© 2000 American Heart Association, Inc.
Integrative Physiology |
Remodeling of Gap Junctional Coupling in Hypertrophied Right Ventricles of Rats With Monocrotaline-Induced Pulmonary Hypertension
From the Department of Circulation (M.U., I.K.), the Department of Humoral Regulation (H.H.), and the Department of Teratology and Genetics (Y.T., L.E.), Research Institute of Environmental Medicine, Nagoya University, Nagoya, Japan; Laboratory of Eukaryotic Molecular Genetics (A.I.M.), National Institute for Medical Research, London, UK; and Department of Cardiac Medicine (N.J.S.), National Heart and Lung Institute, Imperial College School of Medicine, London, UK.
Correspondence to Haruo Honjo, Department of Humoral Regulation, Research Institute of Environmental Medicine, Nagoya University, Nagoya 464-8601, Japan. E-mail honjo{at}riem.nagoya-u.ac.jp
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Abstract |
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Abstract—The present study investigates the remodeling of gap junctional organization in relation to changes in anisotropic conduction properties in hypertrophied right ventricles (RVs) of rats with monocrotaline (MCT)-induced pulmonary hypertension. In contrast to controls that showed immunolocalization of connexin43 (Cx43) labeling largely confined to the intercalated disks, RV myocytes from MCT-treated rats showed dispersion of Cx43 labeling over the entire cell surface. The disorganization of Cx43 labeling became more pronounced with the progression of hypertrophy. Desmoplakin remained localized to the intercalated disks, as in controls. In RV tissues, the proportion of Cx43 label at the intercalated disk progressively decreased. Quantitative analysis of en face views of intercalated disks revealed a significant decrease in the disk gap junctional density in RV tissues of MCT-treated rats (control, 0.18 versus MCT-treated, 0.14 at 2 weeks; control, 0.16 versus MCT-treated, 0.11 at 4 weeks). Conduction velocity in RVs parallel to the fiber orientation was significantly lower (30.2% [n=9]) in MCT-treated rats at 4 weeks than in control rats, whereas there was no significant difference observed in the conduction velocity across the fiber orientation between control and MCT-treated rats. The anisotropic ratio of MCT-treated rats (1.38±0.10) was significantly lower than that of control rats (1.98±0.12). These results suggest that RV hypertrophy induced by pressure overload is associated with both disorganization of gap junction distribution and alteration of anisotropic conduction properties.
Key Words: ventricular hypertrophy • connexin • immunohistochemistry • anisotropy • conduction
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Introduction |
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Ventricular hypertrophy is associated with an increased risk of cardiac arrhythmia and sudden death.1 2 In addition to changes in active membrane properties (ionic currents, pumps, and exchangers), alterations in the passive properties of the myocardium are implicated in the arrhythmogenic substrate of hypertrophied ventricles.3 Central to the determinants of the passive properties are gap junctions that form the low-resistance pathway for propagation of electrical impulse between cardiac cells.4 5 Gap junctions are composed of transmembrane proteins that belong to the connexin family. The principal gap junctional protein expressed in the ventricles of mammalian heart is connexin43 (Cx43).6 7 Immunohistochemical studies using anti-Cx43 antibodies in ischemic heart disease and hypertrophic cardiomyopathy have revealed marked alteration of Cx43 expression and distribution.7 8 9 Remodeling of gap junctions may thus be an important morphological factor in abnormal conduction properties of the diseased heart, but our knowledge of this process in cardiac hypertrophy remains limited.
In rats, a parental dose of monocrotaline (MCT), a pyrrolizidine alkaloid, is known to cause pulmonary hypertension within a few weeks.10 The pressure overload in turn results in right ventricular (RV) hypertrophy, leading to right-sided congestive heart failure within several weeks. This model is suitable for chronological investigation of remodeling of the gap junction because progressive RV hypertrophy can be produced in a short period without any significant pathological changes in the left ventricle (LV).11
In the present study, we investigated changes in gap junction distribution and organization in the RVs of hypertrophied rat hearts 1 to 4 weeks after MCT treatment with the aid of anti-Cx43 antibody labeling and confocal laser scanning microscopy. Both disaggregated myocytes and multicellular tissue preparations were used for the immunohistochemistry. The distribution and organization of desmosomes, junctions responsible for mechanical linkage of intermediate filaments between cardiac cells,12 were examined in parallel using an antidesmoplakin antibody. Morphological remodeling of gap junctions observed in association with ventricular hypertrophy was then correlated with altered anisotropic conduction properties measured in RV myocardium.
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Materials and Methods |
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Animals
MCT (60 mg/kg) was injected in 79 5-week-old male Wistar rats (Chubu-kagaku-shizai, Nagoya, Japan) to produce pulmonary hypertension.11 Saline was injected in 59 rats as controls. Hypertrophy was estimated by the heart-to-body weight ratio and by the weight ratio of the RV free wall to the LV free wall plus interventricular septum. All procedures were conducted in accordance with statutory Japanese regulations.
