A STUDY OF THE ADRENOTROPIC RECEPTORS

RAYMOND P. AHLQUIST
From the Department of Pharmacology, Uniwvsity of Georgia School of Medicine
AUGUSTA, GEORGIA

T
HE adrenotropic receptors are those hypothetical structures or systems
located in, on or near the muscle or gland cells affected by epinephrine.
The concept of a receptive mechanismwas introduced by Langley (I, 2) to
explain the action of curare on skeletal muscle. Dale was probably the first to make
significant use of the receptor concept in connection with the sympathetic nervous
system. In his classicalpaper (3) on the sympatholytic action of the ergot alkaloids,
he recognized that what he called the sympathetic myoneural junction could also be
called ‘the receptive mechanismfor adrenaline’; and he used this mechanismto ex-
plain the fact that the ergot alkaloids prevented only the motor (excitatory) actions
of epinephrine and had no effect on the inhibitory actions of epinephrine or on the
excitatory actions of barium or pituitrin.
The adrenotropic receptors have been considered to be of two classes,those
whoseaction results in excitation and those whoseaction results in inhibition of the
eff ector cells. Experiments described in this paper indicate that although there are
two kinds of adrenotropic receptors they cannot be classifiedsimply as excitatory or
inhibitory since each kind of receptor may have either action depending upon where
it is found. The evidence for these conclusionsis, in brief, that a seriesof six sympa-
thomimetic amines has one order of potency-r, 2, 3, 4, 5, 6-on the following
functions: vasoconstriction, excitation of the uterus and ureters, contraction of the
nictitating membrane, dilation of the pupil and inhibition of the gut. In contrast,
this sameseriesof amineshas an entirely different order of potency-z, 4, 6, 5, 3, I-
on the following functions: vasodilation, inhibition of the uterus and myocardial
stimulation.
The variations in activity could be due to any or all of three factors: a) quantita-
tive differences in potency, b) qualitatively different effects or c) differences due
entirely to the experimental methods used. If the last Tao factors are controlled
as much aspossibleby the selectionof the aminesand by using suitable experimental
techniques, then the variations in activity are presumably due to actual differences
in the receptors involved. Tentatively the first kind of receptor has been called the
alpha adrenotropic receptor and the secondkind the beta receptor. This concept of
two fundamental types of receptorsis directly opposedto the concept of two mediator
substances(sympathin E and sympathin I) as propounded by Cannon and Rosen-
blueth (4) and now widely quoted as a ‘law’ of physiology. Results reported in this
paper indicate that conclusions drawn by Cannon and Rosenblueth are open to
controversy and that there is no known amine which fulfills the requirements for

Received for publication May IO, Ig48.

586

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June 1948 ADRENOTROPIC RECEPTORS 587

either sympathin E or I. The evidence presented strongly supports the concept that
epinephrine is the only sympathetic adrenergic mediator.
The amines used were restricted to those which produced responses in the same
dosage range as epinephrine, for which equivalent doses had similar durations of
action, and which had qualitatively identical actions on the myocardium. Sane
of them produced tachyphylaxis. The following amines were studied (abbreviations
given in italics will be used throughout the remainder of this paper):
I. d-(3, +dihydroxyphenyl) ethanolamine. Arterenol, art.
II. dl-(3, +dihydroxyphenyl) isopropanolamine. Cobefrine, mefhyl-art.
III. and IV. dl, and Z-(3, +dihydroxyphenyl) methyl ethanolamine. Racemic
and levo epinephrine, dl-epi. and I-epi.
V. dl-(3, +dihydroxyphenyl) methyl isopropanolamine. Methyl-epi.
VI. dl-(3, +dihydroxvphenyl)
d isopropyl ethanolamine. N-isopropyl artere-
nol, N-Go-art.
All of these amineshave been studied extensively in the past (5-20) and hence
there is a voluminous literature concerning them. No previous studies, however,
have included all of them together in a quantitative comparison.

