close

Вход

Забыли?

вход по аккаунту

?

Localization of CD44 and hyaluronan in the synovial membrane of the rat temporomandibular joint.

код для вставкиСкачать
THE ANATOMICAL RECORD PART A 288A:646 – 652 (2006)
Localization of CD44 and Hyaluronan
in the Synovial Membrane of the Rat
Temporomandibular Joint
AKIKO SUZUKI,1* KAYOKO NOZAWA-INOUE,1 NORIO AMIZUKA,1,2
KAZUHIRO ONO,3 AND TAKEYASU MAEDA1,2
1
Division of Oral Anatomy, Department of Oral Biological Sciences, Niigata
University Graduate School of Medical and Dental Sciences, Niigata, Japan
2
Center for Transdisciplinary Research, Niigata University, Niigata, Japan
3
Division of Dental Hygiene and Health Promotion, Faculty of Dentistry, Niigata
University, Niigata, Japan
ABSTRACT
Previous studies have pointed out a lack of adhesion structures in the synovial lining layer of the rat temporomandibular joint (TMJ) despite showing an epithelial arrangement. CD44, a major cell adhesion molecule, plays crucial
roles as an anchor between cells and extracellular matrices by binding hyaluronan (HA) for the development of organs
or the metastasis of tumors. The present study examined the localization of CD44 in the synovial membrane of the rat
TMJ by immunocytochemistry for OX50, ED1, and Hsp25, which are markers for the rat CD44, macrophage-like type
A, and fibroblast-like type B synoviocytes, respectively. Histochemistry for HA-binding protein (HABP) was also
employed for the detection of HA. OX50 immunoreactions were found along the cell surface and, in particular,
accumulated along the surface of the articular cavity. Observations by a double immunostaining and immunoelectron
microscopy revealed that all the OX50-immunopositive cells were categorized as fibroblastic type B cells, which had
many caveolae and a few vesicles reactive to intense OX50. However, the macrophage-like type A cells did not have any
OX50 immunoreaction in the synovial lining layer. A strong HABP reaction was discernable in the extracellular matrix
surrounding both OX50-positive and -negative cells in the synovial lining layers, exhibiting a meshwork distribution,
but weak in its sublining layer. This localization pattern of CD44 and HABP might be involved in the formation of the
epithelial arrangement of the synovial lining layer. Furthermore, OX50 immunonegativity in the type A cells suggests
their low phagocytotic activity in the rat TMJ under normal conditions. Anat Rec Part A, 288A:646 – 652, 2006.
©
2006 Wiley-Liss, Inc.
Key words: fibroblast-like type B cell; macrophage-like type A cell; CD44; hyaluronan; temporomandibular joint
The temporomandibular joint (TMJ), which is a bilateral diarthrosis between the temporal bone and the mandibular condyle, develops a synovial membrane. Many
researchers agree with the notion that the synovial lining
layer of the TMJ consists of macrophage-like type A cells
and fibroblast-like type B cells (Graabæk, 1984; NozawaInoue et al., 1998, 2003), as shown in the systemic joints
(for review, see Iwanaga et al., 2000). According to their
ultrastructural configurations, the former cells absorb and
degrade of various extracellular matrices and cell debris
as well as antigens in the synovial fluid and synovial
membrane (Okada et al., 1981; Graabæk, 1984; Athanasou, 1995), whereas the latter cells serve in the synthesis
and secretion of collagen (Visnapuu et al., 2000), fibronectins (Mapp and Revell, 1985), and hyaluronan (HA) (Roy
and Ghadially, 1967), all of which are the components of
the synovial fluid as well as the extracellular matrices
(ECMs) of the synovial membrane. In addition, biochemical studies have also detected HA, albumins, white blood
©
2006 WILEY-LISS, INC.
cells, and debris of the synovial membrane in the synovial
fluid (Berumen-Nafarrate et al., 2002). This viscous synovial fluid makes smooth jaw movements possible, serving
as a lubricant in both the sliding and rotating movement
Grant sponsor: Japanese Ministry of Education, Culture,
Sports, Science and Technology (MEXT); Grant number:
16659498.
*Correspondence to: Akiko Suzuki, Division of Oral Anatomy
Department of Oral Biological Science Niigata University Graduate School of Medical and Dental Sciences, 2-5274 Gakkochodori, Niigata 951-8514, Japan. Fax: 81-25-223-6499.
E-mail: a-suzuki@dent.niigata-u.ac.jp
Received 13 December 2005; Accepted 13 February 2006
DOI 10.1002/ar.a.20331
Published online 3 May 2006 in Wiley InterScience
(www.interscience.wiley.com).
CD44 AND HA IN RAT TMJ
of the condyle, in addition to supplying oxygen and nutrition to the articular cartilage.
