III.

A NOTE ON THE ACTION OF PILOCARPINE, ATRO-

PINE AND ADRENALINE UPON THE TONUS
WAVES IN THE TERRAPIN HEART

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 November 23, 1919

I have found since publishing an article on the action of adre-

naline, piocarpine and atropine on the tonus waves of the terrapin
heart, with Markel recently, that similar work had been done by
Fano and Bodano and Bottazzi. We failed to note their experi-
ments because the titles of the articles in which they appeared
gave no indication of their presence therein. Since, however,
our experiments, and this later series of mine, contradict Fano
and Bodano’s work and enlarge upon those of Bottazzi, their
publication may prove of some value.
Fano and Sciolla (1) in 1887 reported that atropine caused the
tonus waves to disappear with increased fundamental contrac-
tions. This was true, also, of waves aroused by muscarine.
Three years later Fano working with Bodano (2), upon terrapin
(Emys europaea) with the heart in situ, found that a 20 per
cent solution of piocarpine increased tonus waves, whereas
atropine in less than 30 per cent solution decreased tone and
caused the tonus waves to disappear. A 30 per cent solution of
atropine caused an increase in the tone of the muscle. In the
same year Bottazzi (3) noted that atropine decreased the tone
of the muscle, but increased the fundamental contractions. In
the following year he (4) found that tonus waves were not para-
lysed by doses of 0.5 to 1 per cent, but only by stronger solutions.
These results that atropine has no effect upon tonus waves,
were corroborated recently by Gruber and Markel (5).
23
 24 CHARLES M. GRUBER

In 1907, Bottazzi (6) found that adrenaline in solutions of

1: 20,000 caused a disappearance of the tonus waves just as did
sympathetic stimulation. Gruber and Market (7) found that it
was possible to bring about a loss of tonus waves not only with
strong solutions of adrenaline but also with extremely dilute
solutions, 1: 174,000,000 to 1: 200,000,000.

METHOD

The experimental animals were terrapin of the species Chry-

semys cineria, Chrysemys elegans, and Malacoclemmys lesuerli.
The terrapin was pithed, the plastron removed, the heart ex-
posed and excised, leaving a portion of the larger veins attached
to the auricles. The ventricles were cut from the auricles. The
auricl#{233}s were suspended vertically; one connected to an alumi-
num wire writing lever of the heart lever by means of an S-shaped
pin hook and silk thread, and the other fastened to a hook on the
L-shaped metal rod in such a way that the tension was put upon
the side opposite the bulbus arteriosis. The lever was loaded
according to the weight of the muscle preparation. The up
stroke on the records indicates systolic, the down stroke diastolic
contraction.
The entire sino-auricular muscle preparation was immersed in
beaker of Ringer’s solution (0.7 per cent sodium chloride, 0.03
per cent potassium chloride, and 0.042 per cent calcium chloride)
‘at a temperature of 20#{176}
to 22#{176}C. The beaker was supported by
.a small wooden stand, and could be removed and replaced by
another filled with a fresh solution at will. Either air or oxygen
was kept bubbling continuously through the fluid. The drugs
were, added directly to the solution in which the heart was
immersed.
An electro-magnetic signal recording five second intervals was
placed beneath the heart lever on the kymograph surface.

RESULTS

In the main my results with pilocarpine hydrochloride do not

corroborate those of Fano and Bodano’s (2). Of the 37 experi-
.ments performed upon as many animals (30 experiments with Mr.
 TONUS WAVES IN THE TERRAPIN HEART 25

Markel and 7 additional experiments) in only two cases was it

possible to obtain an increase in tonus waves. The solutions
varied in concentration from 0.0005 per cent to 0.05 per cent as
compared with Fano and Bodano’s 20 per cent solutions. In
all cases it produced a marked slowing and a decreased height
of the contractions.
Figure 1 is one of the two records obtained in which piocar-
pine produced an increase in the general tone and tonus waves.

FIG. 1. In this and the following records the upper curve is that of the con-
tracting auricles; the lower that of the time interval in five seconds. The up
stroke is the systolic contraction, the down stroke the diastolic contraction.
1, Pilocarpine hydrochloride, 5 mgm. in 100 mils. of saline; 2, adrenaline chloride,
4 mils. of a 1: 1,000 solution; 3, atropine sulphate, 5 mgm.

