(Investigative Ophthalmology and Visual Science. 2000;41:2432-2437.)
© 2000
by The Association for Research in Vision and Ophthalmology, Inc.
Leakage of Aquaporin 5 in the Tear of Dacryoadenitis Mice
Shin–ichiro Hirai,
Naruhiro Ishida,
Katsuhiko Watanabe and
Shiro Mita
From the Discovery Research Division, Santen Pharmaceutical Co., Ltd., Ikoma, Japan.
 |
Abstract
|
|---|
PURPOSE. The objective of this study was to investigate whether leakage of
aquaporin 5 (AQP5) in tear is associated with damage of lacrimal glands
(LGs) in dacryoadenitis models.
METHODS. Female MRL/lpr (24-week-old), male NOD/Shi Jci (5-, 8-, and
10-week-old), female NFS/s-TX (10-week-old), and lipopolysaccharide
(LPS)-induced dacryoadenitis model mice were used. Tear fluid was
collected by a cotton thread. Tear proteins in the thread were
dissolved in sodium dodecyl sulfate buffer, and AQP5 proteins were
analyzed by the Western blot technique using anti-AQP5 antibody. LGs
were prepared for hematoxylin and eosin staining or immunostaining of
AQP5.
RESULTS. In MRL/lpr, NFS/s-TX, 8- and 10-week-old NOD/Shi Jci mice, AQP5 protein
was detected in the tear by Western blot analysis. Inflammatory
lymphocyte infiltrations were observed in LGs of these dacryoadenitis
model mice. In contrast, AQP5 leakage and damage of LG were not
observed in normal mice. In 5-week-old NOD/Shi Jci mice, infiltration
was not seen in LG, and AQP5 leakage was not detected in the tear. In
LPS-induced dacryoadenitis model mice, either tissue destruction with
inflammation in LG or AQP5 leakage in the tear was observed. AQP5 in
the tear and tissue inflammation in LGs was not found in control mice.
These results indicate that AQP5 is leaked in tears when LGs are
damaged by dacryoadenitis.
CONCLUSIONS. Leakage of AQP5 in the tear was found to be related to LG damage. This
finding suggests that detection of AQP5 in tear is useful for specific
diagnosis of LG disorders with tissue
destruction.
|
Introduction
|
|---|
In chronic dacryoadenitis, Sjögren’s syndrome is a
systemic autoimmune disorder characterized by dry eye and dry mouth.
This dryness results from lymphocyte infiltration into lacrimal and
salivary glands.1
In acute dacryoadenitis, intralobular
edema, lymphocytes, and plasma cell infiltration were observed in the
lacrimal gland.2
In Sjögren’s syndrome, it is
assumed that infiltrated T cells may recognize unknown self-antigens
and produce inflammatory cytokines and autoantibodies.1
Histopathologically, lymphocytes penetrate the epithelium of the
salivary gland ducts and cause cytolysis of lacrimal gland
cells.3
To diagnose these kinds of damage to the lacrimal
gland, an histologic evaluation of biopsy specimens is widely used.
However, this procedure has limitations. The biopsy is not available
for reexamination and causes discomfort to the patients. The evaluation
of reflex tearing is also used for diagnosis of Sicca syndrome. It has
been reported that poor reflex tearing is associated with lymphocyte
infiltrations in the lacrimal gland.4
Computed tomography
or magnetic resonance imaging of the glands is another procedure used
for the diagnosis of lacrimal tumors.5
However, these
methods are not always specific or convenient for diagnosis. Hence, a
potent technique that can obviate biopsy, or specific markers to detect
lacrimal gland damage, is needed.