CD44 is a cell surface transmembrane glycoprotein receptor (Aruffo et al., 1990), categorized in hyaladherins. A
variety of cells exhibit this molecule, which involves cell
adhesion (Lesley and Hyman, 1998), the metastasis of
tumors (Alaniz et al., 2002), cell migration (Lesley and
Hyman, 1998), and development of almost all organs
(Knudson and Knudson, 1993; Ruiz et al., 1995). CD44
binds not only ECMs such as HA (Asari et al., 1995;
Knudson, 2003), types I and IV collagens (Faassen et al.,
1992; Knudson et al., 1996), and chondroitin sulfate, but
also non-ECM ligands including fibronectins, osteopontin,
and serglycin (Aruffo et al., 1990; Lesley et al., 1993).
Therefore, this molecule has been considered to play a role
as a cell-to-cell and cell-to-ECM anchor.
The synovial lining cells in TMJ show an epitheliumlike arrangement without a distinct adhesive structure,
leading us to a possibility that CD44 is a candidate for a
molecule to serve as cell-to-cell adhesion. However, there
has been controversy over the distinct localization of CD44
immunoreaction in the synovial membrane. CD44 immunoreaction has been reported in human knee joints under
normal (Asari et al., 1995) and pathological conditions
(Johnson et al., 1993; Naor and Nedvetzki, 2003). Furthermore, CD44 immunoreaction decreased in number in the
synovial tissue in the human knee joint with rheumatoid
arthritis (RA) (Henderson et al., 1994). Although both type
A and B cells in TMJ were also positive in CD44 immunoreaction (Yamada et al., 2002), these studies failed to
identify the types of CD44-positive cells because they did
not utilize any specific cell marker for each synovial lining
cells.
HA, widely distributed in the extracellular compartment of most tissues, is a large linear glycosaminoglycan
(GAG). This GAG has multiple functions as a scaffold for
various cells in cell migration, immune reactions, phagocytosis, and adherence (Hakansson et al., 1980; Forrester
and Lackie, 1981; Pisko et al., 1983; Hardwick et al., 1992;
Termeer et al., 2003) under both physiological and pathological conditions (Knudson and Knudson, 1993; Goebeler
et al., 1996). These regulatory functions of HA on cellular
activities are due to its ability to interact with the cell
surface hyaladherins and matrix hyaladherins. In addition, since HA is a major component of the synovial fluid
and cartilage (Pitsillides et al., 1994; Tulamo et al., 1994;
Parkkinen et al., 1996), it has been supposed to play
crucial roles in joint lubrication and reduction of cartilage
attrition during joint movements in synovial arthrosis
including TMJ (Ghosh, 1994; Cascone et al., 2002).
The present study was therefore undertaken to investigate the localization of CD44 in the normal rat TMJ by
employing immunocytochemistry for OX50, which can recognize rat CD44 (Paterson et al., 1987; Wirth et al., 1993)
at light and electron microscopic levels. Histochemistry
for HA-binding protein (HABP), which specifically combines with HA (Hascall and Heinegard, 1974; Asari et al.,
1995), was carried out to clarify the topographical relationship between the synovial lining cells and HA in the
synovial membrane. Furthermore, a double staining with
OX50 and several marker proteins for the synovial lining
cells was performed for identifying its immunopositive cell
types.
647
MATERIALS AND METHODS
All experiments were performed under guidelines of the
Niigata University Faculty of Dentistry Intramural Animal Use and Care Committee (approval number 150).
Tissue Preparation
Male Wistar rats (4 weeks of age, 105–120 g, n ⫽ 6; 8
weeks of age, 300 –330 g, n ⫽ 2) were used in this study.
Under anesthesia by an intraperitoneal injection of 8%
chloral hydrate (400 mg/kg), they were perfused with a
fixative containing 4% paraformaldehyde and 0.0125%
glutaraldehyde in a 0.1 M phosphate buffer (pH 7.4). The
removed heads were decalcified with a 5% ethylene diamine tetra-acetic acid disodium (EDTA-2Na) solution for 4
weeks at 4°C. One side of each head was embedded in a
30% sucrose solution at 4°C overnight for cryoprotection
and embedded in an OCT compound (Leica, Nussloch,
Germany). Serial sagittal sections were cut at a thickness
of 8 or 35 ␮m in a cryostat (CM 3050S; Leica) for immunocytochemistry at the levels of light and electron microscopy, respectively, and mounted onto silane-coated glass
slides. Serial paraffin sections were sagittally cut at a
thickness of 5 ␮m, stained with hematoxylin and eosin
(H&E) for histological observations, and processed for immunohistochemistry.
Immunohistochemistry
Dewaxed sections were predigested with 0.1% trypsin
(Wako Chemicals, Tokyo, Japan) in 0.01 M phosphatebuffered saline (PBS; pH 7.4) for 30 min at 37°C as an
unmasking procedure according to a report by Parkkinen
et al. (1996). After the inhibition of endogenous peroxidase
with 0.3% H2O2 in absolute methanol for 30 min, the
sections were incubated for 24 hr at 4°C with an OX50monoclonal antibody (1:1200, Serotec, Oxford, U.K.). The
reaction sites of the antigen-antibody were localized using
biotinylated antimouse IgG (1:100; Vector Lab, Burlingame, CA) for 2 hr at room temperature and subsequently
avidin-conjugated peroxidase (ABC kit; Vector Lab) for 90
min at room temperature. Final visualization employed
0.04% 3,3⬘-diaminobenzidine tetrahydrochloride (DAB)
and 0.0125% H2O2 in a 0.05 M Tris-HCl buffer (pH 7.6).
