PERFUSION OF THE MEDULLA OF THE TURTLE

III. EPINEPHRIN

A. D. BUSH
From the Laboratory of Pharmacology, University of North Dakota

Received for publication April 10, 1920

In the perfusions of the medulla of the striped turtle with

solutions of epinephrin, the tabulated results shown on page 298
were obtained.
In every case where solutions of different strengths . of epi-
nephrin were used, or where a drug succeeded epinephrin or an-
other drug, the whole medullary field was first washed by per-
fusion with 20 mils of frog Ringer, running at a rate of not over
5 mils a minute. The epinephrin (“Adrenalin”) was physio-
logically tested and found potent. From these findings, it would
seem that epinephrin exerts little or no influence on the cardio-
inhibitory center.
Just as these experiments had been concluded, there appeared
in this Journal (vol. 14, no. 1, p. 17) a report on similar work on
epinephrin by Heinekamp (University of Illinois), who found
that in his experiments adrenalin perfused through the medulla
caused definite cardiac slowing and occasional inhibition. Such
diverse findings necessitated a repetition of my work, so that there
might be found a reason for the discrepancy. So, a new lot of
epinephrin was obtained direct from the manufacturer (“Adre-
nalin” 1-1000, Parke, Davis and Company) with the assurance
that the sample was fresh and potent. Its potency was demon-
strated on intestinal strips, dog circulation, and the author’s
nasal mucosa.
In this second set of epinephrin experiments the vagus was
tested each time just before and just after drug perfusion, a
tetanic current being used of a minimal strength to obtain ces-
sation of heart action with an application of one second or less,
297

THE JOUR. OF PHARM. AND EXPER. THERAP., VOL. XV, NO. 4

298 A. D. BUSH

when applied to the right vagus. In all these second experiments

the epinephrin used was in 1-100,000 strength in a vehicle of frog
Ringer. In each case 50 mils were perfused at a uniform rate of

Results obtained with solutions of epinephrin

F. F.
, HO RATE z
STRENGTH OF 4 EFFECT ON EFFECT ON
DRUG MINUTE HEART SEQUENT DRUG HEART

F.

cc. cc.

37 1-100,000 50 3 None Quinine, 0.01% 5 Inhibition

38 1-100,000 50 4 Slight Quinine, 0.01% 5 Slowing
slowing
39a 1-100,000 50 3 Augmented Quinine, 0.01% 4 Inhibition
tone
39b 1-100,000 50 3 None Quinine, 10 Slight
0.005% slowing
40 1-1,000 30 2 None Curare, 0.001% 20 None
Quinine, 0.01%
41 1-1,000 30 2 None Curare, 0.01% 20 None
Quinine, 0.01% 20 None
Strychnine, 15 Slowing
0.003%
42 1-3,000 30 3 None Curare, 0.05% 15 Slowing
Quinine, 0.01% 30 None
43 1-10,000 20 2 None
1-20,000 20 2 None Antipyrin, 25 None
1-50,000 20 2 None 0.2%
1-75,000 20 2 None Strychnine 2 Inhibition
44 1-150,000 50 3 None Antipyrin, 26 Brief inhi-
1-100,000 20 3 None 0.2% bition
1-75,000 20 3 None Prolonged
1-10,000 20 3 None slowing
45 1-50,000 15 2 Slight in- Antipyrin, 15 Prolonged
1-30,000 15 2 creased 0.2% slowing
1-20,000 15 2 systolic Strychnine, 5 Marked
1-10,000 15 2 tone 0.003% slowing
48 1-200,000 35 3 None Urea, 0.6% 35 None
Strychnine 4 Inhibition
49 1-1,500,000 30 3 None Urea, 2% 15 None
Strychnine 16 Inhibition
50 1-400,000 20 2 None Strychnine, . 12 Inhibition
1-200,000 20 4 None 0.003%