Preparation of Samples
For standard microscopy, ventricular sections were
stained with hematoxylin and eosin (H&E). In addition, thin sections
fixed with 2% glutaraldehyde were stained with
toluidine blue. Single myocytes were isolated from
ventricles11 and fixed with 2% paraformaldehyde.
Cryosections of ventricles were prepared from 2%
paraformaldehyde–fixed hearts for tissue immunolabeling.
Immunohistochemistry
For immunodetection of gap junctions and desmosomes, a mouse
monoclonal anti-Cx43 antibody (Chemicon) and rabbit antidesmoplakin
antiserum were used. The specificity of both antibodies has been
demonstrated previously.13 14
After permeabilization (0.3% Triton X-100), quenching (0.1 mol/L NH4Cl), and blocking (3% normal goat serum/5% BSA), samples were incubated with anti-Cx43 antibody (1:200) or a mixture of anti-Cx43 (1:200) and antidesmoplakin (1:100) antibodies overnight. Primary antibody-bound Cx43 complexes were visualized by FITC-conjugated antimouse IgG, and desmoplakin complexes were detected by biotinylated antirabbit IgG and Texas Red–conjugated streptavidin. Samples processed without primary antibody served as negative controls.
The labeled samples were examined using a confocal microscope (BioRad MRC-1024). In addition to single-plane evaluation, optical section series were taken.
Quantitative Image Data Analysis
The proportion of Cx43 immunolabeling at the intercalated disks
relative to overall Cx43 was quantified according to a procedure
described previously.15 Three randomly selected fields
from longitudinally sectioned tissue were analyzed using NIH
Image 1.61 (NIH). The Cx43 gap junctional density in the intercalated
disk area was estimated in projection images of transversely
sectioned tissue using a protocol reported previously.16
Thirty disks were randomly selected in each group.
Western Blotting
The amount of Cx43 was evaluated by Western blotting of RV
homogenates.17 The intensity of Cx43 bands was
quantified by densitometry and normalized to actin.
Electrophysiological Study
Extracellular electrograms were recorded from the epicardial
surface of an arterially perfused isolated RV free wall
through an electrode array (7x7 mm) consisting of 64 pairs of
modified bipolar electrodes at 36°C.18 The endocardial
Purkinje network was ablated by phenol to eliminate the preferential
conduction and spontaneous excitation. The local activation time was
measured under regular stimulation, and maps of excitation spread were
constructed. Conduction velocity (
) was determined by
linear regression of the isochrone distance versus activation time.
Lines parallel and perpendicular to the fiber orientation were defined
as the direction of longitudinal (L) and transverse
(T) propagation, respectively.
Statistics
Descriptive statistics are expressed as mean±SEM. Data were
analyzed using ANOVA or nonparametric procedures
wherever appropriate. Details of the procedures are stated in the
legends of each figure and table. All effects and interactions are
tested at the 0.05 level of significance. Data analysis was
conducted using SAS 6.12 (SAS Institute).
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Results |
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RV Hypertrophy in MCT-Treated Rats
Figure 1
shows a comparison of the
heart weight to body weight (HW/BW) ratio and the tissue weight ratio
of RV free wall to LV free wall plus interventricular
septum (RV/[LV+IVS]) between control and MCT-treated rats. Fifty-six
rats (6 control and 8 MCT-treated rats at 1, 2, 3, and 4 weeks) were
used. The ratios in control rats were constant throughout the
observation period, whereas the ratios in MCT-treated rats increased
progressively. The HW/BW ratio and RV/(LV+IVS) ratio were significantly
larger in MCT-treated rats than controls 3 and 4 weeks after injection.
Ten MCT-treated rats were followed up for 5 weeks after injection. In
the fifth week, 6 MCT-treated rats died, and the remaining rats showed
physical signs of right-sided congestive heart failure.
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,
Control; 
, MCT. Data were analyzed using 2-way ANOVA. The 2
factors were treatment (saline and MCT) and time after treatment (1, 2,
3, and 4 weeks). The least-significant difference procedure was used
for multiple comparisons with Bonferroni adjustment for number of
comparisons determined before the experiment (ie, 10). *Mean of MCT is
significantly different from the mean of the control. 
Mean of that
week is significantly different from the mean of the previous adjacent
week.
Figures 2A and 2B show the
representative change in macroscopic morphology after
MCT treatment. The thickness of the RV free wall in the MCT-treated rat
was markedly increased, whereas the LV wall thickness was
unaffected.
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We also estimated the extent of cell hypertrophy based on cell size data obtained from 6 rats (3 control and 3 MCT-treated rats at 4 weeks). Twenty-five myocytes were randomly selected from a cell suspension of each control rat, and 41 or 42 myocytes were selected from a cell suspension of each MCT-treated rat. The cell width of the MCT-treated RV myocytes (45.0±0.6 µm) was significantly larger than controls (28.9±0.3 µm), whereas there was no significant difference in the cell length between the 2 groups (control, 133.7±2.4 µm versus MCT-treated, 136.7±1.5 µm) (ANOVA for hierarchical classification). The average length-to-width ratio decreased from 4.6 in the control myocytes to 3.0 in the MCT-treated myocytes.
