doi: 10.1161/hc2601.090987
(Circulation. 2001;104:102.)
© 2001 American Heart Association, Inc.
Basic Science Reports |
Isoproterenol Activates Extracellular Signal–Regulated Protein Kinases in Cardiomyocytes Through Calcineurin
From the Department of Cardiovascular Science and Medicine, Chiba University Graduate School of Medicine, Chiba (Y.Z., I.K.); Department of Cardiovascular Medicine, University of Tokyo Graduate School of Medicine, Tokyo (A.Y., W.Z., S.K., Y.H., M.S., H.U., T.T., R.N., Y.Y.); Department of Medicine II, Saitama Medical School, Saitama (O.K.); and Tokyo Metropolitan Institute of Medical Science, Tokyo (F.S.), Japan.
Correspondence to Issei Komuro, MD, PhD, Department of Cardiovascular Science and Medicine, Chiba University Graduate School of Medicine, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan. E-mail komuro-tky{at}umin.ac.jp
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Abstract |
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Background—Extracellular signal–regulated kinases (ERKs) and calcineurin have been reported to play important roles in the development of cardiac hypertrophy. We examined here the relation between calcineurin and ERKs in cardiomyocytes.
Methods and Results—Isoproterenol activated ERKs in cultured cardiomyocytes of neonatal rats, and the activation was abolished by chelation of extracellular Ca2+ with EGTA, blockade of L-type Ca2+ channels with nifedipine, or depletion of intracellular Ca2+ stores with thapsigargin. Isoproterenol-induced activation of ERKs was also significantly suppressed by calcineurin inhibitors in cultured cardiomyocytes as well as in the hearts of mice. Isoproterenol failed to activate ERKs in either the cultured cardiomyocytes or the hearts of mice that overexpress the dominant negative mutant of calcineurin. Isoproterenol elevated intracellular Ca2+ levels at both systolic and diastolic phases and dose-dependently activated calcineurin. Inhibition of calcineurin also attenuated isoproterenol-stimulated phosphorylation of Src, Shc, and Raf-1 kinase. The immunocytochemistry revealed that calcineurin was localized in the Z band, and isoproterenol induced translocation of calcineurin and ERKs into the nucleus.
Conclusions—Calcineurin, which is activated by marked elevation of intracellular Ca2+ levels by the Ca2+-induced Ca2+ release mechanism, regulates isoproterenol-induced activation of ERKs in cardiomyocytes.
Key Words: calcium • calcineurin • myocytes • kinases • isoproterenol
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Introduction |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Cardiac hypertrophy is observed in various cardiovascular diseases, including hypertension, valvular diseases, myocardial infarction, and cardiomyopathy. Clinical studies have demonstrated that cardiac hypertrophy is not only a cause of congestive heart failure but also an independent risk factor for myocardial infarction, arrhythmia, and sudden death.1 Therefore, it is even more important to determine the molecular mechanism of the development of cardiac hypertrophy. A variety of stimuli, such as mechanical stress, ischemia, and neurohumoral factors, can induce cardiac hypertrophy.2 3 These growth stimuli activate multiple intracellular signal transduction pathways, which lead to the reprogramming of gene expressions and an increase in protein synthesis in cardiomyocytes.2 3 A number of intracellular signaling molecules, such as protein kinase C (PKC), tyrosine kinases, the mitogen-activated protein kinase (MAPK) family, and the Janus kinase/signal transducers and activators of transcription (JAK/STAT) family, have been reported to play important roles in the development of cardiac hypertrophy.2 3 In particular, extracellular signal–regulated kinases (ERKs) have been extensively investigated and found to play a pivotal role in hypertrophic responses of cardiomyocytes both in vitro4 5 and in vivo,6 7 although activation of ERKs does not always lead to cardiomyocyte hypertrophy.8
