidic protein
(GFAP; panels A, C and D) and a negative control (panel B). Panel A (and panel C) represents the most posterior (and anterior) side of the gure. Intervals between panels
are 2 mm (AB) and 4.5 mm (BC), respectively. Panel A displays the plate-like cartilage of the helix protruding superiorly. Panel B is a negative control without the primary
antibody. Panel E exhibits cartilages along the inferior wall of the external acoustic meatus (EAM). Panels C and D include the second pharyngeal arch [Reicherts] cartilage
(RC) and the rst pharyngeal arch [Meckels] cartilage (MC), respectively: neither shows GFAP immunoreactivity. Panel F shows a small cluster of GFAP-positive mesenchymal
cells at the incudomalleolar joint. All panels are prepared at the same magnication (scale bar in panel B). IN, incus; M, malleus; PG, parotid gland; TB, tympanic bone; TM,
tympanic membrane.
 Y. Ikari et al. / Annals of Anatomy 195 (2013) 128136 131

sections of 22 fetuses (1530 weeks of gestation; crown-rump

length (CRL) 100270 mm): 6 specimens had a CRL of 100130 mm
(1516 weeks); 6 had a CRL of 170180 mm (1920 weeks); and
10 had a CRL of 250270 mm (around 30 weeks). In each of
these groups, 2 specimens had been processed for sagittal sec-
tions, another 2 for horizontal sections, and the others for frontal
sections. All specimens were part of the large collection kept at
the Embryology Institute of the Universidad Complutense, Madrid,
being the products of urgent abortion, miscarriages or ectopic
pregnancies managed at the Department of Obstetrics of the uni-
versity. Approval for the study was granted by the university ethics
committee. The donated fetuses had been xed in 10% (v/v) for-
malin solution and stocked in the same solution for more than
3 months.
After division into the head and neck, thorax, abdomen and
pelvis, and four extremities, the head specimens were decalcied
by incubating them at 4 C in 0.5 mol/L EDTA solution (pH 7.5;
Decalcifying solution B; Wako, Tokyo) for 24 days, depending on
the size of the material. However, because of advanced ossication,
decalcication of the 10 specimens around 30 weeks was per-
formed during 35 days at room temperature using Plank-Rychlo
solution (AlCl2 /6H2 O, 7.0%, w/v; HCl, 3.6; HCOOH, 4.6; Wako, Tokyo,
Japan). After routine procedures for parafn-embedded histology,
sections 5 or 10 m thick were prepared at intervals of 0.11 mm
depending on the size of the specimen. Most sections were stained
with hematoxylin and eosin (HE) while some (23 sections per
tissue block) were subjected to aldehyde-fuchsin staining for elas-
tic bers with post-xation of the relevant sections in Bouins
xative overnight (Fujita, 1959; Sunami-Kataoka et al., 2001). In
addition, 34 horizontal sections from 2 specimens (1 at 1516
weeks; 1 at 1920 weeks) were used for immunohistochemistry of
glial brillary acidic protein (GFAP). The primary antibody was rab-
bit polyclonal anti-human GFAP (Dako Cytomation, Kyoto, Japan).
The second antibody was labeled with horseradish peroxidase
(HRP), and antigen-antibody reactions were detected using the
HRP-catalyzed reaction with diaminobenzidine (with hematoxylin
counterstaining). Because of the rigorous conditions necessary for
decalcication, immunohistochemistry was not successful in the
large specimens examined (data not shown).

