19

Contractile Properties of Small Arterial Resistance

Vessels in Spontaneously Hypertensive and
Normotensive Rats
MICHAEL J. MULVANY AND WILLIAM HALPERN

SUMMARY The small arteries play an important functional role in establishing the increased peripheral resistance
found in essential hypertension. This paper concerns the direct measurement of the intrinsic mechanical and
contractile properites of two categories of small arterial resistance vessels in the mesenteric bed of 5-month-old
normotensive Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR). The vessels had mean internal
diameters of 246 fim and 153 /*m when relaxed at 100 mm Hg effective transmural pressure. Segments (1 mm) were
mounted in such a way that internal circumference could be controlled and the circumferential wall tension (T)
measured. After mounting, each vessel was maximally activated (by K* depolarization at 37°C in the presence of 5
mM Ca2+) at an internal circumference for which AT was approximately maximal, where AT = T actlve - T relaxed . From
the average values of AT measured we have estimated (on the basis of Laplace's equation) that the SHR vessel would
average values of AT measured we have estimated (on the basis of Laplace's equation) that the SHR vessels would
have been able to contract against 34% greater pressures than the WKY vessels (P < 0.001). Optical measurements
of the dimensions of these vessels showed a 23% greater wall thickness in the SHR vessels (P < 0.02). There were no
significant differences in the calculated active wall stresses of the SHR and WKY vessels; this suggests that the greater
contractility found in the SHR vessels may be due to their having a greater smooth muscle cell content. These vessel
measurements have been examined as well as the rats' blood pressures and heart to body weight ratios. The
comparison points to the possibility that the disturbance to the cardiovascular regulatory system which results in
hypertension produces similar cellular responses in both the myocardium and the peripheral vasculature.

