From the Department of Cardiovascular Medicine, University of Tokyo
Graduate School of Medicine, Tokyo 113-8655,
1st
Department of Internal Medicine, Faculty of Medicine, Tottori
University, Tottori 680-8550, § International Medical Center
of Japan, Tokyo 162-8655, and ¶ Department of Medicine III, Chiba
University School of Medicine, Chiba 260-8670, Japan
gp130, a common receptor for the
interleukin 6 family, plays pivotal roles in growth and survival of
cardiac myocytes. In the present study, we examined the role of gp130
in pressure overload-induced cardiac hypertrophy using transgenic (TG)
mice, which express a dominant negative mutant of gp130 in the heart
under the control of
myosin heavy chain promoter. TG mice were
apparently healthy and fertile. There were no differences in body
weight and heart weight between TG mice and littermate wild type (WT)
mice. Pressure overload-induced increases in the heart weight/body
weight ratio, ventricular wall thickness, and cross-sectional areas of
cardiac myocytes were significantly smaller in TG mice than in WT mice. Northern blot analysis revealed that pressure overload-induced up-regulation of brain natriuretic factor gene and
down-regulation of sarcoplasmic reticulum Ca2+ ATPase
2 gene were attenuated in TG mice. Pressure overload activated ERKs and
STAT3 in the heart of WT mice, whereas pressure overload-induced activation of STAT3, but not of ERKs, was suppressed in TG mice. These
results suggest that gp130 plays a critical role in pressure overload-induced cardiac hypertrophy possibly through the STAT3 pathway.
 |
INTRODUCTION |
Because recent clinical studies have suggested that cardiac
hypertrophy is an independent risk factor of cardiac morbidity and
mortality (1), it has become even more important to clarify the
mechanism of how cardiac hypertrophy is developed. Cardiomyocyte hypertrophy can be induced by a variety of factors such as mechanical stress (2), cathecholamines (3), angiotensin II (4), endothelin-1 (5),
and cytokines (6). Among them, hemodynamic overload, namely mechanical
stress, is clinically most important. We and others (7-9) have
reported that mechanical stress induces cardiomyocyte hypertrophy
through vasoactive peptides such as angiotensin II and
endothelin-1.
Cardiotrophin-1, a member of the interleukin 6 (IL-6)1 family, was isolated
and found to have a potent hypertrophic effect on cultured
cardiomyocytes (10). The IL-6 family of cytokines promotes cell
type-specific pleiotropic effects by engaging multimeric receptor
complexes that share the common affinity converter/signal-transducing subunit gp130 (11-13). Cardiotrophin-1 has been reported to induce hypertrophy of cardiac myocytes in vitro (14). It has been
reported that transgenic mice expressing both IL-6 and soluble IL-6
receptor, in which the gp130 is continuously activated, showed marked
hypertrophy of the ventricular myocardium (15) and that targeted
disruption of gp130 leads to severe anemia and a hypoplastic
ventricular myocardium in the embryo (16). These results suggest that
activation of gp130 induces cardiac hypertrophy, and it is still
unknown whether gp130 mediates load-induced cardiac hypertrophy.
Cardiotrophin-1 has been reported to promote survival of cardiac
myocytes (17). Ventricular-restricted gp130 knockout mice showed
marked ventricular wall dilatation with marked cardiomyocyte apoptosis
and died in a week by pressure overload (18). These results suggest
that gp130 signalings prevent cardiomyocytes from apoptotic cell death during the pressure overload.
In the present study, to determine the physiological significance of
gp130 in load-induced cardiac hypertrophy, we generated transgenic (TG)
mice, which express a dominant negative form of gp130 specifically in
the heart and examined hypertrophic responses by pressure overload
produced by constriction of the abdominal aorta.
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MATERIALS AND METHODS |
Transgene Construction and Generation of TG Mice--
A dominant
negative mutant of gp130 (D.N.gp130) was constructed by converting
cysteine at 702 to a stop codon as described previously (19). D.N.gp130
cDNA was inserted into the unique KpnI site of
p
MHCSA, which carries the mouse
myosin heavy chain (
MHC) promoter (20).
