Download Myocardial High Energy Phosphate Stores in Cardiac Hypertrophy

Myocardial High Energy Phosphate Stores in
Cardiac Hypertrophy and Heart Failure
By Peter E. Pool, M.D., James F. Spann, Jr., M.D., Robert A. Buccino, M.D.,
Edmund H. Sonnenblick, M.D., and Eugene Brounwold, M.D.
With the Technical Assistance of Shirley C. Seogren
Downloaded from http://circres.ahajournals.org/ by guest on June 12, 2017
ABSTRACT
The aim of this study was to determine whether the impairment of contractility in the myocardium obtained from hypertrophied and failing hearts is
due to a decreased store of energy and whether an imbalance exists between
energy production and energy utilization in these hearts in vivo. Right ventricular hypertrophy and right ventricular failure were produced in cats by
constriction of the main pulmonary artery. Concentrations of creatine phosphate
(CP) and adenosine triphosphate (ATP) were determined in samples removed from the right ventricles of these animals during life and in papillary
muscles isolated from these same hearts. The papillary muscles from the cats
with hypertrophy and failure exhibited depressed intrinsic contractility. The
stores of ATP were normal in both the ventricular muscles and the papillary
muscles in animals with hypertrophy and failure. Although the stores of CP
were significantly depressed in the right ventricles of these animals, the stores
had increased toward normal in papillary muscles isolated from these same
hearts in vitro. The finding of normal energy stores in the papillary muscles
from animals with hypertrophy and failure indicates that their intrinsically
depressed contractility cannot be due to reductions of their energy stores
in vivo such as were found in the samples from the ventricular wall. The
differences between high energy phosphate stores in failing heart in vivo and
these stores as they are replenished in vitro also indicate that the in vivo
depressions are secondary to an imbalance between energy production and
energy utilization in the overloaded, hypertrophied and failing heart.
ADDITIONAL KEY WORDS
creatine phosphate
creatine
energy metabolism
energetics
• A major unresolved question concerning
the mechanism of heart failure is whether a
specific biochemical abnormality is responsible for this condition. Numerous investigations of this question have involved the study
of patients with naturally occurring decompensation as well as animals with experimentally produced forms of heart failure. At
various times it has been suggested that defects exist in energy production (1-4), energy
storage (5-9), and energy utilization (10,
11). On the other hand, other studies have
suggested that these three basic metabolic
From the Cardiology Branch, National Heart Institute, Bethesda, Maryland 20014.
Accepted for publication July 17, 1967.
CircuUuon ReiMrcb, Vol. XXI, StpHrattr 1967
adenosine triphosphate
right ventricular wall
cat papillary muscle
functions are entirely normal in heart failure
(10, 12, 13). Furthermore, when evidence
for specific defects in energy metabolism was
obtained, it was unclear whether the observed
defect was causally related to the heart failure state.
Considerable information can be obtained
by measuring the myocardial high energy
phosphate stores, creatine phosphate (CP)
and adenosine triphosphate (ATP). While
such determinations do not provide a complete description of the energy metabolism of
cardiac muscle, they do permit an assessment
of the balance between energy production
and energy utilization. However, until recently, it has been difficult to measure myocar365
366
POOL, SPANN, BUCCINO, SONNENBLICK, BRAUNWALO
Downloaded from http://circres.ahajournals.org/ by guest on June 12, 2017
dial high energy phosphate stores with accuracy because of the great lability of these
stores. The importance of extremely rapid
freezing techniques in the accurate determination of these energy stores was first
demonstrated by Wollenberger and associates
in 1958 (14). Subsequently, modifications of
this technique (15) and improved analytic
methods (16-18) have made possible the accurate determination of these stores.
The development of a technique which allows detailed evaluation of the contractile
function of the myocardium obtained from
the hearts of animals with ventricular hypertrophy and heart failure combined with modern techniques for detennining energy stores
provided a unique opportunity for studying
the question of a specific biochemical defect
in heart failure. It was the aim of this study
to determine whether the recently demonstrated impairment of cardiac contractility in the myocardium obtained from hypertrophied and failing hearts (19) is due to a
decreased store of energy and whether an imbalance between energy production and energy utilization exists in these hearts in vivo.