PROCEDURE

Dogs, cats, rats and rabbits were used in this study. The amines, in the form
of their hydrochlorides, were made up in M/ IOOO stock solutions using isotonic saline
containing 0.1 per cent sodiumbisulfite and o. I per cent chlorobutanol as the solvent.
Higher dilutions were made at the time of use with isotonic saline. The useof other
drugs was kept to a minimum. Sodium pentobarbital, 20 to 30 mgm. per kgm., or
urethane, I gram per kgm., was usedas the anesthetic. The dogsand rabbits were
usually pretreated with morphine sulfate, IO mgm. per kgm. Atropine, 0.5 mgm. per
kgm., was used as the anticholinergic agent. Three sympatholytic agents were
used: ergotoxine, 2 mgm. per kgm., dibenamine (21), 25 mgm. per kgm. and Priscol
( 22. >
Two types of dosagewere employed with the amines:equimolar and equivalent.
Truly equivalent dosesin intact animals were many times unobtainable. For exam-
ple, in attempting to determine the equivalent dosagesnecessaryto inhibit the intact
intestine it was found that the marked cardiovascular effects (somepressorand others
depressor)interfered with or in themselvesproduced changesin the intestinal activity.
For this reasonequivalent doseswere determined chiefly on isolated structures and
equimolar dosescompared in intact animals. In determining the relative activity
of equimolar doses,both the degreeand the duration of the responsewere considered.
The amineswere administered intravenously unlessotherwise stated.
Cardiovascular. There are at least three functions served by adrenotropic
receptors in the cardiovascular system: vasoconstriction, vasodilation and myocardial
stimulation. Arterial pressurechangesin themselvesare not suitable for comparing
the activity of these amines on these receptors becausethe arterial pressurerepre-
sents the resultant of a number of fat tors such as the cardiac output and venous re-
turn, the balance between constriction, dilation and blood viscosity, and the me-
chanical effects produced by the muscular contractions such as occur in the heart,

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intestine or uterus. Therefore in order to study each type of receptor it was necessary
to work with more or less isolated parts of the cardiovascular system and then to
correlate these results with those obtained from arterial pressure studies.
The arterial pressure was recorded by means of either a mercury manometer or
a Hamilton manometer (25) from the carotid or femoral artery. In many of the dogs
the aortic pressure pulse was recorded by means of a high frequency Hamilton ma-
nometer using a metal sound passed through the left common carotid into the aortic
arch. In these latter experiments two recording cameras were used, one running
continuously at low speed for a complete record of the amine response, and the other
at high speed for pulse contours at appropriate times during the amine response.
The blood flow was measured in the important vascular beds by means of flow-
meters introduced into either the venous outflow or the arterial supply. A Shipley
optically recording rotameter (24, 25) or a modified Soskin type (26) ‘bubble’ flow-
meter was used. The ‘bubble’ flowmeter was equipped with a reversing system so
that only a single air bubble was needed for any one experiment. Heparin, IO mgm.
per kgm. every two hours, was used as the anticlotting agent.
The effects of the amines on the vascular receptors were compared by deter-
mining their actions on the vasomotor resistance (VR). The VR represents the
P - 20
pressure-flow relationship calculated by the formula (27), VR = F in which P

is the arterial pressure in mm. Hg and F is the volume flow in cc. per minute. P
- 20 was used since the P/F ratio changes abruptly at a pressure of about 20 mm.
Hg (28), a phenomenon due in part to the presence of the cellular elements in the
blood.
The results on the VR of the renal, mesenteric and femoral beds are shown in
figure I and tabulated in table I. It will be seen that the increase in VR produced by
epi. in these vascular beds bears an inverse relationsh ip to the decrease in VR pro-
duced by N-iso-art. In the bed (renal) which shows the least dilation with N-ho-art.,
dl-epi. is the most active among the racemic amines in producing constriction. In
the femoral bed, in which N-iso-art. produces the greatest dilation, the net constrictor
effect of &epi. is diminished by its own dilator action. These results are interpreted
as showing I) that the ratio of constrictor to dilator receptors varies in the different
vascular beds and 2) that dl-epi. is the most active of the racemic amines on the
constrictor receptor. Art. on the other hand, produces the greatest constriction in
those beds which have the most dilator receptors, since this amine has very little
action on the dilators (see below). The remaining two racemic amines show vari-
able amounts of constrictor as against dilator activity.
This same relationship was shown in the arterial pressure responses to these
amines as shown in figure 2. In the cat, which appears to have many dilator recep-
tors, art. is the most active pressor agent among the racemic amines, with dZ-epi. being
third. In the rabbit, which appears to have few dilator receptors, dl-epi. is more
active than art. Even N-iso-art., which is a depressor in cats and dogs, produces a
slight pressor response in rabbits. Blood flow studies showed that this pressor ac-
tion of N&o-art. in rabbits was due in part to vasoconstriction.
The comparative effects of these amines on the vasodilator receptors were deter-

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June 1948 ADRENOTROPIC RECEPTORS 589

mined by measurementsof the coronary flow in perfused hearts (seebelow) and by

their depressoreffects in intact animalsafter the administration of the sympatholytic
agent Dibenamine. In cats treated with the sympatholytic agent, I-epi. and dZ-epi.