Standard Light Microscopy
Standard light microscopy was used to assess histopathological
features in 10 rats (2 control and 3 MCT-treated rats at 2 and 4 weeks)
(Figures 2C
through 2F). Minimal myofiber disarray was observed
in RV tissue sections stained with toluidine blue, but the general
anisotropic architecture composed of 3 myocardial layers was well
preserved even 4 weeks after MCT injection.
Distribution of Immunolabeled Gap Junctions and Desmosomes in
Isolated Myocytes
RV and LV myocytes isolated from 32 rats (4 control and 4
MCT-treated rats at 1, 2, 3, and 4 weeks) were labeled with anti-Cx43
and antidesmoplakin antibodies to study immunolocalization of gap
junctions and desmosomes (Figure 3). In
control RV myocytes labeled with Cx43 antibody alone, gap junctions
were visualized as aggregates of bright punctate fluorescent
domains at the cell termini, marking the positions of intercalated
disks (Figure 3A). The staining often extended across the full
width of the myocyte, but shorter punctate lines were also observed at
the sites of bifurcations of the main cell body. Double staining for
Cx43 (green) and desmoplakin (red) confirmed coexistence of gap
junctions and desmosomes at the cell termini (Figure 3E). These
patterns were common to myocytes from RVs and LVs.
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The Cx43 staining patterns of RV myocytes from rats treated with MCT
for 2 weeks and longer differed markedly from controls; the gap
junctional labeling was no longer confined to the cell termini but
showed varying degrees of dispersion over the cell surface (Figures 3B and 3C). Double staining (Figures 3F and 3G) clearly
revealed that the dispersed Cx43 immunolabeling on the cell surface was
independent from the disk-like structure at the cell termini composed
of both desmoplakin and Cx43 labeling. The disordered patterns of Cx43
immunostaining tended to become more pronounced with
the progression of hypertrophy as assessed by increased
cell width. The dissociation of Cx43 and desmoplakin labeling was also
remarkable (Figures 3G and 3H). In contrast,
ventricular myocytes isolated from the LVs of MCT-treated
rats showed normal Cx43 labeling patterns (Figure 3D). The
changes described on hypertrophied RV myocytes are particularly clearly
illustrated in double-labeled aggregates (3 myocytes with lateral
contact) obtained from control and MCT-treated (4 weeks) rats (Figures 3I and 3J).
Distribution of Immunolabeled Gap Junctions and Desmosomes in
Tissue Sections
Immunolabeling of Cx43 and desmoplakin in ventricular
tissue sections was carried out in 48 rats (6 control and 6 MCT-treated
rats at 1, 2, 3, and 4 weeks). Figures 4A
through 4D show the distributions of immunolabeled Cx43 gap junctions
and desmosomes in single confocal optical slices through longitudinally
sectioned RV myocardium. In control rats, Cx43-containing
gap junctions are highly organized into clusters of fluorescent
label at the intercalated disks running across the longitudinal axis
(Figure 4A). Double staining of RV sections of control rats
(Figure 4C) confirmed that the desmosomes are also confined to
the disks.
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In rats treated with MCT for longer than 2 weeks, the Cx43 staining was
no longer confined to the intercalated disks (Figure 4B).
Instead, the staining pattern showed varying degrees of dispersion over
the cell surface. It was possible to identify the approximate position
of the intercalated disks by the presence of aggregates of labeled
junctions, but the amount of signal was less and its distribution was
more irregular than controls. Double staining (Figure 4D)
confirmed a marked dissociation of Cx43 from the disks as identified by
desmoplakin labeling. A parallel myofiber arrangement was generally
preserved in the hypertrophied myocardium, but constituent
myocytes showed more complex and irregular configurations than control
myocardium.
We estimated the proportion of Cx43 label in transverse array (at the
position of intercalated disks) over the total label present in the
3 test fields from each RV tissue preparation sectioned longitudinally.
The results obtained from 24 rats (6 control and 6 MCT-treated rats at
2 and 4 weeks) are summarized in Figure 4E. The proportion of
Cx43 label at the intercalated disks was significantly lower in
MCT-treated rats 2 and 4 weeks after injection. The proportion of
desmoplakin label at the intercalated disks in RV
myocardium was always greater than 90% in both control and
MCT-treated rats (data not shown).
Figure 5 shows Cx43 immunolabeling in the
intercalated disk area seen en face in RV tissues sectioned
transversely. Images were taken at 1-µm intervals to cover a full
thickness of the intercalated disk, and the entire series was
projected as a single composite image. In the control tissue, there
was a normal distribution of gap junctional label, with small central
gap junctions surrounded by extensive larger spots of label at the disk
periphery. In the tissues obtained from MCT-treated rats (for 2 to 4
weeks), the larger peripheral gap junctions were preserved,
but there was a striking loss of the central smaller gap junctions,
giving rise to a more empty appearance of the disks. We estimated the
gap junctional density in the intercalated disk in 12 rats (3 control
and 3 MCT-treated rats at 2 and 4 weeks) using a protocol reported by
Kaprielian et al.16 Thirty intercalated disks in 10 fields
were analyzed in each group (Table). The ratio of gap
junctional area to the total disk area was significantly lower in
MCT-treated rats (2 and 4 weeks) than in respective controls.