Catecholamines not only modulate cardiac functions but also induce hypertrophic responses.9 10 Both in vivo and in vitro studies demonstrate that the ß-adrenergic receptor agonist isoproterenol (ISO) induces expression of proto-oncogenes and cardiac hypertrophy.9 11 12 13 We have reported that ISO activates ERKs through both Gs- and Gi-dependent pathways and induces cardiomyocyte hypertrophy.13 Bogoyevitch et al12 indicated that ISO activates ERKs through Ca2+. We also observed that Ca2+ is involved in norepinephrine-induced activation of ERKs.10 Ca2+ regulates a number of cellular events, such as contraction, fertilization, differentiation, growth, and survival.14 In response to stimuli, many cells increase their cytosolic Ca2+ levels.14 Intracellular Ca2+ usually mediates cellular events through Ca2+-binding proteins. Among them, calmodulin (CaM) is a major Ca2+-binding protein present in all eukaryotic cells.15 Overexpression of CaM has been reported to induce proliferative and hypertrophic growth of cardiomyocytes in transgenic mice.16 Ca2+/CaM activates various functional molecules, including Ca2+/CaM-dependent protein kinases (CaMKs) and phosphatases.17 18 CaMKII is a ubiquitous serine/threonine protein kinase that is involved in diverse functions of cells ranging from contraction, secretion, and synaptic transmission to gene expression.17 CaMKII regulates phenylephrine-induced gene expression of atrial natriuretic peptide in cardiomyocytes.19 Calcineurin is a ubiquitously expressed protein phosphatase that plays a pivotal role in neuronal functions and immunoreactions.18 Recently, calcineurin has attracted great attention as a critical molecule that induces cardiac hypertrophy.20 Overexpression of calcineurin and of its downstream transcription factor nuclear factor of activated T cells (NFAT) 3 induced marked cardiac hypertrophy in transgenic mice, whereas calcineurin inhibitors suppressed phenylephrine- and angiotensin II (Ang II)–induced cardiomyocyte hypertrophy in vitro.20 We examined here how Ca2+ is involved in ISO-induced activation of ERKs in cardiomyocytes.
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Methods |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Cell Culture and Transfection
Cardiomyocytes from ventricles of 1-day-old Wistar rats were cultured as previously described.10 13 A cDNA encoding constitutively active mutants of the calcineurin A subunit (CACnA) was constructed by deleting the autoinhibitory (carboxy terminus) and the CaM-binding domains of CnA through introducing a stop codon after N407 amino acids.21 The dominant negative mutants of CnA (DNCnA) were obtained from CACnA by mutating histidine at position 160, a calcineurin active site, to glutamine.21 The DNCaMKII was constructed by substituting alanine for lysine at position 43 of CaMKII
.22 Amounts of 7.5
µg of DNCnA and CACnA in pMep vector and DNCaMKII in pcDNA3 vector or
other plasmids were transfected into cultured
cardiomyocytes with 2.5 µg of hemagglutinin-tagged ERK2
(HA-ERK2) DNA by the calcium phosphate
method.13
Transgenic Mice
HA-tagged DNCnA was subcloned into the 
-myosin
heavy chain promoter–containing expression
vector.20 The linearized DNA
was injected into pronuclei of eggs from BDF1 mice, which were
transferred into the oviducts of pseudopregnant ICR mice. The transgene
was identified by polymerase chain reaction with transgene-specific
primers.23 Northern blot
analysis using cDNA probe corresponding to N1 to 407 of DNCnA
and Western blot analysis using an anti-HA antibody and an
anti-CnA antibody raised against a carboxy terminus peptide of the
CnAß, respectively, revealed that the DNCnA gene and protein were
specifically and abundantly expressed in the hearts of transgenic mice
(manuscript in preparation). There was no significant difference in the
expression levels of endogenous CnA between the transgenic
and wild-type mice. In addition, we also observed that there are no
significant differences in blood pressure, heart weight,
endocardiography, cardiomyocyte size, and myocardial
fibrosis at control state between the transgenic and wild-type mice
(manuscript in preparation). Twelve-week-old DNCnA transgenic mice and
wild-type littermate mice were used in the present study. All
protocols were approved by the guidelines of the University of
Tokyo.