loo

IO

i
----e.- -,
- ---. we- M/l*~o~------.-- ---- ---e-v__B__

I
I* + I I I I I I I I I I I I

I
MESENTERIC
IO

LI I 4 I I I I I I I 1
I I I I

F FEMORAL

Fig. I. COMPARATIVE ACTION OF AMINES on vasomotor resistance in the renal, mesenteric and
femoral vascular beds of dogs. Ordinates: VR plotted logarithmically for convenience only; abscis-
sae: time marks at IO sec. intervals. The amines were injected intra-arterially as 0.1 cc. of the
concentration shown. In the case of the renal and mesenteric curves, lower concentrations of
methyl-epi. and N-Go-art. had no appreciable effect on the VR.

were both very active depressors. On the other hand, art. and methyl-art. were both
poor depressors,with the former being the less active in this respect. The usual
depressoreffects of MethyLepi. and N-Go-art. were slightly augmented. In dogsthe
results were essentially the same,with the exception that art. did not lower the pres-

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590 RAYMOND P. AHLQUIST VoZ26me 153

sure in any animal tested. In rabbits none of these amines produced depressor
responsesafter Dibenamine, with the occasionalexception of N-Go-art.
The relative order of activity of these amineson the vascular receptors can now
be summarized as follows (seetable I). On the dilator receptors N-iso-art. is the
TABLE I. EXPERIMENTAL RESULTS. THE MOST ACTIVE AMINE ON EACH RECEPTOR IS NUMBERED I,
THE NEXTMOSTACTIVE 2,ETC.

ORDEROFRELATIVE ACTIVITY
ADRENOTROPIC RECEPTOR AND METHOD OF EVALUATION

l-epi. dl-epi. art. methyl- methyl- N-iso-

art. epi. art.
-___ ----- --~-_______~ --

CARDIOVASCULAR
Vasoconstrictor
Renal vasomotor resistance. ................... I 2 4 5
Mesenteric “ 66 .................... I 2 4 5
Femoral “ 64 .................... I 3 4 5
Pressor action in cats. ........................ I 4 3 5
Pressor action in dogs. ....................... 1 3 4 5
Pressor action in rabbits. ..................... I 2 4 5 6
Vasodilator
Renal vasomotor resistance. ................... 2
Mesenteric “ (6 .................... 2
Femoral “ 66 .................... 2
3 4
Coronary dilation. ........................... 2 4 6 5 3
Depressor after Dibenamine. .................. 2 4 6 5 3
Myocardial
Perfused rabbit and cat. ...................... 2 4 6 5 3 I
Intact dog .................................. 2 4 6 5 3 I
INTESTINAL (inhibitory)
Isolated rabbit, rat etc .......................... I 2 3 4 5 6
Intact dog, cat and rabbit. ...................... I 2 3 4 5 6
UTERINE
Excitatory
Isolated rabbit. .............................. I 2 3 4 5 6
Intact dog and rabbit. ........................ I 2 3 4 5 6
Inhibitory
Isolated rat, cat, rabbit. ...................... 2 4 6 5 3 I
Intact dog, cat, rabbit. ....................... 2 4 6 5 3 I
URETERAL (excitatory)
Intact rabbit. ................................. I 2 3 4 5 6?
DILATOR PUPII,LAE (excitatory)
Intact cat ..................................... I 2 3 4 5 6
NICTITATING MEMBRANE (excitatory)
Intactcat ..................................... I 2 3 4 5 6?

most active followed in order by l-e@., methyl-e& dl-epi., methyl-art. and art. The
relative activity on the constrictor receptors must be interpreted by correlating their
actions on both the dilator and constrictor receptors with the relative ‘number’ of
each of these receptors in the vascular bed or speciesstudies. The dilator activity
of I-epi. and dl-epi. is so great that this effect diminishesthe net constrictor or pressor
effect in those vascular beds or specieshaving many dilator receptors. Therefore

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Jzme 1948 ADRENOTROPIC RECEPTORS 591
Z-e@. is considered to be the most active amine on the constrictor receptors followed
in order by dl-epi., art., methyl-art., methyl-epi. and N-iso-art.

- 1 CATS Afm DIBENAMI NE

It--\
)A
METHYL-El%
N -iSO-ART.
\ hi/lO,OCXl

I ----a ART. I

n 4BBlTS I -N- W-ART.

Fig. 2. COMPARATIVE ACTION OF AMINES on mean arterial pressure of cats, dogs and rabbits.
Ordinates: pressure in mm. Hg; abscissae: time marks at IO sec. intervals. The amines were in-
jected intravenously.
Cats: average results from 12 determinations in 6 cats. Dosage, 0.1 cc. k1/1ooo solution per
kgm. Cats after dibenamine-average results from 3 determinations in 3 cats. Dosage, 0.1 cc. per
kgm. of the concentrations shown. Lower concentrations of art. did not produce any depressor
responses. Dogs: average results from 12 determinations in 8 dogs. Dosage, 0.05 cc. M/IOOO
solution per kgm. Rabbits: average results from IO determinations in 6 rabbits. Dosage, 0.1 cc.
M/IOOO per kgm. Larger doses of N-Go-art. produced depressor responses.

