THE EFFECT OF EPINEPHRINE AND NOR-EPINEPHRINE ON

APPROACH-AVOIDANCE B E H A V I O R

APPROVED!

f i .

jor Professor ft

Minor Professor

Dean of the School of Education

Dean of the Graduate School

THE EFFECT OF EPINEPHRINE AND NOR-EPINEPHRINE ON

APPROACH-AVOIDANCE BEHAVIOR

T H E S I S

Presented to the Graduate Council of the

North Texas State University in Partial

Fulfillment of the Requirements

For the Degree of

MASTER OF SCIENCE

By

John Wesley Carley III, B. S.

Denton, Texas

June, 1966

TABLE OF CONTENTS

Page

LIST OF TABLES iv

LIST OF ILLUSTRATIONS v

Chapter

I. INTRODUCTION 1

II . METHOD 13

SubJects Apparatus Procedure Injecti ons

III. RESULTS 19

IV. DISCUSSION 26

V. SUMMARY AND RECOMMENDATIONS 34

Summary Recommendations

BIBLIOGRAPHY 36

iii

LIST OF TABLES

Table Page

I. Summary of Analysis of Variance for the 3 X 3 Latin Square Design 21

II. Summary of Analysis of Variance for the Three Treatment Conditions of Epinephrine, Norepinephrine, and Placebo 22

III. Critical Values for Differences Between

Treatment Totals 22

IV. Mean Distance for Each Treatment Group 23

V. Differences Between Totals for the Treatment Conditions 23

iv

LIST OF ILLUSTRATIONS

Figure Page

1. Injection Ratio for Experimental Drugs and Placebo 15

2. Phase 1 Training Trials Without Shack With Mean Running Times Measured in Seconds. . . . 19

3. Phase 2 Training Trials With Mean Distance Measured in Centimeters . . . . . 20

CHAPTER I

INTRODUCTION

Ever since men first began to speculate about human

nature, the question of man's emotions has inevitably arisen.

Throughout the years, psychologists have tried to develop a

valid concept of emotion, but because of the inherent nature

of emotion, many basic problems remain unanswered. However,

despite the difficulties of definition and experimentation

the area of emotional processes has by no means been neglected.

A distinguished theory of emotions affiliated with the

names of William James and C. G. Lange was set forth by them

independently. James first presented his view in 1864}

Lange's monograph appeared in Danish in 1865. James stated

. . the bodily changes follow directly the perception of

the exciting fact, and that our feeling of the same changes

as they occur IS, the emotion" (10, p. 34) . He conti nued by

saying that the common sensational, associ ational and motor

elements explain all. The evidence that James cites for the

theory is that we are aware of our tensions and throbs j we

feel them the moment they occur. He further stated that if

we should take away these bodily symptoms from the picture of

an emoti on, nothi ng would be left. Lange stated ". . . we

2

o w e all t h e e m o t i o n a l s i d e of o u r m e n t a l l i f e , o u r j o y s and

s o r r o w s , o u r h a p p y and u n h a p p y h o u r s , to o u r v a s o m o t o r

s y s t e m " ( 1 1 , p. 3 6 ) . It is e v i d e n t that L a n g e had t h e s a m e

c o n c e p t i o n as J a m e s , but r e s t r i c t e d h i s d e s c r i p t i o n of e m o -

t i o n to c h a n g e s in t h e c i r c u l a t o r y s y s t e m a l o n e .

In 1 9 0 8 M c D o u g a l 1 ( 1 2 ) a d v a n c e d a t h e o r y of e m o t i o n

s u g g e s t i n g a r e l a t i o n s h i p b e t w e e n c e r t a i n i n s t i n c t i v e r e a c -

t i o n s and e m o t i o n s . T h e t e n d e n c y to act w h e n in t h e p r e s e n c e

of a c e r t a i n o b j e c t in t h e e n v i r o n m e n t w a s M c D o u g a l 1 * s e x -

p l a n a t i o n of an i n s t i n c t . T h e t e n d e n c y to f e e l w a s h i s

m e a n i n g of e m o t i o n . H e s t a t e d t h a t t h e e m o t i o n of f e a r w a s

a s s o c i a t e d w i t h t h e i n s t i n c t of f l i g h t , and t h e e m o t i o n of

a n g e r or r a g e w a s a s s o c i a t e d w i t h t h e i n s t i n c t of f i g h t .

H . A. C a r r ( 3 ) o p p o s e d t h e J a m e s - L a n g © t h e o r y s t a t i n g

that t h e r e is a p s y c h o p h y s i c a l n a t u r e of e m o t i o n , and t h a t

e m o t i o n is p a r t l y r e s p o n s i b l e f o r t h e b e h a v i o r a l a c t . H e

d e f i n e s e m o t i o n as a u t o m a t i c o r g a n i c r e a d j u s t m e n t s . In t h e

c a s e of f e a r t h e o r g a n i c r e a d j u s t m e n t r e s u l t s in a m o b i l i z a -

t i o n of e n e r g y . T h e o r g a n i s m ' s p u l s e q u i c k e n s , t h e r e i s r a p i d

r e s p i r a t i o n , and t h e r e is an o v e r - s e c r e t i o n of e p i n e p h r i n e

by t h e a d r e n a l g l a n d . B u t , C a r r d o e s not h o l d t h a t all e m o -

t i o n s a r e b i o l o g i c a l l y u s e f u l . H e f u r t h e r s t a t e d t h a t t h e

n a m e g i v e n to an e m o t i o n d e p e n d s u p o n the s i t u a t i o n in w h i c h

it o c c u r s .

