| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
ARTICLES |
Departments of Pharmacology and Toxicology (M.M., N.H., M.T., H.R.) and Physiology (O.V.), Biocenter Oulu, University of Oulu, FIN-90014 University of Oulu, Finland; and Clinical Research Institute of Montreal (P.P., M.N.), University of Montréal, Montréal H2W 1R7, Canada
Address all correspondence and requests for reprints to: Heikki Ruskoaho, M.D., Ph.D., Department of Pharmacology and Toxicology, Faculty of Medicine, University of Oulu, P.O. Box 5000, FIN-90014 University of Oulu, Finland. E-mail: heikki.ruskoaho{at}oulu.fi
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
-actin as well as the
induction of atrial natriuretic peptide (ANP) and B-type natriuretic
peptide (BNP) gene expression in the ventricle (2, 3, 4, 5). In
normal adult heart, BNP is produced by both atria and ventricles
(6, 7). The induction of BNP gene expression is one of the
earliest cardiac myocyte-specific markers of hemodynamic overload;
indeed, cardiac BNP mRNA and protein are increased rapidly at the onset
of hemodynamic stress well before the development of left ventricular
hypertrophy (8, 9, 10).
Because it is not possible to model in vitro the complex
hemodynamic and neurohumoral stimuli that are associated with
hemodynamic overload in vivo, little is known with respect
to nuclear signaling involved in initiating or maintaining the response
to the hemodynamic stress in the adult heart (11, 12, 13, 14).
Ayogi and Izumo (15) demonstrated that injected
c-fos promoter is regulated by pressure overload stimulus in
the perfused rat heart preparation and that the pressure response
element coincides with a serum response element. In addition, an
activator protein-1 (AP-1)-like element has been reported to be
responsible for conferring pressure overload responsiveness to the ANP
promoter (16, 17), although this result is controversial
(18, 19). Previous study has also demonstrated the
importance of upstream stimulatory factor 1 binding to an E-box motif
as an important transcription factor in the regulation of hemodynamic
mediated changes in -MHC gene (20). Furthermore, recent
work has suggested that GATA binding sites seem to be required for
activation of ß-MHC (19) and angiotensin II type 1a
(AT1a) receptor (21) expression in
response to pressure-overload hypertrophy in rats. Whether these
changes are coupled to increased blood pressure or ventricular
hypertrophy is not clearly established.
In the present study, we used the approach of DNA injection (22) into the myocardium to identify a cis element within the BNP promoter that mediates rapid response to a hemodynamic stress stimulus produced by bilateral nephrectomy in the intact adult rat heart. Nephrectomy is a complex hemodynamic stimulus, producing volume and pressure overload as well as neurohumoral activation, including tissue renin-angiotensin system (23, 24, 25). The BNP promoter is particularly well suited for these studies because, unlike c-fos (which is also expressed in nonmuscular cells), BNP is a cardiac myocyte-specific marker (6) that is rapidly induced by hemodynamic stress at the transcriptional level (26). Our results show that the GATA elements are necessary and sufficient to confer hemodynamic stress responsiveness of the BNP gene.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
In vivo gene transfer
The 10- to 12-wk-old male Sprague Dawley rats were anesthetized
with 250 µg/kg medetomidine hydrochloride, and 50 mg/kg ketamine
hydrochloride ip. Plasmids in 0.9% NaCl were injected directly into
the left ventricular free wall close to the apex under direct
visualization by using a 100-µl Hamilton precision syringe
(22). The injection vol was 100 µl, based on the
previously published studies in various rat models (15, 22). The heart was then repositioned in the chest, the rats were
briefly hyperventilated, and the incision was closed. Surgical and
postoperative mortality was approximately 5% in all groups. At the
time of death, the injection site appeared to be normal cardiac tissue
with no fibrotic or necrotic signs. At the precise site of injection, a
small light spot could be noted. To ensure reliable measurements of
luciferase activities of the short deletion and mutation promoter
constructs 1 wk after the injection, plasmid DNA consisted of 50–100
µg of the reported construct driven by the promoter of interest and
100 µg of a ß-galactosidase expression vector
(pSVß-gal, Promega Corp., San Diego, CA) to
correct for variation in transfection efficiency. Because of the
concern that there might be competition among promoters for
transcription factors (22), these dosages were chosen
based on preliminary experiments showing that the amount of luciferase
activity in cardiac homogenates increases dose-dependently after
injection of a constant 100 µl vol containing 10–100 µg of
plasmids. For p-2200BNPluc construct, luciferase activities were as
follows: 10 µg, 12.5 ± 2.5 light units; 25 µg, 21.7 ±
3.8 light units; and 100 µg, 76.7 ± 7.6 light units (n =
8–9). Furthermore, in the animals subjected to nephrectomy and
injected with 10 µg p-2200BNPluc, the fold increase of luciferase
activity was similar to that after injection of 100 µg of the same
plasmid (data not shown). Samples in which luciferase activity was less
than 1.0 (background averaged 0.1–0.2), the injection failed, and
values were not reported. The variation of the transfection efficiency
estimated by different level of activities was less than 10%. The
experimental design was approved by the Animal Use and Care Committee
of the University of Oulu.