The relative activity of these amines was determined on both the perfused and
intact heart. The isolated heart of the rabbit or cat was perfused with Ringer-
Locke solution by the method of Lagendorff. Oxygen containing 5 per cent carbon
dioxide was used to aerate the solution and to maintain the perfusion pressure
(usually about IOO mm. Hg). Coronary inflow was measuredby the rotameter and

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592 RAYMOND P. AHLQUIST Volw~te 153

the cardiac contractions were recorded with an optical lever system. This method
allowed an absolutely simultaneous record of coronary inflow and myocardial ac-
tivity to be obtained. Figure 3 illustrates the type of record obtained, while table
2 gives the results obtained on 20 rabbit hearts.

Fig. 3. EXAMPLE OF THE ACTION of the amines on the perfused rabbit heart. Cardiac contrac-
tions (myocard.) recorded with an optical lever. Coronary inflow (car.) recorded with a rotameter.
Time marks at 30 sec. intervals. The measurements listed in table z were made at the indicated
points: R & A-I, the increased rate and amplitude at the point of maximal effect; R-z, the increased
rate at a point about z min. after the amine administration (a measure of the duration of action);
D.F., the decreased coronary inflow due to the increased myocardial activity; and I.F., the increased
coronary flow due to the direct action on the coronary vessels. The amines were injected into the
coronary inflow.

TABLE 2. COMPARATIVE EFFECTS OF THE AMINES ON THE PERFUSED RABBIT HEART’

COaONAaY FLOW

D.F.O j I.F.d
,
% 4
,V-iso-art.. ........ IO1 90 II I ;;
l-cpi. ........ 75 70 Ii 15
melllyl-cpi.. ........ 62 67 13 IO

dl-epi.. ...... 57 45 18 4
methyl-art. . . 40 43 16 4
arl. . . . 3g 3.5 20 3

* Averages obtained from zo hearts in which each amine was tested in each heart in a dosage
of 0.1 cc. M/~o,ooo solution injected into the coronary inflow. See figure 3.
* Increase in rate and amplitude at the time of maximum effect.
b Increase in rate about 2 min. after injection.
E Decreased coronary inflow measured at point of maximum decrease.
d Increased coronary inflow measured at point of maximum increase.

Each of the amines increased the rate and amplitude of the cardiac contractions,
the relative order of activity being given in tables I and 2. Simultaneous with the
myocardial stimulation there was a decrease in the coronary inflow which was prob-
ably mechanical in origin (29). The flow then returned to or above the control value.
Since the coronary flow in the perfused heart is determined by the balance between
the myocardial stimulation and the vasodilation, those amines with the least dilator
activity would allow the greatest mechanical decrease in flow. In contrast, those
amines with the greatest dilator action would allow the least mechanical effect. The

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June 1948 ADRENOTROPIC RECEPTORS 593