MHC promoter-D.N.gp130-poly(A) DNA
was excised by XhoI and NotI and microinjected
into the pronuclei of fertilized BDF1 mouse eggs. Offsprings
from eggs microinjected with the DNA were selected by Southern blot
analysis and polymerase chain reaction.
Pressure Overload Model--
Male TG and littermate wild type
(WT) mice of 20 weeks old were used in the present study. Mice were
housed under climate-controlled conditions with a 12-h light/dark cycle
and were provided with standard food and water ad libitum.
All protocols were approved by local institutional guidelines. Pressure
overload was produced by constriction of the abdominal aorta as
described previously in our laboratory (21, 22). Briefly, mice were
anesthetized by intraperitoneal injection of sodium pentobarbital (30 mg/kg). The abdominal aorta was constricted at the suprarenal level
with 7-0 nylon strings by ligation of the aorta with a blunted
27-gauge needle, which was pulled out thereafter.
Echocardiographic Measurement--
Transthoracic
echocardiography was performed with HP Sonos 100 (Hewlett-Packard Co.)
with a 10-MHz imaging transducer as described previously (22, 23). Mice
were anesthetized with ketamine (10 mg/kg intraperitoneal) and xylazine
(15 mg/kg intraperitoneal). After a good quality two-dimensional image
was obtained, M-mode images of the left ventricle were recorded.
Left ventricular end-diastolic internal diameter (LVEDD), left
ventricular end-systolic internal diameter (LVESD), intraventricular
septum thickness, and left ventricular posterior wall thickness were
measured. All measurements were performed by use of the leading
edge-to-leading edge convention adopted by the American Society
of Echocardiography (24). Fractional shortening was calculated as
fractional shortening = ((LVEDD
LVESD)/LVEDD) × 100. Ejection fraction was calculated by the cubed method as follows:
ejection fraction = ((LVEDD)3
(LVESD)3)/LVEDD3.
Histological Analysis--
For histological analysis, hearts
were fixed with 10% formalin by perfusion fixation. Fixed hearts
embedded in paraffin were sectioned at 4-µm thickness and stained
with hematoxylin-eosin. Cross-sectional areas of cardiac myocytes were
measured from 10 sections. Suitable cross-sections were defined as
having nearly circular capillary profiles and nuclei.
RNA Preparation and Northern Blot Analysis--
The left
ventricle was excised, and total RNA (10 µg) was prepared using ZolB
(Biotecx Laboratories, Inc.), fractionated in 1% formaldehyde agarose
gel, and transferred to nylon membrane. The blots were hybridized with
the cDNA fragments of gp130, brain natriuretic factor (BNP), and
sarcoplasmic reticulum Ca2+ ATPase 2 (SERCA2) genes.
Western Blot Analysis of STAT3 Phosphorylation--
Polyclonal
antibody to STAT3 (C-20) was purchased from Santa Cruz
Biotechnology, Inc. The left ventricle was excised and lysed in lysis
buffer (25 mM Tris-HCl, 25 mM NaCl, 1 mM sodium orthovanadate, 10 mM NaF, 10 mM sodium pyrophosphate, 10 nM okadaic acid,
0.5 mM EGTA, and 1 mM phenylmethylsulfonyl
fluoride), and equal amounts (100 µg) of protein were incubated with
1 µg of anti-STAT3 for 1 h at 4 °C. The immune complexes were
precipitated with protein A-Sepharose, and the immuneprecipitates were
separated by SDS polyacrylamide gel electrophoresis and transferred to
nitrocellulose membrane. The membranes were then blocked and incubated
with anti-phosphotyrosine antibody (4G10), and the phosphotyrosine was
detected by ECL (Amersham Pharmacia Biotech).
Assay of Extracellular Signal-regulated Kinases (ERKs)--
The
activities of ERKs were measured using myelin basic protein-containing
gel (25). In brief, lysates of the left ventricles were subjected to
electrophoresis on an SDS polyacrylamide gel containing 0.5 mg/ml
myelin basic protein. ERKs in the gel were denatured in guanidine HCl
and renatured in Tris-HCl (pH 8.0) containing 0.04% Triton X-100 and
2-mercaptoethanol (5 mM). Phosphorylation activities of
ERKs were assayed by incubating the gel with
[
-32P]ATP.
