IV.

THE ANTAGONISTIC ACTIONS OF EPINEPHRIN

AND POTASSIUM CHLORIDE ON THE TONUS AND
TONUS WAVES IN THE EXCISED TERRAPIN
AURICLES
CHARLES M. GRUBER
From the Department of Physiology and Pharmacology and the Henry S. Denison
Research Laboratories in the University of Colorado

Received for publication April 8, 1920

Few men working upon the terrapin heart directed their atten-
tion to the action of the inorganic salts of the blood upon the
general tonus and tonus waves observed in the auricles.
Working upon strips of the vena cava Howell (1) observed
the tonus waves either to disappear or remain unchanged when
the strips were immersed in a calcium chloride solution. On
the other hand the tone was increased when they were immersed
in a 0.2 per cent potassium chloride solution.
Martin (2) in testing the effect of the inorganic salts found
that the auricles were less readily affected by potassium chlor-
ide than were the ventricles. It was observed by Fano and
Bodano (3) that potassium chloride increases the tonus waves
and decreases the contractions in the terrapin auricles. It must
be noted that they used strong solutions 1 to 5 per cent, dropped
upon the intact auricles. Their results were corroborated by
Bottazzi (4).
The effect that epinephrin has upon the increased tonus and
tonus waves produced by potassium has not been investigated
as far as I am aware.

THE METHOD

The method employed in this work was the same as that

described in detail in a previous article (5). The experiments
were performed on auricles immersed and contracting, in some
271
272 CHARLES M. GRUBER

cases in Ringer’s solution, in others in 0.7 per cent sodium

chloride solution. To these solutions were added from 1 to 5
mils. of 10 per cent potassium chloride or calcium chloride solu-
tion. The final dilutions were, therefore, from 0.1 to 0.5 per
cent calcium chloride or of potassium chloride in the latter and
from 0.14 to 0.54 calcium chloride or from 0.13 to 0.53 per cent
potassium chloride in the former.

RESULTS

Potassium chloride in the majority of the experiments de-

creased the rate and height of contraction. In some cases how-
ever there was a temporary acceleration of the rate of the beat.
Increased tone and tonus waves were observed in 55 of the 56
experiments performed. In the 55 successful experiments the
dose varied from a 0.1 per cent to a 0.4 per cent solution of
potassium chloride with an average for the entire group of 0.22
per cent.
This average percentage is interesting in that it is nearly
that (0.23) found by Martin (6) to be the dose at which varia-
tions in tone do not occur in either direction in the ventricles
of the terrapin. He found that the concentration of potassium
chloride necessary to produce complete quiescence in the auricle
is about 0.25 per cent.
In figure 1 (at 1) 2 mils. of potassium chloride was added
to the 98 mils. of normal saline solution making a dilution of
potassium chloride of 0.2 per cent. The tonus waves soon in-
creased in amplitude and rate but the rate of contraction de-
creased. In figure 2 (at 1 and 3), potassium chloride was added
to the Ringer’s solution in which the muscle was contracting.
The strength of the solution was about 0.15 per cent. The
contractions entirely disappeared but the tonus waves became
extremely marked. In figure 3 a sufficient amount of potas-
sium chloride was added to the Ringer’s solution to make a
0.23 per cent potassium chloride solution. Both general tone
and tonus waves were increased.
 TONUS AND TONUS WAVES IN TERRAPIN AURICLES 273

FIG. 1. CHRYSEMYS CINERA

In this and following records the upper record is that of the contracting auricles
and the lower, that of the time in five second intervals. Up-stroke systole, down-
stroke diastole. 1, Potassium chloride 0.2 per cent; S, calcium chloride 0.1 per
cent; 3, adrenalin chloride; 1:100,000 solution.

FIG. 2. CHRYSEMYS CINERA

1, Potassium chloride 0.15 per cent; 2, Ringer’s solution; 3, potassium chloride

0.15 per cent; 4, epinephrin 1:85,000 in the Ringer’s solution containing 0.15 per
cent potassium chloride.

FIG. 3. CHRYSEMYS ELEGANS

1, Potassium chloride 0.2 per cent; 2, adrenalin chloride 1:10,000 solution.