HUMAN essential hypertension is associated with an ele- Although there is evidence to suggest that the various
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vated blood pressure but a normal cardiac output.1 The
total peripheral resistance is therefore elevated. The cause
of this elevation is not clear but a genetic factor seems to
be involved.2-3 As a means to investigate genetic hyper-
tension in detail, a strain of spontaneously hypertensive
rats (SHR) has been developed by Okamoto and his col-
leagues4 from a colony of Wistar-Kyoto rats (WKY).
These SHR reach a stage of established hypertension by
the age of 5 months. At this time their systolic blood
pressure is about 50% higher than that of their WKY
controls although, as in human essential hypertension,
their cardiac output is normal.5
The vascular bed of these rats contains vessels with
diameters ranging from 3 mm in the aorta to about 7 ^m
in the capillaries. Although all these vessels must contrib-
ute to the resistance of the vascular bed to some extent, it
is in general the smaller arterial resistance vessels which
present the greatest resistance, and which are most in-
volved in regulating blood flow and capillary pressure.6
Perfusion studies on isolated vascular beds7"" have pro-
vided valuable insight into their general mode of action.
Such studies are not, however, able to analyze the specific
contribution of the various parts of the vascular bed.
From the Biophysics Institute, Aarhus University, Aarhus, Denmark,
and Department of Physiology and Biophysics, University of Vermont,
Burlington, Vermont.
Supported by National Institutes of Health Grant 5R01 HL-17335.
Address for reprints: M.J. Mulvany, Biophysics Institute, Aarhus Uni-
versity, Universitetsparken, 8000 Aarhus C , Denmark.
Received August 9, 1976; accepted for publication November 24,
1976.
portions of the vasculature react differently,12'13 the work
with isolated vessels has been largely confined to the
aorta,14"16 various conduit arteries,17"19 and smaller arter-
ies having internal diameters greater than 300 jim. !Oi!l
Technical difficulties have hitherto prevented direct data
being obtained from smaller vessels. It is clear however
that such data is of importance in understanding the action
of the vasculature and we22 have therefore developed a
method first proposed by Bevan and Osher23 to make
direct mechanical measurements on intact segments from
small vessels. With this method we have investigated ves-
sels from SHR and WKY having diameters in the range
75-300 ^.m. Our studies have been confined to determin-
ing the intrinsic mechanical and contractile properties of
these vessels. We have therefore limited our experiments
to the examination of vessels either when they were fully
activated or when they were fully relaxed.24
Methods
ANIMALS
The SHR and their WKY controls were bred in our own
colony from a pair provided by the National Institutes of
Health. Systolic blood pressures were measured by the
tail-cuff method. Experiments were performed when the
rats were about 5 months old.
DISSECTION
The arterial resistance vessels used were the first and
second branches off mesenteric arcades.25 The second
branch resistance vessels are those that directly enter the
 20 CIRCULATION RESEARCH
FIGURE 1 Dissection and mounting procedures, a: sketch of
vascular bed with veins dissected away revealing arteries beneath.
The vessel labeled MA is a mesenteric arcade. The vessels labeled 1
and 2 are first and second branch resistance vessels, respectively.
The arterial tree bounded by the dotted line was dissected out after
first removing the surrounding adipose tissue, b: diagram showing
how the proximal end of this arterial tree was then threaded onto a
32- \un tungsten wire (Westinghouse) previously attached to a spec-
imen support (L). The tree than acted as its own guide as it was
pulled along the wire, enabling the wire to be threaded through the
small resistance vessel at the distal end of the tree. This resistance
vessel was then positioned between the jaws of the support, and the
second end of the wire was secured under tension by the second
screw. The unwanted part of the arterial tree was then dissected
away and a second wire was passed through the lumen of the vessel.
This wire was then likewise attached at each end to the second
support (R). c: diagram showing thefinalconfiguration.
intestinal wall; they stem from first branches (Fig. la).
One vessel segment was taken from each rat and threaded
onto two parallel tungsten wires; the wires in turn were
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attached to two specimen supports (Fig. lb and c). During
this process, which took about 1 hour, the vessel was kept
cool (5-15°C) in Ringer's solution. Vessel segments were
about 0.7 mm long (but 1.5 mm long in some experiments
in which larger specimen supports were used). After
mounting, the vessels were equilibrated at an internal
circumference for which the wall tension (see below) was
about 0.2 mN/mm.
APPARATUS
The myograph used is shown in Figure 2. The specimen
supports were mounted, respectively, on a tension trans-
ducer and a displacement device. The latter measured the
circumference to be controlled. A heat exchanger kept the
temperature of the circulated solutions constant to within
±0.2°C. Solution changes were effected by draining the
chamber and then circulating the new solution. The my-
ograph was mounted on the stage of a microscope which
was used to measure the dimensions of the vessels to
within 0.2 (im using a 10x filar micrometer eyepiece
(American Optical, model 426) and a water-immersion
objective. The overall compliance of the apparatus was
measured by observing the movement of the wires after
activation of the vessel. This compliance was 0.5 /xm/mN
at the center of the wires and has not been corrected for.
SOLUTIONS
Vessels were equilibrated in a modified Ringer's solu-
tion26 containing (ITIM): NaCl, 119; NaHCO3, 14.9; KC1,
4.7;KH 2 PO 4 , l.l8;MgSO 4 , 1.17;CaCI2, 1.6; ethylenedi-
aminetetraacetic acid (EDTA), 0.026; glucose, 5.5; and
VOL. 41, No. 1, JULY 1977
activated or relaxed in a K+ solution in which there was an
equimolar substitution of KCI for NaCl and a substitution
of 5 mM CaCl2 (activating solution) or 1 mM ethylene
glycol-bis[/3-aminoethyl ether]-/V,/V'-tetraacetic acid)
(EGTA) (relaxing solution) for the 1.6 mM CaCl2. Graded
activation was obtained by varying the CaCl2 concentra-
tion in the K+ solution. All solutions were adjusted to pH
7.4, oxygenated externally with 95% O2-5% CO2 and
circulated at 8 ml/min at 37°C. EDTA and EGTA were
obtained from Sigma.
VESSEL DIMENSIONS
The following dimensions were measured with the ves-
sel set to internal circumference, L, (see below): mean
wall thickness, w, (average of 5-10 measurements); mean
wire thickness, d (average of six measurements); mean
distance between the inner edges of the wires, f, (average
of three measurements); mean vessel segment length, g,