Recently, functional water channels have been identified in mammals,
and this group of proteins has been referred as the
aquaporins.6
We have shown that aquaporin 5 (AQP5) protein
is specifically localized in the apical membrane of acinar and duct
cells in mouse7
and human (Tsubota K, Hirai
S, et al., unpublished data, 2000) lacrimal gland. In other
ocular tissues, AQP5 expression was not demonstrated in
conjunctiva.8
In contrast to AQP5, membrane channel
proteins, such as ion channels or sodium/potassium
adenosinetriphosphatase, have been shown as present not only in the
lacrimal gland9
10
but also in the corneal
epithelium11
and the conjunctiva.12
To the
best of our knowledge, no specifically localized protein in the
lacrimal gland other than AQP5 has been reported. Based on this
evidence, we hypothesize that AQP5 is specifically leaked into tear
fluid when the lacrimal gland is damaged by lymphocyte infiltrations
and inflammation with tissue destruction. In central diabetes
insipidus,13
impaired water excretion, and
hyponatremia,14
aquaporin 2 (AQP2), which was excreted
into urine, was found to be related to the pathologic state of the
disease. In this study, we demonstrate that in experimental
dacryoadenitis models, the detection of AQP5 in tear fluid is useful
for specifically diagnosing lacrimal gland damage.
|
Methods
|
|---|
All experiments were carried out in accordance with the ARVO
Statement for the Use of Animals in Ophthalmic and Vision Research.
Dacryoadenitis models were female MRL/lpr, male NOD/Shi Jci (Japan
CLEA, Tokyo, Japan), female NFS/s-TX, and lipopolysaccharide
(LPS)-induced dacryoadenitis mice. These mice were maintained in a
specific pathogen-free colony and given food and water ad libitum.
NFS/sld mice were purchased from the Central Institute for
Experimental Animals (Kawasaki, Japan). They were thymectomized on day
3 after birth.15
Thymectomized NFS/sld mice are
referred to as NFS/s-TX mice. As control animals, we purchased female
BALB/c for MRL/lpr, male Jcl:ICR for NOD/Shi Jci, and female
NFS/s-nonTX (non-thymectomized mice) for NFS/s-TX mice, respectively.
MRL/lpr and BALB/c mice were killed at 24 and 7 weeks after birth,
respectively. NFS/s-TX and NFS/s-nonTX mice were killed at 10 weeks.
NOD/Shi Jci and Jcl:ICR mice were killed at 5, 8, and 10 weeks. As
LPS-induced dacryoadenitis model mice, 5-week-old male BALB/c mice were
anesthetized with sodium pentobarbital (60 mg/kg), and injected with
saline-dissolved LPS (25 µg/2 µl; Difco, Detroit, MI) into
the exorbital lacrimal gland. Saline-injected mice were used as control
animals. Mice in this group were killed at 3, 9, and 24 hours after
injection.
For immunoblot detection of AQP5 in the tear fluid, mice were
anesthetized with sodium pentobarbital (60 mg/kg), and the tears were
collected with a cotton thread treated with the phenol red pH indicator
(Showa Yakuhin Kako, Tokyo, Japan) used in Schirmer’s test.
Tear volume was determined by the length of wet thread indicated with
phenol red pH indicator. The unit of length was the millimeter. Tear
proteins in 10 mm of thread were dissolved in 10 µl of a loading
buffer (125 mM Tris–HCl, pH 7.4, 4% sodium dodecyl sulfate, 20%
glycerol, 5% 2-mercaptoethanol) and electrophoresed on a 10% to 20%
gradient polyacrylamide gel. Tear proteins were transferred to
polyvinylidene difluoride membrane (Millipore, Bedford, MA) and
immunolabeled using affinity-purified anti-AQP5 IgG at a 1:1000
dilution. Anti-AQP5 rabbit IgG that recognizes the carboxyl-terminus of
rat AQP5 was prepared as previously described.7
The bound
antibodies were detected using the ECL chemiluminescence method
(Amersham, Buckinghamshire, UK). For positive control of immunoblot
detection of AQP5, a membrane fraction was isolated from BALB/c mouse
lacrimal glands.7
Glands were homogenized in a buffer
solution of 30 mM mannnitol, 10 mM CaCl2, 10 mM
Tris–HCl (pH 7.4), containing 1 mM phenylmethylsulfonyl fluoride. The
homogenate was centrifuged at 3000g for 15 minutes, and the
supernatant was centrifuged at 42,000g for 20 minutes. The
resultant pellet was suspended with 100 mM mannitol and 10 mM Tris–HCl
(pH 7.4) and recentrifuged at 42,000g for 20 minutes; the
final pellet was then resuspended with this buffer and used as a
positive control.