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In the LV tissues, the normal labeling patterns of Cx43 were well preserved in MCT-treated rats even 4 weeks after injection (data not shown).
Immunoblot and Immunoconfocal Analysis
Western blotting was carried out in 12 rats (6 control and 6
MCT-treated rats at 4 weeks). The Cx43 antibodies recognized 3 bands
migrating between 42 and 45 kDa (phosphorylated and
nonphosphorylated states) on immunoblots
from RV tissue homogenates of control and MCT-treated rats
(Figure 6) as demonstrated
previously.19 Densitometric quantification revealed no
significant differences in the amount between control and MCT-treated
rats. In line with the immunoblot analysis, the
quantity of Cx43 signal per unit volume of RV myocyte (a total of 20
cells from each group) measured by quantitative immunoconfocal
microscopy was not significantly different between control and
MCT-treated rats 4 weeks after injection.
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Anisotropic Conduction Properties
Anisotropic conduction properties in the epicardial surface of RV
tissues were examined in 18 rats (9 control and 9 MCT-treated rats at 4
weeks) (Figure 7). Constant stimuli (2.5
Hz) were applied to the middle of the upper edge of the 64-channel
electrode grid (
1 mm below the atrioventricular
groove). In controls (Figure 7A, top), the activation front
proceeded at the highest speed in a direction parallel (longitudinal,
L) to the subepicardial fiber orientation and at the slowest
speed in a direction perpendicular (transverse, T). The
isochrones showed an elliptical activation pattern indicating the
normal uniform anisotropy. In the tissue from a rat treated with MCT
for 4 weeks (Figure 7A, bottom), the elliptical isochrone
pattern was less marked (more circular) because of a moderate slowing
of L propagation. Figure 7B shows for
L and T propagation and their ratios. The
conduction velocity parallel with the fiber orientation
(L) in RV tissues from MCT-treated rats was
significantly less than controls (30.2% on average), but there was no
significant difference in conduction velocity across fiber orientation
(T) between the 2 groups. The
anisotropic ratio of conduction velocity
(L/T) in
MCT-treated rats (1.38±0.10) was significantly lower than controls
(1.98±0.12).
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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In the present study, we investigated remodeling of gap junctional coupling in hypertrophied RVs of rats secondary to MCT-induced pulmonary hypertension. The novel findings are that (1) immunolocalized Cx43 in hypertrophied myocytes is not confined to cell termini (intercalated disks) but shows varying degrees of dispersion over the entire cell surface; (2) there is a loss of the population of small gap junctions in the intercalated disk center of the hypertrophied myocardium, giving rise to a decrease in the disk gap junctional density; and (3) there is a decrease of
L, whereas
T is preserved. A variety of alterations in Cx43 gap junctions have been reported in previous studies on the hypertrophied heart. For example, globally reduced levels of Cx43 protein have been reported in immunoblot studies on ventricular hypertrophy in transgenic hypertensive rats20 and in immunoconfocal studies on chronically pressure-overloaded hypertrophied myocardium in humans undergoing surgical replacement of a stenosed aortic valve.21 On the other hand, no alteration in Cx43 transcript levels was detectable at the 4-week stage in the hypertrophied hearts of rats made hypertensive by renal artery clipping or deoxycorticosterone/salt administration,22 whereas elevated Cx43 protein was reported in the early phase of hypertrophy due to renovascular hypertension in the guinea pig.23 Thus, although altered Cx43 gap junctional expression seems well documented in hypertrophy, no single form of change common to all forms of hypertrophy has been identified. Factors contributing to this apparent lack of uniformity in the findings to date may include the use of different species, sampling periods, and models in which hypertrophy may develop at different rates.
The remarkable dispersion of gap junctions over the lateral surface of RV myocytes reported in the present study was not apparent in any of the above studies. This stage-dependent change in distribution, readily recognized 2 weeks after MCT injection and yet more prominent at 4 weeks, involved a marked and progressive decrease in the measured proportion of Cx43 labeling of intercalated disks but no detectable alteration in the global Cx43 content. This dramatic change in gap junction distribution shows some similarity to that seen in hypertrophic cardiomyopathy24 and at the border zone of the healed human myocardial infarct25 ; but, in contrast to these situations, the changes observed in the present study were not associated with a major alteration in myocyte orientation or myofiber disarray. Because gap junctional disorganization can develop rapidly after acute myocardial infarction,26 27 it might be suggested that the changing gap junction distribution observed in the present study could similarly be attributable to acute cell injury arising directly from the effect of MCT. However, it was shown in our previous experiments that direct application of MCT (1 to 60 mg/L) caused no significant effects on action potential configuration and ionic currents (transient outward current and L-type calcium current) of rat ventricular myocytes.11 The conduction velocity in rat RV epicardial surface was also unaffected by direct application of MCT (Kodama I, Honjo H, Uzzaman M, unpublished observation, 1999). Moreover, the pattern of Cx43 gap junctions in the LV myocytes remained completely normal throughout MCT treatment. Hence, the altered Cx43 gap junction distribution observed in the RV myocardium is interpreted as a component of the extensive hypertropic remodeling that occurs in response to pressure overload (pulmonary hypertension) rather than to direct toxic effects of MCT treatment.