flow of 3 mils a minute, which is somewhat less than the rate of

blood flow in the carotid of a striped turtle of 8 inches length.
There was no registrable change in the rate of the heart in any
 PERFUSION OF MEDULLA OF TURTLE 299

of these 18 experiments, either during the flow of the perfusate

or for the succeeding fifteen minutes and in but one case was there
any change in systolic tone. The vagus influence, as tested by
the induced current, seemed normal after the perfusion. As soon
as the drug flow had ceased, the medulla was perfused with frog
Ringer, and soon afterward the sensitiveness of the medulla was
tested with strychnine solution, 0.003 per cent which gave
prompt inhibition in every case but one, in amounts required
ranging from 1 to 15 mils; the one exception showed slowing from
the strychnine, but not complete inhibition.
These thirty-one experiments would seem to indicate that
epinephrin does not stimulate the vagus center in the striped
turtle. How, then, may Heinekamp’s findings to the contrary
be explained? It would appear there were several divergencies
in method and possibly some of the discrepancies arose from
that fact. In the first place, in only one case did he tie off, or
clamp, the second carotid. Now, (1) whenever solution pressure
is high enough to cause a flow of 3 mils a minute, there will be a
slow backflow throughout an untied carotid sufficient to effect
the systemic circulation; this may readily be demonstrated by
the use of strychnine solution. (2) The color of the untied carot-
id gradually loses its crimson color, becoming blanched as the
solution displaces the blood. (3) Section of the untied carotid
showed perfusate present. (4) Control experiments with the
second carotid untied gave cardiac irregularities in 60 per cent of
tests with epinephrin, with cardiac inhibition in 44 per cent.
(5) Section of the vagi in strychnire inhibition invariably re-
moved the inhibition, but strangely enough failed to do so in
half the epinephrin controls mentioned under 4.
Bearing somewhat on the question of backflow through an
untied second carotid is Heinekamp’s statement that in some
cases he was obliged to break both the humeri and the femurs in
order to minimize spontaneous movements. Inasmuch as some
of his experiments involved the use of strychnine, one is led to
wonder if these so-called spontaneous movements were not
really strychnine convulsions brought on by the back seepage.
Such a consequence has been demonstrated in my laboratory sev-
eral times.
300 A. D. BUSH

Heinekamp pithed the cord of his turtles. No control experi-

ments have been done here to determine the relative reaction to
drugs of a pithed turtle from an unpithed one ; but since pithing
the cord produces a not inconsiderable physiological disturb-
ance, it is probable that a pharmacodynamic correlative must
ensue. Whatever may be the spinal tone influencing the heart
and vaso-constrictors, it would necessarily be altered by spinal
pithing.
Heinekamp cut the cord high in the neck, and then passed a
tight ligature around the remaining cervical tissues, not includ-
ing, of course, the carotids and vagi. In our work here we
followed Peeler’s original plan, and severed all the tissues in
the neck, except the vagi and carotids. Thus, in our work, the
medulla was completely isolated from the rest of the turtle, ex-
cept for the connection of the vagi and the two carotids-one of
which was tied. There was thus no chance for any cardiocipetal
influence to be exerted except through the vagi. Even the sev-
ered tissues were kept separated by means of a glass plate from
the remaining structures-an important precautionary measure,
as many experimenters with strychnine have quickly discovered.
In our experiments no blood vessels were tied except the two
carotids.
Heinekamp used a 0.9 per cent saline solution for his diluent,
while we used frog Ringer; but several control experiments did
not show this to be as important a point as might be supposed,
though the tonicity departure from the reptilian norm must
have introduced a new factor. Heinekamp makes no mention
of having kept his solution oxygenated, and though this may be
a relatively unimportant point yet the steady aeration we gave
our solutions must have kept conditions a little nearer normal.
In fact, it would seem that the conditions of our experiments
were more conducive to uncomplicated medullary reaction.
Therefore for the present, at least, we may feel justified in
making the following

CONCLUSION

Epinephrin does not seem to exert a registrable influence on

the cardio-inhibitory center of the striped turtle.