C a n n o n and Bard (2) d e v e l o p e d an i d e a s u g g e s t i n g t h a t

the h y p o t h a l a m u s was the specific area of the brain responsible

for t h e c o n t r o l of b e h a v i o r a l r e a c t i o n s in e m o t i o n s . T h e y

s t a t e d t h a t e m o t i o n a l e x p e r i e n c e and t h e e x p r e s s i v e r e s p o n s e

o c c u r s i m u l t a n e o u s l y as a r e s u l t of h y p o t h a l a m i c a c t i v i t y .

C a n n o n and B a r d n o t e d t h a t o n e l a r g e g r o u p of s y m p t o m s

p r o m i n e n t in a n g e r and r a g e p r e p a r e t h e o r g a n i s m to f a c e

e m e r g e n c i e s . T h e y e x p l a i ned t h i s b e h a v i o r on a p h y s i o l o g i -

c a l b a s i s . T h e y c o n t i n u e d by s a y i n g t h a t t h e a n g e r and f e a r

of the f l i g h t - f i g h t r e a c t i o n w e r e a d m i x e d , i n s e p a r a b l e , and

always a s s o c i a t e d w i t h the s e c r e t i o n of e p i n e p h r i n e . How-

e v e r , t h e y w e r e u n a b l e to e x p l a i n all of t h e i r f i n d i n g s o n

t h e b a s i s of epi n e p h r i ne a l o n e and p r o p o s e d two o t h e r

syrapathorai neti c s u b s t a n c e s , S y m p a t h i n E and S y m p a t h i n I. In

l a t e r s t u d i e s p h y s i o l o g i s t s h a v e f o u n d t h a t S y m p a t h i n E and

n o r - e p i n e p h r i ne w e r e i d e n t i c a l .

IS. G. W o l f f ( 1 3 ) , in his s t u d i e s of p h y s i o l o g i c a l

changes accompanying various emotional states, a c c u m u l a t e d

c o n f i r m a t o r y e v i d e n c e that t h e f l i g h t - f i g h t emotions w e r e

separable. He s h o w e d t h a t in various o r g a n s s u c h as t h e

s t o m a c h , c o l o n and t h e n o s e , d i f f e r e n t e m o t i o n s e v o k e d dif-

ferent p h y s i o l o g i c a l r e a c t i o n s . W h e n a subject was angry,

the s t o m a c h lining b e c a m e r e d and t h e r e w a s an increase in

its r h y t h m i c c o n t r a c t i o n s and in t h e s e c r e t i o n of h y d r o c h l o -

r i c acid. When the same subject was depressed or f r i g h t e n e d ,

the stomach lining was p a l e in color and t h e r e was a decrease

in peristaltic m o v e m e n t s and in h y d r o c h l o r i c acid s e c r e t i o n .

His e x p e r i m e n t s g a v e e v i d e n c e t h a t the a d r e n a l medulla

secreted two substances rather than one. A. F. Ax studied

the physiological differentiation between fear and anger in

h u m a n s by the use of a p o l y g r a p h . H e a l s o f o u n d d i f f e r e n t

physiological reactions. These experiments suggest that the

flight-fight reaction may be well separated into component

emotions with d i f f e r e n t p h y s i o l o g i c a l a c c o m p a n i m e n t s . T h i s

is c o n t r a r y to C a n n o n * s t h e o r y , w h i c h s t a t e d t h a t a n g e r and

f e a r of t h e f l i g h t - f i g h t r e a c t i o n r e s p o n s e w e r e a d m i x e d and

i n s e p a r a b l e .

In r e c e n t y e a r s psychologists h a v e c o n c l u d e d t h a t t h e

symptoms of e m o t i o n , especially of i n t e n s e e m o t i o n , include

p r o f o u n d c h a n g e s t h r o u g h o u t t h e b o d y , c h a n g e s r e g u l a t e d in

a c o m p l e x way by the divisions of the a u t o n o m i c n e r v o u s

system and by the endocrine glands. Out of the mass of thest

p h y s i c a l c h a n g e s p s y c h o l o g i s t s and p h y s i o l o g i s t s h a v e b e e n

a b l e to p o s t u l a t e t h a t t h e r e exi s t c e r t a i n c o r r e s p o n d i n g

ratios b e t w e e n s p e c i f i c e m o t i o n a l s t a t e s and physiological

changes. T h e m a n y b o d i l y c h a n g e s t h a t o c c u r d u r i n g e m o t i o n

are not u n r e l a t e d p h e n o m e n a , t h e y fit t o g e t h e r into o r g a n i z e d

p a t t e r n s .