ERK Activity
The activity of ERKs was measured by use of the
myelin basic protein (MBP)–containing gel as previously
described.13 The activity of
transfected HA-ERK2 was assayed by use of MBP as a substrate after
immunoprecipitation with an anti-HA antibody (Mitsubishi Biochemical
Laboratories) as previously
described.13
Intracellular Ca2+
Levels
Intracellular Ca2+ levels
were measured with the Ca2+
fluorescent dye indo 1 (Dojin Kagaku) as described
previously.24 The ratio of
400-nm fluorescence to 500-nm fluorescence, which was
collected from the myocytes illuminated by 360-nm light, was used as an
indicator for intracellular Ca2+
concentration.24
Calcineurin Activity
The activity of calcineurin was determined with
phosphorylated GST-RII peptide as a substrate as
previously described,25 with
some modifications. We separated CaM-bound calcineurin (active
calcineurin, >100 kDa) from free calcineurin (inactive calcineurin,
<100 kDa) using Ultrafree-MC centrifugal filter units
(Millipore).
Activation of Src, Shc, and Raf-1
Kinase
Src and Shc were immunoprecipitated with anti-Src and
anti-Shc antibodies (Santa Cruz Biotechnology), respectively. The
immune complexes were subjected to SDS-PAGE, and the blotted membranes
(Millipore) were hybridized with an anti-phosphotyrosine antibody
(4G10) (Santa Cruz Biotechnology). Immunoreactivity was detected with
an enhanced chemiluminescence (ECL) reaction system
(Amersham) according to the manufacturer’s
directions. Raf-1 kinase activity was determined by use of a specific
substrate rMAPKK as described
previously.10
Immunocytochemistry
Cardiomyocytes cultured on glass cover slides in
serum-free DMEM for 24 hours were incubated with phalloidin-TRITC,
anti–-actinin, anti-CnAß, or anti–phospho-ERK antibodies (Santa
Cruz Biotechnology), and then with secondary antibodies according to
the manufacturer’s directions.
Statistical Analysis
Statistical comparison was carried out within 3
independent experiments by 1-way ANOVA and Dunnett’s
t test. Values of
P<0.05 were considered
statistically significant.
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Results |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Involvement of ERKs in ISO-Induced Cardiac Hypertrophy
We first determined in the present study whether activation of ERKs is involved in ISO-induced cardiac hypertrophy. Stimulation with ISO (1 µmol/L for 48 hours) induced a significant increase in the cell size and improved the organization of myofibrils (Figure 1B
) in cultured cardiomyocytes compared
with saline incubation
(Figure 1A), whereas addition of ISO together with PD98059
(50 µmol/L), a specific inhibitor of MEK1, to the culture
medium did not induce hypertrophy
(Figure 1C). Injection of ISO (1 mg · kg body
wt-1 · d-1
IP) for 7 days increased left ventricular mass
(Figure 1E) and transverse diameter of cardiac myocytes
(Figure 1H) compared with saline injection in mice
(Figure 1D and 1G). Injection of PD98059 (100 mg · kg body
wt-1 · d-1
IP) significantly suppressed the ISO-induced morphological changes in
the heart
(Figure 1F and 1I). These results suggest that ERKs play an
important role in ISO-induced cardiac
hypertrophy.
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Involvement of Ca2+
in Activation of ERKs in Cardiomyocytes
We next examined whether
Ca2+ is involved in the activation of ERKs
by ISO in cultured cardiomyocytes. ISO (10 µmol/L for 8
minutes) strongly activated ERKs, as reported
before.13 The activation of
ERKs was completely suppressed by pretreatment with EGTA (5 mmol/L
for 10 minutes), an extracellular Ca2+
chelator, or nifedipine (1 µmol/L for 30 minutes), an
L-type Ca2+ channel antagonist
(Figure 2A), suggesting that the
Ca2+ influx from extracellular space through
L-type Ca2+ channels is required for
ISO-induced activation of ERKs. To elucidate whether
Ca2+-induced Ca2+
release (CICR) is involved in the activation of ERKs, intracellular
Ca2+ stores were depleted by pretreatment
with thapsigargin (2 µmol/L for 24 hours), an inhibitor
of sarcoplasmic reticulum Ca2+-ATPase.
Thapsigargin significantly suppressed ISO-induced activation of ERKs
(Figure 2A). Pretreatment of cardiomyocytes with
EGTA, nifedipine, or thapsigargin alone did not affect
basal activity of ERKs
(Figure 2B).