Some immunostained sections were counterstained with
0.03% methylene blue.
For immunocytochemistry at the electron microscopic
level, immunostained cryostat sections without counterstaining were postfixed in 1% osmium tetroxide reduced
with 1.5% potassium ferrocyanide for 90 min at 4°C, dehydrated in ascending ethanols, and finally embedded in
epoxy resin (Epon 812, Taab, Berkshire, United Kingdom). One ␮m thick sections were stained with 0.03%
methylene blue in order to check the locations under a
light microscope. Ultrathin sections (70 nm thick) were
prepared in a Leica Ultracut R (Leica Microsystems, Wetzlar, Germany) with a diamond knife and briefly stained
with lead citrate. They were examined in a Hitachi
H-7000 transmission electron microscope (Hitachi, Tokyo,
Japan).
For the detection of HA, deparaffinized sections were
reacted with biotin-labeled HABP (2 ␮g/ml; Seikagaku,
Tokyo, Japan) for 24 hr at 4°C, followed with avidinconjugated peroxidase (ABC kit; Vector Lab) for 90 min at
room temperature. They were developed with DAB solution mentioned above.
648
SUZUKI ET AL.
Double Labeling With HABP and OX50
Cryostat sections were primarily incubated with a biotinylated HABP at a concentration of 4 ␮g/ml for 24 hr at
4°C and subsequently with fluorescein isothiocyanate
(FITC)-conjugated Avidin D (1:300, Vector Lab) for 1 hr at
room temperature. After the inhibition of avidin and biotin according to the manufacturer’s instruction (Vector
Lab), these reacted sections were incubated with an OX50
antibody in the same way mentioned above. Following two
consecutive incubations of biotinylated antimouse IgG
and rhodamine isothiocyanate (RITC)-conjugated Avidin
D (1:300; Vector Lab), the double-labeled sections were
mounted in the Vectashield mounting medium with 4⬘,6diamidino-2-phenylindole (DAPI; Vector Lab). They were
examined with a fluorescent microscope (Axiophot, Carl
Zeiss, Jena, Germany).
Double Labeling Immunocytochemistry for
OX50 With Either ED1 or Hsp25
The detection of ED1 or heat shock protein 25 (Hsp25)
was carried out by immunofluorescence using FITC-conjugated Avidin D (1:300; Vector Lab). The ED1-monoclonal antibody (1:500; Serotec) and antiserum against
Hsp25 (1:5,000; Stressgen Biotechnologies, Victoria, Canada) used here can recognize monocyte/macrophage lineages (Dijkstra et al., 1985) and fibroblastic type B cells
(Nozawa-Inoue et al., 1999a; Andoh et al., 2001) in the
TMJ, respectively. OX50 immunoreaction was made visible by an incubation of RITC-conjugated Avidin D (1:300;
Vector Lab).
Immunohistochemical or Histochemical
Controls
Immuohistochemical controls of OX50 were performed
by replacing the primary antibodies with nonimmune sera
or PBS and omitting the antimouse or rabbit IgG, or the
ABC conjugated with peroxidase. For the HA reaction,
negative controls were prepared by treating paraffin sections with Streptomyces hyalurolyticus hyaluronidase
(400 turbidometric units/ml; Seikagaku) for 2 hr at 60°C
prior to incubation with the biotinylated HABP. Immunocontrol experiments with ED1 and Hsp25 have been described in our previous studies (Nozawa-Inoue et al.,
1999a; Andoh et al., 2001).
Fig. 1. Sagittal sections of the synovium of the rat TMJ. OX50 (a–c),
OX50 and Hsp25 (d), and OX50 and ED1 immunoreaction (e) in 4-week(a, b, d, and e) and 8-week-old (c) rats. a: The anterior portion of the
synovial membrane is a thick synovial lining cell layer. OX50 immunoreaction is intense around the synovial lining cells (arrowheads). Some
OX50-positive cells with slender profiles (arrows) are scattered in the
sublining layer. b: At the posterior region of the synovial fold, the synovial
lining cells form a sparse cell layer and the OX50 immunoreactions are
observed as a line along the surface of the synovial membrane. c: The
anterior portion of the synovial membrane at 8 weeks. Immunolocalization pattern of CD44 does not differ between 4- (a) and 8-week-old (c)
rats. Arrowheads indicate OX50-positive synovial lining cells. d: OX50
immunoreactions (red) appear along the cellular outlines (arrows) while
Hsp25 immunoreactions (green) are diffused in the cytoplasm. The articular cavity is lined with the cytoplasmic processes of OX50-positive
cells with Hsp25 immunoreaction (arrowheads). e: The red OX50 immunoreaction is detectable at the periphery of the cells, whereas ED1
reactions take on a granular appearance in the cytoplasm, appearing
green (arrows). The ED1-positive cells lack OX50 immunoreaction. Scale
bars ⫽ 20 ␮m (a– d).