Terminal Deoxynucleotidyl Transferase Assay (TUNEL)--
The
4-µm thickness paraffin sections were deparaffinized by immersing in
xylene, rehydrated, and incubated with proteinase K (20 µg/ml). Next,
the sections were incubated in methanol with 3%
H2O2 to inactivate endogenous peroxidases,
washed in phosphate-buffered saline, and incubated with terminal
deoxynucleotidyl transferase and fluorescein isothiocyanate-dUTP for 90 min and horseradish peroxidase-conjugated anti-fluorescein
isothiocyanate for 30 min at 37 °C using an apoptosis detection
kit (Takara Biochemicals). The sections were stained with
diaminobenzine and hematoxylin and mounted for light microscopic observations.
Statistics--
Differences within groups were compared by the
one-way analysis of variance (ANOVA) and Dunnett's t test.
The accepted level of significance was p < 0.05.
 |
RESULTS |
Generation of TG Mice--
The carboxyl-terminal region of gp130
containing box3 is considered to play a critical role in gp130-mediated
biological responses (26). D.N.gp130 TG mice were generated by
overexpression of a box3-deleted form of gp130 under the control of
MHC promoter (Fig. 1). Six founders
containing the transgene were identified, and three transgenic lines,
in particular number 3, were used in this study. TG mice were
apparently healthy and fertile, and there was no difference in the
heart weight to body weight ratio between TG mice and WT mice, at
baseline (Fig. 2).

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Fig. 1.
Diagram of the transgene construct and
expression in the heart. A, schematic structure
of the MHC promoter-D.N.gp130-poly(A) transgene construct. A
2.4-kilobase of truncated murine gp130 cDNA was
subcloned between the murine MHC promoter and poly(A). The truncated
form of gp130 contains the membrane-proximal 63-amino acid part
of the cytoplasmic region of gp130 but lacks box3. B,
expression of the truncated form of gp130. 10 µg of total RNA from
the hearts were hybridized with labeled gp130 cDNA. TM,
transmembrane.
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Fig. 2.
Heart weight/body weight ratio. Pressure
overload was produced by constriction of the abdominal aorta for 4 weeks. Mean ± S.E. of the heart weight/body weight ratio was
shown (n = 3 in sham-operated mice, n = 4 in constricted mice). *p < 0.05; N.S.,
not significant.
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Cardiac Hypertrophy Induced by Pressure Overload--
Constriction
of the abdominal aorta is an established method to produce pressure
overload-induced cardiac hypertrophy (21, 22). 4 weeks after the
operation, all mice were healthy, and the blood pressure monitored at
right carotid artery was elevated as reported before in our laboratory
(22) without any differences between TG mice and WT mice (data not
shown). Echocardiography at 4 weeks after the operation revealed that
left ventricular wall thickness was markedly increased
(intraventricular septum thickness, sham 0.50 ± 0.03 versus constriction 0.81 ± 0.03; left ventricular
posterior wall thickness, sham 0.54 ± 0.04 versus constriction 0.83 ± 0.08) in WT mice; however, TG mice showed less increase in left ventricular wall thickness (intraventricular septum thickness, sham 0.48 ± 0.03 versus constriction
0.59 ± 0.00; left ventricular posterior wall thickness, sham
0.52 ± 0.00 versus constriction 0.64 ± 0.04) (Table I). Furthermore,
constriction of the abdominal aorta for 4 weeks increased the heart
weight/body weight ratio by 40 ± 4% in WT mice and 15 ± 3% in TG mice (Fig. 2). The left ventricle of WT mice showed more
marked hypertrophy than that of TG mice (Fig.
3A). Microscopic analysis
showed that cross-sectional areas of cardiac myocytes of WT mice were
more enlarged by pressure overload than those of TG mice (Fig.
3B). These results suggest that cardiac hypertrophy induced
by pressure overload was attenuated in D.N.gp130 TG mice.
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Table I
Analysis of cardiac size and function
LVESD, left ventricular end-systolic internal diameter; IVST,
intraventricular septum thickness; LVPWT, left ventricular posterior
wall thickness; FS, fractional shortening; EF, ejection fraction.
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Fig. 3.
Histological analysis. A,
transverse sections of the hearts at papillary muscle level at 4 weeks
after constriction of the abdominal aorta. Data are representative of
three or four independent experiments with nearly identical results.