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Roles of
Ca2+-Binding Proteins in Activation of
ERKs
We further examined which
Ca2+-binding proteins are involved in
ISO-induced activation of ERKs in cardiomyocytes. The CaM
inhibitor W7 strongly inhibited ISO-evoked ERK activation
(Figure 3A). Cyclosporin A (CsA), a highly specific
calcineurin inhibitor, also abolished the activation of
ERKs, whereas the CaMKII inhibitor KN93 had no effect
(Figure 3A). FK506, another calcineurin
inhibitor, showed the same inhibitory effects
on ISO-induced activation of ERKs (data not shown). We also examined
the role of calcineurin in ERK activation induced by other ligands. CsA
had marginal effects on phenylephrine- and Ang II–induced
activation of ERKs in cardiomyocytes
(Figure 3B), suggesting that the role of calcineurin in ERK
activation is different among ligands.
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To confirm the pharmacological results, we used the genetic
strategy. We transfected the cDNA plasmids of DNCnA, CACnA, or DNCaMKII
with HA-ERK2 into cardiomyocytes. There were no significant
differences in expression levels of HA-ERK2 among samples
(Figure 4A). Although overexpression of DNCnA had no effect
on the basal ERK2 activity, it strongly suppressed ISO-induced
activation of ERK2
(Figure 4B). To the contrary, overexpression of DNCaMKII did
not affect ISO-induced activation of ERK2. Overexpression of CACnA
significantly increased the activity of ERK2. These results
collectively suggest that calcineurin, but not CaMKII, plays a pivotal
role in ISO-induced ERK activation in
cardiomyocytes.
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We also addressed the role of calcineurin in vivo using
wild-type mice treated with FK506 and DNCnA transgenic mice, which
overexpress DNCnA specifically in the heart
(Figure 5A).23
Injection of ISO (0.5 µg · kg body
wt-1 ·
min-1 IV) into the left femoral
vein for 10 minutes significantly activated 42-kDa ERK in the
heart of wild-type mice
(Figure 5B). When the mice were pretreated with injection of
FK506 (1 mg · kg body wt-1 ·
d-1 IM), the ERK activation was
significantly suppressed. Moreover, ISO did not increase the activity
of ERKs in the heart of DNCnA transgenic mice
(Figure 5B).
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Elevation of Intracellular
Ca2+ Levels and Activation of
Calcineurin
We examined whether ISO increases intracellular
Ca2+ levels and activates
calcineurin in cultured cardiomyocytes. We measured
intracellular Ca2+ levels using the
Ca2+ fluorescent dye indo 1.
ISO (100 nmol/L) significantly increased the beating rate of
cardiomyocytes and induced a rapid and significant increase
in cytosolic Ca2+ levels at both the
systolic and diastolic phases
(Figure 6A).
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ISO (10 µmol/L) rapidly increased the activity of
calcineurin
(Figure 6B
). The activity of calcineurin was significantly
increased from 2 minutes, peaked at 5 minutes, and decreased
thereafter. An increase in the activity of calcineurin was detected at
10 nmol/L of ISO, and the maximum activation was observed at 10
µmol/L of ISO
(Figure 6C). Similarly, intravenous injection of
ISO (0.5 µg · kg-1 ·
min-1) for 5 minutes activated
calcineurin in the heart of wild-type but not of DNCnA transgenic mice
(Figure 6D).
Involvement of Calcineurin in ISO-Induced
Activation of Src, Shc, and Raf-1 Kinase
We have reported that ISO activates ERKs
through both Gs and Gi
proteins and that the Gi pathway involves Src,
Shc, Ras, and Raf-1 kinase in
cardiomyocytes.13
We examined whether calcineurin is involved in activation of these
molecules. ISO rapidly induced tyrosine phosphorylation
of Src and Shc and activation of Raf-1 kinase
(Figure 7). The phosphorylation and
activation of these proteins were significantly attenuated by
pretreatment with CsA. These results suggest that calcineurin regulates
the ISO-induced activation of ERKs possibly through the Src/Shc/Raf-1
kinase pathway.