3. Results

In the small specimens examined (CRL 110130 mm, or 1516

weeks), external ear cartilages are composed of three groups: (1) a
ring-like cartilage at the putative tragus, (2) two or three bar-like
cartilages along the inferoposterior wall of the external acoustic
meatus, and (3) a plate-like cartilage in a skin fold for the putative
helix of the auricle (Figs. 1 and 2). In this order (1-2-3), the three
groups of cartilages are arranged from the anterior to the posterior
side of the external acoustic meatus. The helix cartilage is longest of
all and protruded superiorly in the auricular skin fold (Fig. 2). All of
these external ear cartilages are positive for GFAP (Fig. 2A and E), in
contrast to other cartilages such as the rst and second pharyngeal Fig. 3. Triangular cartilage bars developing along the inferior wall of the external
arch cartilages and the otic capsule cartilage, enclosing the inner meatus: horizontal sections of a CRL 180-mm specimen. HE staining (panels AD)
ear (Fig. 2C and D), which are GFAP negative. In addition, a small and aldehyde-fuchsin staining (panel E). Panel A (and panel D) represents the most
superior (and inferior) side of the gure. Interval between the panels is 0.5 mm.
cluster of mesenchymal cells expresses GFAP immunoreactivity at
Panels AD are prepared at the same magnication (scale bar in panel A). Cartilages
the incudomalleolar joint (Fig. 2F). are distributed along the inferior wall of the external acoustic meatus (EAM) and
The ring-like cartilage at the tragus shows a constant mor- are joined by ligament-like structures (star): each of the cartilages appears triangu-
phology, although it is more evident in frontal sections than in lar in shape at the bottom of the meatus (panel C), but its anterior and posterior
other sectional planes. The ring incompletely surrounds the lat- apices curve and extend superiorly along the meatus wall. The superior wall is
unaccompanied by cartilages (not shown). Panel E, corresponding to the square
eral end of the external acoustic meatus, and bears two notches:
in panel B, displays elastic bers in the center of each of the cartilages, as well as
a narrow superior one and a wide inferior one (Fig. 1B). The ring in the ligament-like structure between the cartilages (star). The tragus cartilage is
issues a fascia-like structure to the second pharyngeal arch carti- increased in size at this stage (not shown). M, malleus; TM, tympanic membrane;
lage. The inferomedial part of the ring issues a cartilage process TMJ, temporomandibular joint.
that extended medially: this corresponds to the most lateral of
132 Y. Ikari et al. / Annals of Anatomy 195 (2013) 128136

Fig. 4. Superior wall of the external acoustic meatus with accompanying bar-like cartilages: frontal sections of a CRL 270-mm specimen. HE staining. Panel A (and panel E)
represents the most anterior (and posterior) side of the gure: the distance between panels A and F is 17 mm. The ring-like cartilage at the tragus (panel A) continues to the
superior and inferior chains of bar-like cartilages along the external acoustic meatus (EAM; panels B and C). The superior and inferior chains communicate via short cartilage
bars along the anterior wall of the meatus (arrowheads in panel C) as well as along the posterior wall (arrowheads in panel E). The lateral part of the chain is connected to
the plate-like cartilage of the helix (panel D). The plate-like cartilage is very long and shows deep waves beneath the skin (panel E). IN, incus; M, malleus; PG, parotid gland;
TM, tympanic membrane.

the bar-like cartilages along the inferior wall of the meatus. The
superior wall of the meatus does not accompany any cartilages at
this stage. The chain of the bar-like cartilage reaches the lateral
one-third of the tympanic membrane. Notably, a fascial structure
is seen connecting between the medial end of the cartilage chain
and the second pharyngeal arch cartilage (Fig. 1F), but it is negative
for GFAP (data not shown). The most posterior group of the exter-
nal ear cartilages, i.e., the cartilage plate for the putative helix, is
slightly curved, irregularly wavy, and extends beneath the dermis
with a few sweat glands (Fig. 2). Ceruminous glands are not evi-
dent along the external meatus at the early stage. Likewise, at this
stage, elastic bers are not identied by aldehyde-fuchsin staining
(data not shown). Although the sections examined are not serial, the
structures observed suggest that a connection between the bar-like
cartilages is unlikely along the inferior wall of the meatus.
In the middle-sized specimens examined (CRL 170180 mm, or
1920 weeks), each of the bar-like cartilages along the inferior wall
of the external acoustic meatus changes into a triangular or J-shape.
Each of the triangular cartilages issues a process extending upward
along the lateral wall of the meatus (Fig. 3): however, no process
reaching the superior side of the meatus is found. Elastic bers
appear in the center of the thickest part of the triangular carti-
lages, and are also seen in the ligament-like structure between the
triangular cartilages (Fig. 3E). The ring-like cartilage at the tragus
increases in size but maintained its shape and thickness. The waves
of the helix plate-like cartilage become deeper than at the early
 Y. Ikari et al. / Annals of Anatomy 195 (2013) 128136 133