B, cross-sectional areas of cardiac myocytes from 10 sections. Normalized values in WT mice without constriction are
arbitrarily expressed as 1.0. *p < 0.05.
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Expression of Cardiac-specific Genes--
Pressure overload by the
constriction of the abdominal aorta up-regulates fetal-type cardiac
genes and down-regulates SERCA2 gene (27). We therefore examined the
expression of BNP and SERCA2 in the hearts of WT and TG mice at the
early and late phase after constriction of the abdominal aorta. BNP
gene was slightly up-regulated at 2 days after pressure overload and
markedly at 28 days in WT mice (Fig. 4).
In TG mice, although BNP gene was also up-regulated by pressure
overload, the expression levels were quite low compared with those of
WT mice. In WT mice, SERCA2 gene was down-regulated from 2 days by
pressure overload (Fig. 4). The down-regulation of SERCA2 gene by
pressure overload was also attenuated in TG mice.

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Fig. 4.
Expression of cardiac genes. Hearts were
excised from WT and TG mice at 2 and 28 days after constriction of the
abdominal aorta. Expression of cardiac-specific genes were examined by
Northern blot analysis. A, a representative autoradiogram
was shown. Ethidium bromide staining of 18 S RNA was shown
below. B, The band intensities of BNP, SERCA2,
and 18 S RNA were quantified by densitometer. Each histogram represents
the fold of BNP and SERCA2 compared with 18 S RNA from three
independent experiments (mean ± S.E.). *p < 0.05 versus control.
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Activation of STAT3 and ERKs--
Activation of gp130 evokes two
distinct pathways, Janus kinase-STAT pathway and Ras-ERKs
pathway (28). We examined which pathway is important in the pressure
overload-induced cardiac hypertrophy (Fig.
5). Pressure overload activated both
STAT3 and ERKs in the heart of WT mice. However, in TG mice, activation of STAT3 was barely detectable. In contrast, there was no difference in
activation of ERKs between TG and WT mice. These results collectively suggest that pressure overload-induced activation of STAT3, but not of
ERKs, is dependent on gp130 and that STAT3 may play a critical role in
pressure overload-induced cardiac hypertrophy.

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Fig. 5.
Activation of ERKs and STAT3 by pressure
overload. The abdominal aorta was constricted for indicated
periods of time, and the activities of ERKs and STAT3 were measured as
described under "Materials and Methods." Each histogram represents
the fold of controls from three independent experiments (mean ± S.E.). *p < 0.05.
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No Increase in a Number of TUNEL-positive Cardiomyocytes--
It
has been reported that activation of gp130 promotes survival of cardiac
myocytes (17) and that ventricular-restricted gp130 knockout mice
showed marked cardiomyocyte apoptosis and marked ventricular wall
dilatation by pressure overload (18). We therefore examined whether TG
mice showed cardiomyocyte apoptosis during pressure overload. As shown
in Fig. 6, there was no increase in the
number of TUNEL-positive cardiomyocytes in the heart of TG mice at the
basal and by pressure overload compared with those of WT mice.

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Fig. 6.
TUNEL assay of the heart. The abdominal
aorta was constricted for 4 weeks, and cardiomyocyte apoptosis was
examined by TUNEL as described under "Materials and Methods." 500 cardiomyocytes were counted from each side. Each histogram represents
the number of TUNEL-positive cells from five independent experiments
(mean ± S.E.). Pretreatment with micrococcal DNase I (1 mg/ml) to
induce DNA strand breaks was carried out as positive controls.
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 |
DISCUSSION |
It has been reported that gp130 is implicated in regulating cell
growth, differentiation, and cell death in response to external stimuli
in various tissues. In the present study, the TG mice were apparently
healthy with no cardiac abnormalities at basal condition, suggesting
that gp130 is not necessary for the development and the physiological
function of the heart after birth. In contrast, gp130 plays a critical
role in pressure overload-induced cardiac hypertrophy. By constriction
of the abdominal aorta, TG mice showed less increase in the heart
weight/body weight ratio and less changes in expression of BNP and
SERCA2 genes compared with WT mice. We used number 3 line of transgenic
mice, which expressed D.N.gp130 most abundantly. Mice of number 1 and
number 2 lines also showed similar results with mild degree (data not
shown). These results suggest that D.N.gp130
dose-dependently suppresses cardiac hypertrophy.