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Localization of Calcineurin and Its
Translocation
We finally examined the localization of calcineurin and
its translocation by ISO in cultured cardiomyocytes by
immunocytochemistry. Immunostaining with the anti-CnA
antibody showed prominent signals on band-like structures
(Figure 8Aa and 8Ac), which were also stained with the
anti–-actinin antibody
(Figure 8Ab and 8Ac). These results suggest that calcineurin
is localized in the Z band of cardiomyocytes. There were no
positive signals in unstimulated cells immunostained with
the anti–phospho-ERK antibody (data not shown). When
cardiomyocytes were incubated with ISO for 10 minutes,
calcineurin was translocated into and around the nucleus
(Figure 8Ba), and activated ERKs were also observed
around the nucleus
(Figure 8Be). Pretreatment with CsA significantly attenuated
the ISO-induced translocation of calcineurin and ERKs
(Figure 8Bb and 8Bf). Conversely,
12-O-tetradecanoylphorbol-13-acetate
(TPA), a PKC activator, induced activation and
translocation of ERKs
(Figure 8Bg); however, it did not induce translocation of
calcineurin
(Figure 8Be). Pretreatment with CsA did not affect
TPA-induced ERK translocation
(Figure 8Bh).
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Discussion |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Although it has been reported that ISO activates ERKs and induces cardiac hypertrophy,9 11 12 13 the role of ERKs in the development of cardiac hypertrophy remains unclear. In the present study, ISO-induced cardiac hypertrophy was suppressed by inhibition of ERK activation, suggesting that ERKs play an important role in ISO-induced cardiac hypertrophy. We also observed that ISO-induced cardiac hypertrophy was also significantly suppressed by pretreatment with CsA both in cultured cardiomyocytes and in mice and that ISO induced less cardiac hypertrophy in the DNCnA transgenic mice than in wild-type mice (unpublished results), suggesting an involvement of the Ca2+-dependent phosphatase calcineurin in ISO-induced cardiac hypertrophy.
Ca2+ influx from the extracellular space through L-type Ca2+ channels and Ca2+ release from intracellular Ca2+ stores were critically involved in ISO-induced activation of ERKs in cardiomyocytes. Many studies have reported that ISO activates L-type Ca2+ channels through Gs/PKA in cardiomyocytes.26 27 We recently reported that ISO activates ERKs through both Gs/PKA- and Gi/Ras-dependent pathways.13 Although some reports indicate that activation of Gi increases Ca2+ entry via T-type Ca2+ channels in adrenal glomerulosa cells28 and that Gi protein directly regulates inositol 1,4,5-triphosphate–dependent Ca2+ release in smooth muscle,29 it remains to be determined whether the Gi pathway is involved in Ca2+ influx in cardiomyocytes.
Ca2+ usually regulates cellular events by binding to Ca2+-binding proteins, such as CaM.15 We observed here that inhibition of CaM by W7 attenuated ISO-induced ERK activation, indicating that Ca2+ mediates ISO-induced activation of ERKs through binding to CaM in cardiomyocytes. The Ca2+/CaM complex exerts its functions through downstream effectors, such as CaMKII and calcineurin.17 18 Calcineurin has been reported to be involved in the activation of ERKs in cultured mouse M1 myeloid leukemic cells.30 In the present study, pretreatment of cultured cardiomyocytes with calcineurin inhibitors and overexpression of DNCnA in cardiomyocytes inhibited ISO-induced activation of ERKs. Moreover, inhibition of calcineurin in vivo by the inhibitor or overexpression of the DNCnA also suppressed ISO-induced activation of ERKs in the heart. These results clearly indicate that calcineurin is required for ISO-induced ERK activation. The calcineurin inhibitor did not strongly suppress phenylephrine- and Ang II–induced ERK activation, suggesting that phenylephrine and Ang II activate ERKs through calcineurin-independent mechanisms. Overexpression of CACnA activated ERKs in cardiomyocytes, suggesting that activation of calcineurin is enough to activate ERKs. It was recently reported that ERKs are activated in the heart of hypertrophic calcineurin transgenic mice.31 CaMKII has been reported to be involved in Ang II–induced ERK activation in smooth muscle cells.22 In this study, however, CaMKII was not involved in ISO-induced ERK activation. Although we do not know at present why calcineurin but not CaMKII mediates ISO-induced ERK activation in cardiomyocytes, stimuli and cell types may determine which Ca2+-dependent molecules mediate the activation of ERKs.