relative order of coronary dilator activity in the perfused heart will then be seento
be the sameas describedabove for the other dilator receptors.
The relative order of activity of these amines on the intact myocardium was
determined by two methods. Wiggers (30), Krop (31) and Remington and Hamilton
(32) have shown that epinephrine shortens the time of systole (T,) even though the
heart rate may be unchanged. This shortening of T, which probably results from
the myocardial stimulation was also produced by the other amines studied. The
T, was measuredon aortic pressure pulse contours of dogs from the beginning of
systolic upstroke to the incisura. The apparent order of activity of theseamines to
shorten the T, was found to be the sameas their relative order on the isolated myo-
cardium.
The blood pressureeffects of these amines in dogs pretreated with ergotoxine
can be used as a measureof their cardiac action since this sympatholytic agent, in
common with dibenamine and Priscol, does not prevent their myocardial actions.
Although ergotoxine unmasks the vasodilator effects of the amines it also produces
enough vasoconstriction in somedogsto diminish the depressoractions of the amines.
This effect allows the aminesto produce an increasein arterial pressurein these ani-
mals by myocardial stimulation, an effect confirmed by cardiac output measurements
by the contour method of Hamilton and Remington (33). The comparative activity
of theseamineson the myocardium astested by this method wasfound to be the same
as found on the perfused heart.
The relative order of activity of these amines as myocardial stimulants will be
seen (table I) to be the sameas that found for their vasodilator actions. This in-
dicates that the myocardial receptor is related to the vasodilator receptor rather than
to the vasoconstrictor receptor.
Intestine. The contractions of the isolated ileum of rabbits, cats and rats were
recorded in the usual manner using a 40 cc. musclechamber and Ringer-Locke solu-
tion. Three types of dosagewere used: a) equimolar concentrations applied to the
same segment of the ileum, b) equivalent concentrations to the samesegment and
c) equivalent concentrations to different segmentsfrom the sameanimal (only one
amine to each segment). The results were the samefor each method and for each
speciesand are tabulated in table I.
The contractions of the intact ileum of anesthetized dogs, cats and rabbits were
recorded with a sensitive Hamilton manometer by meansof a small, water-filled bal-
loon inserted through a stab wound in the intestinal wall. In all of theseanimals the
arterial pressurewas also recorded and in many of them other records such as mesen-
teric or femoral blood flow, or activity of the uterus or nictitating membrane were
alsoobtained. Only equimolar dosesof the amineswere compared sincethe vascular
effects of these aminesinterfered with their intestinal effects. The order of relative
activity was found to be the sameas that found on the isolated ileum. The most
active amine was l-e@., followed in order by dl-epi., art., methyl-art., methyl-epi. and
N&o-art. This order of activity indicates that the intestinal inhibitory recep-
tor must be related to the vasoconstrictor receptor rather than to the vasodilator
receptor.
Uterus. This organ presentsa peculiar problem becauseof its well-known varia-

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594 RAYMOND I’. AHLQUIST I~olfillre 15.3

tions of response to epinephrine. The findings in the literature, and those reported
herein, indicate that all uteri have two adrenotropic receptors, one excitatory and
the other inhibitory in function, and that the response to epinephrine is determined by
which receptor is predominant. The cat uterus, for example, is usually inhibited
when nonpregnant and excited when pregnant. The isolated rabbit uterus, on the
other hand, is always excited. The intact uteri of dogs, rabbits and humans (preg-
nant or nonpregnant) exhibit a diphasic response, excitation followed by inhibition.
The responses of the isolated nonpregnant rat uterus (inhibitory) and the rabbit
uterus (excitatory) were recorded as described above for the isolated intestine. The
comparative effects of the amines on these two species are given in table I. The
order of inhibitory action is seen to be the same as found on the vasodilator and
myocardial receptors: N-iso-arl. is the most active, followed in order by 1-epi.,
methyl-epi., dl-epi., methyl-art. and art. The order of excitatory activity is the same

Fig. 4. EFFECT OF INTRA-ARTERIALLY INJECTED EPIxwmfRINE on the uterine activity and blood
tlow. Calibrations: arterial pressure (A.P.) and intrauterine pressure (U.P.) in units of 50 mm.
Hg; uterine blood tlow (F.) in units of IO cc. per min. Time marks at xo-sec. intervals. During the
control period the uterus was contracting every 3 to 4 min. At the signal, 0.1 cc. M/rooo dl-epi. was
injected into the uterine artery. Note that during the control contraction the decrease in blood flow
occurs simultaneously with the uterine contraction. Following the dl-epi., the flow diminished first,
then one uterine contraction appeared, followed by complete inhibition. The zero flow and uterine
inhibition persisted for more than 30 min.

as found on the vasoconstrictor and intestinal inhibitory receptors, 1-epi., the most
active, followed by dl-epi., art., methyl-art., methybepi. and N-iso-art.
When the isolated rabbit uterus was pretreated with Priscol, a sympatholytic
agent, the amines lost their excitatory effects and produced only inhibition, with the
order of activity being the same as found on the rat uterus. Due to a limited supply,
only 2 virgin cat uteri were tested and these gave results identical to those found on
the rat uterus.
The responses of the intact uteri of pregnant and nonpregnant dogs, cats and
rabbits were recorded as described above for the intact intestine. The results ob-
tained here were difficult to interpret, not only because of the diphasic effects that
occurred but also because the muscular effects were complicated by the vascular ef-
fects. This can best be described by the following example. The uterine blood
flow and the intrauterine pressure were measured simultaneously in a dog a few days
post-partum (34). The intra-arterial injection of 0.1 cc. of M/moo dl-epi. produced
a complete cessation of blood flow lasting about 30 minutes. One uterine contrac-
tion was produced and this was followed by complete inhibition also lasting for about
30 minutes (fig. 4.). In contrast to this result, the administration of an equimolar
dose of N-Go-art. produced a slight increase in flow together with a slight inhibition