(average of two measurements). As the vessel wall was
found to be flat between the wires the internal circumfer-
ence, L,, was calculated from:
L, = (77 + 2)d + 2f,.
Dimensions were measured with the vessel in Ringer's
solution after it had been activated at least twice. The
internal circumference when the vessel was stretched or
released from L, was determined from the displacement
transducer output. The segment length was not found to
change with stretch or release or following activation.
FIGURE 2 View of myograph. The supports (L, R) shown in
Figure Ic were mounted, respectively, on a force transducer (DSC
6, Kistler-Morse, lowest resonant frequency with support mounted-
400 Hz) and a piezoelectric vibrator (PZ40, Burleigh; frequency
range, 0-2,000 Hz). The latter was mounted on a micrometer
translator. An eddy-current displacement transducer (KD 2300.5
SU, Kaman Sciences) monitored movement of R. The chamber
was heated by thermostatically controlled water circulating through
the myograph block. Solutions entered and left the chamber as
indicated by the arrows. The myograph was constructed of Teflon-
coated brass and was mounted on the stage of a microscope
(Universal, Zeiss). Transmission illumination was used, the light
from the condenser (IVIz- 7, Zeiss) entering the chamber through a
small glass window inserted beneath the supports. The working
distance of the objective (40x, water immersion, NA = 0.75,
Zeiss) and condenser were 1.2 mm and 7 mm, respectively, and
sufficient to allowfreecirculation of the solution around the prepa-
ration.
 CONTRACTILITY OF SHR RESISTANCE VESSELS/Mulvany and Halpern
NOMENCLATURE
From the measurements of both of these dimensions
and of the force, F, exerted by vessels on the tension
transducer, the following parameters have been derived:
1. Wall tension, T: Circumferential wall force per unit
length given by:
T = F/2 g l .
2. Active wall tension, AT: Change in wall tension
upon isometric activation given by:
AT = 1a c l j V e , — T r e i a x e d ,
where TactiVf. and Treiaxe(i are the wall tensions in activating
and relaxing solutions, respectively.
3. Active wall stress, ACT: Active wall tension per unit
wall thickness given by:
ACT = AT/w,
where w is the wall thickness.
Under physiological conditions the in situ vessels must
have been under a transmural pressure and have had an
approximately circular cross section. On this basis the
following "effective" parameters were calculated:
1. Effective lumen diameter, 1: The calculated effective
diameter corresponding to the determined internal cir-
cumference, L, given by:
1 = L/TT.
2. Effective transmural pressure, p: Effective pressure
calculated (on the basis that the vessel wall was sufficiently
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thin for Laplace's equation to apply, and for it to be
unaffected by the curvature imposed by the wires) from
the equation:
p = 277-T/L,
where L is the internal circumference corresponding to
wall tension T. In other words p is an estimate of the
transmural pressure which would have been required to
maintain the in situ vessel at the same internal circumfer-
ence and wall tension under the same activation or relaxa-
tion conditions.
3. Effective active pressure, Ap: Change in effective
transmural pressure upon isometric activation given by:
Ap = Pactive ~ Preiaxed,
where pacuve a r | d Preiaxed a r e the effective transmural pres-
sures in activating and relaxing solutions, respectively;
4. Contractility: This term is here used synonymously
with effective active pressure, indicating the ability of a
vessel to contract against a given pressure.
RESTING TENSION: INTERNAL CIRCUMFERENCE
RELATION
Vessels were immersed in relaxing solution and
stretched in steps of about 10 /i.m until the wall tension
was about 1 mN/mm. The vessels were then released in
similar steps until the tension was about zero. Vessels were
held at each length for 1 minute and the wall tension
immediately before the next step was taken as the wall
tension at the internal circumference concerned. The
points obtained could be fitted by an exponential curve.27
21
NORMALIZATION OF INTERNAL CIRCUMFERENCE
Vessel dimensions were normalized by using the resting
wall tension-internal circumference curve determined as
described above. Lloo was defined as the internal circum-
ference corresponding to the point on the fitted exponen-
tial curve for which the effective transmural pressure was
100 mm Hg. Thus l100 = L1Oo/i" is an estimate of the lumen
diameter the vessel would have had in situ when relaxed
and under a transmural pressure of 100 mm Hg. For
reasons given below the experiments described in this
paper were performed at an internal circumference L, =
0.8 L100, except where stated otherwise.
ELECTRON MICROSCOPY AND HISTOLOGY
Vessels were fixed at L,. Before fixation vessels were
equilibrated in the chamber in relaxing solution for at least
15 minutes. The chamber was then drained and refilled
with prefixative (2.5% glutaraldehyde, 7% sucrose in 75
miu cacodylate, 37°C) and left overnight at room tempera-
ture. The solution was then changed to cacodylate buffer
and the wires were carefully removed from the vessel. The
vessel was then postfixed in 1% OsO4, blockstained with
uranyl acetate, and embedded in Epon.28 Thick sections
(0.5 fj.m) were cut from the block and stained with tolui-
dine blue for light microscopic measurements of medial
thickness. Thin sections were cut for electron microscopy
to determine the proportion of the media occupied by
smooth muscle cells. This factor was obtained from cross-
sectional photographs by planimetry.
CRITERION FOR REJECTION OF RESULTS
Experiments were rejected if the effective active pres-
sure at Lj was less than 100 mm Hg. Out of the 62 vessels
which were successfully mounted, results from 13 of these
were thus rejected (9 WKY, 4 SHR).
STATISTICS
Results were analyzed for possible significant differ-
ences between groups by the Student's /-test. Results are
presented as means ± SE (n = number of experiments), SE
bars are shown on the figures.
Results
CHARACTERISTICS OF K+ CONTRACTURES
K+ Response
Typical responses to activating solution (5 HIM Ca2+ in
K -Ringer), relaxing solution (EGTA in K+-Ringer), and
+
normal Ringer's solution are shown in Figure 3. The
response to activating solution has two parts. During the
first, about 80% of final tension is developed within about
10 seconds; in the second the tension rises more slowly to
the final value and reaches it after about 1 minute. The
response to relaxing solution consists of a transient tension
increase (caused by membrane depolarization) followed
by a decay to the resting tension within about 1 minute (as
the Ca2+ is withdrawn). In Ringer's solution the vessel
normally relaxed to the same resting tension found in
relaxing solution. Occasionally, however, toward the end
of experiments the relaxation in Ringer's solution was
incomplete, and this was attributed to membrane failure.
 22 CIRCULATION RESEARCH VOL. 41, No. 1, JULY 1977