The mechanism of how calcineurin activates ERKs is unknown at present; several lines of evidence, however, have suggested the existence of a cross-talk between the calcineurin and MAPK pathways.31 32 33 Calcineurin has been reported to regulate the activity of JNK, a member of MAPK family, in concert with PKC.31 It has been reported that calcineurin potentiates the formation of cAMP in adrenal glomerulosa cells32 and that calcineurin is required for cyclic stretch–induced Src activation in endothelial cells.33 Because both cAMP and Src are upstream activators of ERKs in cardiomyocytes,13 calcineurin may be involved in the activation of ERKs through cAMP and Src. Our present study showed that inhibition of calcineurin significantly suppressed tyrosine phosphorylation of Src and Shc and activation of Raf-1 kinase, suggesting that calcineurin is involved in ISO-induced activation of the Src/Shc/Raf-1 kinase pathway. It remains to be determined how calcineurin activates this pathway in cardiomyocytes.
ISO induced a rapid and significant increase in cytosolic Ca2+ levels during both the systolic and diastolic phases and significantly increased the activity of calcineurin. It has been reported that ISO activates PKA, which phosphorylates L-type Ca2+ channels, leading to an enhanced Ca2+ influx.26 Ca2+ influx through L-type Ca2+ channels induces a large amount of Ca2+ release from intracellular Ca2+ stores by the CICR mechanism.34 35 It has been unknown how calcineurin is activated in cardiomyocytes, in which intracellular Ca2+ levels go up and down at every contraction-relaxation cycle. The release of Ca2+ from intracellular Ca2+ stores was necessary for ISO-induced activation of calcineurin (data not shown), suggesting that a large increase in intracellular Ca2+ levels is required for activation of calcineurin in cardiomyocytes. Calcineurin was localized in the Z band of cardiomyocytes. In cardiomyocytes, the Z band is close to the T tubules.36 37 Because voltage-dependent Ca2+ channels are abundant in the T tubules and Ca2+-releasing channels are also localized in the membrane of sarcoplasmic reticulum, which is close to the T tubules,37 38 the local Ca2+ concentration around the Z band might be quite high. Calcineurin has been reported to be activated by sustained increase in Ca2+.18 Because ISO strongly elevated both systolic and diastolic Ca2+ levels by a Ca2+ influx from the extracellular space through L-type Ca2+ channels and by CICR, free Ca2+ levels around calcineurin in the Z band might be high enough to activate calcineurin. ISO induced rapid translocation of calcineurin as well as ERKs into the nucleus. Although activated calcineurin has been reported to form complexes with NFAT and to translocate into the nucleus,21 the role of translocation of calcineurin in the activation of ERKs remains to be determined.
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Acknowledgments |
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This work was supported by a Grant-in-Aid for Scientific Research, Developmental Scientific Research, and Scientific Research on Priority Areas from the Ministry of Education, Science, Sports, and Culture and by the Program for Promotion of Fundamental Studies in Health Sciences of the Organization for Drug ADR Relief, R&D Promotion and Product Review of Japan (to Dr Komoru). Dr Zou is a recipient of a postdoctoral fellowship from the Japan Society for the Promotion of Science. We wish to thank Dr M. Karin for HA-ERK2 plasmids and Dr T. Shimizu for cell culture.
Received December 21, 2000; revision received January 28, 2001; accepted January 29, 2001.
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References |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
1. Levy D, Garrison RJ, Savage DD, et al. Prognostic implications of echocardiographically determined left ventricular mass in the Framingham heart study. N Engl J Med. 1990;322:1561–1566.[Abstract]
2. Komuro I, Yazaki Y. Control of cardiac gene expression by mechanical stress. Annu Rev Physiol. 1993;55:55–75.[Medline] [Order article via Infotrieve]
3. Chien KR, Grace AA, Hunter JJ. Molecular biology of cardiac hypertrophy and heart failure. In: Chien KR, ed. Molecular Basis of Cardiovascular Disease. Philadelphia, Pa: WB Saunders Co; 1998.
4.