In contrast to the appearance of extensive lateral Cx43 staining in remodeling RVs after MCT treatment, a reduction in lateral cell-cell contact has been reported in remodeling LV canine myocardium after infarction.28 The changes reported in the latter model were identified 3 to 10 weeks after infarction in selected areas of myocardium around the infarct scar that maintained a well-arrayed cell orientation, and the resulting predicted increase in resistance to transverse current flow was considered consistent with earlier electrophysiological findings.29 On this basis, it might correspondingly be predicted that increased lateral Cx43 staining, as observed in the present study, might lead to an increase in transverse conduction velocity. Our finding that the transverse conduction velocity remained unchanged in practice emphasizes that Cx43 protein distribution may not always directly reflect the distribution of functional gap junction channels. One possible explanation for the presence of extensive seemingly nonfunctional lateral Cx43 could relate to processes of gap junction disassembly and degradation, perhaps consequent to enhanced gap junction turnover30 during rapid myocardial remodeling. Lateral Cx43 staining in myocytes at the border zone of human infarcts is due at least in part to segments of internalized gap junction membrane,25 and our preliminary immunogold electron microscopic observations in the MCT model suggest that a proportion of the lateral Cx43 label is not associated with trilaminar gap junction structures. The presence of such features in the MCT model but not other models of hypertrophy may be due to the comparative rapidity and severity of the hypertrophic response in the former.
Cable theory predicts that as cell diameter increases, as occurs in cardiac hypertrophy, conduction velocity will increase. However, using isolated hypertrophied human myocardium, it has been found that a reduction of conduction velocity accompanies cell enlargement, and it was proposed that this is because of an additional increase of intracellular resistivity (Ri). Cooklin et al3 measured the impedance to current flow in the intracellular compartment of guinea pig–hypertrophied LV myocardium prepared by aortic constriction. Their results revealed that an extensive LV hypertrophy is associated with an increased Ri, which can be attributed solely to an increase of the junctional resistance between adjacent cells.3 They explained the decrease in conduction velocity of the animal model by an inhibition of intercellular electrical coupling.4 Notwithstanding the need for caution in relating Cx43 distribution to electrophysiological properties, as highlighted above, one predicted effect of the decreased gap junctional density in the intercalated disk observed in the present study might be to reduce local current flow parallel with the myocardial fiber orientation. That reduced levels of Cx43 can result in slowed conduction has been demonstrated from studies on mice heterozygous for a null mutation of the Cx43 gene.31 In the hypertrophied RV free wall, in fact, the longitudinal conduction velocity decreased significantly, but the transverse conduction velocity was unchanged, giving rise to a significant reduction of anisotropic ratio in comparison with control preparations.
In conclusion, we have shown that RV hypertrophy induced by pressure overload is associated with alterations of anisotropic properties and altered distribution of Cx43 gap junctions. Whether a causal relationship exists between these changes requires further investigation.
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Acknowledgments |
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The authors thank Dr Tobias Opthof (University Medical Center, Utrech, The Netherlands) for his comments on the manuscript.
Received July 20, 1999; accepted February 7, 2000.
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References |
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|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
1. Kannel WB, Doyle JT, McNamara PM, Quickenton P, Gordon T. Precursors of sudden coronary death: factors related to the incidence of sudden death. Circulation. 1975;51:606–613.
2. Levy D, Anderson KM, Savage DD, Balkus SA, Kannel WB, Castelli WP. Risk of ventricular arrhythmias in cardiac failure and hypertrophy: the Framingham heart study. Am J Cardiol. 1987;60:560–565.[Medline] [Order article via Infotrieve]
3.
Cooklin M, Wallis WRL, Sheridan DJ, Fry CH. Changes in
cell-to-cell electrical coupling associated with left
ventricular hypertrophy. Circ Res. 1997;80:765–771.
4.
Shaw RM, Rudy T. Ionic mechanisms of propagation in
cardiac tissue: roles of the sodium and L-type calcium currents during
reduced excitability and decreased gap junction coupling. Circ
Res. 1997;81:727–741.
5.
Rohr S, Kucera JP, Kléber AG. Slow conduction in
cardiac tissue, I: effects of a reduction of excitability versus a
reduction of electrical coupling on microconduction. Circ
Res. 1998;83:781–794.
6. Gros DB, Jongsma HJ. Connexins in mammalian heart function. Bioessays. 1996;18:719–730.[Medline] [Order article via Infotrieve]
7. Severs NJ, Dupont E, Kaprielian RR, Yeh H-I, Rothery S. Gap junctions and connexins in the cardiovascular system. In: Yacoub MH, Carpentier AF, Pepper J, Fabini J-N, eds. Annual of Cardiac Surgery. London, England: Rapid Science Publishers; 1996:31–44.