Exorbital lacrimal glands were removed from the strains of mice and
fixed with 4% phosphate-buffered paraformaldehyde at pH 7.2. After
fixation, lacrimal glands were dehydrated with ethanol and embedded in
paraffin. Tissue sections (4-µm-thick) were prepared for hematoxylin
and eosin (HE) staining or immunohistochemical staining of AQP5.
Immunohistochemistry was performed on lacrimal gland sections of
NOD/Shi Jci and Jcl:ICR mice using anti-AQP5 guinea pig IgG that
recognizes the extracellular domain of rat AQP5.7
Sections
were rinsed with phosphate-buffered saline (PBS) at pH 7.2 and
incubated with 0.1% trypsin in PBS for 30 minutes. After incubation,
sections were rinsed with PBS and treated with methanol containing
0.3% H2O2 for 15 minutes.
Sections were rinsed and incubated with blocking agent (15 µl/ml
normal goat serum, 1% bovine serum albumin, 0.5% Triton-X in PBS, pH
7.2) for 1 hour at room temperature. After incubation with anti-AQP5
antibody at a 1:1000 dilution overnight at 4°C, sections were rinsed
with PBS 3 times for 5 minutes and treated with biotinylated
anti–guinea pig IgG for 1 hour, then rinsed with PBS 3 times. For
immunofluorescence detection, the antibody-incubated slides were
treated with PBS containing avidin–fluorescein isothiocyanate for 30
minutes and mounted using an anti-fade solution to retard fluorescence
photobleaching. HE-stained or immunostained sections were photographed
by Fuji super HG-100 film (Fuji Photo Film, Tokyo, Japan) using an
Olympus light microscope (Olympus Optical, Tokyo, Japan) or by
a digital image analyzing system using a CSU-10 confocal scanning
microscope (Yokogawa Electric, Tokyo, Japan).
|
Results
|
|---|
MRL/lpr and NFS/s-TX Mice
Histologic examination revealed destructive lymphocyte
infiltration in lacrimal glands of 24-week-old MRL/lpr (Fig. 1A
) and 10-week-old NFS/s-TX mice (data not shown). Secretory lobules
were extensively replaced by aggregations of lymphocytes, especially
around the secretory ducts. Acinar and duct cells of the gland showed
destructive changes, including destruction of the cell membrane and
degeneration. On the other hand, lymphocyte infiltration was not found
in the lacrimal glands of 7-week-old BALB/c (Fig. 1B)
and 10-week-old
NFS/s-nonTX mice. In MRL/lpr and NFS/s-TX mice, Western blot analysis
revealed that AQP5 protein had leaked into the tear fluid. As shown in
Figure 1C , 28-kDa bands were specifically stained with
affinity-purified anti-AQP5 IgG. AQP5 was not detected in the tear
fluid of normal mice (Fig. 1C)
.
View larger version (85K):
[in this window]
[in a new window]
|
Figure 1. Histologic appearance of lacrimal gland of 24-week-old MRL/lpr
(A) and 7-week-old BALB/c (B) mice. Note the
severe lymphocyte infiltrations in MRL/lpr mouse lacrimal gland. HE
stained. Scale bar, (A, B) 20 µm.
(C) Western blot analysis of AQP5 protein in the tear of
24-week-old MRL/lpr (lane 1), 10-week-old NFS/s-TX
(lane 2), 7-week-old BALB/c (lanes 3 and
5), and 10-week-old NFS/s non-TX (lane 4) mice.