Glennon PE,
Kaddoura S, Sale EM, et al. Depletion of mitogen-activated
protein kinase using an antisense oligodeoxynucleotide
approach downregulates the phenylephrine-induced
hypertrophic response in rat cardiac myocytes.
Circ Res. 1996;78:954–961.
5. Thorburn J, Carlson M, Mansour SJ, et al. Inhibition of a signaling pathway in cardiac muscle cells by active mitogen-activated protein kinase kinase. Mol Biol Cell. 1995;6:1479–1490.[Abstract]
6.
Izumi Y, Kim S,
Murakami T, et al. Cardiac mitogen-activated protein kinase
activities are chronically increased in stroke-prone hypertensive rats.
Hypertension. 1998;31:50–56.
7. Takemoto Y, Yoshiyama M, Takeuchi K, et al. Increased JNK, AP-1 and NF-kappa B DNA binding activities in isoproterenol-induced cardiac remodeling. J Mol Cell Cardiol. 1999;31:2017–2030.[Medline] [Order article via Infotrieve]
8.
Post GR, Goldstein
D, Thuerauf DJ, et al. Dissociation of p44 and p42
mitogen-activated protein kinase activation from
receptor-induced hypertrophy in neonatal rat
ventricular myocytes. J
Biol Chem. 1996;271:8452–8457.
9. Zimmer HG. Catecholamine-induced cardiac hypertrophy: significance of proto-oncogene expression. J Mol Med. 1997;75:849–859.[Medline] [Order article via Infotrieve]
10.
Yamazaki T,
Komuro I, Zou Y, et al. Norepinephrine induces the raf-1
kinase/mitogen-activated protein kinase cascade through both
1- and ß-adrenoceptors.
Circulation. 1997;95:1260–1268.
11. Brand T, Sharma HS, Schaper W. Expression of nuclear proto-oncogenes in isoproterenol-induced cardiac hypertrophy. J Mol Cell Cardiol. 1993;25:1325–1337.[Medline] [Order article via Infotrieve]
12. Bogoyevitch MA, Andersson MB, Gillespie BJ, et al. Adrenergic receptor stimulation of the mitogen-activated protein kinase cascade and cardiac hypertrophy. Biochem J. 1996;314:115–121.
13.
Zou Y, Komuro I,
Yamazaki T, et al. Both Gs and
Gi proteins are critically involved in
isoproterenol-induced cardiomyocyte
hypertrophy. J Biol
Chem. 1999;274:9760–9770.
14. Berridge MJ, Bootman MD, Lipp P. Calcium: a life and death signal. Nature. 1998;395:645–648.[Medline] [Order article via Infotrieve]
15. Means AR, VanBerkum MF, Bagchi I, et al. Regulatory functions of calmodulin. Pharmacol Ther. 1991;50:255–270.[Medline] [Order article via Infotrieve]
16.
Gruver CL, DeMayo
F, Goldstein MA, et al. Targeted developmental overexpression of
calmodulin induces proliferative and hypertrophic growth of
cardiomyocytes in transgenic mice.
Endocrinology. 1993;133:376–388.
17. Braun AP, Schulman H. The multifunctional calcium/calmodulin-dependent protein kinase: from form to function. Annu Rev Physiol. 1995;57:417–445.[Medline] [Order article via Infotrieve]
18.
Klee CB, Ren H,
Wang X. Regulation of the calmodulin-stimulated protein
phosphatase, calcineurin. J Biol
Chem. 1998;273:13367–13370.
19.
Ramirez MT, Zhao
XL, Schulman H, et al. The nuclear deltaB isoform of
Ca2+/calmodulin-dependent
protein kinase II regulates atrial natriuretic factor gene
expression in ventricular myocytes.
J Biol Chem. 1997;272:31203–31208.
20. Molkentin JD, Lu JR, Antos CL, et al. A calcineurin-dependent transcriptional pathway for cardiac hypertrophy. Cell. 1998;93:215–228.[Medline] [Order article via Infotrieve]
21. Shibasaki F, Price ER, Milan D, et al. Role of kinases and the phosphatase calcineurin in the nuclear shuttling of transcription factor NF-AT4. Nature. 1996;382:370–373.[Medline] [Order article via Infotrieve]
22.