The intracellular signaling pathways evoked by gp130 activation include
the Janus kinase-induced STAT pathway and the Ras-ERKs pathway (28).
Activated STAT3 has been reported to form a homodimer that subsequently
forms cis-inducing factor complexes and induces hypertrophy of cardiac myocytes (29). It has been reported that STAT3
plays a critical role in generating the hypertrophic signal (30) and
that mice overexpressing STAT3 showed marked cardiac hypertrophy (31).
In the present study, pressure overload activated ERKs and STAT3 in the
heart of WT mice, whereas pressure overload-induced activation of
STAT3, but not of ERKs, was suppressed in TG mice. These results
suggest that pressure overload-induced activation of STAT3, but not of
ERKs, is dependent on gp130 and that STAT3 may play a critical role in
pressure overload-induced cardiac hypertrophy.
Hirota et al. (18) have reported that ventricular-restricted
knockout mice of gp130 showed dilatation of left ventricular wall and
died within a week by constriction of transverse aorta without
progression of adaptive hypertrophy of left ventricle (18). Although
cardiac hypertrophy was attenuated, there was no sign of heart failure
such as dilatation of left ventricular wall and reduced cardiac
function nor death by constriction of the abdominal aorta in the
D.N.gp130 TG mice. There are several possibilities for this
discrepancy. First, the methods applied to produce the pressure
overload were different. Transverse aortic constriction, applied in
their experiment, induces stronger pressure overload than constriction
of the abdominal aorta. It is possible that if the pressure overload is
too strong, heart failure, but not adaptive cardiac hypertrophy, would
be induced. To determine the role of gp130 in pressure overload-induced
cardiac hypertrophy, we used a mild pressure overload model of
abdominal aortic constriction. Second, they used gp130 null mice,
whereas we used D.N.gp130-overexpressing mice. Because pressure
overload-induced activation of STAT3 was abolished in the heart of
D.N.gp130 TG mice, endogenous gp130 activity is thought to be strongly
suppressed by overexpression of D.N.gp130. However, there is a
possibility that the two mice have different gp130 activity. The
transgene lacks box3 but has box1 and box2 regions of the cytoplasmic
domain of gp130. Although box3 is indispensable for the Janus
kinase-induced activation of STAT3 (26), there is a possibility that
box1 and box2 have some biological functions. There is a possibility
that the two mice have different ERK activities, which they did not
examine (18). It has been reported that the ERK signaling pathway is important for the gp130-dependent cell survival of cardiac
myocytes (17) and that the tyrosine-containing motif,
Tyr116-Xaa-Xaa-Val, of the cytoplasmic domain of
gp130 is indispensable for the activation of SHP-2, a key molecule for
the gp130-mediated signaling pathway leading to ERKs (26, 32). Although
the transgene lacks this motif, pressure overload induced activation of
ERKs also in TG mice. These results suggest that some other factors such as angiotensin II and endothelin-1, which exert their effects through G protein-coupled receptors, play a predominant role in the
activation of ERKs and cardiomyocyte survival in the TG mice. Although
it remains to be determined how pressure overload induces production of
the IL-6 family of cytokines in the heart, the present study suggests
that gp130 plays a critical role in pressure overload-induced cardiac
hypertrophy possibly through the STAT3 pathway.
The abbreviations used are:
IL, interleukin;
TG, transgenic;
D.N., dominant negative;
MHC, myosin heavy
chain;
WT, wild type;
LVEDD, left ventricular end-diastolic internal
diameter;
LVESD, left ventricular end-systolic internal diameter;
BNP, brain natriuretic factor;
SERCA, sarcoplasmic reticulum
Ca2+ ATPase;
STAT, signal transducers and activators of
transcription;
ERK(s), extracellular signal-regulated kinase(s);
TUNEL, terminal deoxynucleotidyl transferase assay.
1.
|
Levy, D.,
Garrison, R. J.,
Savage, D. D.,
Kannel, W. B.,
and Castelli, W. P.
(1990)
N. Engl. J. Med.
322,
1561-1566[Abstract]
|
2.
|
Komuro, I.,
Katoh, Y.,
Kaida, T.,
Shibazaki, Y.,
Kurabayashi, M.,
Takaku, F.,
and Yazaki, Y.