Methods
Right ventricular hypertrophy and right ventricular failure were produced in adult cats by
constriction of the main pulmonary artery. Twenty-one to 90 days following operation, circulatory
dynamics was evaluated by means of right heart
catheterization under barbiturate anesthesia. On
the basis of measurements of the right ventricular
end-diastolic pressure, cardiac output, and arteriomixed venous O2 difference, as well as the findings provided by autopsv, it was possible to separate animals with hypertrophy from those with
overt heart failure (19).
Following hemodynamic study in the intact
animal, the chest was opened widely and samples
of right and left ventricular myocardium were
obtained with a 6-inch mastoid rongeur, while
intermittent positive pressure ventilation was
provided via a tracheostomy. Care was taken to
insure a stable hemodynamic state and an adequate O2 supply at the time of the biopsy; at this
time the arterial blood O ; content was not different from that of normal cats, and in the few cats
in which it was measured, arterial (X saturation
was greater than 90%. The sample from the second
ventricle was analyzed only if obtained within 3
sec of the sample from the first ventricle. The
samples were obtained from the free wall of each
ventricle, and those from the right ventricle wert
within 5 mm of the base of the papillary muscle.
All of the samples were through the ventricular
wall and usually tissue from the right ventricle
was removed first. The specimens, weighing an
average of 22 mg, were frozen in liquid nitrogen
within 0.5 sec. After these specimens were obtained, the hearts were rapidly extirpated and
right ventricular papillary muscles were isolated.
These were placed in a myograph in Krebs solution at 26°C (17), continuously aerated via a
sintered glass gas dispersion tube with 95% O25% CO2, and allowed to equilibrate for 30 min
while being stimulated to contract isometrically at
a frequency of 12/min at low resting tensions.
Following determination of the active length-tension curve, each muscle was stimulated to contract isometrically at the top of this curve for an
additional 30 min, and then was rapidly frozen
between contractions by quickly removing the
muscle bath and replacing it with a beaker of
2-methylbutane (isopentane) previously cooled in
liquid nitrogen to —150° to —160°C. This procedure freezes the muscle without further contraction.
The papillary muscles and samples removed
from the myocardium were stored in liquid
nitrogen. On the day of assay, each tissue was pulverized at —50°C as previously described (18),
and perchloric acid extracts were prepared. CP
and inorganic phosphate (Pj) were determined
by the phosphomolybdate method of Fiske and
Subbarow (20) as modified by Furchgott and
De Gubareff (16), creatine concentrations by the
a-naphthol-diacetyl method of Ennor and Rosenberg (21, 22), and ATP by a modification of the
firefly luminescence method of Strehler and McElroy (23). Modifications of these procedures
have been described previously in detail (17).
Statistical tests of the significance of the difference between group means and paired samples
(right ventricle vs. papillary muscle) were made
by the appropriate f-tests (24).
Results
The extent of the elevation of right ventricular weight, right ventricular systolic and
end-diastolic pressures and the absence of an
increase in ventricular water content in the
animals with right ventricular hypertrophy
and heart failure are presented in detail elsewhere (19).
SAMPLES OF VENTRICULAR
DURING LIFE
MUSCLE OBTAINED
Right Ventricle—In. specimens of right venRtitfcb,
Vol. XXI, Stpttmht 1967
367
MYOCARDIAL ENERGY STORES IN HEART FAILURE
CREATINE
ATP
PHOSPHATE
• - Control - 17
E 3 - RVH - 10
E 3 - CHF - 8
.2?
o
E
o
E
Downloaded from http://circres.ahajournals.org/ by guest on June 12, 2017
,02 J -p<.05 J
p<.01
-N.'s. -
'
FIGURE 1
In vivo right ventricular high energy phosphate stores in normal cats and cats with right ventricular hypertrophy (RVH) and congestive heart failure (CHF). Units for ordinates are fimoles
per gram wet weight of heart muscle. Vertical lines with cross bars = ± 1 SEM.
tricle obtained from cats in congestive heart
failure (CHF), levels of ATP and Pf did not
differ significantly from those found in right
ventricular tissue obtained from normal animals or animals with right ventricular hypertrophy (RVH) (Fig. 1). On the other hand,
the concentrations of both total creatine and
CP were significantly reduced; the total
creatine averaged 15.5±0.6 (SEM), 12.5±
0.9 and 9.7 ±0.6 /nmoles/g respectively in the
normal, RVH and CHF groups, while the CP
averaged 7.22 ±0.49, 5.42 ±0.45 and 4.15 ±
0.40 fimoles/g in these three groups. The
fraction of total creatine present as CP was
essentially identical in all three groups, averaging 46%, 43%, and 43% respectively (Table 1).