Abraham ST,
Benscoter HA, Schworer CM, et al. A role for
Ca2+/calmodulin-dependent
protein kinase II in mitogen-activated protein kinase signaling
cascade of cultured rat aortic vascular smooth muscle cells.
Circ Res. 1997;81:575–584.
23.
Saito S, Hiroi Y,
Zou Y, et al. ß-Adrenergic pathway induces apoptosis through
calcineurin activation in cardiac myocytes.
J Biol Chem. 2000;275:34528–34533.
24. Kinugawa K, Takahashi T, Kohmoto O, et al. Ca2+-growth coupling in angiotensin II-induced hypertrophy in cultured rat cardiac cells. Cardiovasc Res. 1995;30:419–431.[Medline] [Order article via Infotrieve]
25.
Shimoyama M,
Hayashi D, Takimoto E, et al. Calcineurin plays a critical role in
pressure overload–induced cardiac hypertrophy.
Circulation. 1999;100:2449–2454.
26.
Yatani A, Brown
AM. Rapid ß-adrenergic modulation of cardiac calcium channel currents
by a fast G protein pathway.
Science. 1989;245:71–74.
27. Hartzell HC, Mery PF, Fischmeister R, et al. Sympathetic regulation of cardiac calcium current is due exclusively to cAMP-dependent phosphorylation. Nature. 1991;351:573–576.[Medline] [Order article via Infotrieve]
28.
Lu HK, Fern RJ,
Luthin D, et al. Angiotensin II stimulates T-type
Ca2+ channel currents via activation of a G
protein, Gi.
Am J Physiol. 1996;271:C1340–C1349.
29. Neylon CB, Nickashin A, Tkachuk VA, et al. Heterotrimeric Gi protein is associated with the inositol 1,4,5-trisphosphate receptor complex and modulates calcium flux. Cell Calcium. 1998;23:281–289.[Medline] [Order article via Infotrieve]
30.
Lotem J, Kama R,
Sachs L. Suppression or induction of apoptosis by opposing
pathways downstream from calcium-activated calcineurin.
Proc Natl Acad Sci
U S A. 1999;96:12016–12020.
31.
De Windt LJ, Lim
HW, Haq S, et al. Calcineurin promotes protein kinase C and c-Jun
NH2-terminal kinase activation in the heart: cross-talk between cardiac
hypertrophic signaling pathways. J
Biol Chem. 2000;275:13571–13579.
32.
Baukal AJ,
Hunyady L, Catt KJ, et al. Evidence for participation of calcineurin in
potentiation of agonist-stimulated cyclic AMP formation by the
calcium-mobilizing hormone, angiotensin II.
J Biol Chem. 1994;269:24546–24549.
33. Naruse K, Sai X, Yokoyama N, et al. Uni-axial cyclic stretch induces c-src activation and translocation in human endothelial cells via SA channel activation. FEBS Lett. 1998;441:111–115.[Medline] [Order article via Infotrieve]
34.
Nabauer M,
Callewaert G, Cleemann L, et al. Regulation of calcium release is gated
by calcium current, not gating charge, in cardiac myocytes.
Science. 1989;244:800–803.
35.
Sham JS, Cleemann
L, Morad M. Functional coupling of Ca2+
channels and ryanodine receptors in cardiac myocytes.
Proc Natl Acad Sci
U S A. 1995;92:121–125.
36.
Schultheiss T,
Choi J, Lin ZX, et al. A sarcomeric alpha-actinin truncated at the
carboxyl end induces the breakdown of stress fibers in PtK2 cells and
the formation of nemaline-like bodies and breakdown of myofibrils in
myotubes. Proc Natl Acad Sci
U S A. 1992;89:9282–9286.
37.
Soeller C,
Cannell MB. Examination of the transverse tubular system in living
cardiac rat myocytes by 2-photon microscopy and digital
image-processing techniques. Circ
Res. 1999;84:266–275.
38. Carl SL, Felix K, Caswell AH, et al. Immunolocalization of sarcolemmal dihydropyridine receptor and sarcoplasmic reticular triadin and ryanodine receptor in rabbit ventricle and atrium. J Cell Biol. 1995;129:672–682.
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