Fig. 5. Circular arrangement of the bar-like cartilages: sagittal sections of a CRL 250-mm specimen. HE staining (panels AE) and aldehyde-fuchsin staining (panels F and
G). Panel A (and panel E) represents the most lateral (and medial) side of the gure: the distance between panels A and E is 5 mm. Panels AE are prepared at the same
magnication (scale bar in panel A). Note that the bar-like cartilages along the external acoustic meatus (EAM) seen in frontal sections (e.g., Fig. 4) are connected mutually by
processes extending along the lateral walls of the meatus (arrowheads in panels A, B and C). The ring-like cartilage is absent at a site 5 mm lateral to the tympanic membrane
(panel E). Panel F, showing elastic bers in a ligament between the cartilage bars, shows a higher-magnication view of the circle in panel B. Panel G displays elastic bers
in the cartilage bar shown in panel B. PG, parotid gland; MXA, maxillary artery; PTA, posterior tympanic artery; SML, sphenomandibular ligament; STA, supercial temporal
artery; TMJ, temporomandibular joint.

stage. Numerous ceruminous glands develop along the entire exter-
nal meatus. At this stage, no fascial structure connecting between
the triangular cartilage and the second pharyngeal arch cartilage is
evident.
In the large specimens examined (CRL 250270 mm, or around
30 weeks), cartilages are characteristically distributed along the
entire aspects of the external acoustic meatus. Thus, in frontal sec-
tions, bar-like cartilages are seen cut on both the superior and
inferior sides of the meatus (Fig. 4). In sagittal sections where
the external meatus was cut transversely, each of the supe-
rior and inferior cartilages is irregularly shaped with a bar-like
process winding around the external meatus (Fig. 5). The two pro-
cesses provide communication between the superior and inferior
cartilages (Figs. 4C, E and 5AC), and are connected by a liga-
ment that contained abundant elastic bers oriented in random
directions (Fig. 5F). The ring-like cartilage at the tragus is located
1020 mm anterior to the plate-like cartilage of the helix. However,
the ring-like tragus is connected with the plate-like helix via a bar-
like cartilage. Numerous ceruminous glands are well developed,
but show no insertion into the cartilage.
Variations are found in (1) the shape of the cartilage at the tragus
and (2) the site of communication between the superior and inferior
cartilage bars along the external meatus. In 4 of 10 large specimens,
the tragus cartilages do not form a ring but a U-shaped arrange-
ment (Fig. 6). In 2 of the 4 specimens with this U-shaped tragus, the
communicating cartilages between the superior and inferior bars
134 Y. Ikari et al. / Annals of Anatomy 195 (2013) 128136

Fig. 6. Anteriorly restricted communications between the superior and inferior cartilage bars along the external meatus: sagittal sections of a CRL 255-mm specimen. HE
staining. Panel A (and panel F) represents the most anterior (and posterior) side of the gure: the distance between panels A and F is 14 mm. Panels AF are prepared at the
same magnication (scale bar in panel F), while panel G is a higher-magnication view of the circle in panel D. The cartilage at the tragus is not ring-like, but composed of
bar-like cartilages arranged in a U-shape. Note that the superior and inferior bar-like cartilages along the external acoustic meatus (EAM) are connected by two anteriorly
located cartilage bars (arrowheads in panel B). Numerous ceruminous glands are well developed, but do not insert into the cartilage (panel G). IN, incus; M, malleus; PG,
parotid gland; TM, tympanic membrane.