Left Ventricle.—As in the right ventricle,
average concentrations of ATP and Ps were
similar in the left ventricles of animals in all
three groups. Both total creatine and CP were
significantly lower in the CHF group than
in the normal and RVH groups (P < .05).
However, no significant differences in creatine and CP concentrations were observed
between the normal and RVH groups. The
fraction of total creatine present as CP was
again similar in all three groups, averaging
Circulation Research, Vol. XXI, September 1967
45% in hearts of control animals, 52% in the
RVH group and 49% in the CHF group.
PAPILLARY MUSCLES
Other papillary muscles from the same
groups of animals as those used for biochemical analysis were subjected to detailed analysis of myocardial mechanics (19); the maximum velocity of shortening of the unloaded
muscle (Vmilx), the rate of development of
tension and the peak isometric tension were
depressed. However, it was considered necessary to obtain an index of the mechanical performance of the muscles actually subjected to
analysis of high energy phosphate stores. The
isometric tension at the peak of the active
length-tension curve (P o ) was therefore determined in each muscle. This variable averaged 5.2 ±0.8 g/mm2 in the muscles removed
from 7 control animals, and was significantly
lower in the 9 muscles obtained from animals
with hypertrophy, in which it averaged 1.8 ±
0.4 g/mm2, and in the 7 muscles obtained
from animals with heart failure, in which it
averaged 1.1 ± 0.5 g/mm2.
As observed in the biopsies of the right
ventricular wall, there were no significant dif-
368
POOL, SPANN, BUCCINO, SONNENBLICK, BRAUNWALD
TABLE 1
Rkht ventricle
Ounolei/g)
Eipt
8
g
10
n
12
Downloaded from http://circres.ahajournals.org/ by guest on June 12, 2017
13
14
25
26
27
28
29
30
31
57
58
60
61
62
65
MEAN
±1SE
4
7
8
9
11
12
14
17
31
32
34
MEAN
±lSE
5
6
10
13
16
20
21
28
29
30
MEAN
±lSE
Cr
CP
ATP
17.8
16.4
13.8
18.6
17.5
18.4
21.4
15.1
15.0
13.6
12.5
14.2
14.0
15.0
.16.9
14.4
17.2
13.8
12.4
12.8
15.5
0.6
7.35
7.70
6.76
10.20
9.36
4.61
4.61
3.22
3.67
4.91
Control
4.43
3.42
5.50
6.12
6.40
11.80
9.13
5.85
4.94
4.34
7.29
5.75
4.96
5.14
4.73
4.95
5.51
3.53
3.16
4.52
2.78
4.23
3.34
6.45
7.54
6.55
5.40
6.26
5.83
7.04
5.71
6.27
5.26
2.94
4.37
2.22
6.11
4.05
5.89
7.22
0.49
5.02
5.13
0.22
2.58
4.10
0.32
9.6
8.2
11.0
14.4
13.4
13.6
12.5
14.7
8.4
15.8
16.4
12.5
0.9
3.52
3.71
4.55
6.04
6.35
5.69
Hypertrophy
3.62
3.70
4.54
3.98
4.55
3.96
5.86
5.16
3.94
5.86
5.21
5.59
5.52
4.71
6.11
8.01
5.42
0.45
5.08
5.41
5.41
7.54
5.13
0.37
7.69
3.66
3.76
3.47
4.67
0.45
3.73
5.99
Failure
6.48
4.61
4.21
3.21
4.29
5.65
2.26
4.45
4.99
3.21
4.15
0.40
3.67
6.56
2.62
5.69
5.05
6.68
5.06
0.55
5.54
10.60
3.22
3.36
2.79
3.19
4.80
1.02
12.7
9.6
9.9
10.0