8. Severs NJ. Pathophysiology of gap junctions in heart disease. J Cardiovasc Electrophysiol. 1994;5:462–475.[Medline] [Order article via Infotrieve]
9. Peters NS. New insights into myocardial arrhythmogenesis: distribution of gap-junctional coupling in normal, ischaemic and hypertrophied human hearts. Clin Sci (Colch). 1996;90:447–452.[Medline] [Order article via Infotrieve]
10.
Miyauchi T, Yorikane R, Sakai S, Sakurai T, Okada M,
Nishikibe M, Yano M, Yamaguchi I, Sugita Y, Goto K. Contribution of
endogenous endothelin-1 to the progression of
cardiopulmonary alterations in rats with monocrotaline-induced
pulmonary hypertension. Circ Res. 1993;73:887–897.
11.
Lee J-K, Kodama I, Honjo H, Anno T, Kamiya K, Toyama J.
Stage-dependent changes in membrane currents in rats with
monocrotaline-induced right ventricular
hypertrophy. Am J Physiol. 1997;272:H2833–H2842.
12. Buxton RS, Magee AI. Structure and interactions of desmosomal and other cadherins. Semin Cell Biol. 1992;3:157–167.[Medline] [Order article via Infotrieve]
13. Vozzi C, Dupont E, Coppen SR, Yeh H-I, Severs NJ. Chamber-related differences in connexin expression in the human heart. J Mol Cell Cardiol. 1999;31:991–1003.[Medline] [Order article via Infotrieve]
14.
Angst BD, Khan LUR, Severs NJ, Whitely K, Rothery S,
Thompson RP, Magee AI, Gourdie RG. Dissociated spatial patterning of
gap junctions and cell adhesion junctions during postnatal
differentiation of ventricular myocardium.
Circ Res. 1997;80:88–94.
15.
Peters NS, Severs NJ, Rothery SM, Lincoln C, Yacoub MH,
Green CR. Spatiotemporal relation between gap junctions and fascia
adherens junctions during postnatal development of human
ventricular myocardium. Circulation. 1994;90:713–725.
16.
Kaprielian RR, Gunning M, Dupont E, Sheppard MN,
Rothery SM, Underwood R, Pennell DJ, Fox K, Pepper J, Poole-Wilson PA,
Severs NJ. Downregulation of immunodetectable connexin43 and decreased
gap junction size in the pathogenesis of chronic hibernation in the
human left ventricle. Circulation. 1998;97:651–660.
17.
Coppen SR, Dupont E, Rothery S, Severs NJ. Connexin 45
expression is preferentially associated with the
ventricular conduction system in mouse and rat heart.
Circ Res. 1998;82:232–243.
18.
Yamamoto M, Honjo H, Niwa R, Kodama I. Low-frequency
extracellular potentials recorded from the sinoatrial node.
Cardiovasc Res. 1998;39:360–372.
19.
Musil LS, Goodenough DA. Biochemical analysis
of connexin43 intracellular transport, phosphorylation,
and assembly into gap junctional plaques. J Cell Biol. 1991;115:1357–1374.
20.
Bastide B, Neyses L, Ganten D, Paul M, Willecke K,
Traub O. Gap junction protein connexin40 is preferentially expressed in
vascular endocardium and conductive bundles of rat
myocardium and is increased under hypertensive conditions.
Circ Res. 1993;73:1138–1149.
21.
Peters NS, Green CR, Poole-Wilson PA, Severs NJ.
Reduced content of connexin43 gap junctions in ventricular
myocardium from hypertrophied and ischemic human
hearts. Circulation. 1993;88:864–875.
22. Haefliger JA, Castillo E, Waeber G, Bergonzelli GE, Aubert JF, Sutter E, Nicod P, Waeber B, Meda P. Hypertension increases connexin43 in a tissue-specific manner. Circ Res. 1997;95:1007–1014.
23. Peters NS, del Monte F, MacLeod KT, Green CR, Poole-Wilson PA, Severs NJ. Increased cardiac myocyte gap-junctional membrane early in renovascular hypertension. Am J Cardiol. 1993;21:59A. Abstract.
24.
Sepp R, Severs NJ, Gourdie RG. Altered patterns of
cardiac intercellular junction distribution in hypertrophic
cardiomyopathy. Heart. 1996;76:412–417.
25. Smith JH, Green CR, Peters NS, Rothery S, Severs NJ. Altered patterns of gap junction distribution in ischemic heart disease: an immunohistochemical study of human myocardium using laser scanning confocal microscopy. Am J Pathol. 1991;139:801–821.[Abstract]
26. Matsushita T, Takamatsu T. Ischemia-induced temporal expression of connexin43 in rat heart. Virchows Arch. 1997;431:453–458.[Medline] [Order article via Infotrieve]
27.