Note leakage of AQP5 was observed in the tear of MRL/lpr and NFS/s-TX
mice. Lane 6, positive control (apical membrane fraction
isolated from BALB/c mouse lacrimal gland as described in the Methods
section). The purified antibody to AQP5 carboxyl-terminus was used.
|
|
NOD/Shi Jci Mice
In NOD/Shi Jci mice, histologic examination showed lymphocyte
infiltration with tissue destruction in lacrimal glands at 8 and 10
weeks but not 5 weeks (Fig. 2A
). This observation shows that lymphocyte infiltration progressed with
age. Immunohistochemical staining revealed the presence of AQP5 in the
lacrimal glands of 5-, 8-, and 10-week-old NOD/Shi Jci and normal
Jcl:ICR mice. Immunolocalization of AQP5 was observed in secretory duct
cells of the lacrimal gland (Figs. 2A
2B)
. By Western blot analysis,
leakage of AQP5 into tear fluid was detected in 8- and 10-week-old
NOD/Shi Jci mice (Fig. 2C)
. In contrast, AQP5 was not detected in the
tear fluid of 5-week-old NOD/Shi Jci mice whose lacrimal glands showed
any infiltration (Figs. 2A
2C)
. In normal Jcl:ICR mice (5-, 8-, and
10-week-old), neither leakage of AQP5 into the tear fluid nor
inflammatory changes in the lacrimal gland were observed (Figs. 2B
2D)
. These results indicate that leakage of AQP5 in the tear is
associated with age-dependent progression of lymphocyte infiltration in
the lacrimal gland.
View larger version (76K):
[in this window]
[in a new window]
|
Figure 2. Immunohistochemical (IHC) detection of AQP5 and
HE (H.E.) staining in lacrimal glands of NOD/Shi Jci (A) and
Jcl:ICR (B) mice at the age of 5, 8, and 10 weeks. Note the
lymphocyte infiltrations in 8- and 10-week-old NOD/Shi Jci mice. AQP5
expression was observed in duct cells of both NOD/Shi Jci and Jcl:ICR
mice. In IHC, the purified antibody to the extracellular domain of AQP5
was used. Scale bar, (A, B) 30 µm.
(C) Western blot analysis of AQP5 protein in the tear of
NOD/Shi Jci mice. Lane 1, at the age of 5 weeks; lane
2, 8 weeks; lane 3, 10 weeks. Lane 4,
positive control (apical membrane fraction isolated from BALB/c mouse
lacrimal gland as described in the Methods section). Note the leakage
of AQP5 protein was observed in the tear of 8- and 10-week old mice.
(D) Western blot analysis of AQP5 protein in the tear of
Jcl:ICR mice. Lane 1, at the age of 5 weeks; lane
2, 8 weeks; lane 3, 10 weeks. Lane 4,
positive control. The purified antibody to AQP5 carboxyl-terminus was
used.
|
|
LPS-Induced Dacryoadenitis Model Mice
Figure 3A
shows the development of inflammation in the lacrimal glands of
LPS-induced dacryoadenitis model mice. At 3 hours after injection of
LPS into the gland, inflammatory changes were not observed in lacrimal
glands. At 9 hours, the lacrimal glands showed histopathologic features
of acute inflammation. Inflammatory monocytes with neutrophils were
found throughout the secretory lobules and ducts. At 24 hours, the
inflammation of the lacrimal gland was developed with numerous
neutrophils among the acinar and duct cells showing destruction or
degeneration. By Western blot analysis, the AQP5 protein was found to
have leaked into the tear fluid of mice killed at 9 and 24 hours (Fig. 3C)
. On the other hand, lacrimal glands were not damaged and AQP5 was
not detected in the tears of either mice killed at 3 hours or
saline-injected control mice (Figs. 3B
3D)
. These observations
indicate that the leakage of AQP5 protein in the tear is closely
associated with lacrimal gland damage caused by LPS-induced acute
inflammation.
View larger version (120K):
[in this window]
[in a new window]
|
Figure 3. Histopathology of LPS-induced dacryoadenitis (A) and
saline-injected (B) lacrimal glands at 3, 9, and 24 hours
after LPS or saline injection. Note inflammation with neutrophils in
9- and 24-hour–killed LPS-induced dacryoadenitis model mice. HE
stained. Scale bar, (A, B) 20 µm.