11.1
8.7
5.5
9.0
10.8
9.6
9.7
0.6
Pi
Cr
22.3
15.8
15.5
17.8
14.1
19.8
20.9
18.4
17.6
18.0
13.6
16.5
17.0
16.0
18.4
18.2
16.9
14.7
18.7
17.4
0.5
12.1
16.1
14.9
14.0
18.0
14.9
13.8
25.0
15.8
22.7
20.4
17.0
1.3
13.1
11.2
14.8
11.2
12.3
11.8
12.7
12.2
12.7
9.8
12.2
0.4
Left v(mtrlcle
(junol e»/g)
ATP
CP
Pi
8.09
10.90
9.26
7.97
8.82
11.10
10.40
5.24
6.13
3.93
4.42
6.10
5.50
6.91
3.94
4.89
6.77
3.76
4.71
5.16
7.70
5.49
4.98
5.16
4.74
7.30
5.77
4.29
5.74
5.26
6.26
5.87
6.73
6.36
4.46
7.23
7.85
0.68
5.76
0.31
5.32
0.38
6.13
8.99
4.30
6.13
3.56
8.59
7.79
8.81
4.81
7.20
2.66
5.09
14.20
8.46
7.74
7.57
6.59
6.08
7.09
8.78
1.06
6.09
6.26
0.55
5.04
5.37
0.80
4.69
7.77
5.95
5.32
6.80
5.06
4.36
5.10
4.58
5.74
8.81
4.79
4.92
6.34
8.66
3.79
6.55
11.50
3.38
5.91
0.75
5.17
6.04
0.55
4.66
5.79
1.09
Cr = creatine; CP = creatine phosphate; ATP = adenosine triphosphate; P, = inorganic
phosphate.
CircuUuon Rtsetrcb, Vol. XXI, Septtmbtr 1967
MYOCARDIAL ENERGY STORES IN HEART FAILURE
369
10 r
• - Control-7
2 - RVH - 5
E3 - CHF - 7
8
6
O
Downloaded from http://circres.ahajournals.org/ by guest on June 12, 2017
0
PAPILLARY
MUSCLE
RIGHT
VENTRICLE
FIGURE 2
ATP stores in the right ventricle in vivo are compared with those found in papillary muscles
isolated from the same hearts and studied in vitro. Abbreviations and symbols as in Figure 1.
10
D - Control - 7
E3 - RVH - 5
E 3 - CHF - 7
§
ih
2
5
RIGHT
VENTRICLE
PAPILLARY
MUSCLE
DIFFERENCE
(PM-RV)
FIGURE 3
Creatine phosphate stores in the right ventricle in vivo are compared iirith those in papillary
muscles isolated from the same hearts and studied in vitro. Difference is between CP concentration in the papillary muscle and that in the right ventricle. PM =: papillary muscle; RV =
right ventricle; other abbreviations and symbols as in Figure 1.
Circulation Research, Vol. XXI, September 1967
370
POOL, SPANN, BUCCINO, SONNENBLICK, BRAUNWALD
TABLE 2
Expt
CP
(^moles/g)
RV
PM
TABLE 3
CP
ATP
RV
PM
PM
Kipt.
Control
A
B
C
D
E
F
G
MEAN
±lSE
9.13
7.29
6.55
6.46
5.89
7.04
5.10
6.78
0.52
9.27
7.44
8.00
8.08
5.60
7.40
6.26
7.44
0.50
4.96
5.51
5.71
5.26
5.02
5.75
5.16
5.34
0.13
5.00
5.45
6.18
5.25
4.23
6.52
4.46
5.30
0.34
4.5
6.4
3.5
6.0
7.3
6.9
1.8
5.2
0.8
Hypertrophy
Downloaded from http://circres.ahajournals.org/ by guest on June 12, 2017
H
I
J
K
L
MEAN
±lSE
6.42
4.71
6.11
2.46
4.95
4.93
0.78
9.98
6.65
7.28
4.37
8.35
7.33
1.04
5.54
5.41
5.41
5.16
4.89
5.28
0.13
8.35
5.85
6.01
4.46
5.10
5.95
0.59
4.3
0.8
2.2
1.7
3.0
2.4
0.7
5.05
6.60
5.29
4.32
4.26
5.61
5.38
5.22
0.33
5.97
5.84
4.87
4.82
3.58
4.90
6.15
5.16
0.37
0.3
2.7
0.5
0.5
0.7
0.1
3.1
1.1
0.5
Failure
M
N
O
P
Q
R
S
MEAN
±lSE
4.99
6.36
3.87
3.45
4.85
4.65
2.69
4.41
0.49
7.76
12.30
7.06
5.06
8.03
554
7.52
7.61
0.96
RV = right ventricle; PM = papillary muscle.