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June 1948 ADRENOTROPIC RECEPTORS 595

of activity; these effects lasted only 3 minutes. These results show that the vascular
effects so modify the duration of action of the muscular effects (constriction increas-
ing and dilation decreasing) that a true comparison of activity becomes almost im-
possible. With intravenous injections the arterial pressure changes added still
another complicating factor
The apparent relative activity of these amines on the intact uteri is given in table
I. The order of activity on the uterine inhibitory receptor (derived from the primary
effect in the non-pregnant cat and the secondary effect in dogs and rabbits) is seen to
be the same as found for the isolated rat uterus. The order of excitatory activity
(derived from the primary effect in dogs and rabbits) was the same as described for
the isolated rabbit uterus.
IrZ:s and Nictitating Membrane. The comparative effects of the amines on these
structures were determined in anesthetized cats. The responses of the dilator
pztpillae were determined by direct observation, using an ordinary 6o-watt lamp at I 2
inchesfor illumination. The movementsof the nictitating membrane were recorded
with a simplestring and lever system. Someof the eyeswere acutely denervated by
section of the cervical sympathetics but this procedure had no apparent influence on
the results. In someof the cats the amineswere injected into the common carotid
to avoid their blood pressureeffects as much as possible.
Equimolar and equivalent doseswere tested. The relative order of activity of
these amineswas found to be the sameon both structures as shown in table I. The
most active amine was Z-e& with dl-epi. about one-half as active, art. about one-
sixth, methyZ-art. one-fifteenth, and methydepi. one-fortieth. Only occasionally did
N-iso-art. have any effect on these structures and then only when given in a very
high dosagedirectly into the artery.
Ureter. The actions of the amines were compared on the ureter of the anes-
thetized rabbit. The ureter was exposedat the renal pelvis and cannulated. Saline
was perfused under a pressureof about 25 cm. of water, the rate of flow being re-
corded with a drop counter. The bladder was incised to allow free escapeof the
perfusate. All of the amines, except N-iso-art., decreasedthe flow when injected
intravenously. The relative order of activity as determined by the amount of flow
decreaseproduced by equimolar dosesis shownin table I. N-iso-art. had practically
no effect on the flow except when given in very high dosageat which time it increased
the flow rate.
The adrenotropic receptor of the rabbit ureter appears to be mainly excitatory
in nature and related to the uterine excitatory and vasoconstrictor receptors.

DISCUSSION

The relative order of activity of these sympathomimetic amineson the various

adrenotropic receptors is summarized in table 3. There are at least two distinct
general types of thesereceptors. One type of receptor is associatedwith most of the
excitatory functions and with at least one of the inhibitory functions (intestine).
The other type is associatedwith most of the inhibitory functions and with one im-
portant excitatory function (myocardium). The relative order of activity of dl-epi.
and art. on the vasoconstrictor receptor is opposite to that previously reported in

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596 RAYMOND P. AHLQUIST Volume 153

the literature. This difference is probably due to the previous methods of compari-
son, in which the pressor effects only were used as the criteria of activity. The fact
that dl-epi. is more active than art. is more in accord with their relative activities on
someof the other excitatory receptors.
Becauseof the opposite effects associatedwith each type of receptor, the cus-
tomary signs, E (excitatory) and I (inhibitory), cannot be applied. Therefore, for

TABLET. SUMMARY OF THE RELATIVE ORDER OF ACTIVITY OF THE AMINES

ORDER OF ACTIVITY
RECEPTOR
Most active --* Least active

Vasoconstrictor . . .. . . . I-epi. dl-epi. art. methyl-art. methyl-epi. N-iso-art.

Uterine excitatory. . .. I-epi. dl-epi. art. methyl-art. methyl-epi. N-iso-art.
Nictitating membrane
excitatory. . . . . .. . . . I-epi. dl-epi. art. methyl-art. methyl-epi. N-iso-art.
Dilator pupillae excit-
atory. . . . . . . . . . . . . . I-epi. dl-epi. art. methyl-art. methyl-epi. N-iso-art.
Ureteral excitatory. .. . I-epi. dl-epi. art. methyl-art. methyl-epi. N-iso-art.
Intestinal inhibitory. .. I-epi. dl-epi. art. methyl-art. methyl-epi. N-iso-art.

Vasodilator . . . . . .. . . . N-iso-art. I-epi. methyl-epi. dl-epi. methyl-art. art.

Uterine inhibitory. . .. . N-iso-art. I-epi. methyl-epi. dl-epi. methyl-art. art.
Myocardial excitatory. N-iso-art. I-epi. methyl-epi. dl-epi. methyl-art. art.