(C) Western blot analysis of AQP5 protein in the tear fluid
of LPS-induced dacryoadenitis model mice. Lane 1, 3 hours;
lane 2, 9 hours; lane 3, 24 hours after injection
of LPS. Lane 4, positive control (apical membrane fraction
isolated from BALB/c mouse lacrimal gland as described in the Methods
section). Note the AQP5 leakage in the tears of mice killed at 9 and 24
hours. (D) Western blot analysis of AQP5 protein in the tear
fluid of saline-injected mice. Lane 1, 3 hours; lane
2, 9 hours; lane 3, 24 hours after saline injection.
Lane 4, positive control. The purified antibody to AQP5
carboxyl-terminus was used.
|
|
|
Discussion
|
|---|
In this study, we first demonstrated that AQP5 protein leaks into
the tears of dacryoadenitis model mice. Lacrimal glands of these model
mice showed lymphocyte infiltration and inflammation. In chronic
dacryoadenitis, such as Sjögren’s syndrome, a characteristic
finding is lymphocyte infiltration into lacrimal and salivary
glands.1
In acute dacryoadenitis, biopsy specimens of the
lacrimal gland show intralobular edema, lymphocytes, and plasma cell
infiltration.2
Most lacrimal gland inflammations (acute,
subacute, or chronic) are typified by lymphocytic and plasmacytic
infiltrates.2
In Sjögren’s syndrome, proteases or
cytokines produced from T cells are thought to have direct cytotoxic or
destructive effects on the lacrimal gland.16
Histopathologic studies revealed that the salivary glands show
intraepithelial penetration of lymphocytes around the
ducts.3
In the lacrimal gland, vacuolized and edematous
cells showing cytolysis were identified using electron
microscopy.3
In this study, lacrimal cells of
dacryoadenitis mice showed destructive changes. These results indicate
that infiltrating lymphocytes destroy the cell membrane in lacrimal
glands with Sjögren-like inflammation. In this study, we detected
AQP5, which is normally localized in the apical membrane of acinar and
duct cells, in dacryoadenitis mouse tear fluid. This evidence suggests
that AQP5 protein leaks from damaged membranes of acinar and duct cells
into the tear fluid of dacryoadenitis models.
For detection of the damage to the lacrimal gland, several methods have
been proposed. Histologic evaluation of biopsy specimens of the
lacrimal gland is often used to detect lacrimal gland damage.
Evaluation of experimental results using computed tomography (CT) and
magnetic resonance imaging of the lacrimal gland5
is
another technique. In CT, the variability of tissues within the mixed
tumor correlates with the radiologic features.5
It has
been reported that evaluation of reflex tearing with nasal stimulation
could identify two groups of patients with dry eye. Histologic
evaluation of the lacrimal gland revealed that patients with poor
reflex tearing were more likely to have lymphocyte infiltrations in the
gland than patients with good reflex tearing.4
This
finding suggests that the measurement of reflex tearing and the
histologic evaluation of the gland are important for the diagnosis of
Sicca syndrome. In the present study, we demonstrated that leakage of
AQP5 protein into the tear fluid is closely associated with damage of
the lacrimal gland. This evidence indicates that detection of AQP5 in
tear fluid can be helpful in the diagnosis of lacrimal gland damage,
thereby avoiding lacrimal biopsy. In the case of severe dry eye (e.g.,
Sjögren’s syndrome), it might be difficult to collect a
sufficiently large volume of tear fluid for AQP5 detection, by reason
of inadequate tear secretion. Having a highly sensitive method that can
detect AQP5 in tear fluid will resolve this problem. In addition, we
have established an enzyme-linked immunosorbent assay system for AQP5
detection in tear fluid. According to our method, the amount of AQP5 in
tear fluid of dacryoadenitis mice was large enough to evaluate the
damage of lacrimal gland (Tsubota K, Hirai S, et al.,
unpublished data, 2000).