D. H. F u n k e n s t e i n (5) c o n d u c t e d a s t u d y w i t h p s y c h o t i c

p a t i e n t s . H e w a s testing Rado's h y p o t h e s i s t h a t t h e p h y s i -

o l o g i c a l c o n c o m i t a n t s of f e a r and of a n g e r d i r e c t e d t o w a r d

t h e s e l f w o u l d b e similar. F u n k e n s t e i n, in o r d e r to t e s t

t h i s hypothesis, studied c o l l e g e s t u d e n t s ' r e a c t i o n s during

a l a b o r a t o r y stress-inducing si t u a t i o n . His results confirmed

the hypotheses that (a) The changes In physiological pat-

terns of those subjects who reported anger outward from the

s e l f differed from those who reported feelings of anger

directed toward the self; (b) T h e physiological patterns of

those subj ects who reported anger directed ou t w a r d away from

the self differed from those who reported feelings classified

as anxiety; and (c) The changes in the physiological patterns

o f those s ubj ects who r epor t ed their f e e l i n g s as anger

directed toward the self were similar to those feelings

reported as anxiety. The evidence from t h i s experiment

i n d i c a t e s that the anger and fear of the flight-fight re-

sponse may be separable o n a physiological and psychological

level. On the physiological level the data from the "anger

out" was associated with nor-epinephrine-like substances, and

anxiety was associated with epi nephri ne-like s u b s t a n c e s .

Fu nkens t ei n (6) cites further evidence supporting the

p r o p o s i t i o n that the flight-fight reaction can be separated

into c o m p o n e n t s . He related that U. S. von Eu l e r , in his

experiments c o n c e r n i n g stimulation of the hypothalamus and

concentration of nor-epi nephri ne in wild animals, found that

stimulation of certai n areas of the hypothalamus caused the

a d r e n a l gland to secrete nor-epi nephri ne, w h e r e a s stimulation

of other areas caused it to secrete epi nephri ne. He also has ^ ]

evidence suggesting that aggressive animals have an e x c e s s i v e ,

amount of nor-epi nephri ne i n their adrenals, and that e a s i l y

frightened animals have an excessive amount of epinephrine. j

In 1955 F u n k e n s t e i n ret es ted the hypothesis that anger

directed outward from the self is associated with the secre-

tion of nor-epi nephri ne while anxiety or feur is associated

with the secretion of epinephrine. The nor-epinephrine/

epinephrine ratio was determined by the raecholyl test. He

noted the r e a c t i o n s of subjects when they were injected with

epi nephri ne and nor-epi nephri ne and quantitatively determined

e p i n e p h r i n e - l i k e and nor-epi nephrine r e a c t i o n s . T h e h y p o t h -

esis was c o n f i r m e d .

T h e elevation of blood pressure under stress has been

known for a long time. H i c k a m , C a r g i 1 1 , and G o l d e n (9) found

that students reacted in different patterns in response to

psychological stress. The patterns were associated with an

elevated blood pressure and the mechanism of each rise was

different. In one of these there was associated with t h e ^

rise in blood pressure, a decreased peripheral resistance,

an increased cardiac output, and an increased pulse rate.

The other blood pressure rise Involved an increased periph-

eral resistance, an unchanged or lowered cardiac output and

a lowered pulse. Goldenberg, Pines, Baldwin, Greene, and

Roh (7) studied the effects of nor-epi nephri ne and epi neph-

rine. T h e y found that the infusion of epinephrine p r o d u c e d

the following pattern: increased blood pressure, increased

pulse, decreased p e r i p h e r a l resistance, and increased cardiac

o u t p u t . The infusion of nor-epi nephri ne resulted in increased

blood pressure, unchanged or lowered pulse, increased

peripheral resistance, and unchanged or decreased cardiac

output. It is evident from these two studies that when an

elevated blood pressure was one of the responses to psycho-

logical stress, at least two mechanisms could account for

the rise. These two mechanisms were similar physiologically

to those produced by the intravenous infusion of epinephrine

and nor-epinephri ne.

Funkenstein and Meade (4) proposed that one type of

elevated systolic blood pressure in response to psychiatric

illness or to psychological stress in healthy norraatensives

was associated with epinephrine, and the other type of

elevated blood pressure with nor-epinephrine. The type of

blood pressure elevation was deterrained on the basis of the

subject's blood pressure reaction following an injection of

raecholyl. In all of the subjects whose blood pressure was

elevated by epinephrine, following the injection of raecholyl,

the "time of homeostasis" was always greater than twenty-five

minutes. When blood pressures were elevated by a constant

intravenous infusion of nor-epinephrine, an entirely different

reaction took place. The "time of homeostasis" was reached

in all cases during the observation period. The mean was

6.3 minutes. Funkenstein stated that these reactions were a

reliable index of the clinical reaction to shock therapy.