are restricted to the anterior side of the meatus (Fig. 6B), although
they are usually evident along both of the anterior and posterior
walls (Fig. 4C and E). Thus, the variation in cartilages along the
meatus accompanies that of the tragus. The distribution of the car-
tilage bars (or plate) is dense on the anterior and inferior sides of
the meatus, but the shape and density of the bars appear to vary
between specimens. Overall, a chain of bar-like cartilages in the
early stage changes into a complex of circular but irregularly shaped
plates in the late stage, via a middle stage showing triangular
cartilages. Fig. 7 shows a schematic representation of the hypothet-
ical developmental steps from a chain of bar-like cartilages to a
complex of irregularly shaped plates. We do not nd any clear
gender difference in the morphology of the external ear cartilages.
4. Discussion
The present study demonstrates that (1) cartilage development
of the helix and tragus is followed by that along the external
acoustic meatus, (2) bar-like cartilages rst appears along the infe-
rior wall of the meatus, and (3) there are individual variations
in the morphology of the superior extension of the meatus carti-
lage bars or plates. Immunohistochemistry for GFAP suggests that
 Y. Ikari et al. / Annals of Anatomy 195 (2013) 128136
Fig. 7. Schematic representations of external ear cartilage development. Panel A
displays the early stage: the tragus cartilage develops at the anterior site, bar-like
cartilages extend along the inferior wall of the external acoustic meatus (EAM) at
the middle site, and the helix cartilage is located posteriorly. The bar-like cartilages
along the EAM appear to change to a complex of H-shaped cartilages (panel C) in the
late fetal stage via an intermediate morphology comprising J-shaped or triangular
cartilages (panel B). To show the cartilage distribution, the later-developing superior
bars are omitted in the models. A ring-like cartilage at the tragus and the plate-like
cartilage at the helix are not included in panels B and C.
these external ear cartilages differentiate into elastic cartilage until
15 weeks of gestation, whereas elastic bers appear at the later
stage. GFAP immunoreactivity is absent in nearby cartilages such
as the second pharyngeal arch cartilage and the otic capsule carti-
lage enclosing the inner ear. This stage, at and before 15 weeks, is
concurrent with the development of the epiglottic cartilage (Katori
et al., 2011). However, development of the epiglottic cartilage dif-
fers from that of the external ear cartilages in that the latter shows
(1) no invasion of glands into the cartilage, and (2) no GFAP-positive
mesenchymal cells accompanying the cartilage. Thus, the exter-
nal ear cartilages may not contain glands because GFAP-positive
mesenchymal cells are absent around them.
In the process of development from a chain of bar-like cartilages
along the inferior wall of the meatus to a complex of circular but
135
irregularly shaped plates, there may be many sources of possible
individual variations, especially at the stages when communica-
tions develop. The triangular cartilages seen in the intermediate
step may correspond to the triangular apophysis described in the
textbook of Poirier and Charpy (1912). However, their apophysis
seems to attach to the tympanic bone or the mastoid process of the
temporal bone. In spite of the presence of many patch-like areas
lacking coverage by cartilages along the meatus, because of the
medial locations, these fetal notches seem not to correspond to any
of the incisura anterior, the incisura intertragica and the incisura
terminalis auricularis. These notches, which are well known in
adults, seem to develop after birth. The irregular and individual
morphology of the cartilages along the external meatus is likely
to allow a variety of cystic lesions to develop between the peri-
chondrium and surface skin, although these would become evident
after birth (Zhu and Wang, 1992; Secor et al., 1999). Despite these
variations, however, the basic arrangement of the cartilages is con-
sistent: the ring-like cartilage of the tragus (most anterior) and the
plate-like cartilage of the helix (most posterior) are interrupted
by a middle-positioned bar-like cartilage for the external meatus
(Fig. 7A).
It is well known that the outer epithelium of the tympanic
membrane develops from the external acoustic meatus. On the
basis of histology, expression analysis and an in vitro study of ear
ossicles, Mallo et al. (2000) demonstrated that the meatus plays
an essential role in establishment of a suitable attachment of the