The quantity of pilocarpine hydrochloride added (at 1) was just

sufficient to produce a 0.005 per cent solution. Simultaneously
with the increased tone there is a marked slowing of the rate
and a decrease in the height of the contractions. In figure 2
(at 1) pilocarpine hydrochloride 0.002 per cent had no effect
upon the tonus waves or general tonus. It decreased the rate
26 CHARLES M. GRUBER

and height of contraction, hut the tonus waves remained un-

changed. This result is similar to those noted by Gruber and
Markel (5) (see curves 1 and ). In figure 3 pilocarpine 0.005
per cent (at I) produced similar results. In all those experiments
in which the tonus waves were not present )iloCarpifle in small

FIG. 2. 1, Pilocarpin. hydrochloride, 2 mgm. in 100 mils. of Ringer’s solution;

2, adrenaline chloride, 1: 30,000 solution.

Fio. 3. 1, Pilocarpine hydrochloride, 5 mgm. in 100 mils. of Ringer’s solution;

2, adrenaline chloride, 2 mils. of a 1: 1000 solution.

quantities, although effective in causing a decreased rate and

height of contraction, was unable to bring about tonus waves.
This is in direct contradiction to Fano’s results upon muscarine
(1) and pilocarpine (2).
It was demonstrated by Bottazzi (4) that small doses of at-
ropine sulphate do not affect the tonus waves. The only effect
that atropine has following the administration of pilocarpine is
 TONUS WAVES IN THE TERRAPIN HEART 27

to cause to disappear very quickly the decreased rate and height

of contraction (see Gruber and Markel (5), figs. 1 and 2).
The effect of adrenaline upon the tonus waves was first studied
by Bottazzi (6) and later again by Gruber and Markel (7). It
was found by the latter observers that adrenaline caused to
disappear very quickly the tonus waves and general tonus and
to increase the height of contraction.
The effect of adrenaline was studied in connection with pile-
carpine. In the two cases in which pilocarpine caused an increase
in the general tone and tonus waves, adrenaline caused a rapid
loss of tone and a disappearance of the tonus waves. Figure 1
illustrates this very well. At (2) 4 mils. (1: 1000) of adrenaline
was added to the 100 mils. of Ringer’s solution to which the
pilocarpine had previously been added. There was, simultane-
ously, with the loss of tone and tonus waves an increase in the
rate and height of contraction. This effect of adrenaline upon
the rate and height of contraction is better illustrated in figures 2
and 3 (at 2). A sufficient quantity of a 1: 1000 solution of
adrenaline chloride was added to make in figure 2 a 1: 30,000, and
in figure 3 a 1: 50,000 solution. In figure 2 the rate quickly
changed from about 4 to 18 beats per minute. In figure 3 the
rate changed from 10 to 27 contractions per minute. The action
is somewhat similar to that caused by atropine after pilocarpine
inhibition, but slower in development. The rapidity at which
atropine acts can be seen in figure 1 at 3 and in figure 1 in the
paper by Gruber and Markel (5). These results seem to sub-
stantiate Bottazzi’s and Sch#{228}ffer’s (8) views that adrenaline acts
irpon the sympathetic mechanism in the heart.

SUMMARY

In my experiments pilocarpine hydrochloride in small doses

0.0005 to 0.05 per cent produced an increase in the tonus and
tonus waves in less than 6 per cent of the experiments performed.
These results are contrary to Fano and Bodano’s findings. The
difference is probably due to a difference in the strength of the
solutions employed. The latter observers used 20 per cent
solutions.
28 CHARLES M. GRUBER

Atropine sulphate has no action on the tonus waves. These

results confirm those of Bottazzi.
Adrenaline chloride decreased the increased tone set up by
piocarpine and simultaneously increased the height and rate of
the contraction. In six other experiments performed, in which
piocarpine produced only a slowing of the heart rate and de-
creased the height of contraction, adrenaline antagonized the
pilocarpine action.

REFERENCES

(1) FANO AND ScI0LLA: Archives Italiennes de Biologic, 1887-88, ix, 61.
(2) FANO AND BODANO: Archives Italiennes de Biologic, 1900, xxxiv, 301.
(3) Borr.&zzi: Archives Italicnncs de Biologic, 1900, xxxiv, 25.
(4) BorrAzzi: Archiv f#{252}r
Physiologic, 1901, 404.

(5) GRUBER AND MARKEL: This Journal, 1918, xii, 53.

(6) BOTAZZI: Zcitschr. f. Allgein. Physiol., 1907, vi, 4170.
(7) GRITBER AND MARKEL: This Journal, 1918, xii, 43.
(8) SCHAFFER: Endocrine Organs, New York, 1916, 84.