its length. Figure 5 shows the relationship obtained for

A,R, three WKY and four SHR first branch vessels. Two points
•• may be noted. First, the resting wall tension-internal cir-
cumference relationships of these SHR and WKY vessels

-i-A (V are very similar. Second, the SHR vessels produced con-
siderably more active wall tension at any particular inter-
nal circumference than the WKY vessels. These points
FIGURE 3 Exact tracing of tension record showing responses of were tested in a larger experiment that included both first
spontaneously hypertensive rat (SHR) first branch resistance vessel and second branch resistance vessels. The details for the
to activating solution (at A,, A,) and relaxing solution (at E). The
vessel was initially in Ringer's solution and the solution was
changed back to Ringer's at R,, R2. The upper trace indicates the
time markers. The internal circumference was held constant
throughout at L, except over the bars where a square wave oscilla-
tion (amplitude = 0.003 Ll00; frequency = 75 Hz) was imposed.
Vessel parameters: 1 l0U = 213 \xm; segment length = 1.39 mm,
wall thickness at L, = 30.5 yjn.

Phasic spontaneous activity was never observed. Histolog-

ical examination of the vessels used here22'2if showed that
all the smooth muscle cells are orientated circumferen-
tially, and therefore that they all contribute to the active
tension measured. The resting tension measured arises
largely, if not entirely, from the properties of the passive
elements in the wall.
Dynamic Stiffness
During the second active response shown in Figure 3 the
dynamic stiffness was measured by imposing square wave
length changes of amplitude 0.003 L,Oo- These were ana-
lyzed by using signal averaging techniques.22 In 11 experi-
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0 2.5 io
Co" Concentration (mM)
FIGURE 4 Ca++ dose-response curves obtained from nine sponta-
neously hypertensive rat (SHR) vessels (%) and 12 normotensive
Wistar-Kyoto rat (WKY) vessels (O) equally divided between pri-
mary and secondary resistance vessels. Vessels were first activated
with a test solution (K+-Ringer with the test concentration ofCa2+)
and then with normal activating solution (K+-Ringer with 5 mM
Ca2+). Responses are expressed as the ratio of the response in the
test solution to the immediately following response in activating
solution. Vessels were held in Ringer's solution between tests.