malleal manubrium to the tympanic membrane. They found two
quite opposite activities of the external acoustic meatus: one was
to induce chondrogenesis, and the other to repress it. Because
we used semiserial sections of fetuses at three limited stages, we
are unable to deny a possibility that the superior bars develop
de novo. However, we consider that the inferior bars are most
likely to extend superiorly. In fact, the adult meatal cartilage is
usually decient posterosuperiorly, the gap being occupied by a
sheet of collagen (Williams, 1995). Notably, the inferior bars seem
to develop along a fascia-like structure that connected to the second
pharyngeal arch cartilage and also limited the superior extension
of the parotid gland. Such a fascia is not evident along the supe-
rior wall of the meatus, although, if present, it would connect
to the rst pharyngeal arch cartilage. When the external mea-
tus induces chondrogenesis of the bar-like, inferior cartilages, the
fascia-like structure (and possibly the second pharyngeal arch car-
tilage) seems to provide a cue for suitable positioning.
Although we do not nd any clear gender differences, we can-
not rule out the possibility that individual variation may mask
such gender difference, if present. The osseus part of the exter-
nal acoustic meatus develops after birth (see Section 1). This later
development seems to be the reason for the sexual difference in the
morphology of the bony covering (Dahm et al., 1993). Overall, the
external ear cartilages seem to develop earlier and independently
of the osseus part from the tympanic ring.
Acknowledgment
We are grateful to Mr. Hiroyuki Oosugi (Department of
Anatomy, Okayama University School of Medicine) for his assis-
tance with elastic ber staining.
References
Dahm, M.C., Shepherd, R.K., Clark, G.M., 1993. The postnatal growth of the temporal
bone and its implication for cochlear implantation in children. Acta Otolaryngol.
Suppl. 505, 139.
Fujita, T., 1959. Histological studies on the neuro-insular complex in the pancreas
of some mammals. Z. Zellforsch. 50, 94109.
Katori, Y., Takeuchi, H., Rodrguez-Vzquez, J.F., Kitano, H., Murakami, G., Kawase,
T., 2011. Fetal development of the human epiglottis revisited: appearance of
136 Y. Ikari et al. / Annals of Anatomy 195 (2013) 128136
GFAP-positive mesenchymal cells and brous connections with other laryngeal
and lingual structures. Ann. Anat. 193, 149155.
Kepes, J.J., Perentes, E., 1988. Glial brillary acidic protein in chondrocytes of elastic
cartilage in the human epiglottis: an immunohistochemical study with polyva-
lent and monoclonal antibodies. Anat. Rec. 220, 296299.
Mallo, M., Schrewe, H., Martin, J.F., Olson, E.N., Ohnemus, S., 2000. Assembling a
functional tympanic membrane: signals from the external acoustic meatus coor-
dinate development of the malleal manubrium. Development 127, 41274136.
Masumoto, H., Katori, Y., Kawase, T., Cho, B.H., Shibata, S., Murakami, G., Matsubara,
A., 2010. False positive reactivity of a substance P-antibody in the epithe-
lial/ectodermal plugs of the nose, ear, eye and perineum of the human and mouse
fetuses. Okajimas Folia Anat. Jpn. 87, 3340.
Michaels, L., Soucek, S., 1989. Development of the stratied squamous epithelium
of the human tympanic membrane and external canal: the origin of the auditory
epithelial migration. Am. J. Anat. 184, 334344.
Nishimura, Y., Kumoi, T., 1992. The embryonic development of the human external
auditory meatus. Preliminary report. Acta Otolaryngol. 112, 496503.
Poirier, P., Charpy, A., 1912. Les Organes des Sens. In: Trait DAnatomie Humaine,
Tome Cinquime, Fascicule II. Masson, Paris, pp. 735736.
Secor, C.P., Farrell, H.A., Haydon III, R.C., 1999. Auricular endochondral pseudocysts:
diagnosis and management. Plast. Reconstr. Surg. 103, 14511457.
Sunami-Kataoka, Y., Akagi, H., Nishizaki, K., Taguchi, T., Murakami, T., Oht-
suka, A., 2001. Chondritin sulfate proteoglycan at the basal lamina beneath
high endothelial cells in human palate tonsils: a light and electron micro-
scopic study using the cationic colloidal iron method. Arch. Histol. Cytol. 64,
535543.
Viale, G., Doglioni, C., DellOrto, P., Zanetti, G., Iuzzolino, P., Bontempini, L., Coggi, G.,
1988. Glial brillary acidic protein immunohistochemistry in human respiratory
tract cartilages and pulmonary chondromatous hamartoma. Am. J. Pathol. 133,
363373.
Williams, P.L., 1995. Grays Anatomy. Churchill Livingstone, London, pp. 593, 1369.
Zhu, L., Wang, X., 1992. Histological examination of the auricular cartilage and
pseudocyst of the auricle. J. Laryngol. Otol. 106, 103104.