Induction of hemodynamic stress
On the sixth day after injection, the rats were anesthetized
with fentanyl citrate (0.26 mg/kg), fluanisone (8.25 mg/kg), midazolam
(4.1 mg/kg ip), and subjected to either bilateral nephrectomy or sham
operation. Nephrectomy was accomplished by tying a 3-0 silk suture
securely around the porta renalis containing the artery, the vein, and
the ureter. In sham-operated animals, the kidneys were exposed, but no
ligature was placed.
Blood pressure monitoring
Rats were anesthetized with 250 µg/kg medetomidine
hydrochloride and 50 mg/kg ketamine hydrochloride ip, and instrumented
with a catheter in the descending aorta coupled with a sensor and
transmitter (TA11PA-C40; Data Sciences, St. Paul, MN) for telemetric
monitoring of blood pressure. Blood pressure and heart rate were
measured every 10 min and averaged every 6 h. On the sixth day
after implantation, the rats were subjected to either bilateral
nephrectomy or sham operation, as described above.
Tissue preparation
Rats were decapitated 7 d after injection of plasmid
constructs (6, 12, or 26–28 h post nephrectomy). The basal one-third
of the left ventricle was homogenized in 1 ml homogenization buffer (20
mM Tris-acetate, pH 7.5, 5 mM EDTA, 20
mM KCl, 2 mM Mg-acetate, and 0.5 mM
dithiothreitol) with an Ultra-Turrax T25-tissue homogenizer (Janke und
Kunkel, Stauffen, Germany). Homogenates were centrifuged at 6000
x g for 10 min at +4 C, and the supernatants were divided
into two samples. Luciferase samples were kept in room temperature, and
samples for ß-galactosidase assay were frozen at -20 C
for subsequent analysis.
Reporter gene assays
Both luciferase and ß-galactosidase activities were
assayed in the same supernatant of left ventricular homogenates. Using
a luminometer (model RS, Labsystems Luminoskan, Helsinki, Finland),
luciferase activity was measured in 20-µl aliquots (2.0–2.5% of the
total volume) of the supernatant, using commercially available
Luciferase Assay System (Promega Corp.).
ß-Galactosidase activity was assayed in 20-µl samples of
the supernatant, using commercially available Luminescent
ß-galactosidase Detection Kit II (CLONTECH Laboratories, Inc., Palo Alto, CA).
Gel mobility shift assays
Nuclear extracts were prepared from ventricular tissue of
sham-operated or nephrectomized rats as described by Deryckere and
Gannon (28). Protein concentration was determined using
the Bio-Rad Laboratories, Inc. Protein Assay, and aliquots
were frozen at -70 degree C until use. Double-stranded synthetic
oligonucleotides containing GATA (5'-TGTGTCT
GATAAATCAGAGATAACCCCACC-3') motifs of the rat BNP
promoter were labeled with
[-32P]dCTP. Binding reactions contained 30
µg crude nuclear extract and 2 µg
poly-(dI-dC).(dI-dC) in a buffer containing 10
mM HEPES (pH 7.9), 1 mM
MgCl2, 50 mM KCl, 1
mM dithiothreitol, 0.1 mM
EDTA, 10% glycerol, 0.025% NP-40, 0.25 mM PMSF,
and 1 µmM of each aprotinin, leupeptin and pepstatin, and when
appropriate, various molar excesses of unlabeled double-stranded
oligonucleotides. Reactions were carried out at room temperature for 20
min, and protein-DNA complexes were separated by electrophoresis on 5%
polyacrylamide gel in 0.5x Tris-borate-EDTA buffer at +4 C. Nonlabeled
double-stranded oligonucleotides corresponding to GATA binding sites of
the BNP promoter and a GATA consensus sequence (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) were used as
specific competitor DNAs. Nonspecific competitor DNAs included a
double-stranded oligonucleotide carrying the mutated binding site for
GATA4 (5'-TGTGTCTGGTAAATCAGAGGTAACCCCACC-3') and
Oct-1 as nonrelated DNA. To confirm that each reaction contained the
equal amount of nuclear protein, the labeled Oct-1 oligonucleotide
probe was used. For supershift experiments, 1 µg goat polyclonal
GATA4, GATA5, or GATA6 affinity-purified IgG (Santa Cruz Biotechnology, Inc.) were used.