(1991)
J. Biol. Chem.
266,
1265-1268[Abstract/Free Full Text]
|
3.
|
Simpson, P.
(1985)
Circ. Res.
56,
884-894[Abstract]
|
4.
|
Baker, K. M.,
and Aceto, J. F.
(1990)
Am. J. Physiol.
259,
H610-H618[Abstract/Free Full Text]
|
5.
|
Shubeita, H. E.,
McDonough, P. M.,
Harris, A. N.,
Knowlton, K. U.,
Glembotski, C. C.,
Brown, J. H.,
and Chien, K. R.
(1990)
J. Biol. Chem.
265,
20555-20562[Abstract/Free Full Text]
|
6.
|
Yamamori, T.,
Fukada, K.,
Aebersold, R.,
Korsching, S.,
Fann, M. J.,
and Patterson, P. H.
(1989)
Science
246,
1412-1416[Medline]
[Order article via Infotrieve]
|
7.
|
Yamazaki, T.,
Komuro, I.,
Kudoh, S.,
Zou, Y.,
Shiojima, I.,
Hiroi, Y.,
Mizuno, T.,
Maemura, K.,
Kurihara, H.,
Aikawa, R.,
Takano, H.,
and Yazaki, Y.
(1996)
J. Biol. Chem.
271,
3221-3228[Abstract/Free Full Text]
|
8.
|
Yamazaki, T.,
Komuro, I.,
Kudoh, S.,
Zou, Y.,
Shiojima, I.,
Mizuno, T.,
Takano, H.,
Hiroi, Y.,
Ueki, K.,
and Tobe, K.
(1995)
Circ. Res.
77,
258-265[Abstract/Free Full Text]
|
9.
|
Sadoshima, J.,
Xu, Y.,
Slayter, H. S.,
and Izumo, S.
(1993)
Cell
75,
977-984[Medline]
[Order article via Infotrieve]
|
10.
|
Pennica, D.,
King, K. L.,
Shaw, K. J.,
Luis, E.,
Rullamas, J.,
Luoh, S. M.,
Darbonne, W. C.,
Knutzon, D. S.,
Yen, R.,
Chien, K. R.,
Baker, J. B.,
and Wood, W. I.
(1995)
Proc. Natl. Acad. Sci. U. S. A.
92,
1142-1146[Abstract]
|
11.
|
Hilton, D. J.
(1992)
Trends Biochem. Sci.
17,
72-76[CrossRef][Medline]
[Order article via Infotrieve]
|
12.
|
Taga, T.,
and Kishimoto, T.
(1997)
Annu. Rev. Immunol.
15,
797-819[CrossRef][Medline]
[Order article via Infotrieve]
|
13.
|
Hirano, T.
(1998)
Int. Rev. Immunol.
16,
249-284[Medline]
[Order article via Infotrieve]
|
14.
|
Wollert, K. C.,
Taga, T.,
Saito, M.,
Narazaki, M.,
Kishimoto, T.,
Glembotski, C. C.,
Vernallis, A. B.,
Heath, J. K.,
Pennica, D.,
Wood, W. I.,
and Chien, K. R.
(1996)
J. Biol. Chem.
271,
9535-9545[Abstract/Free Full Text]
|
15.
|
Hirota, H.,
Yoshida, K.,
Kishimoto, T.,
and Taga, T.
(1995)
Proc. Natl. Acad. Sci. U. S. A.
92,
4862-4866[Abstract]
|
16.
|
Yoshida, K.,
Taga, T.,
Saito, M.,
Suematsu, S.,
Kumanogoh, A.,
Tanaka, T.,
Fujiwara, H.,
Hirata, M.,
Yamagami, T.,
Nakahata, T.,
Hirabayashi, T.,
Yoneda, Y.,
Tanaka, K.,
Wang, W. Z.,
Mori, C.,
Shiota, K.,
Yoshida, N.,
and Kishimoto, T.
(1996)
Proc. Natl. Acad. Sci. U. S. A.
93,
407-411[Abstract/Free Full Text]
|
17.
|
Sheng, Z.,
Knowlton, K.,
Chen, J.,
Hoshijima, M.,
Brown, J. H.,
and Chien, K. R.