ments with WKY vessels the mean value of the dynamic

stiffness was found to be 56 ± 6 AT,/Li where AT, was the
active wall tension at L,. These small vibrations never
produced any drop in tension as has been reported for
larger amplitude vibrations.30
Graded Responses
The active response was found to be dependent on the
concentration of Ca2+ in the K+-Ringer. Figure 4 shows
the determined Ca2+ dose-response curves for SHR and
WKY vessels. The responses are expressed as a proportion
of the response with 5 mM Ca2+. Higher concentrations
were not used because this resulted in calcium precipita-
tion in the K+-Ringer. Hansen and Bohr19 used different
buffer systems for K+ solutions containing high Ca2+ con-
centrations, but found that increasing the Ca2+ concentra- 0.6 0.8 1.0
tion above 5 mM resulted in a depression of the response. Internal Circumference (I_/Lioo)
We also conducted pilot experiments with epinephrine, FIGURE 5 Resting and active wall tension-internal circumference
norepinephrine, and electrical stimulation as Herlihy and relationships for four spontaneously hypertensive rat (SHR)
Murphy24 did in their investigation of hog carotid artery. (closed symbols) and three normotensive Wistar-Kyoto rat (WKY)
These tests confirmed that a maximal response was given (open symbols) first branch resistance vessels. Vessels were alter-
with 5 mM Ca2+ in K+-Ringer. Thus for both WKY and nately placed in activating solution (for about 2 minutes) and
SHR vessels the response to our activating solution has relaxing solution or Ringer's solution (for about 15 minutes, dur-
been taken as a measure of the maximal contractile re- ing which time the internal circumference was changed to a new
value). The active wall tension (triangles) was taken as the maxi-
sponse of the vessel.
mum measured tension in activating solution less the resting wall
tension (circles) immediately before activation. To facilitate com-
COMPARATIVE RESPONSES OF SHR AND WKY
parison the responses have been grouped, the numbers beside each
VESSELS
point showing the total number of responses measured in each
As in striated muscle both the active isometric response group. The order of internal circumference settings was quasi-
and the resting tension of smooth muscle are dependent on random.
 CONTRACTILITY OF SHR RESISTANCE VESSELS/Mu/vany and Halpern 23

rats used and the experiment plan are shown in Table 1. It TABLE 1 Characteristics of Rats Used in This Investigation
will be noted that the SHR had a 52% higher systolic and Experiment Plan
blood pressure and a 21% higher heart-body weight ratio
SHR WKY
than the WKY. Their age, weight, and sex distribution
were similar. Number 18 29
Systolic blood pressure 195 ± 7* 128 ± 3
Resting Wall Tension Heart/body weight (mg/g) 3.4 ± 0.1* 2.8 ± 0.04
Age (weeks) 21 ± 1 21 ± 1
The resting wall tension-internal circumference relation Weight (g) 316 ± 13 314 ± 13
of all vessels was measured as described in Methods. The Sex (% male) 83 79
values of the effective normalized diameter, l10o, obtained Resistance vessel type used
from the exponential curve fitted to the points obtained 1st branch 11 15
2nd branch 7 14
are shown in Table 2. These indicate that l100 is not de-
pendent on whether the vessel was taken from an SHR or SHR = spontaneously hypertensive rats; WKY = normotensive Wistar-
WKY. The mean l100 value for first branch resistance Kyoto rats.
* Different from WKY, P < 0.001.
vessels was 61% larger than that for the second branch
vessels. The exponential curve obtained for each vessel
was then expressed in the form: Active Wall Tension
T = T100 exp{[(L - L100)/L100]/y3}, The active wall tension-internal circumference curves
(Fig. 5) show that for both SHR and WKY vessels the
where T100 is the resting wall tension at L100 and /3 is the maximum wall tension is obtained at between 0.9 L100 and
proportional change in internal circumference required to L100. In this region the resting wall tension is between 30%
change the resting wall tension by a factor exp(l). The and 50% of the active wall tension. Thus, precise active
mean values found for /3 are shown in Table 2. There was wall tension measurements are to some extent masked
a wide variation in the values obtained within each group, here by the resting wall tension. For this reason the active
but there was no indication of any differences between responses were compared at L, = 0.8 L100, where the
groups. resting wall tension was 10-15 % of the active wall tension.