Histology
Hearts were washed and stopped in diastole by perfusion with PBS
containing 50 mM KCl and then fixed by perfusion with Bouin
solution (29). Specimens were embedded in paraffin and cut
into 5-µm sections for immunostaining with rabbit polyclonal
anti-GATA4 antibody (streptavidin/biotin immunoperoxidase method) as
described previously (29).
Isolation and analysis of RNA
Total RNA was isolated from left ventricles by the guanidine
thiocyanate-CsCl method (30). For the RNA Northern blot
analysis, 20-µg samples of the RNA from the left ventricles were
separated by electrophoresis on agarose gel and transferred to nylon
membranes. A 390-bp fragment of rat BNP cDNA (31), cDNA
probes for rat GATA4 (1417 bp) and GATA6 (1175 bp) made by the RT-PCR
technique, and a full-length cDNA probe complementary to rat
glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) were labeled
(32), and the membranes were hybridized and washed as
described previously (10).
BNP RIA
The BNP RIA was performed as previously described
(10). The sensitivity of the assay was 2 fmol/tube, and
50% displacements of the respective standard curve occurred at 25
fmol/tube. The intra- and interassay variations were less than 10% and
15%, respectively.
Statistics
Values are mean ± SEM. For the comparison of
statistical significance between two groups, t test was
used. A P value less than 0.05 was considered statistically
significant.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
). This -114 bp fragment contains two
major cis-acting elements, an AP-1-like element and a GATA
motif (27, 33). Deletion of the AP-1-like element
(construct -100 bp to +75 bp) decreased promoter activity over
10-fold, and deletion of the distal GATA motifs (construct -76 bp to
+75 bp) reduced promoter activity further by 4-fold (1.7% of -2200 bp
construct promoter activity, Fig. 1). Mutation of the AP-1-like element
reduced BNP expression to 32%, and mutation of both GATA motifs at
-90 bp decreased BNP expression to 57% of BNP-114 bp construct. The
contribution of the GATA box at -90 bp to the ventricular-specific
activity of the BNP promoter was further confirmed by the fact that a
38-bp oligonucleotide containing this GATA motif displayed
transcriptional activity (2.1% of BNP-2200luc construct) when inserted
upstream of the minimal BNP -60 promoter (Fig. 1). The mean activity
of shortest construct (at light units) was about 10-fold higher than
that of the promoterless background vector and was within the linear
range of the luminometer (data not shown).
|
). Also, endogenous ANP mRNA
levels were 2.2-fold higher in nephrectomized rats than in
sham-operated rats (P < 0.001, data not shown). The
increase in ventricular BNP mRNA levels was accompanied with a
significant increase in immunoreactive BNP levels in the left ventricle
(15.9 ± 1.3 fmol/mg vs. 8.9 ± 0.8 fmol/mg,
P < 0.001), and there was a tendency for plasma
immunoreactive BNP levels to be higher in nephrectomized rats than in
sham-operated rats (24.3 ± 3.7 fM
vs. 13.2 ± 2.6 fM, NS). The
index of ventricular hypertrophy, ventricular weight to body weight
ratio, was equal in sham-operated and nephrectomized rats (3.73 ±
0.09 mg/g, n = 6 vs. 3.69 ± 0.08 mg/g, n
= 6).
|
). The
expression level of pSVß-galactosidase was similar in the
left ventricles of sham-operated and nephrectomized animals (7.1
± 0.5 U/ml vs. 8.3 ± 0.5 U/ml), showing that
increases in the expression of p-2200BNPluc are attributable to
augmentation of the transcriptional activity of the BNP promoter
sequences. The effect was sequence-specific, as illustrated by the
absence of hemodynamic stress-stimulated increases in transcription
directed by the proximal ANP promoter (p-135ANPluc, data not
shown).