Peters NS, Coromilas J, Severs NJ, Wit AL. Disturbed
connexin43 gap junction distribution correlates with the location of
reentrant circuits in the epicardial border zone of healing canine
infarcts that cause ventricular tachycardia.
Circulation. 1997;95:988–996.
28. Luke RA, Saffitz JE. Remodeling of ventricular conduction pathways in healed canine infarct border zones. J Clin Invest. 1991;87:1594–1602.
29.
Dillon SM, Allessie MA, Ursell PC, Wit AL. Influences
of anisotropic tissue structure on reentrant circuits in the epicardial
border zone of subacute canine infarcts. Circ Res. 1988;63:182–206.
30.
Beardslee MA, Laing JG, Beyer EC, Saffitz JE. Rapid
turnover of connexin43 in the adult rat heart. Circ Res. 1998;83:629–635.
31. Guerrero PA, Schuessler RB, Davis LM, Beyer EC, Johnson CM, Yamada KA, Saffitz JE. Slow ventricular conduction in mice heterozygous for a connexin 43 null mutation. J Clin Invest. 1997;99:1991–1998.[Medline] [Order article via Infotrieve]
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 J. Qu, F. M. Volpicelli, L. I. Garcia, N. Sandeep, J. Zhang, L. Marquez-Rosado, P. D. Lampe, and G. I. Fishman Gap Junction Remodeling and Spironolactone-Dependent Reverse Remodeling in the Hypertrophied Heart Circ. Res., February 13, 2009; 104(3): 365 - 371. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. J. Severs, A. F. Bruce, E. Dupont, and S. Rothery Remodelling of gap junctions and connexin expression in diseased myocardium Cardiovasc Res, October 1, 2008; 80(1): 9 - 19. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Sayed, S. Rane, J. Lypowy, M. He, I.-Y. Chen, H. Vashistha, L. Yan, A. Malhotra, D. Vatner, and M. Abdellatif MicroRNA-21 Targets Sprouty2 and Promotes Cellular Outgrowths Mol. Biol. Cell, August 1, 2008; 19(8): 3272 - 3282. [Abstract] [Full Text] [PDF]
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|
|
|
|
|
T. Nakagami, H. Tanaka, P. Dai, S.-F. Lin, T. Tanabe, H. Mani, K. Fujiwara, H. Matsubara, and T. Takamatsu Generation of reentrant arrhythmias by dominant-negative inhibition of connexin43 in rat cultured myocyte monolayers Cardiovasc Res, July 1, 2008; 79(1): 70 - 79. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Sato, T. Ohkusa, H. Honjo, S. Suzuki, M.-a. Yoshida, Y. S. Ishiguro, H. Nakagawa, M. Yamazaki, M. Yano, I. Kodama, et al. Altered expression of connexin43 contributes to the arrhythmogenic substrate during the development of heart failure in cardiomyopathic hamster Am J Physiol Heart Circ Physiol, March 1, 2008; 294(3): H1164 - H1173. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Amino, K. Yoshioka, T. Tanabe, E. Tanaka, H. Mori, Y. Furusawa, W. Zareba, M. Yamazaki, H. Nakagawa, H. Honjo, et al. Heavy ion radiation up-regulates Cx43 and ameliorates arrhythmogenic substrates in hearts after myocardial infarction Cardiovasc Res, December 1, 2006; 72(3): 412 - 421. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. D. Veenstra Gap junction heterogeneity in reentrant ventricular tachycardia Cardiovasc Res, November 1, 2006; 72(2): 196 - 197. [Full Text] [PDF]
|
|
|
|
|
|
R. J.P. Musters Atrial gap junction remodeling: Looking for lost gaps and orphaned connexins in three dimensions Cardiovasc Res, October 1, 2006; 72(1): 5 - 6. [Full Text] [PDF]
|
|
|
|
 |
|
 H. Endo, M. Miura, M. Hirose, J. Takahashi, M. Nakano, Y. Wakayama, Y. Sugai, Y. Kagaya, J. Watanabe, K. Shirato, et al. Reduced Inotropic Effect of Nifekalant in Failing Hearts in Rats J. Pharmacol. Exp. Ther., September 1, 2006; 318(3): 1102 - 1107. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Boissiere, M. Gautier, M.-C. Machet, G. Hanton, P. Bonnet, and V. Eder Doppler tissue imaging in assessment of pulmonary hypertension-induced right ventricle dysfunction Am J Physiol Heart Circ Physiol, December 1, 2005; 289(6): H2450 - H2455. [Abstract] [Full Text] [PDF]
|
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|
|
 |
|
 T. A.B. van Veen, H. V.M. van Rijen, M. J.A. van Kempen, L. Miquerol, T. Opthof, D. Gros, M. A. Vos, H. J. Jongsma, and J. M.T. de Bakker Discontinuous Conduction in Mouse Bundle Branches Is Caused by Bundle-Branch Architecture Circulation, October 11, 2005; 112(15): 2235 - 2244. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. E.J. Teunissen, H. J. Jongsma, and M. F.A. Bierhuizen Regulation of myocardial connexins during hypertrophic remodelling Eur. Heart J., November 2, 2004; 25(22): 1979 - 1989. [Abstract] [Full Text] [PDF]
|
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|