RESULTS
Immunolocalization of OX50 in Rat TMJ
Double Immunostaining With OX50 and ED1 or
Hsp25
Immunostaining with the OX50 antibody was able to
demonstrate an intense immunoreaction in the synovial
lining layer of the rat TMJ (Fig. 1). The immunolocalization pattern in the rats aged 4 weeks (Fig. 1a) was identical to that in 8-week-old animals (Fig. 1c). These immunoreactions were localized along both the upper and lower
articular cavities. At the anterior region of the synovial
membrane where the synovial lining cells showed a thick
cell layer, OX50 immunoreactivity was discernable
around the synovial lining cells, assuming a meshwork
distribution (Fig. 1a). At the posterior portion, OX50 immunoreactions were observed as a line along the surface of
the articular cavities because the synovial lining layer was
thin (Fig. 1b). Some slender cells in the sublining layer
also reacted with the OX50 antibody (Fig. 1a).
A double immunostaining clearly succeeded in distinguishing the synovial lining cells according to their immunostaining patterns: synoviocytes coexpressed Hsp25 and
OX50 immunoreactions (Fig. 1d), and ED1-positive synoviocytes lacked OX50 reaction (Fig. 1e). In a double immunofluorescent staining, OX50, ED1, and Hsp25 immunoreactions respectively took on a cellular outline,
cytosolic granular appearance, or diffused cytosolic distribution. Those cells double positive for OX50 and Hsp25
extended their cytoplasmic projections toward the intercellular spaces or the surface of the articular cavities (Fig.
1d). Their immunoexpression patterns suggested that the
OX50-immunopositive and -negative lining cells were fibroblast-like type B cells and macrophage-like type A
cells, respectively.
CD44 AND HA IN RAT TMJ
649
Fig. 2. Immunoelectron micrographs showing OX50-positive (a, c,
and d) and -negative (b) cells in the synovial lining layer (a– d) and
sublining layer (e) of the rat TMJ. Preembedding method using DAB
development. a: Two types of the synovial cells are recognizable in the
synovial lining layer: cells with (B) and without (A) OX50 immunoreactions
and cells. The OX50-positive cells (B) assemble at the upper portion of
the synovial membrane. Their cytoplasmic projections cover the surface
of the articular cavity. b: The OX50-negative cell in the synovial lining
layer has lysosomes (arrowheads), well-developed filopodia-like pro-
cesses (arrows), and a heterochromatin-rich dark nucleus. c: The OX50positive cells in the synovial lining layer have a well-developed rough
endoplasmic reticulum and bright large nuclei. d: Higher magnification of
the boxed area in c. The OX50-positive cell has numerous caveolae with
intense (arrowheads) and weak (open arrowheads) immunoproducts. In
addition to caveolae, there are some immunopositive vesicles (arrows) in
its cytoplasm. e: The OX50-positive cell in the sublining layer displays a
serrate outline and has a large irregular nucleus and poor organelle in its
cytoplasm. Scale bars ⫽ 5 ␮m (a); 2.5 ␮m (b, c, and e); 500 nm (d).
Immunoelectron Microscopic Observation for
OX50-Immunopositive Cells
tained resultant vesicles, which revealed an OX50 immunoreactivity, implying an internalization of the extracellular matrix. From their cell bodies, the OX50-positive
cytoplasmic processes extend toward the articular cavity
and took a transverse course to align with the articular
cavity surface (Fig. 2a). These cellular processes appeared
almost continuous; thus, few portions of the sublining
layer were exposed to the articular cavity.
On the other hand, immunoelectron microscopy confirmed the existence of some OX50-positive cells in the
sublining layer. The positive cells in the sublining layer
could be divided into two types according to their ultrastructural configurations. One was a large round cell that
had an invaginated cell surface and a large irregular nucleus with rich heterochromatins, suggesting its categorization as a leukocyte (Fig. 2e). The other type was a
In immunoelectron microscopy, immunoreactive products for OX50 were observed as electron-dense deposits.
They were confined to the area around the cell membrane
of the synovial lining cells (Fig. 2a). In particular, the
immunoreactive products appeared to gather at the surface of the synovial lining cell facing the articular cavity.
The OX50-immunopositive cells in the synovial lining
layer possessed well-developed rough endoplasmic reticula, a bright large nucleus with less heterochromatins,
and numerous caveolae with small smooth outlines (Fig.
2c and d). Although the OX50 immunoreaction in the
caveolae showed variable intensities, no immunonegative
caveolae existed in the OX50-positive synovial lining cells
(Fig. 2d). Furthermore, the OX50-positive lining cells con-
650
SUZUKI ET AL.
therefore, the colocalization of HABP/OX50 was intensely
found in the synovial lining layer. The extracellular matrix around some cells in the sublining layer showed a
faint reactivity for HABP and OX50.
DISCUSSION
Fig. 3. HABP reaction (a and b), negative control (c), and double
fluorescent reaction with OX50 and HABP (d). a: The synovial membrane
has an intense HABP reaction (arrowheads) as compared with the articular disk (D). HABP reactions are diffusely distributed in the articular
cavities. b: The higher magnification boxed in a. The intense HABP
reactions are found in synovial lining layer, compared with sublining
layer. c: The equivalent portion to b. The sections treated with hyaluronidase lack any HABP reactivity. d: HABP reactions (green) are found in
the extracellular matrix around both the OX50-positive (red) and -negative cells in the synovial lining layer while the sublining layer contains
weak HABP reactions. T, temporal bone; C, condyle. Scale bars ⫽ 600
␮m (a); 20 ␮m (b– d).
slender cell that possessed a clear flat or ovoid nucleus,
well-developed rough endoplasmic reticula, and a long
thin cytoplasmic process, indicating a putative synovial
fibroblast (data not shown).
OX50-immunonegative cells also existed in both the
synovial lining and sublining cell layers (Fig. 2a). They
were ultrastructurally characterized by the presence of
lysosomes, well-developed filopodia-like processes, and a
dark nucleus with rich heterochromatins (Fig. 2b). Taken
together with their immunoelectron microscopic observation and immunoexpression patterns, these findings indicate that the OX50-immunopositive and -negative lining
cells were fibroblast-like type B cells and macrophage-like
type A cells, respectively.
Histochemistry for HABP
Immunoexpression pattern and histochemistry for HA
did not differ in the TMJ between 4- and 8-week-old rats.
Intense HABP reactions were found in the synovial
membrane but were weak in the articular disk (Fig. 3a
and b). No reactivity for HABP was recognized in the
cytoplasm of the synovial lining cells. The sections pretreated with hyaluronidase did not show any specific reaction (Fig. 3c).
Double Staining With HABP and OX50
Reactions
Double staining was able to confirm the presence of an
HABP reaction (Fig. 3d). The HABP reactions were localized in the extracellular matrix but not in the cytoplasm;
they surrounded both the OX50-positive and -negative
synoviocytes in the synovial lining layer. On the other
hand, weak HABP reactions existed in the sublining layer;
The present immunocytochemical study was clearly
able to demonstrate a precise localization of CD44 and HA
in the synovial membrane of the rat TMJ: the HA reaction
was abundantly distributed in the synovial lining layer,
while CD44 was localized on the cell membrane of the
fibroblast-like type B synoviocytes, not the macrophagelike type A synoviocytes.
The cell type expressing the CD44 molecule remains
controversial. In the human knee joint, some researchers
suggested that both type A and B synoviocytes expressed
CD44 in the healthy and inflammatory conditions (Asari
et al., 1995; Naor and Nedvetzki, 2003). In addition to in
vivo studies, the cultured type B cells in RA showed a
CD44 immunoreaction (Wibulswas et al., 2002; Tolboom
et al., 2004). However, it is well known that the cultured
synovial lining cells change their features, unlike the original synoviocytes (Gadher and Woolley, 1987; Vandenabeele et al., 2003). These controversial findings may be
caused by the absence of any specific cellular markers to
identify the cell type of the positive synoviocytes both in
vivo and in vitro. Current double immunolabeling and
immunoelectron microscopy have revealed an intense
CD44 immunoreactivity in the normal fibroblast-like type
B synovial cells. As far as we know, this is the first report
to disclose the presence of CD44 in the fibroblast-like type
B cells and not the type A cells.
The synovial lining layer shows an epithelium-like arrangement in spite of the absence or incompleteness of
adhesive structures between synovial type B cells
(Nozawa-Inoue et al.,1998, 1999b), giving rise to the question of how the lining cells form and keep their epitheliumlike arrangement. The present immunostaining showed
intense CD44 immunoreaction in these type B cells whose
peripheral ECM displayed HABP reaction. Since CD44,
known as a primary receptor for HA, mediates both cellto-cell and cell-to-matrix interactions (Aruffo et al., 1990;
Underhill, 1992; Lesley et al., 1993; Lesley and Hyman,
1998; Knudson, 2003), it is easily supposed that CD44
serves as an adhesion molecule among the type B cells in
the synovial lining layer both to make and to maintain an
epithelial-like arrangement.
The fibroblast-like type B cells are characteristic of welldeveloped caveolae whose functional significance remains
unknown (Nozawa-Inoue et al., 1998). Current immunostaining demonstrated the localization of CD44 immunoexpression in numerous caveolae and some vesicles with
various intensities in the type B cells. Our careful observations, however, failed to find CD44-positive phagosomes
and coated pits in both typed synovial lining cells, comparable with a previous study on cultured skin fibroblasts
(Culty et al., 1992). Since the uptake of HA is performed
by the membranous indentations with CD44 receptors,
the CD44-immunopositive caveolae in the type B cells has
been considered to participate in this process (Tammi et
al., 2001). Taken together with the fact that the type B
cells synthesize HA (Vuorio et al., 1982; Smith and Ghosh,
1987; Anggiansah et al., 2003), we can easily suppose the
involvement of fibroblast-like type B cells in the HA metabolism.
CD44 AND HA IN RAT TMJ
It is noteworthy that the macrophage-like type A cells in
the TMJ were devoid of CD44 immunoreaction. The expression of CD44 immunoreactivity has been found in
other cell lines of ED1-positive monocyte/macrophages lineages such as osteoclasts and alveolar macrophages
(Green et al., 1988; Culty et al., 1994; Nakamura et al.,
1995; Vivers et al., 2002), which have an ability to phagocytose matrix HA through CD44. This discrepancy may be
explained by the difference in the endocytotic activity
between these CD44/ED1-positive cells and synovial type
A cells. Indeed, type A cells do not develop phagosomes in
their cytoplasm and express a very faint reactivity for acid
phosphates, a marker for phagocytotic activity, under normal conditions (Nozawa-Inoue et al., 1998; Iwanaga et al.,
2000). Under inflammatory conditions, on the other hand,
the type A cells and macrophages with a CD44 immunoreaction on their cell membrane migrate to the synovial
membrane from blood vessels (DeGrendele et al., 1996;
Johnson et al., 2000). These migrated type A cells actively
phagocytose various ECMs or antigens through the CD44
receptor. Taken together with our current observations,
this lack of CD44 immunoreaction in the macrophage-like
type A cells indicates lower and/or no phagocytotic activity
under normal conditions, although they can absorb and
degrade various extracellular components derived from
the synovial fluid and synovial membrane under pathological conditions.
In conclusion, the cell-specific expression and distribution of CD44 and HA in the synovial membrane of rat TMJ
suggest a close relation between type B cell with CD44
immunoreaction and the epithelial-like arrangement of
the synovial lining layer.
ACKNOWLEDGMENTS
The authors thank M. Hoshino and K. Takeuchi, Division of Oral Anatomy, Department of Oral Biological Science, Niigata University Graduate School of Medical and
Dental Sciences, for their technical assistance.
LITERATURE CITED
Alaniz L, Cabrera PV, Blanco G, Ernst G, Rimoldi G, Alvarez E, Hajos
SE. 2002. Interaction of CD44 with different forms of hyaluronic
acid: its role in adhesion and migration of tumor cells. Cell Commun
Adhes 9:117–130.
Andoh E, Kawano Y, Ajima H, Nozawa-Inoue K, Kohno S, Maeda T.
2001. Expression of 25 kDa heat shock protein by synovial type B
cells of the mouse temporomandibular joint. Arch Oral Biol 46:947–
954.
Anggiansah CL, Scott D, Poli A, Coleman PJ, Badrick E, Mason RM,
Levick JR. 2003. Regulation of hyaluronan secretion into rabbit
synovial joints in vivo by protein kinase C. J Physiol 550:631– 640.
Aruffo A, Stamenkovic I, Melnick M, Underhill CB, Seed B. 1990.
CD44 is the principal cell surface receptor for hyaluronate. Cell
61:1303–1313.
Asari A, Miyauchi S, Kuriyama S, Machida A, Kohno K, Uchiyama Y.
1995. Localization of hyaluronic acid in human articular cartilage.
J Histochem Cytochem 42:513–522.
Athanasou NA. 1995. Synovial macrophages. Ann Rheum Dis 54:392–
394.
Berumen-Nafarrate E, Leal-Berumen I, Luevano E, Solis FJ, MunozEsteves E. 2002. Synovial tissue and synovial fluid. J Knee Surg
15:46 – 48.
Cascone P, Fonzi Dagger L, Aboh IV. 2002. Hyaluronic acid’s biochemical stabilization function in the temporomandibular joint. J
Craniofac Surg 13:751–754.
Culty M, Nguyen HA, Underhill CB. 1992. The hyaluronan receptor
(CD44) participates in the uptake and degradation of hyaluronan.
J Cell Biol 116:1055–1062.
651
Culty M, O’Mara TE, Underhill CB, Yeager H Jr, Swartz RP. 1994.
Hyaluronan receptor (CD44) expression and function in human
peripheral blood monocytes and alveolar macrophages. J Leukoc
Biol 56:605– 611.
DeGrendele HC, Estess P, Picker LJ, Siegelman MH. 1996. CD44 and
its ligand hyaluronate mediate rolling under physiologic flow: a
novel lymphocyte-endothelial cell primary adhesion pathway. J Exp
Med 183:1119 –1130.
Dijkstra CD, Dopp EA, Joling P, Kraal G. 1985. The heterogeneity of
mononuclear phagocytes in lymphoid organs: distinct macrophage
subpopulations in the rat recognized by monoclonal antibodies ED1,
ED2 and ED3. Immunology 54:589 –599.
Faassen AE, Schrager JA, Klein DJ, Oegema TR, Couchman JR,
McCarthy JB. 1992. A cell surface chondroitin sulfate proteoglycan,
immunologically related to CD44, is involved in type I collagenmediated melanoma cell motility and invasion. J Cell Biol 116:521–
531.
Forrester JV, Lackie JM. 1981. Effect of hyaluronic acid on neutrophil
adhesion. J Cell Sci 50:329 –344.
Gadher SJ, Woolley DE. 1987. Comparative studies of adherent rheumatoid synovial cells in primary culture: characterisation of the
dendritic (stellate) cell. Rheumatol Int 7:13–22.
Ghosh P. 1994. The role of hyaluronic acid (hyaluronan) in health and
disease: interactions with cells, cartilage and components of synovial fluid. Clin Exp Rheumatol 12:75– 82.
Goebeler M, Kaufmann D, Brocker EB, Klein CE. 1996. Migration of
highly aggressive melanoma cells on hyaluronic acid is associated
with functional changes, increased turnover and shedding of CD44
receptors. J Cell Sci 109:1957–1964.
Graabæk PM. 1984. Characteristics of the two types of synoviocytes in
rat synovial membrane: an ultrastructural study. Lab Invest 50:
690 –702.
Green SJ, Tarone G, Underhill CB. 1988. Aggregation of macrophages
and fibroblasts is inhibited by a monoclonal antibody to the hyaluronate receptor. Exp Cell Res 178:224 –232.
Hakansson L, Hallgren R, Venge P. 1980. Regulation of granulocyte
function by hyaluronic acid. In vitro and in vivo effects on phagocytosis, locomotion, and metabolism. J Clin Invest 66:298 –305.
Hardwick C, Hoare K, Owens R, Hohn HP, Hook M, Moore D, Cripps
V, Austen L, Nance DM, Turley EA. 1992. Molecular cloning of a
novel hyaluronan receptor that mediates tumor cell motility. J Cell
Biol 117:1343–1350.
Hascall VC, Heinegard D. 1974. Aggregation of cartilage
proteoglycans: II, oigosaccharide competitors of the proteoglycanhyaluronic acid interaction. J Biol Chem 249:4242– 4249.
Henderson KJ, Edwards JC, Worrall JG. 1994. Expression of CD44 in
normal and rheumatoid synovium and cultured synovial fibroblasts. Ann Rheum Dis 53:729 –734.
Iwanaga T, Shikichi M, Kitamura H, Yanase H, Nozawa-Inoue K.
2000. Morphology and functional roles of synoviocytes in the joint.
Arch Histol Cytol 63:17–31.
Johnson BA, Haines GK, Harlow LA, Koch AE. 1993. Adhesion molecule expression in human synovial tissue. Arthritis Rheum 36:
137–146.
Johnson P, Maiti A, Brown KL, Li R. 2000. A role for the cell adhesion
molecule CD44 and sulfation in leukocyte-endothelial cell adhesion
during an inflammatory response? Biochem Pharmacol 59:455–
465.
Knudson CB, Knudson W. 1993. Hyaluronan-binding proteins in development, tissue homeostasis, and disease. FASEB J 7:1233–1241.
Knudson W, Aguiar DJ, Hua Q, Knudson CB. 1996. CD44-anchored
hyaluronan-rich pericellular matrices: an ultrastructural and biochemical analysis. Exp Cell Res 228:216 –228.
Knudson CB. 2003. Hyaluronan and CD44: strategic players for cellmatrix interactions during chondrogenesis and matrix assembly.
Birth Defects Res Part C 69:174 –196.
Lesley J, Hyman R, Kincade PW. 1993. CD44 and its interaction with
extracellular matrix. Adv Immunol 54:271–335.
Lesley J, Hyman R. 1998. CD44 structure and function. Front Biosci
3:D616 –D630.
Mapp PI, Revell PA. 1985. Fibronectin production by synovial intimal
cells. Rheumatol Int 5:229 –237.
652
SUZUKI ET AL.
Nakamura H, Kenmotsu S, Sakai H, Ozawa H. 1995. Localization of
CD44, the hyaluronate receptor, on the plasma membrane of osteocytes and osteoclasts in rat tibiae. Cell Tissue Res 280:225–233.
Naor D, Nedvetzki S. 2003. CD44 in rheumatoid arthritis. Arthritis
Res Ther 5:105–115.
Nozawa-Inoue K, Takagi R, Kobayashi T, Ohashi Y, Maeda T. 1998.
Immunocytochemical demonstration of the synovial membrane in
experimentally induced arthritis of the rat temporomandibular
joint. Arch Histol Cytol 61:451– 466.
Nozawa-Inoue K, Ohshima H, Kawano Y, Yamamoto H, Takagi R,
Maeda T. 1999a. Immunocytochemical demonstration of heat shock
protein 25 in the rat temporomandibular joint. Arch Histol Cytol
62:483– 491.
Nozawa-Inoue K, Ajima H, Takagi R, Maeda T. 1999b. Immunocytochemical demonstration of laminin in the synovial lining cell layer
of the rat temporomandibular joint. Arch Oral Biol 44:531–534.
Nozawa-Inoue K, Amizuka N, Ikeda N, Suzuki A, Kawano Y, Maeda
T. 2003. Synovial membrane in the temporomandibular joint: its
morphology, function and development. Arch Histol Cytol 66:289 –
306.
Okada Y, Nakanishi I, Kajikawa K. 1981. Ultrastructure of the mouse
synovial membrane: development and organization of the extracellular matrix. Arthritis Rheum 24:835– 843.
Parkkinen JJ, Häkkinen TP, Savolainen S, Wang C, Tammi R, Ågren
UM, Lammi MJ, Arokoski J, Helminen HJ, Tammi MI. 1996. Distribution of hyaluronan in articular cartilage as probed by a biotinylated binding region of aggrecan. Histochem Cell Biol 105:187–
194.
Paterson DJ, Jefferies WA, Green JR, Brandon MR, Corthesy P,
Puklavec M, Williams AF. 1987. Antigens of activated rat T lymphocytes including a molecule of 50,000 Mr detected only on CD4
positive T blasts. Mol Immunol 24:1281–1290.
Pisko EJ, Turner RA, Soderstrom LP, Panetti M, Foster SL, Treadway WJ. 1983. Inhibition of neutrophil phagocytosis and enzyme
release by hyaluronic acid. Clin Exp Rheumatol 1:41– 44.
Pitsillides AA, Worrall JG, Wilkinson LS, Bayliss MT, Edwards JC.
1994. Hyaluronan concentration in non-inflamed and rheumatoid
synovium. Br J Rheumatol 33:5–10.
Roy S, Ghadially FN. 1967. Synthesis of hyaluronic acid by synovial
cells. J Pathol Bacteriol 93:555–557.
Ruiz P, Schwarzler C, Gunthert U. 1995. CD44 isoforms during differentiation and development. Bioessays 17:17–24.
Smith MM, Ghosh P. 1987. The synthesis of hyaluronic acid by human
synovial fibroblasts is influenced by the nature of the hyaluronate
in the extracellular environment. Rheumatol Int 7:113–122.
Tammi R, Rilla K, Pienimaki JP, MacCallum DK, Hogg M, Luukkonen M, Hascall VC, Tammi M. 2001. Hyaluronan enters keratinocytes by a novel endocytotic route for catabolism. J Biol Chem
276:35111–35122.
Termeer C, Sleeman JP, Simon JC. 2003. Hyaluronan: magic glue for
the regulation of the immune response? Trends Immunol 24:112–
114.
Tolboom TC, Huidekoper AL, Kramer IM, Pieterman E, Toes RE,
Huizinga TW. 2004. Correlation between expression of CD44 splice
variant v8 –v9 and invasiveness of fibroblast-like synoviocytes in an
in vitro system. Clin Exp Rheumatol 22:158 –164.
Tulamo RM, Heiskanen T, Salonen M. 1994. Concentration and molecular weight distribution of hyaluronate in synovial fluid from
clinically normal horses and horses with diseased joints. Am J Vet
Res 55:710 –715.
Underhill CB. 1992. CD44: the hyaluronan receptor. J Cell Sci 103:
293–298.
Vandenabeele F, De Bari C, Moreels M, Lambrichts I, Dell’Accio F,
Lippens PL, Luyten FP. 2003. Morphological and immunocytochemical characterization of cultured fibroblast-like cells derived from adult
human synovial membrane. Arch Histol Cytol 66:145–153.
Visnapuu V, Peltomaki T, Saamanen AM, Ronning O. 2000. Collagen
I and II mRNA distribution in the rat temporomandibular joint
region during growth. J Craniofac Genet Dev Biol 20:144 –149.
Vivers S, Dransfied I, Hart SP. 2002. Role of macrophage CD44 in the
disposal of inflammatory cell corpses. Clin Sci (Lond) 103:441– 449.
Vuorio E, Einola S, Hakkarainen S, Penttinen R. 1982. Synthesis of
underpolymerized hyaluronic acid by fibroblasts cultured from
rheumatoid and non-rheumatoid synovitis. Rheumatol Int 2:97–
102.
Wibulswas A, Croft D, Pitsillides AA, Bacarese-Hamilton I, McIntyre
P, Genot E, Kramer IM. 2002. Influence of epitopes CD44v3 and
CD44v6 in the invasive behavior of fibroblast-like synoviocytes
derived from rheumatoid arthritic joints. Arthritis Rheum 46:
2059 –2064.
Wirth K, Arch R, Somasundaram C, Hofmann M, Weber B, Herrlich
P, Matzku S, Zoller M. 1993. Expression of CD44 isoforms carrying
metastasis-associated sequences in newborn and adult rats. Eur J
Cancer 29A:1172–1177.
Yamada K, Kakudo K, Sako J, Sasaki N, Iseki T, Shirasu R. 2002.
Expression of Hyaluronic acid binding protein (HABP) and CD44 in
murine temporomandibular joint synovium. Jpn J Oral Maxillofac
Surg 48:349 –354.
Документ
Категория
Без категории
Просмотров
2
Размер файла
251 Кб
Теги
hyaluronic, temporomandibular, cd44, joint, localization, rat, membranes, synovial
1/--страниц
Пожаловаться на содержимое документа