|
GATA motif is essential for hemodynamic stress induction of the BNP
gene
To identify which DNA sequences mediate hemodynamic
overload-stimulated increases in BNP transcription, the ability of
nested 5' deletions to respond to nephrectomy was evaluated. The 114-bp
fragment of rat BNP promoter was shown to be sufficient for inducible
expression in hemodynamic stress, because deletion to -114 bp resulted
in no significant decrease in inducibility in response to nephrectomy
(Fig. 3
). In fact, p-114BNPluc showed the highest level of inducibility
(7.2-fold) among the constructs tested. Hemodynamic stress
significantly increased (over 5-fold) the expression of the -114 bp
constructs containing mutation or deletion of the AP-1-like element. In
contrast, mutation of both GATA motifs at -90 bp, as well as deletion
of the -90 GATA-sites (construct -76 bp to +75 bp), resulted in
almost complete loss of inducible expression (Fig. 3). The contribution
of the GATA box at -90 bp to the inducible expression of the BNP
promoter was confirmed by using a 38-bp oligonucleotide containing this
site, inserted upstream of the minimal BNP -60 promoter. Hemodynamic
stress significantly (3.5-fold, P = 0.013) increased
the expression of this construct similarly to that exhibited by the
longest -2200 bp BNP-luciferase construct (Fig. 3).
Hemodynamic stress up-regulates left ventricular GATA binding
activity
Gel mobility shift assays were used to analyze the
DNA-binding activities that interact with the GATA motifs of the BNP
promoter (Fig. 4). When left ventricular
nuclear extracts were incubated with the 30-bp double-stranded
oligonucleotide containing the -90 BNP GATA sites, a specific
complex was obtained (Fig. 4A, lane 2). To determine the specificity of
ventricular GATA binding activity, competition analyses were performed.
The formation of complexes with the rBNP-90 GATA probe was effectively
inhibited by the unlabeled self (Fig. 4A, lanes 3 and 4) and GATA
consensus DNA (Fig. 4A, lanes 7 and 8), indicating that the DNA-protein
complex was the result of a specific interaction. The binding was
unaffected by an excess of oligonucleotides corresponding to the
nonrelated competitor DNA Oct-1 (Fig. 4A, lane 5) or the mutated BNP
GATA site (Fig. 4A, lane 6). To confirm further that the complex bound
to the BNP GATA site contains GATA proteins, supershift assays were
carried out by using GATA4 (Fig. 4B, lanes 2 and 6), GATA5 (Fig. 4B, lanes 3 and 7), and GATA6 (Fig. 4B, lanes 4 and 8) antibodies.
Experiments in which ventricular extracts from sham-operated or
nephrectomized rats were used clearly showed antibody-induced
supershift of GATA4 but not GATA5 or GATA6 complexes. Bilateral
nephrectomy for 26–28 h produced a 1.93-fold increase
(P < 0.01) in BNP GATA binding activity (Fig. 4C). At
6 and 12 h post nephrectomy, there was a tendency for BNP GATA
binding activity to increase, but these changes (1.33- and 1.35-fold,
respectively, data not shown) were not statistically significant.
|
). There was no observable difference
in GATA4 immunoreactivity between the ventricles of the nephrectomized
and control rats as assessed by immunohistochemical staining (data not
shown). |
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The typical response to hemodynamic stress is the rapid activation of the immediate early gene c-fos (3, 36). This results in the accumulation of c-fos protein, which interacts with Jun family members to constitute AP-1 activity (37). The AP-1-like element is highly conserved across species in the BNP gene (27). Mutation of the AP-1-like element decreased BNP promoter activity; and when the AP-1-like site was deleted, basal promoter activity was reduced over 10-fold. In vitro results with nested 5' deletions at bp -100 have previously shown about 4-fold decrease in BNP promoter activity (27); thus, both in vitro and in vivo results indicate a role for this site in basal transcription. However, mutation or deletion of the AP-1-like motif had no significant effect on the hemodynamic stress-induced increase in BNP promoter activity. This result is different from those obtained with ANP promoter by von Harsdorf et al. (17), who studied the effects of aortic banding for 7 d in dogs injected with rat ANP reporter constructs. Site-specific mutation of the ANP AP-1-like element abrogated the response to pressure overload, showing that this site is necessary for the induction of the ANP gene (17). It has also been reported that the AP-1 site and CRE-site were sufficient to induce ANP promoter activity after acute elevation of cardiac wall stress (16). Although several factors may account for the divergent results, the data suggest that different transcriptional factors may regulate the induction of ANP and BNP genes in response to hemodynamic overload in the intact adult heart. The response of the two gene products to mechanical stretch is also different. Both in vitro and in vivo, ANP gene expression is unresponsive to short-term wall stretch, whereas BNP is rapidly activated (8, 9, 38).