ferences among the ATP concentrations in
the papillary muscles obtained from the three
groups of animals, these values averaging 5.30
±0.34, 5.95 ±0.59 and 5.16 ±0.37 ^.moles/g
in muscles obtained from control cats and
those with hypertrophy and heart failure respectively (Fig. 2). However, in contrast to
the differences in concentrations of CP observed in right ventricular wall, no differences
in these concentrations were observed in the
three groups of papillary muscles (Table 2)
(Fig. 3).
COMPARISONS BETWEEN VENTRICULAR WALLS AND
PAPILLARY MUSCLES
No significant differences were observed
between the ATP concentrations of the right
ventricular wall and the papillary muscles
A
B
C
D
MEAN
±lSE
ATP
l
RV
PM
Hypertrophy-Resting
7.10
11.40
2.53
7.59
3.78
5.90
10.60
6.42
4.96
8.90
1.24
1.48
RV
PM
5.75
6.65
5.21
5.54
5.79
0.36
5.83
5.56
5.90
4.54
5.46
0.36
RV = right ventricle; PM = papillary muscle.
obtained from the same ventricles (Fig. 2).
However, the CP concentrations were significantly higher in the papillary muscles; in
the control animals this difference averaged
0.7 ±0.3 /xmoles/g, which was equivalent to
]0$ of the right ventricular concentration
(Fig. 3). In the animals in the RVH and CHF
groups the differences between CP concentration in papillary muscles andrightventricles were significantly greater than in the
control group. In the RVH group this difference (2.4 ±0.5 ^moles/g) averaged 49$ of
the right ventricular concentration, while in
the animals with CHF, this difference (3.2 ±
0.7 //.moles/g) averaged 73$ of therightventricular concentration (Table 2 and Fig. 3).
To determine whether the energy stores
were related to the mechanical activity of the
papillary muscles, the right ventricles and papillary muscles from a separate group of four
animals with hypertrophy were studied. In
this group, however, the papillary muscles,
following 30 min of equilibration in vitro with
stimulation 12 times/min, were allowed to
rest for 30 min instead of being stimulated at
a rate of 12/min. In this experiment, there
was an even greater difference between the
CP concentrations in the papillary muscles
and the right ventricles (3.9 ±0.7 /xmoles/g)
(Table 3) than that which was found when
the contracting papillary muscles from cats
with hypertrophy were compared to their
right ventricles (2.4 ± 0.5 ^imoles/g) (Table 2).
Discussion
The objective of this investigation was to
determine whether (1) an abnormality in
Circtlation Rtiarcb, Vol. XXI, Stfitmttr 1967
MYOCARDIAL ENERGY STORES IN HEART FAILURE
Downloaded from http://circres.ahajournals.org/ by guest on June 12, 2017
myocardial energy stores occurs in cardiac
hypertrophy and congestive heart failure;
(2) the impairment of myocardial contractility in these states can be attributed to a depression of myocardial energy stores; and
(3) an imbalance exists between energy
production and energy utilization in failing
and hypertrophied hearts.
While ATP stores were maintained, a significant depression of CP concentrations occurred in the right ventricles of cats in the
RVH and CHF groups. Although in one earlier study, such depressions of energy stores
in chronic experimental congestive heart failure were not observed (10), the study was
made before the general availability of the
present, more accurate methods for measuring these compounds. However, the recent
findings of Fox et al. (8) in dogs with chronic
congestive heart failure and those of Feinstein (7) in the failing heart of the guinea
pig are in general accord with ours. It is interesting, however, that in these studies (7,
8), modest depressions of ATP stores were
also reported, while we found no changes in
the concentrations of this compound, despite
the presence of moderately severe ventricular
hypertrophy and failure. Perhaps in the presence of more severe or prolonged stresses,
these stores might be reduced. Fox et al. (8)
found no changes in CP, ATP or creatine in
the right ventricles of dogs with right ventricular hypertrophy without failure; we observed a significant depression both of total
creatine and CP in cats with simple right ventricular hypertrophy, and Minton et al. (11)
made similar observations in rats with cardiac
hypertrophy. It is possible that the absence of
significant changes in energy stores in the
hypertrophied hearts reported by Fox et al.
was due to a lesser degree of hypertrophy,
although this was not evaluated in their study.