TABLE 4. STRUCTURES OR FUNCTIONS CONTAINING OR ASSOCIATED WITH EACH OF THE TWO TYPES
OF ADRENOTROPIC RECEPTORS'
ALPHA RECEPTOR BETA RECEPTOR

Vasoconstriction Vasodila tion

Viscera Skeletal muscle
Skin Coronary
Nictitating membrane Viscera (few)
Uterus (excitatory) Myocardium
Rabbit Uterus (inhibitory)
Dog Rat
Human Cat
Intestine Dog
Ureter Human
Dilator pupillae Bronchi (20)

l The following structures or functions have not as yet been completely tested: spleen, erectores
pilorum, urinary bladder, glands and glycogenolysis.

convenience they have been designatedas the alpha adrenotropic receptors and the
beta receptors. Table 4 lists the structures or functions associatedwith each type of
receptor.
This concept of two fundamental types of adrenotropic receptors is directly op-
posed to the concept of two adrenergic mediators. Epinephrine has all of the chem-
ical and physical properties, and, as shown in table 3, all of the physiological proper-
ties necessaryto be the only adrenergic mediator. It is the most active substanceon
the alpha receptors and almost the most active on the beta receptors. It is, therefore,
the one amine which is both the best excitatory agent and the best inhibitory agent

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JUTE Ig@ ADRENOTROPIC RECEPTORS 597

on the effector cells thus far tested. It is fundamentally the most logical substance
to be the sympathetic neuro-hormone since all histological and embryological evi-
dence points to the similarity of the adrenal medulla and the adrenergic post-gang-
lionic nerves.
Cannon and Rosenblueth basedtheir belief in two kinds of sympathin on three
fundamental observations (4) : a) ergotoxine affects releasedsympathin and injected
epinephrine differently, b) the combined effect of simultaneously releasedsympathin
from two different sourcesappears to be greater than expected and c) the substance
releasedby the stimulation of the hepatic nerves (liver sympathin) has physiological
actions different from those of injected epinephrine. In order to support the theory
of a single mediator and two receptors, it is necessaryto challenge the validity of
the conclusionsdrawn from these observations.
A comparisonof the effects of intravenously injected epinephrine and sympathin
releasedby stimulation of the lower abdominal sympathetics is not valid becauseof
the difference in the manner of delivery of the two substancesto the effector cells.
A constrictor substancepresented intra-arterially, or releaseddirectly into a vascular
bed by nerve stimulation, tends to hold itself in the periphery by its own constricting
action. This can be readily shown by injected epinephrine into the femoral artery
while measuring the femoral blood flow. Ergotoxine does not prevent this effect
since it, in common with the other known sympatholytics, is not absolute in its
sympatholytic or adrenolytic action. Therefore, a large amount of sympathin
formed in the periphery by nerve stimulation (the figure reproduced by Cannon and
Rosenblueth shows that prolonged nerve stimulation was done) would, even in the
presenceof ergotoxine, result in contraction of the vesselsof that region, which in
turn would hold the sympathin and not allow free circulation to other sites. The
constriction would therefore be restricted and prolonged. Epinephrine, on the
other hand, administered intravenously, would be delivered to all parts of the body,
and therefore in low enoughconcentration to produce vasodilation by selective action
upon the adrenotropic dilator receptors.
The apparent difference between sympathins from two sources (the cardio-
accelerator and splanchnic nerves) as interpreted by Cannon and Rosenblueth is
questionable, since the cardiovascular effects were not considered. The effects (on
the nictitating membrane) of the sympathins were separately quantitated against
intravenously injected epinephrine. Simultaneous stimulation of the two sources
then produced a greater responsethan expected from the original quantitation.
The amount of sympathin delivered to the nictitating membraneis a function of the
amount formed at the remote ending and of the blood flow from that region to the
membrane. The blood flow was undoubtedly increasedduring the double stimula-
tion (especially by the cardio-acceleration) and tended to force (against the holding
effect described above) more of the splanchnic sympathin to the membrane. This
n-odd render the original quan tita tion invalid. The results may then be explained
on the basisof delivery of different amounts of a single substancerather than by a
potentiated effect produced by two different substances.
The substanceproduced by stimulation of the hepatic nerves (liver sympathin)
has been the subject of much study. It is evident that liver sympathin is not epine-