A major finding of the present study was that the regulatory elements
conferring induction of the BNP gene during hemodynamic overload map to
the GATA motif. GATA4, -5, and -6, which are expressed in the
developing heart (27, 39, 40, 41), play important roles in the
transcriptional regulation of cardiac genes and heart morphogenesis
(42, 43). Induction of GATA4 precedes expression of
cardiac marker genes and appearance of beating cells (34).
Moreover, inhibition of GATA4 expression with GATA4 antisense construct
specifically blocked cardiac muscle gene expression (34, 35). In vitro, transfection studies have established
that GATA4 is a potent activator of BNP (27), ANP
(44, 45), cardiac troponin C (46), -MHC
(47), cardiac troponin I (48), and m2
muscarinic acetylcholine receptor (49). Mutation of GATA
elements in -MHC and myosin light chain decreases their
transcriptional activities in vivo (47, 50). In
agreement with these reports, mutation of GATA motifs located in the
BNP promoter between positions -95 bp and -85 bp decreased promoter
activity about 40%, showing that the GATA motif has a slight effect on
basal ventricular-specific BNP gene expression. Furthermore, in
vivo injection experiments showed that after mutating or deleting
both proximal GATA-sites, the inducibility to nephrectomy was
completely abolished. Finally, the GATA sequences were sufficient
to convert a neutral promoter to one which is hemodynamic
stress-responsive.
Recently, it has been shown that the GATA sequences are required for the induction of ß-MHC (19) and AT1a receptor expression (21) in response to pressure overload hypertrophy. In the latter study, the authors detected reproducible binding on AT1a GATA sites in extracts of hypertrophied (but not control) hearts (21), suggesting that GATA binding activity is enhanced in the hypertrophied myocardium. The present study extends those findings in demonstrating that left ventricular BNP GATA4, but not GATA5 and GATA6 binding, is activated at an early stage of hemodynamic overload, well before the development of left ventricular hypertrophy. Furthermore, no significant differences in the levels of GATA4 and GATA6 mRNA or GATA4 immunoreactivity were observed, suggesting that the increase in GATA binding activity may involve posttranslational mechanisms.
The present in vivo transfection experiments demonstrate
that the rat GATA motif transduces the hemodynamic stress stimulus
26–28 h post nephrectomy. However, it is possible that other
cis elements may also contribute to hemodynamic stress
response. In vitro, other transcription factors, such as
nuclear factor-
B (NF-B) (51) and nuclear factor of
activated T cells 3 (NF-AT3) (52), are also important for
BNP gene expression, and recent work has suggested that ANP expression
may be regulated by cooperative interaction of cis elements
with GATA4 and Csx/Nkx2.5 (45). The potential interaction
of these transcription factors with GATA4, in promoting the changes in
cardiac gene expression in response to hemodynamic stress in the intact
rat heart, remains to be studied.
In conclusion, our study demonstrates that acute hemodynamic stress produced by bilateral nephrectomy increases BNP reporter expression through a GATA4-dependent pathway. These results identify, for the first time, a tissue-specific pathway that is involved in the adaptive response of the adult heart to acute hemodynamic overload. Whether GATA4 mediates the cardiac response to other stimuli that induce both BNP gene expression and hypertrophy (such as angiotensin II, endothelin-1, or noradrenaline) remains to be tested, especially because bilateral nephrectomy induces neurohumoral activation (23, 24, 25).
|
Acknowledgments |
|---|
|
Footnotes |
|---|
Abbreviations: ANP, Atrial natriuretic peptide; AP-1,
activator protein-1; BNP, B-type natriuretic peptide; GAPDH,
glyceraldehyde-3-phosphate-dehydrogenase; MHC, myosin heavy chain;
NF-AT3, nuclear factor of activated T cells 3; NF-B, nuclear factor
B.