In an earlier study from this laboratory Chidsey and associates (25) reported that cardiac
CP and ATP stores were not depressed in
patients with congestive heart failure. However, the interpretation of this finding is limited by the fact that the hypertrophied right
CircmUHtm Rutrcb,
VoL XXI, Stpumbtr
1967
371
ventricles in patients with tetralogy of Fallot
served as controls.
The depressions of CP and creatine stores
noted in the left ventricles of the animals
with primary right ventricular failure suggests( that the left ventricle may also be susceptible to the biochemical stresses imposed
on the right ventricle if they are severe enough.
Similar effects of right ventricular failure on
the left ventricle have already been demonstrated both, hemodynamically and biochemically (10, 26-28).
From the results of our studies on papillary
muscles, it is clear that the depression of
myocardial contractility found in hypertrophy
and congestive heart failure and defined as a
depression in both the maximum force (P o )
and intrinsic speed of contraction (V mai ) of
the myocardium (19) is not a function of
decreased energy stores. This finding correlates with the observation of normal oxidative phosphorylation in mitochondria isolated
from these hearts studied in vitro (29). Although the mechanical function of these muscles obtained from failing and hypertrophied hearts was depressed throughout the
experiment, energy stores were "restored"
from the low in vivo levels to levels equal to
those observed in papillary muscles obtained
from normal cats (Fig. 3). Although it was
found impossible to determine high energy
phosphate stores in the papillary muscle in
vivo because the muscle cannot be excised
rapidly under physiologic conditions, it is not
likely that its in vivo energy stores would be
different from those found in the specimen of
the right ventricle, which was taken within
5 mm from the origin of the papillary muscle.
In addition, since it has been shown that the
papillary muscles obtained from hypertrophied and failing hearts undergo the characteristic anatomic and physiologic changes
of these conditions (19), it is expected that
they also participate in the biochemical
changes occurring in the remainder of the
hypertrophied and failing heart.
In hypertrophied and failing hearts the
small normal differences between CP concentrations in the in vivo right ventricle and
372
POOL, SPANN, BUCCINO, SONNENBLICK, BRAUNWALD
Downloaded from http://circres.ahajournals.org/ by guest on June 12, 2017
isolated papillary muscle are significantly
larger. Hochrein and Doling (30) have
shown that increasing work alone in the intact heart may lower CP stores. In the in
vitro papillary muscles in the present study,
CP levels are higher than in vivo, which is
consonant with the reduced work load in
vitro as compared to that found in vivo. This
reduction in work load in the hypertrophied
and failing muscle is accompanied by a
greater increase in CP levels than is found in
normal muscles. Taken together these findings would indicate that there is an imbalance
between energy production and energy utilization in the heart, and that this may be
augmented in the presence of hypertrophy
and failure. Whether this is due to an excessive increase in energy utilization or an inadequate increase in energy production cannot be answered at this time. Nevertheless,
this imbalance occurs despite the normal in
vitro function of mitochondria prepared from
these hearts (29).
In conclusion, it was observed in this investigation that reductions in myocardial energy stores exist in the presence of moderately severe degrees of ventricular hypertrophy
in vivo and that these reductions are more
marked in the presence of heart failure. The
papillary muscles from these same hearts exhibited depressed contractility. However, the
finding of normal energy stores in these papillary muscles indicate that their intrinsically
depressed contractility cannot be due to the
reductions of their energy stores. The differences between high energy phosphate stores
in failing heart muscle studied in vivo and
in vitro also indicates that the in vivo depressions are secondary to an imbalance between
energy production and energy utilization in
the overloaded, hypertrophied and failing
heart.
Acknowledgment
The assistance of Nancy Dittemore, Robert M.