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593 RAYMOND P. AHLQUIST Volume r53

phrine. There is, however, no absolute evidence that it is the neuro-hormone

(sympathin) or sympathin E. According to Cannon and Rosenblueth, liver sympa-
thin contracts the nictitating membrane, increases arterial pressure, is not ‘reversed
by ergotoxine’ and does not dilate the pupil or relax the pregnant cat’s uterus. Fur-
ther studies by Greer et al. (13), and Gaddum and Goodwin (I 7) have confirmed, in
the main, these observations, but have also shown that it doesrelax the intestine?
and may dilate the pupil and relax the cat’s uterus.
Liver sympathin is related to epinephrine, and several investigators (9, IO, 13,
17) have suggestedthat it might be art. Euler has obtained a sympathomimetic
substanceby extraction of mammalianhearts and hassuggested(19) that it might be
the epinephrine isomer, methyl-art. Of the aminesused in this study art. appearsto
be the most similar to liver sympathin.
Cannon and Rosenblueth consideredthe lack of pupillary dilator action of liver
sympathin as a paradox, on the basis that this substance was the hypothetical
sympathin E. Actually however, the stimulation of the nictitating membrane by
liver sympathin is the paradox, if liver sympathin is art. It has been shown that
art. is almost equally effective on the iris and the nictitating membrane. Therefore,
an amount of liver sympathin, if it is art. that would contract the membrane, should
also dilate the pupil. As Gaddum and Goodwin (I 7) have pointed out, the nictitat-
ing membrane is very responsive to many substancesother than sympathomimetic
amines. It is possible that stimulation of the hepatic nerves could produce small
amounts of histamine or acetylcholine which might account for the marked action of
liver sympathin on the nictitating membrane.
Assuming that liver sympathin is art., does this amine possessthe necessary
qualifications to be called sympathin E? The answer must be no, since the results
reported herein (based on the racemic forms) demonstrate conclusively that art.
is not the most active agent on any of the excitatory adrenotropic receptors. Just
why sympathin from the liver should be chiefly excitatory has never been explained.
The source must either be the nerve endings in the liver cells or the endings in the
smooth muscleof the hepatic blood vessels. Evidence has been obtained by direct
flow studies that epinephrine can produce vasodilation in the hepatic vesselsin a.
manner a.nalogousto its action on other vascular beds (35). Therefore, if there are
two kinds of sympathin there is no good reasonwhy an inhibitory substanceshould
not also be formed by hepatic nerve stimulation.
Considering all of the facts available, it is apparent that liver sympathin is a
unique product, possibly a modification of the neuro-hormone chemically changed
by the liver substanceafter formation; and that no other substancelike it has ever
been conclusively demonstrated asbeing formed by stimulation of other sympathetic
nerves. It should therefore be consideredas an exception and not be used as the
entire basisfor the proof of the existence of two kinds of sympathin.
Although little can be said at the present time as to the fundamental nature of
the adrenotropic receptor and the difference between the alpha and beta types, this
concept should be useful when studying the various actions of epinephrine, the ac-
tions and interactions of the sympathomimetic agents, and the effects of sympathetic
nerve stimulation. Useof the terms sympathin E and I should be discouraged,and

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Jane 1948 ADRENOTROPIC RECEPTORS 599

the term sympathin should be used to distinguish between the neuro-hormone pro-
duced by nerve stimulation and exogenous epinephrine. Fortunately . in the case of
the cholinergic nerves there has never been any suggestion that there might be two
mediators, although both excitatory and inhibitory effects are produced. The di-
verse effects of the cholinergic mediator, acetylcholine, have always been ascribed to
differences in the receptors upon which it acts.
SUMMARY

There are two distinct types of adrenotropic receptors as determined by their

relative responsiveness to the series of racemic sympathomimetic amines most closely
related structurally to epi.nephrine. The alptza adrenotropic receptor is associated
with most of the excitatory functions (vasoconstriction, and stimulation of the uterus,
nictitating membrane, ureter and dilator pupillae) and one important inhibitory
function (intestinal relaxation). The beta adrenotropic receptor is associatedwith
most of the inhibitory functions (vasodilation, and inhibition of the uterine and
bronchial musculature) and one excitatory function (myocardial stimulation).
Racemic epinephrine (and therefore levo-epinephrine which is about twice as active)
is the one amine which is the most active on both the alpha and beta receptors.
The results support the theory that there is only one adrenergic neuro-hormone,
or sympathin, and that sympathin is identical with epinephrine. The so-called
liver sympath in (the sympathin E of Cannon and Rosenblueth) is regarded as a
modified form of sympathin, chemically changed by the liver after formation and
liver sympathin is also considereda unique product and should not be regarded as
sympathin E or even sympathin.
The amines were kindly furnished by the following: Drs. K. K. Chen, M. L. Tainter and Mel
ville Sahyun. The heparin was generously supplied by Lederle Laboratories, the dibenamine by
Givauden-Delawanna, Inc. and the Priscol by Ciba Pharmaceutical Products, Inc. The author is
especially grateful to his colleagues of the Departments of Pharmacology and Physiology, who have
lIeen most generous and helpful with their criticisms and suggestions.

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