(1997)
J. Biol. Chem.
272,
5783-5791[Abstract/Free Full Text]
|
18.
|
Hirota, H.,
Chen, J.,
Betz, U. A.,
Rajewsky, K.,
Gu, Y.,
Ross, J. J.,
Muller, W.,
and Chien, K. R.
(1999)
Cell
97,
189-198[Medline]
[Order article via Infotrieve]
|
19.
|
Kumanogoh, A.,
Marukawa, S.,
Kumanogoh, T.,
Hirota, H.,
Yoshida, K.,
Lee, I. S.,
Yasui, T.,
Yoshida, K.,
Taga, T.,
and Kishimoto, T.
(1997)
Proc. Natl. Acad. Sci. U. S. A.
94,
2478-2482[Abstract/Free Full Text]
|
20.
|
Ng, W. A.,
Grupp, I. L.,
Subramaniam, A.,
and Robbins, J.
(1991)
Circ. Res.
68,
1742-1750[Abstract]
|
21.
|
Komuro, I.,
Kurabayashi, M.,
Takaku, F.,
and Yazaki, Y.
(1988)
Circ. Res.
62,
1075-1079[Abstract]
|
22.
|
Harada, K.,
Komuro, I.,
Shiojima, I.,
Hayashi, D.,
Kudoh, S.,
Mizuno, T.,
Kijima, K.,
Matsubara, H.,
Sugaya, T.,
Murakami, K.,
and Yazaki, Y.
(1998)
Circulation
97,
1952-1959[Abstract/Free Full Text]
|
23.
|
Kojima, M.,
Shiojima, I.,
Yamazaki, T.,
Komuro, I.,
Zou, Z.,
Wang, Y.,
Mizuno, T.,
Ueki, K.,
Tobe, K.,
and Kadowaki, T.
(1994)
Circulation
89,
2204-2211[Abstract]
|
24.
|
Sahn, D. J.,
DeMaria, A.,
Kisslo, J.,
and Weyman, A.
(1978)
Circulation
58,
1072-1083[Abstract]
|
25.
|
Yamazaki, T.,
Tobe, K.,
Hoh, E.,
Maemura, K.,
Kaida, T.,
Komuro, I.,
Tamemoto, H.,
Kadowaki, T.,
Nagai, R.,
and Yazaki, Y.
(1993)
J. Biol. Chem.
268,
12069-12076[Abstract/Free Full Text]
|
26.
|
Ernst, M.,
Novak, U.,
Nicholson, S. E.,
Layton, J. E.,
and Dunn, A. R.
(1999)
J. Biol. Chem.
274,
9729-9737[Abstract/Free Full Text]
|
27.
|
Komuro, I.,
and Yazaki, Y.
(1993)
Annu. Rev. Physiol.
55,
55-75[CrossRef][Medline]
[Order article via Infotrieve]
|
28.
|
Ernst, M.,
Oates, A.,
and Dunn, A. R.
(1996)
J. Biol. Chem.
271,
30136-30143[Abstract/Free Full Text]
|
29.
|
Kodama, H.,
Fukuda, K.,
Pan, J.,
Makino, S.,
Baba, A.,
Hori, S.,
and Ogawa, S.
(1997)
Circ. Res.
81,
656-663[Abstract/Free Full Text]
|
30.
|
Kunisada, K.,
Tone, E.,
Fujio, Y.,
Matsui, H.,
Yamauchi, T. K.,
and Kishimoto, T.
(1998)
Circulation
98,
346-352[Abstract/Free Full Text]
|
31.
|
Kunisada, K.,
Negoro, S.,
Tone, E.,
Funamoto, M.,
Osugi, T.,
Yamada, S.,
Okabe, M.,
Kishimoto, T.,
and Yamauchi, T. K.
(2000)
Proc. Natl. Acad. Sci. U. S. A.
97,
315-319[Abstract/Free Full Text]
|
32.
|
Takahashi, T. M.,
Yoshida, Y.,
Fukada, T.,
Ohtani, T.,
Yamanaka, Y.,
Nishida, K.,
Nakajima, K.,
Hibi, M.,
and Hirano, T.
(1998)
Mol. Cell. Biol.
18,
4109-4117[Abstract/Free Full Text]
|