TABLE 2 Summary of Vessel Parameters Obtained in This Study

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First branch resistance vessel Second branch resistance vessel P values from (-test*

SHR vs. WKY 1st vs. 2nd

1st and
SHR WKY SHR WKY 1st 2nd 2nd SHR WKY

1. Effective 236 ± 5 (11) 257 ± 9 (15) 152 ± 9 (7) 154 ± 8 (14)

normalized
diameter,

2. Wall tension 3.0 ± 0 . 2 (11) 2.5 ± 0.1 (15) 1.8 ± 0 . 2 (7) 1.3 ± 0.1 (14)
(mN/mm)
3. Effective ac- 249 ± 1 6 ( 1 1 ) 199 ± 12 (15) 266 ± 18 (7) 157 ± 12 (14)
tive pressure
at L, (mm
Hg)
4. Wall thick- 17 ± 2 (11) 13 ± 1 (15) 16 ± 2 (7) 14 ± 1 (14)
ness at Llt>0

5. Wall stress 153 ± 8 (11) 148 ± 7 ( 1 5 ) 104 ± 17 (7) 76 ± 6 (14)

at L, (mN/
2
mm )
6. Smooth 0.41 ± 0.02 (2) 0.36 ± 0.06 (2) 0.32 ± 0.03 (2) 0.25 ± 0.03 (4)
muscle cell
content
(proportion)
7. Smooth 373 411 325 304
muscle
stress (mN/
mm2)t
8. Length con- 0.13 ± 0.01 (11) 0.12 ± 0.01 (15) 0.12 ± 0.01 (7) 0.11 ± 0.01 (14)
stant /3
(L,oo)
Values are means ± SE. SHR = spontaneously hypertensive rats; WKY = normotensive Wistar-Kyoto rats. Numbers in parentheses denote number of rats.
* Symbols have the following meanings: + + + ,/>< 0.001; + + ,P < 0.005; +,/ > <0.02; - , not significant (P > 0.05). Blank spaces indicate that the (-test
is not applicable.
t Evaluated by the line 5/line 6; mean value only.
 24 CIRCULATION RESEARCH
Repeated activations at L, never showed any potentiation
in the response and often showed a cumulative small
decline in the response measured. This decline was attrib-
uted to a failure of the smooth muscle cells and, as we
sought to compare the maximal response of the vessels, we
have compared the initial responses of the vessels at L,.
The mean of these responses is shown in Table 2 and in
Figure 6. These results show that the SHR vessels are able
to contract about 34% more strongly than the correspond-
ing WKY vessels. The second branch resistance vessels in
both cases produced less tension than the first branch
resistance vessels, but if their contractility is normalized by
using Laplace's equation to determine the pressure against
which they would just be able to contract (effective active
pressure, Table 2) the difference is less marked.
Active Wall Stress
Table 2 also shows the wall thickness measured at L100.
Here we found that there was no difference between the
wall thickness of first and second branch vessels, but that
on average the SHR vessels had a 23% greater wall thick-
ness than the WKY vessels. These measurements were
used to compute the active wall stress of each vessel (Table
2). There was no significant difference in the active wall
stress of SHR and WKY vessels, but in each case the
active wall stress of second branch resistance vessels was
lower than that of first branch resistance vessels.
The medial thickness and smooth muscle cell content of
some of the vessels was determined from histological sec-
tions taken from vessels after fixation at L,. Both the
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medial thickness and the smooth muscle cell content of the
second branch vessels are smaller than those of the first
branches. Thus the active force per unit area of smooth
muscle cell (on average, 353 mN/mm2) was about the
same in all cases. We have not, however, made sufficient
measurements to test these results (summarized in Table
2) rigorously, nor have we been able to determine whether
the increased active wall tension of the SHR vessels is due
.1
I Our finding that the wall thickness of the SHR vessels
I ,.o
O 100 200 300
Effective Diameter (/im)
FIGURE 6 Active wall tension developed at L, by first branch
(circles) and second branch (triangles) resistance vessels taken
from spontaneously hypertensive rats (SHR) (closed symbols) and
normotensive Wistar-Kyoto rats (WKY) (open symbols). Ordinate
shows mean active wall tension developed for each group. Abscissa
is the mean effective diameter of the vessels when tested.
VOL. 41, No. 1, JULY 1977
to an increased smooth muscle cell content, although our
results suggest that this may be the case.
Discussion
Previous investigations into the contractile properties of
the vasculature of genetic hypertensive rats have included
studies of complete vascular beds and also of isolated
vessels, although the latter studies have been confined to
the aorta and arteries and veins having lumen diameters
greater than 500 fxm. These studies have, however, given
rather conflicting results (see Bohr" for a review). Thus
helical strips from SHR aorta14'15 and femoral artery19
were found to have lower contractility than their WKY
controls. In contrast others16'17 found no differences be-
tween the contractile properties of aortic strips from their
SHR and WKY. Studies of the pressor response to activa-
tion of complete vascular beds7"11 indicate, however, that
taken as a whole the vascular bed of SHR contracts more
strongly than that of WKY. Thus it has seemed that the
increased contractility (defined here as the ability to con-
tract against a given transmural pressure) must reside in
the smaller vessels. This conclusion is now supported by
the major finding of this study, i.e., that the contractility
of first and second branch arterial resistance vessels is
about 34% higher in SHR than in WKY.
In comparing our results with those obtained from
larger vessels it must be remembered that the preparative
technique we have used is very different. For the following
reasons, however, we consider that our method is inher-
ently less traumatic to the tissue than the conventional
method of cutting helical strips. In our technique the
medial layer is protected throughout by the internal elastic
lamina and the adventitial layer. At no time during dissec-
tion or mounting is the vessel stretched beyond about L,
and, provided that the ends are cleanly cut, the end effects
are minimal because of the circumferential orientation of
the smooth muscle cells. The responses we measured,
when expressed as force per unit smooth muscle cell area
(Table 2), are very similar to that found in the media of
hog carotid artery (370 mN/mm2).26 The dynamic active
elastic stiffness measured (56 AT,/L,) is similar to that
previously measured in activated smooth muscle (58 AT,/
L,).32 Last, the "effective active pressure" measured was
in excess of the systolic blood pressure of the rats con-
cerned. We therefore believe that the forces we have
measured are a good estimate of the maximal in vivo
performance of the vessels.
was about 23% greater than that of the corresponding
WKY vessels is in agreement with the findings of other
investigators concerning the morphological changes asso-
ciated with hypertension.33"35 Our histological evidence
suggests (Table 2) that this increased wall thickness may
be associated with an increased smooth muscle cell con-
tent, and that it is this which accounts for the increased
contractility we found in the SHR vessels. Folkow and his
colleagues7'8 have suggested, however, that hypertrophy
of the arterial wall of SHR could in itself be a primary
cause of hypertension if the increased wall thickness en-
croaches on the lumen. This hypothesis is supported by
their findings that the resistance of relaxed vascular beds is
 CONTRACTILITY OF SHR RESISTANCE VESSELS/Mulvany and Halpern
elevated in 7-month-oId SHR. Finch and Haeusler10 con-
firmed this but found no differences in the relaxed vascular
resistance in 3-month-old rats. Our results (from 5-month-
old rats), show no significant difference in the effective
lumen diameters between corresponding SHR and WKY
vessels. Furthermore, the form of the resting wall tension-
internal circumference curve is the same for both WKY
and SHR vessels. Therefore our results show no differ-
ences in the passive properties of the SHR and WKY
vessels we tested which could account for increased vascu-
lar resistance.
One of the purposes of investigations concerning the
vasculature of hypertensive animals has been to obtain
evidence as to whether increased vascular resistance is a
primary cause of increased blood pressure or whether it is
a secondary effect. Our findings suggest to us, however,
that the changes in both the vasculature and blood pres-
sure are secondary effects. The 52% greater systolic blood
pressure found in our SHR indicates that their mean blood
pressure was about 35% greater than that of the WKY (B.
Folkow, personal communication). This may be compared
with the 30% greater vessel contractility, the 23% greater
wall thickness, and the 21% greater heart-body weight
ratio found in the SHR. We are therefore inclined to
interpret our results as indicating that both the myocar-
dium and the vasculature have reacted equally to some
disturbing influence in the cardiovascular regulatory sys-
tem. The possible site of this disturbance must for us be a
matter of speculation. The evidence of other workers has
suggested that the site may be neurogenic,36'37 in the
smooth muscle cell membrane, ' 38> 39 or in the barore-
Downloaded from http://ahajournals.org by on September 19, 2023
ceptors.40'41 All these sites are no doubt involved and, as
the systems analysis approach of Guyton and his col-
leagues indicates, a full understanding of the causes of
hypertension requires an understanding of all the many
factors involved in the control of the circulation.
Another approach to determining the prime cause of
hypertension may be an evolutionary one. It is clear that
blood pressure is directly related to the two most impor-
tant functions of the cardiovascular system, namely, blood
flow and the maintenance of capillary pressure. In SHR
the cardiac index5 (and the blood flow index in the supe-
rior mesenteric artery43) is the same as in WKY. Although
the capillary pressure of SHR with established hyperten-
sion has not been measured, in genetically hypertensive
cats44 it is not more than 10% greater than that of the
normotensive controls. Thus in both genetically hyperten-
sive and normotensive animals the two most important
functions of the cardiovascular system are apparently simi-
lar. It seems likely, however, that the cardiovascular sys-
tem has evolved in such a way that the expenditure of
energy required to perform these functions is a minimum.
Why then is 195 mm Hg systolic blood pressure required
in 5-month-old SHR when only 128 mm Hg is required in
WKY? We suggest that a detailed analysis of the con-
straints that determine optimal blood pressure may help to
identify those factors which are primarily involved in the
etiology of essential hypertension.
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The Influence of the Time Interval between

Coronary Artery Occlusion and the Administration
of Hyaluronidase on Salvage of Ischemic
Myocardium in Dogs
L. DAVID HILLIS, MICHAEL C. FISHBEIN, EUGENE BRAUNWALD, AND PETER R. MAROKO
Downloaded from http://ahajournals.org by on September 19, 2023

SUMMARY The purpose of this study was to determine the time interval following coronary artery occlusion
during which the administration of hyaluronidase exerts a significant protective effect on injured myocardium. Forty-
eight open-chest dogs with coronary artery occlusion were studied. Fourteen were untreated (controls). Hyaluroni-
dase (500 NF units/kg, iv) was administered 20 minutes (12 dogs), 3 hours (8 dogs), 6 hours (8 dogs), or 9 hours (6
dogs) after occlusion. Epicardial electrograms, recorded from 10 to 16 sites on the anterior surface of the left ventricle
before, 15 minutes after, and 24 hours after coronary occlusion were analyzed for S-T segment elevation and changes
in QRS morphology. Transmural specimens, excised 24 hours after occlusion from the sites at which the electrograms
were recorded, were analyzed for creatine phosphokinase (CPK) activity and histological appearance. In all five
groups, myocardial CPK depression, histological evidence of the extent of necrosis, and changes in QRS configuration
correlated well with one another. In the controls, S-T segment elevation 15 minutes after occlusion (ST15m) correlated
well with myocardial CPK depression, histological extent of necrosis, and changes in the QRS complex 24 hours later.
When hyaluronidase was given 20 minutes, 3 hours, or 6 hours after coronary occlusion, myocardial salvage was
reflected in significantly less myocardial CPK depression for any given ST]5rn, less histological evidence of infarction,
and less extensive changes in QRS configuration than in the untreated dogs, although there was a progressive
reduction in tissue salvage as the time interval between occlusion and drug administration lengthened. Hyaluronidase
administered 9 hours after occlusion had no demonstrable effect on the development of myocardial necrosis,
suggesting that ischemic injury is totally irreversible by this time.

MYOCARDIAL necrosis following coronary artery oc-
clusion can be limited by certain interventions designed to
improve the balance between oxygen supply and de-
mand.1'2 Numerous interventions — hemodynamic, meta-
From the Departments of Medicine and Pathology, Harvard Medical
School and Peter Bent Brigham Hospital, Boston, Massachusetts.
Supported in part by Contract N01-HV-53000 under the Cardiac Dis-
eases Branch, Division of Heart and Vascular Diseases, National Heart,
Lung, and Blood Institute, National Institutes of Health. Dr. Hillis is the
recipient of a postdoctoral fellowship award (No. 1 F 32 HL 05147-01)
from the National Heart, Lung, and Blood Institute.
Address for reprints: Peter R. Maroko, M.D., Harvard Medical School,
Building A, 25 Shattuck Street, Boston, Massachusetts 02115.
Received September 7, 1976; accepted for publication December 9,
1976.
bolic, and pharmacological —have been shown to reduce
ischemic injury.2-3 In most of these studies the intervention
under investigation has been applied either immediately
before coronary artery occlusion or shortly thereafter
(i.e., 30 minutes after occlusion), at a time when most
cells still are reversibly injured. A few observations have
been made in which interventions have been applied sev-
eral hours after occlusion, and a beneficial effect has, in
general, been observed.2"7 However, a systematic study
designed to examine the relative efficacy of an interven-
tion administered at various time intervals after coronary
artery occlusion has not been performed.
Hyaluronidase has been shown to limit myocardial ne-