274 CHARLES M. GRUBER

The results obtained in the experiments with calcium chloride

are somewhat similar to those observed by Howell (1) upon
the strips of vena cava. Calcium chloride in the majority of
the cases had no effect upon the tonus waves. In a few experi-
ments the results were so variable that no conclusion could be
drawn. There seems to be a cooperative action between po-
tassium chloride and calcium chloride. The general tone and
tonus waves always seemed to be more marked when the potas-
sium chloride was added to the Ringer’s solution in which the
auricles were contracting or to sodium chloride solution con-
taming some calcium chloride.
In figure 1 (at 1) potassium chloride was added to the nor-

mal saline to make a 0.2 per cent solution. The tonus waves
here produced were about of equal height. At (2) 0.1 per
cent of calcium chloride was added. There was immediately a
marked increase in both general tone and tonus waves. It
has been found by the author that calcium chloride is insuffi-
cient of itself to bring about such a result. This would seem
to suggest some synergic action of these two salts. This is in
harmony with Howell’s findings when experimenting with the
ventricles (7).
In previous articles (8) it was shown that epinephrin de-
creased the general tone and simultaneously caused the tonus
waves to disappear, in normal muscles, and in muscles treated
with atropine sulphate and pilocarpine hydrochloride. All the
figures here published demonstrate that it also produces a de-
crease in general tone and a disappearance of the tonus waves in
muscles affected by potassium chloride.
In figure 1 (at 3) adrenalin chloride, in sufficient quantity to
make 1: 100,000 solution was added to the solution containing
0.7 per cent sodium chloride, 0.2 per cent potassium chloride
and 0.1 per cent calcium chloride. The effects of these salts-
marked increase of general tone and tonus waves-were entirely
counteracted by the adrenalin. Not only did it counteract the
increased tone but it also increased the rate of contraction from
21 to 30 contractions per minute. In figure 2 (at 4) epinephrin
(1: 85,000 final dilution) was added to Ringer’s solution con-
 TONUS AND TONUS WAVES IN TERRAPIN AURICLES 275

taming the 0.15 per cent potassium chloride. The tonus disap-
peared immediately, and the contractions which had disappeared
as a result of the potassium chloride returned.
In figure 3 (at 2) a sufficient quantity of adrenalin chloride
was added to the Ringer’s solution containing 0.2 per cent potas-
sium chloride to make a 1 ; 10,000 solution. The tone and tonus
waves disappeared at once. The rate of contraction increased
from about 11 to 21 contractions per minute.

DISCUS5ION

Fano (9), Fano and Bodano (3), Bottazzi (10), Soraku Oinuma
(11) and Gault (12) observed that stimulation of the vagus
causes an increase in the tonus waves but that sympathetic
stimulation causes a disappearance of the tonus waves in the
terrapin heart. Rosenzweig (13) obtained negative results.
Bottazzi (10) observed tonus waves in the sinus venosus.
These waves were increased by vagus stimulation and decreased
upon sympathetic stimulation.
In 1905 Howell (14) observed that an increase in the amount
of potassium salt augments the sensitiveness of the heart to
vagus inhibition until the amount of potassium becomes suffi-
ciently large to cause marked inhibitory effect. Under the
latter conditions the effect of vagus inhibition is greatly les-
sened. A complete lack of potassium salts is attended by a
diminution or an entire loss of vagus control of the different
chambers of the heart in terrapin and frog.
Upon perfusing the heart with a solution containing sodium
chloride only, there was complete loss of vagus control. In-
creased calcium chloride 0.025 to 0.138 per cent establishes an
independent rhythm between the auricles and ventricles with
increased auricular tonus. In these cases stimulation of the
vagus caused inhibition of t.he auricles only.
Miss Brine (15) recently corroborated Howell’s observations.
She found that calcium free solutions did not interfere with
vagus activity in the terrapin. The fact that potassium is
essential in vagus inhibition was further studied by Howell
 276 CHARLES M. GRUBER