|
 |
|
 C. Forster, S. Kietz, K. Hultenby, M. Warner, and J.-A. Gustafsson Characterization of the ER{beta}-/-mouse heart PNAS, September 28, 2004; 101(39): 14234 - 14239. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. J. Severs, S. R. Coppen, E. Dupont, H.-I Yeh, Y.-S. Ko, and T. Matsushita Gap junction alterations in human cardiac disease Cardiovasc Res, May 1, 2004; 62(2): 368 - 377. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Kostin, S. Dammer, S. Hein, W. P Klovekorn, E. P Bauer, and J. Schaper Connexin 43 expression and distribution in compensated and decompensated cardiac hypertrophy in patients with aortic stenosis Cardiovasc Res, May 1, 2004; 62(2): 426 - 436. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. R. de Groot, T. Veenstra, A. O. Verkerk, R. Wilders, J. P.P. Smits, F. J.G. Wilms-Schopman, R. F. Wiegerinck, J. Bourier, C. N.W. Belterman, R. Coronel, et al. Conduction slowing by the gap junctional uncoupler carbenoxolone Cardiovasc Res, November 1, 2003; 60(2): 288 - 297. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Nygren, C. Kondo, R. B. Clark, and W. R. Giles Voltage-sensitive dye mapping in Langendorff-perfused rat hearts Am J Physiol Heart Circ Physiol, March 1, 2003; 284(3): H892 - H902. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. G. Petrich, X. Gong, D. L. Lerner, X. Wang, J. H. Brown, J. E. Saffitz, and Y. Wang c-Jun N-Terminal Kinase Activation Mediates Downregulation of Connexin43 in Cardiomyocytes Circ. Res., October 4, 2002; 91(7): 640 - 647. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Kostin, G. Klein, Z. Szalay, S. Hein, E. P Bauer, and J. Schaper Structural correlate of atrial fibrillation in human patients Cardiovasc Res, May 1, 2002; 54(2): 361 - 379. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. J. Barker, R. L. Price, and R. G. Gourdie Increased Association of ZO-1 With Connexin43 During Remodeling of Cardiac Gap Junctions Circ. Res., February 22, 2002; 90(3): 317 - 324. [Abstract] [Full Text] [PDF]
|
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|
|
|
|
H. Honjo, M. R Boyett, S. R Coppen, Y. Takagishi, T. Opthof, N. J Severs, and I. Kodama Heterogeneous expression of connexins in rabbit sinoatrial node cells: correlation between connexin isotype and cell size Cardiovasc Res, January 1, 2002; 53(1): 89 - 96. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Chen, X. T. Gan, J. V. Haist, Q. Feng, X. Lu, S. Chakrabarti, and M. Karmazyn Attenuation of Compensatory Right Ventricular Hypertrophy and Heart Failure following Monocrotaline-Induced Pulmonary Vascular Injury by the Na+-H+ Exchange Inhibitor Cariporide J. Pharmacol. Exp. Ther., August 1, 2001; 298(2): 469 - 476. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. A.B. van Veen, H. V.M. van Rijen, and T. Opthof Cardiac gap junction channels: modulation of expression and channel properties Cardiovasc Res, August 1, 2001; 51(2): 217 - 229. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M.W Veldkamp, A.O Verkerk, A.C.G van Ginneken, A Baartscheer, C Schumacher, N de Jonge, J.M.T de Bakker, and T Opthof Norepinephrine induces action potential prolongation and early afterdepolarizations in ventricular myocytes isolated from human end-stage failing hearts Eur. Heart J., June 1, 2001; 22(11): 955 - 963. [Abstract] [PDF]
|
|
|
|
|
|
S. P. Thomas, L. Bircher-Lehmann, S. A. Thomas, J. Zhuang, J. E. Saffitz, and A. G. Kleber Synthetic Strands of Neonatal Mouse Cardiac Myocytes : Structural and Electrophysiological Properties Circ. Res., September 15, 2000; 87(6): 467 - 473. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. J. Jongsma and R. Wilders Gap Junctions in Cardiovascular Disease Circ. Res., June 23, 2000; 86(12): 1193 - 1197. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. J. Barker, R. L. Price, and R. G. Gourdie Increased Association of ZO-1 With Connexin43 During Remodeling of Cardiac Gap Junctions Circ. Res., February 22, 2002; 90(3): 317 - 324. [Abstract] [Full Text] [PDF]
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|
|
|
|
|
T. Koura, M. Hara, S. Takeuchi, K. Ota, Y. Okada, S. Miyoshi, A. Watanabe, K. Shiraiwa, H. Mitamura, I. Kodama, et al. Anisotropic Conduction Properties in Canine Atria Analyzed by High-Resolution Optical Mapping: Preferential Direction of Conduction Block Changes From Longitudinal to Transverse With Increasing Age Circulation, April 30, 2002; 105(17): 2092 - 2098. [Abstract] [Full Text] [PDF]
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