When the blood pressure reaction to raecholyl was classified

as "marked" and the prei nj ection level was not reached during

8

the t w e n t y - f i v e m i n u t e o b s e r v a t i o n p e r i o d , the r e s p o n s e to

electric shock was good. In contrast to this, when the blood

p r e s s u r e r e s p o n s e to m e c h o l y l was c l assified as " m o d e r a t e or

mi Id" and the prei nj ection level was reached w i t h i n the

t w e n t y - f i v e m i n u t e o b s e r v a t i o n p e r i o d , the p a t i e n t s usually

did not respond to electric shock t r e a t m e n t . T h e e x p e r i - L/

mental evidence offered suggested that the m e d i a t i n g s u b s t a n c e

in one type of elevated blood p r e s s u r e in a s s o c i a t i o n with

p s y c h i a t r i c illness and in n o r m a t e n s i v e students under

p s y c h o l o g i c a l stress was an e p i n e p h r i n e - l i k e s u b s t a n c e , and

in the other type of elevated blood p r e s s u r e a nor-epi nephri ne-

like s u b s t a n c e . The blood p r e s s u r e r e a c t i o n following m e c h o l y l

seemed a r e l i a b l e i n d i c a t i o n of the type of elevated blood

p r e s s u r e which was present in a given case.

A d d i t i o n a l studies yielded results which showed that

students who responded to a stressful s i t u a t i o n by anger

directed outward away from the self had p h y s i o l o g i c a l r e a c -

tions similar to those produced by an i n j e c t i o n of nor-

e p i n e p h r i n e . H o w e v e r , those who responded with d e p r e s s i o n

or anxiety had p h y s i o l o g i c a l r e a c t i o n s similar to those of

epi nephri ne. T h e s e f i n d i n g s raised the question : Does the

same i ndividual secrete unusual amounts of nor-epi nephri ne

when angry and of e p i n e p h r i n e when frightened ? A. F. Ax (1)

designed e x p e r i m e n t s to study this q u e s t i o n . His results

revealed that when a subject was angry at o t h e r s , the p h y s -

iological r e a c t i o n s were like those induced by the i n j e c t i o n

of n o r - e p i nephr i ne; when t h e same s u b j e c t was f r i g h t e n e d ,

t h e r e a c t i o n s were l i k e t h o s e of e p i n e p h r i n e . Th i s i n d i c a t e d

t h a t t h e phys io logy was s p e c i f i c f o r t he emotion r a t h e r than

f o r t h e p e r s o n . \

\ W. R. Hess (8) conc ludes t h a t n o r - e p i n e p h r i n e and |

i \

e p i n e p h r i n e a r e s e c r e t e d by d i f f e r e n t c e l l s i n the adrena l?

m e d u l l a , and when t h e s e c e l l s a r e s t i m u l a t e d they s e c r e t e | 1

t h e i r r e s p e c t i v e hormones. From t h e e x p e r i m e n t a l d a t a I I

a v a i l a b l e i t was assumed t h a t n o r - e p i n e p h r i n e was a major J \

p h y s i o l o g i c a l f a c t o r i n the emotion of r age or a n g e r , and •

t h a t e p i n e p h r i n e was a major p h y s i o l o g i c a l f a c t o r in t h e

emotion f e a r .

I t was t h e purpose of t h e p r e s e n t s tudy to compare the

e f f e c t of i n t r a p e r i t o n e a l i n j e c t i o n s of t h e f o l l o w i n g drugs

on a condi t i o n e d app roach -avo idance r e s p o n s e in mice . These

drugs were e p i n e p h r i n e and n o r - e p i nephr i ne . The avo idance

c o n f l i c t c r e a t e d in t h i s s tudy was used to measure f e a r or

anger d i r e c t e d toward t h e s e l f . This measure was used on the

assumpt ion t h a t a f e a r r e a c t i o n to an a v e r s i v e s i t u a t i o n would

lead to avo idance b e h a v i o r . Aggress ion d i r e c t e d away from

t h e s e l f was measured by t h e d i s t a n c e each mouse ran beyond

t h a t which was r eco rded wh i l e he was under t he i n f l u e n c e of

e p i n e p h r i n e or a p l a c e b o . Funkens te i n has impl ied t h a t anger

d i r e c t e d inward or f e a r i s a s s o c i a t e d wi th e p i n e p h r i n e - l i k e

s u b s t a n c e s and anger d i r e c t e d away from the s e l f i s a s s o c i a t e d

with n o r - e p i n e p h r i n e - l i k e s u b s t a n c e s . Havi ng made the

10

assumptions t h a t ep inephr ine w i l l e l i c i t a f e a r response and

t ha t no r -ep inephr ine w i l l e l i c i t an a g g r e s s i v e response

d i r e c t e d away from the s e l f , the fo l lowing hypotheses were

proposed s

A, Sub jec t s r e c e i v i n g an i n j e c t i o n of ep inephr ine wi 11

show a s i g n i f i c a n t l y g r e a t e r f e a r response than those sub-

j e c t s r e c e i v i n g e i t h e r an i n j e c t i o n of nor -ep i nephri ne or an

i n j e c t i o n of a p l acebo .