and Duke (16). These observers, experimenting upon dogs, rabbits

and cats observed that stimulation of the vagus nerve caused
an increase in the potassium content of the circulating medium.
The increase may amount to as much as 29 per cent and each
stimulation of one-half to one minute is capable of liberating
from 0.4 to 0.5 mgm. of potassium. They observed no change
in the calcium content of the medium. Upon stimulation of
the sympathetic nerve there is no increase in the potassium
content of the circulating fluid. The potassium salts concerned
in vagus action are situated according to Burridge (17) at the
“Ner’e-endings” in the muscle.
In comparing these findings on the organ in situ and my
results with potassium in solution on the excised auricles it may
be assumed that the tonus waves produced upon vagus stimula-
tion are due, not to the nerve impulse per se, but to the potas-
sium thus liberated.
A number of experiments previously referred to showed that
stimulation of the sympathetic nerves caused a disappearance
of the tonus waves either when normally present or when origi-
nated by vagus stimulation. Bottazzi (10) and Sch#{228}ffer (18)
believe that epinephrin acts upon the sympathetic nerve endings.
This would seem to be supported by the fact that the addition
of epinephrin to the solutions in which the auricles are con-
tracting brings about the same response as does sympathetic
stimulation.
Unless the potassium content was too great (0.4 to 0.5 per
cent) epinephrin in all the experiments performed caused a
rapid disappearance of the general tonus and tonus waves. It
also caused in the majority of cases an increased rate and force
of contractions after they had almost disappeared from the
effects of the potassium.
 TONUS AND TONUS WAVES IN TERRAPIN AUBICLES 277

SUMMARY

Potassium chloride causes an increase in the general tone

and tonus waves and may cause a disappearance of the contrac-
tions in the excised terrapin auricles. The strength of potas-
sium necessary to produce this change varies in different auricles
but the average found for 55 experiments was 0.22 per cent.
The results with calcium chloride were so variable that no
deductions could be drawn.
No antagonistic action but rather a synergistic action exists
between potassium chloride and calcium chloride upon the
general tone and tonus waves in the auricles of terrapin.
Epinephrin in proper doses causes a disappearance of the
general tone and tonus waves aroused by potassium chloride
alone or in combination with calcium chloride, causes a reap-
pearance of the contractions if they have disappeared and in-
creases the rate and the strength of contractions when still
present.

REFERENCES

(1) HOWELL: Amer. Journ. of Physiol., 1898-99, ii, 47.

(2) MARTIN: Amer. Jour. of Physiol., 1904, xi, 370.
(3) FANO AND BODANO: Archiv. Ital. de Biol., 1900, xxxiv, 301.
(4) BorrAzzi: Arch. f. Physiol., 1901, 392.
(5) GRUBER: This Journal, 1920, xv, 23.
(6) MARTIN: Amer. Journ. of Physiol., 1904, xi, 375.
(7) HOWELL: Amer. Journ. of Physiol., 1901-02, vi, 187.
(8) GRUBER AND MARKEL: This Journal, 1918, xii, 43; ibid., 53; Gruber: ibid.,
1920, xv, 23.
(9) FANO: Beitrage z#{252}r Physiologie, Festschrift zu Ludwig Gewidmet, Leipzig,
1887, 287.
(10) BorrAzzi: Archiv. Ital. de Biol., 1900, xxxiv, 25; ibid., 1901, xxxvi, 277;

Zeitschr., f. Ailgem. Physiol., 1907, vi, 140.

(11) SORAKU OINuisA: Pfluger’s Archiv. f. Physiol., 1910, cxxxiii, 500.
(12) GAULT: Amer. Journ. of Physiol., 1917, xliii, 32.
(13) R08ENzwEIG: Archiv. f. Physiol., 1903, Suppl., 192.

(14) HOWELL: Amer. Journ. of Physiol., 1905-06, xv, 280.

(15) BRINE: Amer. Journ. of Physiol., 1917, xliv, 171.
(16) HOWELL AND DUKE: Amer. Journ. of Physiol., 1908, xxi, 51.
(17) BURRIDGE: Journ. of Physiol., 1917, li, 45.
(18) SCHAFFER: Endocrine Organs, New York, 1916, 84.