Lewis, and Richard McGill is gratefully acknowledged.
tion in sarcosomes from experimentally-induced
failing rat heart. Proc. Soc. Exptl. Biol. Med.
117: 380, 1964.
2.
3.
4.
5.
6.
7.
1. ARGUS, M. F., ARCOS, J. C, SABDESAI, V. M., AND
OVERBY, J. L.: Oxidative rates and phosphoryla-
Some metabolic characteristics of mitochondria
from chronically overloaded, hypertrophied
hearts. Exptl. MoL Pathol. 2: 251, 1963.
SCHWARTZ, A., AND LEE, S.: Study of heart mitochondria and glycolytic metabolism in experimentally induced cardiac failure. Circulation
Res. 10: 321, 1962.
GEBTLER, M. M.: Differences in efficiency of energy transfer in mitochondrial systems derived
from normal and failing hearts. Proc. Soc. Exptl.
Biol. Med. 106: 109, 1961.
FLECKENSTEIN, A.: Die Bedeutung der energiereichen Phosphate fur Kontraktilitat und Tonus
des Myokards. Verhandl. Deut. Ges. Inn. Med.
70: 81, 1964.
FURCHGOTT, R. F., AND LEE, K. S.: High energy
phosphates and the force of contraction of cardiac muscle. Circulation 24: 416, 1961.
FEINSTEEN, M. B.: Effects of experimental congestive heart failure, ouabain, and asphyxia on
the high-energy phosphate and creatine content of the guinea pig heart. Circulation Res.
10: 333, 1962.
8. Fox, A. C., WIKI.ER, N. S., AND REED, G. E.:
High energy phosphate compounds in the myocardium during experimental congestive heart
failure: Purine and pyrimidine nucleotides, creatine, and creatine phosphate in normal and
in failing hearts. J. Clin. Invest. 44: 202,
1965.
9. FURCHGOTT, R. F., AND DEGUBAREFF, T.: High
energy phosphate content of cardiac muscle
under various experimental conditions which
alter contractile strength. J. Pharmacol. Exptl.
Therap. 124: 203, 1958.
10. OLSON, R. E.: Myocardial metabolism in congestive heart failure. J. Chronic Diseases 9: 442,
1959.
11. MLNTON, P. R., ZOLL, P. M., AND NORMAN, L. R.:
Levels of phosphate compounds in experimental cardiac hypertrophy. Circulation Res. 8:
924, 1960.
12. BUCKLEY, N. M., AND TSUBOT, K. K.: Cardiac
nucleotides and derivatives in acute and chronic
ventricular failure of the dog heart. Circulation
Res. 9: 618, 1961.
13. BING, R. J.: Cardiac metabolism. Physiol. Rev.
45: 171, 1965.
14.
References
WOLLENBERGER, A . , KLEITKE, B . , AND R A A B E , G . :
WOLLENBEHGER, A., KRAUSE, E . G., AND W A H L E R ,
B. E.: Orthophosphatund Phosphokreatingehalt des Herzmuskels. Naturwissenschaften 45:
294, 1958.
CircuUsio* Rvurck, Vol. XXI, Septtmbf 1967
MYOCARDIAL ENERGY STORES IN HEART FAILURE
373
Kaplan, New York, Academic Press, vol. 3,
p. 871, 1957.
15. POOL, P. E., COVELL, J. W., CHIDSEY, C. A., AND
BRAUNWALD, E.: Myocardial high energy phosphate stores in acutely induced hypoxic heart
failure. Circulation Res. 19: 221, 1966.
24.
A New Approach. Brooklyn, The Free Press of
Glencoe, Inc., 1956, p. 420.
16. FUHCHCOTT, R. F., AND DEGUBAREFF, T.: Deter-
mination of inorganic phosphate and creatine
phosphate in tissue extracts. J. Biol. Chem.
223: 377, 1956.
17.
25.
28.
ture extraction of small samples of tissue.
Chemist-Analyst. 56: 38, 1967.
Downloaded from http://circres.ahajournals.org/ by guest on June 12, 2017
of cardiac muscle obtained from cats with experimentally produced ventricular hypertrophy
and heart failure. Circulation Res. 21: 341,
1967.
20.
27.
28.
tine. J. Biol. Chem. 81: 629, 1929.
29.
ENNOR, A. H.: Determination and preparation of
N-phosphates of biological origin. In Methods
in Enzymology, edited by S. P. Colowick and
N. O. Kaplan, New York, Academic Pre.ss,
vol. 3, p. 850, 1957.
23. STREHLER, B. L., AND MCELROY, W. G.: Assay
of adenosine triphosphate. In Methods in Enzymology, edited by S. P. Colowick and N. O.
OrctthzHon Rtitircb, Vol. XXI, Stptmhtr 1967
CHIDSEY, C. A., KAISEH, G. A., SONNENBLICK, E.
H., SPANN, J. F., AND BRAUNWALD, E.: Cardiac
norepinephrine stores in experimental heart
failure in the dog. J. Clin. Invest. 43: 2386,
1964.
21. ENNOR, A. H., AND ROSENBERG, H.: Determina-
22.
BARGER, A. C , ROE, B. B., AND RICHARDSON,
G. S.: Relation of valvular lesions and of exercise to auricular pressure, work tolerance, and
to the development of chronic congestive failure in dogs. Am. J. Physiol. 169: 384, 1952.
FISKE, C. H., AND SuBaABOw, Y.: Phosphocrea-
tion and distribution of phosphocreatine in
animal tissues. Biochem. J. 51: 606, 1952.
CHANDLER, B. M., SONNENBLICK, E. H., SPANN,
J. F., JR., AND POOL, P. E.: Myofibrillar ATPase
activity in experimental right ventricular hypertrophy and right ventricular failure. Physiologist
10: 140, 1967.
18. POOL, P. E., AND SEACHEN, S. C : LOW Tempera-
19. SPANN, J. F., JR., BUCCINO, R. A., SONNENBLICK,
E. H., AND BRAUNWALD, E.: Contractile state
CHIDSEY, C. A., WEJNBACH, E. C , POOL, P. E.,
AM> MORROW, A. G.: Biochemical studies of
energy production in the failing human heart.
J. Clin. Invest. 45: 40, 1966.
POOL, P. E., AND SONNENBLICK, E. H.: Mechano-
chemistry of cardiac muscle: I. The isometric
contraction. J. Gen. Physiol. 50: 951, 1967.
WALLIS, W. A., AND ROBERTS, H. V.: Statistics:
SOBEL, B. E., SPANN, J. F., JR., POOL, P. E.,
SONNENBLICK, E. H., AND BRAUNWALD, E.:
Normal oxidative phosphorylation in mitochondria from the failing heart. Circulation Res.
21: 355, 1967.
30.
HOCHREIN, H.. AND DORING, H. J.: Die ener-
giereichen Phosphate des Myokards bei Variation der Belastungsbedingungen. Pfluegers Arch.
Ges. Physiol. 271: 548, 1960.
Myocardial High Energy Phosphate Stores in Cardiac Hypertrophy and Heart Failure
PETER E. POOL, JAMES F. SPANN, Jr., ROBERT A. BUCCINO, EDMUND H.
SONNENBLICK, EUGENE BRAUNWALD and Shirley C. Seogren
Downloaded from http://circres.ahajournals.org/ by guest on June 12, 2017
Circ Res. 1967;21:365-374
doi: 10.1161/01.RES.21.3.365
Circulation Research is published by the American Heart Association, 7272 Greenville Avenue, Dallas, TX 75231
Copyright © 1967 American Heart Association, Inc. All rights reserved.
Print ISSN: 0009-7330. Online ISSN: 1524-4571
The online version of this article, along with updated information and services, is located on the
World Wide Web at:
http://circres.ahajournals.org/content/21/3/365
Permissions: Requests for permissions to reproduce figures, tables, or portions of articles originally published in
Circulation Research can be obtained via RightsLink, a service of the Copyright Clearance Center, not the
Editorial Office. Once the online version of the published article for which permission is being requested is
located, click Request Permissions in the middle column of the Web page under Services. Further information
about this process is available in the Permissions and Rights Question and Answer document.
Reprints: Information about reprints can be found online at:
http://www.lww.com/reprints
Subscriptions: Information about subscribing to Circulation Research is online at:
http://circres.ahajournals.org//subscriptions/