B. Sub jec t s r e c e i v i n g an i n j e c t i o n of no r - ep inephr ine

w i l l show a s i g n i f i c a n t l y g r e a t e r aggres s ive response than

those s u b j e c t s r e c e i v i n g e i t h e r an i n j e c t i o n of ep inephr ine

or an i n j e c t i o n of a p lacebo .

CHAPTER BIBLIOGRAPHY

1. Ax, A. F., "The Physiological Differentiation Between

Fear and Anger," Psychosomatic Medicine. XV (1953), 433.

2. Cannon, W. B., Bodily Changes in Pain. Mm§£* i&MX., and Rage. New York and London, D. Appleton and Company, 1929.

3. Carr, H. A., h Study of Mental Activity. New York, Longmans, Green, 1925.

4. Funkenstei n, 0. H. and L. N. Meade, "Nor-Epi nephri ne-like and Epi nephri ne-like Substances and the Elevation of Blood Pressure During Acute Stress," Journal of Nervous and Mental Disorders. CXIV (1954), 380-397.

5. Funkenstein, D. H., S. H. King, and M, Drolette, "The Direction of Anger During a Laboratory Stress-Inducing Situation," Psychosomatic Medicine. XVI (1954), 404-413.

6. Funkenstein, D. H., "The Physiology of Fear and Anger," Scientific American. CXCII (1955), 74-80.

7. Goldenberg, M., K. L. Pines, E. Baldwin, D. G. Greene, and C. E. Roh, "The Hemodynamic Response of Man to Nor-Epi nephri ne and Epi nephri ne and Its Relation to the Problem of Hypertension," American Journal of,

cine. V (1948), 792.

8. Hess, W. R. and K. Albert, "Experimental Data on the

Role of the Hypothalamus in Mechanism of Emotional Behavior," Archives & £ Nfmrpfrftgy M EAMMMIX, LXXIII (1955), 127-131.

9. Hickam, J. B., W. H. Cargill, and A. C. Golden, "Cardiovascular React ions to Emotional Stimuli, Effect on the Cardi ac Output, A-V Oxygen Difference, and Peripheral Resistance," Journal of Clinical Investi-gation, XXVI (1947) , 1.

10. James, William, Psychology. New York, Harper and Row, 1961.

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11. James, Will iam arid C. 6 . Lange, T&& E m o t i o n s . B a l t i m o r e , Wil l iams and Wilkin*, 1922.

12. McDougall, Wi l l i am, Introduction to S o c i a l Psycho logy . London, D. Appleton and C o m p a n y , 1908.

13. Wol f f , H. G. , L i f e D i s t r e s s e s and D i s e a s e . B a l t i m o r e , Wil l iams and W i l k i n s , 1950.

CHAPTER II

METHiD

Subj ects

In this experiment thirty naive, male, black mice from

the colony C57BL/6J at the Jackson Memorial Laboratories

were used. All subjects were between thirty-five and forty-

five days of age at the time of testing. The subjects were

maintained on jyi libiturn food and water. This strain was

selected because of its wide use in studying physiological

and behavioral changes.

Apparatus

A rectangular three compartment linear maze was used.

The maze was a plywood box, 172 centimeters long, 9.0 centi-

meters wide, and 9.0 centimeters deep. It consisted of a

12 centimeter long start box at one end, and at the opposi te

end a goal box 12 centimeters long. A divided copper screen

grid 12 centimeters by 9.0 centimeters was placed directly

in front of the entrance to the goal box. An R.C.A. forty-

five volt dry cell battery was attached to the copper grid

to provide shock for avoidance conditioning. The start box

and the goal box were each separated from the runway by a

13

14

guillotine door. These doors were operated manually from

the outside. A hinged plexiglass top covered the maze. This

top w a s calibrated and marked in centimeters, a l l o w i n g for

measurement of the distance which each subject ran.

Procedure

In the prelimi nary training, the mice were familiarized

with the routine of the maze. This was accomplished by al-

lowing the mice to explore the maze for five minutes each day

for three days.

There were two phases to the training. In the first

phase all mice were trained to an approach response. The

s u b j e c t s were on a twenty-four hour food deprivation schedule

which served as the motivation for action. Each subject

received a .5 gram food reward in the goal box. Each animal

was given a total of fifteen trials, three trials a day for

five days. T h i s criterion was e s t a b l i s h e d in a pilot study

using twelve mice. The running speed for each subj ect was

recorded in seconds after each trial.

In the second phase all mice were trained to shock

avoidance. As the animal approached the goal, he passed

across the divider copper grid and was shocked with forty-

five volts. Each subj ect was g i v e n a total of fifteen trials,

three trials a day for five days. This criterion was also

established in the above ment i o ned pilot study.

Af te r the approach-avoidance t r a i n i n g was completed

each animal was randomly a s s igned , by the use of a t a b l e of

random numbers, to one of t h r e e groups of ten s u b j e c t s each.

This was done to t e s t the order of p r e s e n t a t i o n .

I n j e c t i o n i

I n j e c t i o n of e p i n e p h r i n e , no r -ep inephr i ne, and the

placebo were made according to the r a t i o suggested by Barn®®

£& &JL* Figure 1 g r a p h i c a l l y i l l u s t r a t e s t h i s r a t i o f o r

e p i n e p h r i n e , nor -ep i nephri ne, and the p l acebo .

1.0

.9

.8 m u m *» .7

m •*4 *6 01 m • 5 Q O .4 JO »

* 3

*2

A

10 11 12 18 14 15 16 Tf

Weight in g ran t

1® 19 20

F ig . 1 — I n j e c t i o n r a t i o f o r exper imenta l drugs and p lacebo .

16

Injections were ad m i n i s t e r e d with a one cubic centimeter

tuberculin syringe and needle. The drugs used were a 1x1000

solution of adrenalin chloride in mammalian Ringer's solu-

tion, a 1:1000 solution of nor-epi nephri ne in mammalian

Ri nger's solution, and a placebo of mammalian R i n g e r ' s solu-

tion. All subjects served under all three experimental

conditions of injectionj these drugs were epinephrine (Parke-

Davis Laboratories), nor-epi nephri ne bi tartrate (Nutritional

Biochemical Corporation), and a placebo. Injections were

administered intraperitoneally, allowing for a retarded onset

of the drug effects (1).

A 3 X 3 Latin Square design was used to test for order

of effects (2) and permitted each subject to serve as his own

control. The order of injection for Group 1 was as follows:

epi nephri ne, nor-epinephrine, and p l a c e b o . The order for

Group 2 w a s placebo, epi nephri ne, and nor-ep i nephri ne} w h i l e

the order for Group 3 was nor-epi nephri ne, placebo, and

epi nephri ne.

Extraneous v a r i a b l e s such as time, temperature, and

place were held constant, and every effort was made to keep

the interexperimental intervals identical for all animals.

A one day i nterval between i nj ections was allowed to permit

complete drug dissipation.

A standard procedure was fo1 lowed for all injections (1)

The experimental drugs and placebo were inj ec ted i ntra-

peritoneally over a period of thirty seconds. The volume

17

relationship was the same for both drugs and placebo. The

subj e c t remained in the start box for five m i n u t e s after

i n j e c t i o n to allow the drug to ta k e effect.

In the test trials, after each s u b j e c t was i n j e c t e d and

had stayed in the start box for five minutes, the start box

door was opened, and the animal ran the maze. The point at

which the s u b j e c t ^topped was marked and this distance from

the start box door was recorded in centimeters. The s u b j e c t

was taken from the maze at the point where he first stopped

and returned to his home c a g e .

CHAPTER BIBLIOGRAPHY

1. Barnes, C. D. and L. G. E i t h e r i n g t o n , Drug Dosage tn L a b o r a t o r y Animals. Berkeley and Los Angeles, U n i v e r s i t y of C a l i f o r n i a , 1 9 6 4 .

2. Winer, B. J . , Statistical Principles in Experimental Design. New York, McGraw - H i l l Book Company, Inc., 1 9 6 2 .

18

CHAPTER I I I

RESULTS

Endeavoring to measure the effects which specific d r u g s

have on approach-avoidanee behavior, thirty male, black mice

from the colony C57BL/6J at the Jackson Memorial Laboratories

were a d m i n i s t e r e d epinephrine, nor-epinephrine, and a placebo.

F i g u r e 2 represents the mean r u n n i n g time of all mice for

f i f t e e n t r i a l s i n the f i r s t p h a s e of t r a i n i n g . The approach

conditioning appears to have s t a b i l i z e d . This i s shown by

the decrease in the r u n n i n g time with each s u c c e s s i v e trial.

20 19 18 IT 16 15 14 13 12

» 11 a 10 S 9 £ 8

7 6 5 4 3 2 1

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 T r i a l

Fig. 2—First phase t r a i n i n g w i t h o u t shock w i t h mean running times m e a s u r e d i n seconds.

19

20

Figure 3 represents the mean distance run in centimeters

by all mice for fifteen trials in the second p h a s e of train-

ing.

200 190 180 170 160 150 140

Z 130 2 120 ® 110 ® 100 t 90 ® 80 ° 70

60 50 40 30 20 10

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Trial

F i g . 3 — S e c o n d p h a s e training t r i a l s w i t h distance m e a s u r e d in centimeters.

It is s h o w n that the a v o i d a n c e c o n d i t i o n i n g had s t a b i l i z e d .

This is indicated by t h e continuing decrease in centimeters

run on each successive trial.

H e t e r o g e n e i t y of variance for t h e t r e a t m e n t e f f e c t s w a s

tested by an Ffflax test and was found not to be significant.

Since the p o p u l a t i o n s a m p l e was homogeneous further statisti-

cal analvais was undertaken.

21

Table I represents a 3 X 3 Latin Square design which

was used to test for the effects of the order of presenta-

tion.

TABLE I

SUMMARY OF ANALYSIS OF VARIANCE FOB THE 3 X 3 LATIN SQUARE DESIGN

Source of Vari ability

Sum 5Q df Mean SO F

Groups 2482.77 2 1241.38 5.50®

Order 2051.36 2 1025.68 4.54*®

Drugs 161785.27 2 80892.63 358.51*

Residual 2346.99 2 1173.49 5.20*

Within 18276.72 81 225.63 NS

•Significant at .01, ••Significant at .05

It is revealed by Table I that the order and groups were

found to be significant. The F value for the residual effects

was also found significant,and this indicates that there was

some degree of group and order interaction.

Since the treatment effects were found to be highly

significant, further analysis was done by a si ngle classifica-

tion analysis of vari ance with repeated measures. Thi s

analysis is shown in Table II.

TABLE II

SUMMARY OF ANALYSIS OF VARIANCE FOB THE THREE TREATMENT CONDITIONS OF EPINEPHRINE,

NOR-EPINEPHRINE, AND PLACEBO

22

Source of Variability

Sum SQ df Mean SQ F

Between Subjects 6245.5 29 215.36

Within Subjects 198858.5 90 2209.53

Drugs 178868.74 3 59622.91 259.5

Residual 19989.76 87 229.76

Total 205103.74 119 • • * • • •

Because a significant F value for the treatment effect

was found, further analysis was done by an Newraan-Keuls test

to determine if the treatment conditions were significantly

different from one another. Table III represents the cri tical

values for differences between treatment totals.

TABLE III

CRITICAL VALUES FOR DIFFERENCES BETWEEN TREATMENT TOTALS

Truncated Range r 2 3 4

q.99 ( r.87) 2. GO 3.40 3.74

q* ̂ (r.67) /nMS error.. 233.24 283.22 308.21

2 3

T a b l e IV r e p r e s e n t s the m e a n distance r u n for e a c h

treatment group.

TABLE IV

MEAN DISTANCE FOR EACH TREATMENT GROUP

Groups Mean. Distance

Non-i nj ection 818.0

Epi n e p h r i ne 3 2 2 . 0

N o r - e p i n e p h r i ne 3 3 4 8 . 0

Hacebo 1194.0

T a b l e V s h o w s the differences b e t w e e n t o t a l s for the

t r e a t m e n t conditions of Group 1, n o n - i n j e c t i o n j G r o u p 2,

epinephrine! G r o u p 3, n o r - e p i n e p h r i n e $ and G r o u p 4, p l a c e b o .

TABLE ¥

DIFFERENCES BETWEEN TOTALS FOR THE TREATMENT CONDITIONS

Order 1 2 3 4

Treatments in Order of Totals Group 2 Group 1 Group 4 G r o u p 3

Group 2 ft ft ft 496.0** 8 7 2 . 0 ® * 3026.0**

Group 1 ft ft ft • • • 3 7 6 . 0 * ® 2 5 3 0 . 0 * ®

Group 4 • ft ft • • • • • • 2 1 5 4 . 0 ® *

G r o u p 3 ft ft ft • • • • • • • e •

S i g n i f i c a n t at .01

24

This d a t a r e v e a l e d t h a t a l l treatment c o n d i t i o n s were s i g n i f i -

c a n t l y d i f f e r e n t from one another at a l e v e l g r e a t e r than .01}

t h e r e f o r e , a l l t he c o n d i t i o n s appeared to have d i f f e r e n t i a l

e f f e c t s .

Since the order of presenta t ion was found to be s i g n i f i -

cant , a x, t e s t was done between the l a s t t r i a l of avoidance

condi t ioning and a po s t - treatment t r i a l . The p o s t - t r e a t m e n t

t r i a l was d e f i n e d as a period of two days a f t e r the l a s t

treatment t r i a l . This was to allow for complete drug d i s s i -

p a t i o n . The jt value was not found to be s i g n i f i c a n t , and t h i s

appears to i n d i c a t e t h a t t h e t r e a t m e n t e f f e c t s were to some

degree independent of t he e x p e r i e n c e in the maze. This

f i n d i n g further supports the assumption that the dependent

var iab le was a r e s u l t of the treatment condi t ion and not

merely the order of p r e s e n t a t i o n .

CHAPTER BIBLIOGRAPHY

1. Barnes, C. 0. and L. G. Eltherington, Drug Dosage Jji Laboratory Animals. Berkeley and Los Angeles, University of California, 1964.