In ocular tissues other than the lacrimal gland, AQP5 is expressed in
cell membranes of the corneal epithelium17
but not in the
bulbar conjunctival epithelium or stroma.8
In the corneal
epithelium, immunoreactivity was localized in the cell membrane of the
superficial, wing, and columnar basal cells by immunohistochemistry and
immunoelectron microscopy.17
It is possible that AQP5
leaked from injured corneal epithelium into tear fluid. However, in the
present study, AQP5 was not detected in the tear fluid of control mice
whose lacrimal gland showed no inflammation. Moreover, a histologic
analysis of the corneal epithelium of dacryoadenitis model mice did not
show any inflammation or tissue destruction (data not shown). These
results suggest that AQP5 in the tear fluid is specifically leaked from
the lacrimal gland and is associated with the damage of the gland. In
addition, standard procedures (e.g., rose bengal or fluorescein
staining of the ocular surface) may detect the corneal epithelium
damage, enabling specific evaluation of the lacrimal gland damage. On
the other hand, the presence of membrane proteins, such as sodium,
chloride, and potassium channels or sodium/potassium
adenosinetriphosphatase, has been reported not only in the lacrimal
gland9
10
but also in the conjunctiva12
and
the corneal epithelium.11
Moreover, to the best of our
knowledge, proteins that show specific localization in the lacrimal
gland have not been reported. We therefore think that proteins other
than AQP5 are not useful as specific markers of lacrimal gland damage.
It has been reported that AQP2 protein excreted into urine is
associated with certain disorders of the kidney. In central diabetes
insipidus, urinary excretion of AQP2 occurred at a low concentration
compared with healthy controls as determined by radioimmunoassay using
an anti-AQP2 antibody.13
In patients with impaired water
excretion and hyponatremia, the urinary AQP2 concentration was higher
than that in control patients under ad libitum water
drinking.14
These results indicate that urinary AQP2 is a
potent marker for diagnosing the pathologic state of central diabetes
insipidus,13
impaired water excretion, and
hyponatremia.14
The present study demonstrates that AQP5
leaks into the tear when lacrimal glands are damaged. In addition,
immunohistochemical studies have shown that AQP5 is localized in the
apical membrane of acinar and duct cells of the lacrimal gland in
humans (Tsubota K, Hirai S, et al., unpublished data,
2000). Thus, we believe that detection of AQP5 in tear fluid is
useful for simply and safely diagnosing lacrimal gland damage. And this
method of AQP5 detection has good specificity and potential for
reexamination. Moreover, AQP5 detection in the tear fluid could be used
as an evaluation method for the therapeutic effects of drugs for
lacrimal gland disorders with tissue destruction. However, further
studies are necessary to determine whether the quantity of AQP5 protein
in the tear fluid is correlated with the progression or healing of
lacrimal gland damage.
In conclusion, leakage of AQP5 in the tear fluid was found to be
related to lacrimal gland damage. This finding suggests that detection
of AQP5 in tear fluid is useful for the specific diagnosis of lacrimal
gland disorders with tissue destruction.
|
Acknowledgements
|
|---|
The authors thank Miyako Kawarasaki of KAC Ltd. for expert
technical assistance.
|
Footnotes
|
|---|
Submitted for publication July 20, 1999; revised January 13 and March 30, 2000; accepted April 19, 2000.
Commercial relationships policy: E.
Corresponding author: Shin–ichiro Hirai, Ophthalmic Research Division, Santen Pharmaceutical Co., Ltd., 8916-16 Takayamacho, Ikoma, 630-0101, Japan. shirai{at}mahoroba.ne.jp
|
References
|
|---|
-
Fox, RI, Kang, Ho–II (1992) Pathogenesis of Sjögren’s syndrome Rheum Dis Clin North Am 18,517-538[Medline][Order article via Infotrieve]
-
Jakobiec, FA, Bilyk, JR, Font, RL (1996) Lacrimal gland tumors William, HS eds. Ophthalmic Pathology: An Atlas and Text Book 4,2485-2525 Saunders Philadelphia.