Received April 2, 2001.
Accepted for publication July 12, 2001.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
-myosin heavy chain gene transcription by a contractile-responsive
E-box binding protein. J Mol Cell Cardiol 30:87–95[CrossRef][Medline]
-myosin heavy-chain gene. Mol Cell Biol 14:4947–4957This article has been cited by other articles:
 |
|
 |
|
  A. M. Richards Natriuretic Peptides: Update on Peptide Release, Bioactivity, and Clinical Use Hypertension, July 1, 2007; 50(1): 25 - 30. [Full Text] [PDF]
|
|
|
|
 |
|
 K.-i. Watanabe, M. Ma, K.-i. Hirabayashi, N. Gurusamy, P. T. Veeraveedu, P. Prakash, S. Zhang, A. J. Muslin, M. Kodama, and Y. Aizawa Swimming stress in DN 14-3-3 mice triggers maladaptive cardiac remodeling: role of p38 MAPK Am J Physiol Heart Circ Physiol, March 1, 2007; 292(3): H1269 - H1277. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. G. Gardner, S. Chen, D. J. Glenn, and C. L. Grigsby Molecular Biology of the Natriuretic Peptide System: Implications for Physiology and Hypertension Hypertension, March 1, 2007; 49(3): 419 - 426. [Full Text] [PDF]
|
|
|
|
 |
|
 J. Wang, P. Paradis, A. Aries, H. Komati, C. Lefebvre, H. Wang, and M. Nemer Convergence of Protein Kinase C and JAK-STAT Signaling on Transcription Factor GATA-4 Mol. Cell. Biol., November 15, 2005; 25(22): 9829 - 9844. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Pikkarainen, H. Tokola, R. Kerkela, and H. Ruskoaho GATA transcription factors in the developing and adult heart Cardiovasc Res, August 1, 2004; 63(2): 196 - 207. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. Tenhunen, B. Sarman, R. Kerkela, I. Szokodi, L. Papp, M. Toth, and H. Ruskoaho Mitogen-activated Protein Kinases p38 and ERK 1/2 Mediate the Wall Stress-induced Activation of GATA-4 Binding in Adult Heart J. Biol. Chem., June 4, 2004; 279(23): 24852 - 24860. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Aries, P. Paradis, C. Lefebvre, R. J. Schwartz, and M. Nemer Essential role of GATA-4 in cell survival and drug-induced cardiotoxicity PNAS, May 4, 2004; 101(18): 6975 - 6980. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Pikkarainen, H. Tokola, T. Majalahti-Palviainen, R. Kerkela, N. Hautala, S. S. Bhalla, F. Charron, M. Nemer, O. Vuolteenaho, and H. Ruskoaho GATA-4 Is a Nuclear Mediator of Mechanical Stretch-activated Hypertrophic Program J. Biol. Chem., June 20, 2003; 278(26): 23807 - 23816. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Akazawa and I. Komuro Roles of Cardiac Transcription Factors in Cardiac Hypertrophy Circ. Res., May 30, 2003; 92(10): 1079 - 1088. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. H. Freitag, M. G. Larson, D. Levy, E. J. Benjamin, T. J. Wang, E. P. Leip, P. W.F. Wilson, and R. S. Vasan Plasma Brain Natriuretic Peptide Levels and Blood Pressure Tracking in the Framingham Heart Study Hypertension, April 1, 2003; 41(4): 978 - 983. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. van Rooij, P. A. Doevendans, C. C. de Theije, F. A. Babiker, J. D. Molkentin, and L. J. De Windt Requirement of Nuclear Factor of Activated T-cells in Calcineurin-mediated Cardiomyocyte Hypertrophy J. Biol. Chem., December 6, 2002; 277(50): 48617 - 48626. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Pan, B. Hinzmann, W. Yan, F. Wu, J. Morser, and Q. Wu Genomic Structures of the Human and Murine Corin Genes and Functional GATA Elements in Their Promoters J. Biol. Chem., October 4, 2002; 277(41): 38390 - 38398. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Suo, N. Hautala, G. Foldes, I. Szokodi, M. Toth, H. Leskinen, P. Uusimaa, O. Vuolteenaho, M. Nemer, and H. Ruskoaho Posttranscriptional Control of BNP Gene Expression in Angiotensin II-Induced Hypertension Hypertension, March 1, 2002; 39(3): 803 - 808. [Abstract] [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK |
