227

J. Physiol. (I 95 I) I"5, 227-236

EFFECT OF CURRENT FLOW ON THE MEMBRANE

POTENTIAL OF CARDIAC MUSCLE
BY SILVIO WEIDMANN*
From the Physiological Laboratory, University of Cambridge
(Received 4 June 1951)
Microcapillaries with an external tip diameter of 0 5p. (Ling & Gerard, 1949)
can be inserted into single fibres of the cardiac syncytium and may be used to
record resting and action potentials. The technique has so far been applied to
the frog ventricle (Woodbury, Hecht & Christopherson, 1951) and to 'false
tendons' of the dog and kid heart (Draper & Weidmann, 1951). The conductive
system of the kid contains typical Purkinje fibres which are larger in diameter
(40-lOOup.) than ordinary heart muscle fibres and which are only slightly
contractile. In this preparation it is possible to insert two microelectrodes
into the same fibre and to leave them intracellular for many minutes. Polarizing
current can be applied through one electrode and the change in membrane
potential -resulting from current flow recorded through the other electrode.
The present paper contains an account of experiments designed to investigate
the cardiac action potential by applying suitable pulses of current at various
stages in the cardiac cycle.
METHOD
The method was the same as that described in an earlier paper (Draper & Weidman, 1951).
Fig. 1 shows the essential elements of the recording and polarizing circuit.

RESULTS
Impedance changes in the course of the cardiac cycle
The relative magnitude of the membrane resistance was measured in the
following way: square pulses of polarizing current were applied through a first
pair of electrodes placed on opposite sides of the surface membrane (Fig. 1).
Potential changes resulting from current flow were recorded by a second pair
of electrodes in a similar position. No voltage changes were observed when
the tip of the recording microelectrode was just extracellular. This indicates
that the voltage changes recorded from the inside of a cell were localized at
* Present address: Department of Physiology, University of Berne, Switzerland.
228 SILVIO WEIDMANN
the fibre membrane. Their amplitude was used to estimate changes in membrane
resistance.
The action potential of Purkinje tissue of the kid consists of an initial spike
followed by a plateau and a phase of repolarization (Draper & Weidmann,
1951). During the period of mechanical rest there is generally some slow
depolarization. In the present series of experiments the upstroke of the action
potential was used to trigger the time base so that successive traces could be
superimposed. The initial spike then appeared extended over the flyback of
the sweep cycle while the plateau of the action potential, the phase of repolariza-
tion and the changes during diastole could be seen on the time base in the usual
manner.
0-90 V

as5Mf

<, XLIt3~~~~~~~~
Rfl
Ag-AgCl
Agar-Ringer
electrode

Square pulse
generator D~Eclaycrc]i
D.

Fig. 1. Diagram of experimental arrangement. PE =polarizing electrode. RE =recording

electrode. DCA =push-pull cathode follower and differential d.c. amplifier. R =high speed
relay (Carpenter 3G2). P=potentiometer with 10 steps of 10kQ.

A typical experiment is illustrated in Fig. 2 which was made by exposing

the film for twenty cycles of activity. During this period rectangular pulses
of anodal current lasting 40 msec. were applied to the membrane at a frequency
of about 12 cyc./sec. These pulses were not synchronized with the action
potential and therefore occurred at many different times in the cardiac cycle.
The upper edge of the superimposed traces corresponds to an action potential
in the absence of applied current while the lower edge represents action
potentials displaced by anodal current. During diastole the process of
charging and discharging the membrane was half complete in about 20 msec.
At the crest of activity the half time was shorter than 1 msec. The breadth of
the band in Fig. 2 was used as a qualitative index of the membrane resistance.
The interpretation of this experiment is as follows: (1) During diastole the
slow depolarization is associated with an increase in membrane resistance.
(2) At the height of the spike the membrane resistance is greatly decreased.
 POLARIZATION OF CARDIAC MUSCLE 229
(3) The resistance recovers during the few milliseconds between the crest of
the spike and the beginning of the plateau. (4) During the plateau the
resistance is comparable to that in diastole and increases as the phase of
repolarization is approached. (5) The shortening of the time constant suggests
that cardiac muscle is similar to other excitable tissues in that the membrane
capacity changes less than the membrane resistance (cf. Cole & Curtis, 1939).

Fig. 2. Impedance changes in the course of the cardiac action potential. Voltage calibration in
steps of 10 mV. Time marks for left-to-right movement of the cathode ray at intervals of
100 msec. Duration of flyback of sweep cycle approx. 12 msec.

A rough estimate of the change in membrane resistance may be obtained

-from the equations of cable theory. For this purpose a relation used by
Hodgkin & Huxley (1947) was employed in the following form
lOg0 [=~~ee f +(f-)(1
_VB toie log,,
where VB = amplitude of the electrotonic potential
I distance between leading-in and recording electrode
A =,space constant of the fibre in diastole
2= ratio of membrane resi-stance in dia-stole and at crest of spike.
In practice it was convenient to use the ratio of current strengths required to
produce the same potential change instead of the ratio of potentials produced
by current of the same strength. In the experiment illustrated by Fig. 3 the
230 SILVIO WEIDMANN
ratio of current strengths was 22 and the electrode distance 0 33 mm. The
space constant was not determined on this fibre but averaged about 2 mm. in
other experiments (Weidmann, in preparation). With these figures equation (1)
indicates a resistance ratio of 56: 1. This ratio is subject to the following sources of
error: (a) The measurements were not made on a uniform fibre of infinite length
but on one which fused into other fibres of the syncytium at a distance of one or

Fig. 3. Potential changes due to square pulses of polarizing current applied in the middle of
diastole. The record shows on the left the phase of repolarization, on the right the upstroke
of the action potential. Seventeen traces are superimposed. Relative strength of polarizing
current reading from bottom to top: 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, O, -1, -2, -3, -4, -5, -6. Time
marks at intervals of 100 msec.

two diastolic space constants. (b) Equation (1) applies to a fibre which is thin
compared to its space constant. This approximation was entirely satisfactory
in diastole but may have introduced some error at the crest of the spike since
the space constant is then only about twice the fibre diameter. (e) Charging
of the membrane capacity by the current pulses was only about 90 % complete
during diastole. These three sources of error tend to make the estimated ratio
too low.
 POLARIZATION OF CARDIAC MUSCLE 231
Subthreshold potentials
Fig. 3 shows a family of electrotonic potentials produced by square pulses
of constant current which were timed to fall at a constant phase of the cycle.
Successive traces were superimposed by synchronizing the sweep circuit with the
upstroke of the action potential. It can be seen that anodal currents of
increasing strength give changes in membrane potential smaller than those
expected on the assumption of a constant membrane resistance. On the other
hand the depolarization produced by a given increment of cathodal current
increased continuously as the threshold was approached. The strongest
cathodal current initiated an extrasystole. The cathodal part of the record
looks strikingly similar to the curves obtained on invertebrate nerve (Hodgkin
& Rushton, 1946) and skeletal muscle (Katz, 1948). One respect in which
it differs is that the time course of the potential after the break of the
current exhibits the phenomena of postcathodal depression and postanodal
enhancement.
Peculiarities of the pacemaker region
The pacemaker was localized by recording simultaneously from two intra-
cellular electrodes and by shifting the one nearest the point of origin of the
spike until it gave the earliest upstroke. At this point the action potential
was regularly found to have a slow upward curvature preceding the most
abrupt part of the rising phase. This feature was absent or less marked in
other parts of the fibre. It was also found that, in this region, application of
a square pulse of subthreshold cathodal current increased the duration of
diastole, presumably as a result of postcathodal depression, while the opposite
effect was observed after the break of an anodal current. If strong enough
the cathodal current evoked an extrasystole. These features are illustrated in
Fig. 4.
Propagated wave of repolarization
In the experiment illustrated in Fig. 5 square pulses of anodal current were
applied during the upstroke of the action potential and terminated at the
beginning of the plateau. The current strength was increased in seven steps
of equal increment. After the break of steps 1-4 the cathode-ray tube beam
followed its usual course. With strength 5 a short spike preceded the return to
the plateau. This overshoot became slightly larger after the break of strength 6
and appeared with a longer latency. An unexpected event appeared when
a current of strength 7 was broken. The membrane potential did not return
to the plateau but stayed near its resting level. The height of the short spike
could be altered continuously by changing the current strength over a limited
range. On the other hand, the reversal of the plateau seen in 7 was an all-or-
nothing event which had a definite threshold. This all-or-nothing repolarization
was observed regularly in nine different kid hearts. The conditions for obtaining
232 SILVIO WEIDMANN

Fig. 4. Superimposed traces from the pacemaker region showing changes in membrane potential
produced by square pulses of cathodal current (relative strength 0, -1, -2, -3, -4). Time
marks at intervals of 100 msec.

Fig. 5. Potential changes produced by square pulses of anodal current applied at the beginning
L of the action potential. Recording electrode close to the polarizing electrode (0-2 mm.).
Relative strength of current from above downwards: 0, 1, 2, 3, 4, 5, 6, 7. Time marks at
intervals of 200 msec.
 POLARIZATION OF CARDIAC MUSCLE 233
it were that the current pulses should be of sufficient strength and that they
should last for more than the first quarter of the action potential.
In the experiment illustrated in Fig. 6 the distance separating the recording
from the leading-in electrode was 4-2 mm. instead of 02 mm. as in Fig. 5.
Square pulses of polarizing current were timed to coincide with the middle of
the plateau. If these were less than a certain strength, they produced small
and graded repolarizations, presumably by passive cable-like spread. How-
ever, above a certain strength a maximal repolarization was produced at
a definite threshold. A comparison of records obtained from various distances

Fig. 6. Effect of polarizing current on a region at some distance from the leading-in electrode
(4-2 mm.). Square pulses of anodal current lasted from a to b. Ten traces are superimposed;
relative strengths of current from above downwards: 0, 1, 2, 3, 4, 5, 6, 7, 8, 9. Time marks
at intervals of 100 msec.

indicated that the subthreshold potentials spread with a decrement while the
all-or-nothing repolarization was propagated without decrement as an 'off-
response'. Attempts to determine the velocity of this off-response were not
entirely successful but indicated that it was much lower than that of the normal
spike. It is therefore to be expectedl that the 'off-response' would disappear
at a large conduction distance since it would be overtaken by the normal
process of repolarization which occurs at a fixed time after the beginning of
the spike. However, this point could not be verified since the total length of
the fibres was insufficient for such an experiment.
DISCUSSION
The finding that the spike is associated with a profound decrease of the
membrane resistance agrees with similar results obtained on the alga Nitella
(Blin s, 1936; Cole & Curtis, 1938), the giant axon of the squid (Cole & Curtis,
234 SILVIO WEIDMANN
1939), skeletal muscle (Katz, 1941) and myelinated nerve (Tasaki & Mizuguchi,
1949). Previous impedance measurements on cardiac muscle have led to
controversial results. Rapport & Ray (1927) recorded an impedance fall
during the isometric contraction of a cannulated tortoise ventricle while
Rosenblueth & del Pozo (1943) measured a systolic impedance rise in strips
of turtle ventricle contracting isometrically. It is felt that the results obtained
with hearts of cold-blooded animals should not be compared with the findings
reported here for a mammalian preparation. For in Purkinje fibres of the kid
the impedance fall is limited to the initial spike while there seems to be no
such spike in the monophasic action potential of the turtle ventricle (Eyster &
Meek, 1942), nor in that of the frog ventricle (Woodbury et al. 1951).
Recent experiments with nerve and muscle indicate that the rising phase of
the action potential is brought about by an increase in permeability which
allows sodium ions to enter the fibre at a high rate (for references see Hodgkin,
1951; Huxley & Stampfii, 1951). It has also been suggested that repolarization
during the falling phase occurs for two reasons (Hodgkin, Huxley & Katz, 1949;
Hodgkin & Huxley, 1950). In the first place sodium entry is reduced by an
inactivation process which decreases the permeability to this ion. In the
second, the exit of potassium ions is accelerated by a rise in potassium perme-
ability. Experiments described in a previous paper (Draper & Weidmann,
1951) suggest that the rising phase in cardiac muscle depends on an increase
in sodium permeability and this conclusion is consistent with the present
finding of a large increase in membrane conductance during the spike. The
falling phase of the cardiac action potential is so different from that of nerve
that it is doubtful if a hypothesis developed for the one tissue can be applied
to the other. But it is interesting to consider briefly whether the general
mechanism suggested for nerve is applicable to the present results. In order
to explain the prolonged phase of depolarization one may suppose either that
there is a long delay in the process which inactivates sodium permeability
(PNa) or that the rise of potassium permeability (PK) is greatly retarded. It is
also possible that one or other of these processes may be entirely absent, or
that one or other may occur in two stages. During the plateau the membrane
potential is not far from zero, indicating that PNa is approximately the same
as PK. This situation might arise because PK has remained at its resting level
whereas PNa has fallen from the high value which it attained at the crest of
the spike. Or it might arise because PK has risen to the high value attained
by PNa during the spike. The first suggestion would be consistent with a rapid
but incomplete inactivation and a slow rise in PK, the second with a slow
inactivation and a rapid rise in PK. It can be seen that only the first of these
suggestions is compatible with the impedance measurements reported in this
paper. For the membrane conductance has been shown to fall to a low level
at the end of the spike and to remain at a low level during the plateau. This
 POLARIZATION OF CARDIAC MUSCLE 235
could not occur if repolarization from spike to plateau depended on an increase
in PK but is quite consistent with its being brought about by a decrease in
PNa. Beyond this point it seems unwise to speculate further. It is perhaps
worth adding that Purkinje fibres in a sodium-free saccharose medium showed
no sign of any 'delayed rectification' (Hodgkin et al., 1949) such as might be
expected if there were a rise in potassium permeability. However, these experi-
ments were not conclusive since the unsteadiness of the electrotonic potentials
indicated that the application of relatively strong currents to a preparation
in saccharose solution may have damaged the membrane in some way.
The ability of cardiac muscle to propagate an all-or-nothing wave of
repolarization is a new but not entirely unexpected finding. In the ordinary
way an excitable fibre starts with a high membrane potential and propagates
a negative wave because local circuit currents depolarize the membrane to
a threshold potential beyond which it tends to attain a new active level.
Granted a suitable relation between threshold and amplitude of the propagated
wave there seems to be no reason why an excitable fibre should not start with
a low membrane potential and propagate a wave of repolarization by local
circuit action. The velocity of propagation would not be the same as that for
the wave of depolarization, one of the reasons being that the relation between
threshold and amplitude would be different in the two cases.

SUMMARY
1. Polarizing current was caused to flow through the surface membrane of
cardiac muscle by means of a microcapillary inserted into the myoplasm of
a single Purkinje fibre of the kid. The membrane potential was recorded by
means of a similar electrode inserted into the same fibre.
2. The amplitude of the electrotonic potential produced by a relatively
weak square pulse of current was used to estimate the electrical resistance of
the surface membrane. Impedance changes were recorded simultaneously
with the action potential. During the greater part of systole (plateau of the
action potential) the membrane resistance had values comparable to those
found in diastole. But a profound fall in resistance was observed during the
short spike at the onset of activity.
3. 'Graded activity' could be demonstrated by applying cathodal current
pulses of variable strength during diastole. The action potential of the pace-
maker region was found to be characterized by a slow upward curvature
preceding the steep part of the rising phase.
4. The action potential could be 'switched off' by applying a relatively
strong anodal current pulse. The off-response resembled the familiar on-response
in that there was a definite threshold and that the response was propagated
without decrement away from the stimulating electrode.
PH. CXV. 16
236 SILVIO WEIDMANN
5. The findings are discussed on the hypothesis that the rhythmical changes
in membrane potential result from a succession of permeability changes to
ions, particularly sodium.
This work was greatly helped by advice and criticism from Mr A. L. Hodgkin and Mr A. F.
Huxley. To Dr R. S. Comline and collaborators I am indebted for kid hearts. The costs of the
investigation were defrayed by the Rockefeller Foundation and the work was done during the
tenure of a scholarship awarded by the Stiftung fur Biologisch-Medizinische Stipendien, Basel.

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