-
Chomette, G, Auriol, M, Liotet, S. (1986) Ultrastructural study of the lacrimal gland in a case of Sjögren’s syndrome Scand J Rheumatol 61,71-75
-
Tsubota, K, Xu, K–P, Fujihara, T, et al (1996) Decreased reflex tearing is associated with lymphocytic infiltration in lacrimal gland J Rheumatol 23,313-320[Medline][Order article via Infotrieve]
-
Karcioglu, ZA (1994) Space-occupying lesions of the lacrimal fossa Curtis, EM eds. Diagnostic Problems in Clinical Ophthalmology ,70-75 Saunders Philadelphia.
-
Chrispeels, MJ, Agre, P. (1994) Aquaporins: water channel proteins of plant and animal cells Trends Biochem Sci 19,421-425[Medline][Order article via Infotrieve]
-
Ishida, N, Hirai, S, Mita, S. (1997) Immunolocalization of aquaporin homologs in mouse lacrimal glands Biochem Biophys Res Commun 238,891-895[Medline][Order article via Infotrieve]
-
Hamann, S, Zeuthen, T, LaCour, M, et al (1998) Aquaporins in complex tissues: distribution of aquaporins 1–5 in human and rat eye Am J Physiol 274,C1332-C1345[Abstract/Free Full Text]
-
Alexander, JH, vanLennep, EW, Young, JA (1972) Water and electrolyte secretion by the exorbital lacrimal gland of the rat studied by micropuncture and catheterization techniques Pflügers Arch 337,299-309[Medline][Order article via Infotrieve]
-
Dartt, DA, Møller, M, Poulsen, JH (1981) Lacrimal gland electrolyte and water secretion in the rabbit: localization and role of (Na+ + K+)-activated ATPase J Physiol 321,557-569[Abstract/Free Full Text]
-
Klyce, SD, Crosson, CE (1985) Transport processes across the rabbit corneal epithelium: a review Curr Eye Res 4,323-331[Medline][Order article via Infotrieve]
-
Maurice, DM (1973) Electrical potential and ion transport across the conjunctiva Exp Eye Res 15,527-532[Medline][Order article via Infotrieve]
-
Saito, T, Ishikawa, S-E, Ando, F, et al (1998) Exaggerated urinary excretion of aquaporin-2 in the pathological state of impaired water excretion dependent upon arginine vasopressin J Clin Endocrinol Metab. 11,4034-4040
-
Saito, T, Ishikawa, S-E, Sasaki, S, et al (1997) Urinary excretion of aquaporin-2 in the diagnosis of central diabetes insipidus J Clin Endocrinol Metab. 6,1823-1827
-
Haneji, N, Hamano, H, Yanagi, K, Hayashi, Y. (1994) A new animal model for primary Sjögren’s syndrome in NFS/sld mutant mice J Immunol 153,2769-2777[Abstract]
-
Fox, PC, Speight, PM (1996) Current concepts of autoimmune excrinopathy: immunologic mechanisms in the salivary pathology of Sjögren’s syndrome Crit Rev Oral Biol Med 7,144-158[Abstract/Free Full Text]
-
Funaki, H, Yamamoto, T, Koyama, Y, et al (1998) Localization and expression of AQP5 in cornea, serous salivary glands, and pulmonary epithelial cells Am J Physiol 275,C1151-C1157
This article has been cited by other articles:
|
|
|
|
 
D. S. Vinay, J. D. Kim, T. Asai, B. K. Choi, and B. S. Kwon
Absence of 4 1BB Gene Function Exacerbates Lacrimal Gland Inflammation in Autoimmune-Prone MRL-Faslpr Mice
Invest. Ophthalmol. Vis. Sci.,
October 1, 2007;
48(10):
4608 - 4615.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. H. Grus, V. N. Podust, K. Bruns, K. Lackner, S. Fu, E. A. Dalmasso, A. Wirthlin, and N. Pfeiffer
SELDI-TOF-MS ProteinChip Array Profiling of Tears from Patients with Dry Eye
Invest. Ophthalmol. Vis. Sci.,
March 1, 2005;
46(3):
863 - 876.
[Abstract]
[Full Text]
[PDF]
|
|
|
Top
Abstract
Introduction