2. Winer, B. J., Statistical Principles in Experimental Design. New York, McGraw-Hill Book Company, Inc., 1962.

25

CHAPTER IV

DISCUSSION

The results in Table V, as presented in Chapter I I I ,

revealed that there was a significant difference between

treatment t o t a l s . This shows that each group, n o n - i n j e c t i o n ,

e p i n e p h r i n e , n o r - e p i n e p h r i n e , and p l a c e b o , was s i g n i f i c a n t l y

d i f f e r e n t from each other at a level greater than .01. C o n -

s e q u e n t l y , the h y p o t h e s i s that subjects r e c e i v i n g an injec-

tion of e p i n e p h r i n e will show a significantly g r e a t e r f e a r

r e s p o n s e than those subj ects receivi ng ei ther an injection of

nor-epi nephri ne or a placebo was c o n f i r m e d . Also, the

h y p o t h e s i s that subjects receivi ng an injection of nor-

epi nephrine will show a s i g n i f i c a n t l y greater r e s p o n s e of

anger directed away from the self than those subj ects

recei vi ng ei ther an i nj ection of epinephrine or a p l a c e b o

was c o n f i r m e d . T h i s o b s e r v a t i o n is c o m p a t i b l e with F u n k e n -

stein's theory (2) whi ch contends that epi nephri ne is a major

p h y s i o l o g i c a l cause for the emotion fear, and that nor-

epinephrine is a major p h y s i o l o g i c a l cause f o r rage or anger

directed away from the self. This theory proposes that a

subject recei vi ng an i n j e c t i o n of epi nephri ne would show a

greater fear r e s p o n s e in a stressful situation than would

26

27

be shown without this injection. It also proposes that a

subject receiving an injection of nor-epi nephri ne would show

a greater response of anger directed away from the self than

would be shown without this injection.

The results of the present study are also in accord

with those found by Ax (1). He contrived laboratory stress-

ful situations which were successful in producing on one

occasion anger and on another occasion fear in the same sub-

jects. Hi8 results showed that when a subject was angry at

others, the physiological reactions were like those induced

by the i nj ection of nor-epi nephri ne\ when the same subject

was frightened, the reactions were like those of epinephrine.

Studies undertaken by Kosman and Gerard (4), Sines and

Keefe (5), and Wurtraan and his co-workers (6) are in

agreement with the hypotheses of this study, stating that

epi nephri ne will cause a fear response. They reported that""?

/

epinephrine suppresses the output of a learned response. /

Funkenstei n (2) states that the emotion of fear and

anger of the "flight-fight" response have been separated on

both physiological and psychological levels by accounting

for the direction of the emotion. As shown in Chapter I,

epinephrine is proposed to underlie the emotion fear and nor-

epinephrine to underli e the emotion of anger. Funkenstei n

states that fear is aggression directed toward the self while

aggression directed away from the self is rage or anger.

Therefore, the stress situation of the present study was

28

constructed so as to allow each subject to show both an

inwardly aggressive response which was interpreted as fear

and an outwardly aggressive response which was interpreted

as anger.

The present study, and the related experiments concern-

ing the effects of epinephrine and nor-epi nephri ne, yielded

data which can also be understood in terms of various obser-

vations .

An observation which can be related to the results of

this study was demonstrated by Hokfelt (3). 11 is known that

anger directed outward is more characteristic of an early

stage of childhood than is anxiety or anger directed toward

the self. The latter two emotions are the result of the

socialization of the child. Hokfelt's experiments yielded

results indicating that at an early age the adrenal medulla

has more nor-epi nephri ne, but later epinephrine becomes

domi nant. This implies that the physiological development

of the child parallels its psychological development.

Clinical observations concerning the psychological

characteristics of paranoid and depressed psychotics reveal

that paranoids show a greater degree of regression than do

depressed psychotics. Many investigators i nterpret regres-

sion as an attempt to return to an earlier stage of life.

Funkenstei n (2) has attempted to relate the secretion of

epinephrine and nor-epi nephri ne to depression and paranoi d

reactions. He states that on a physiological level patients

29

with an excessive secretion of nor-epi nephri ne, which repre-

sents a secretion characteristic of an earlier stage of

development, are similar to those patients classified as

paranoid. He further states that patients who are classified

as psychotieally depressed secrete an excessive amount of

epinephrine, which is characteristic of a later period of

development. As stated above, two emotions are associated

with this later period of development^ they are anger directed

toward the self or anxiety.

One of the important psychological differences between

the "sick" and the "well" is the ability to master the situa-

tion. Healthy subjects whose reaction to acute stress is

depression are able, as time passes, to master the situation;

the emotion subsides, and the physiology returns to its pre-

stress level. On the other hand, depressed patients show a

failure in mastery so that the reaction becomes sustained,

the physiology increases in intensity and the clinical reac-

tion type becomes recognizable. On a physiological level,

evidence of excessive secretion of nor-epinephrine and

epinephrine can be found in both psychotic patients and in

healthy subjects. The evidence of excessive secretion of

these hormones can be found in the patients on the wards

without external stress, whereas, in the healthy subj ects,

excessive secretion is found only when the subject is under

stress. For example, depressed patients on a ward will show

evidence of excessive secretion of epinephrine, whereas

30

students whose style of reacting during acute stress is

depression w i l l show excessive secretion of e p i n e p h r i n e only

during such acute stressful times. This finding also sup-

ports Funkenstein*s theory that an excessive secretion of

epinephrine is present during depression.

E s s e n t i a l l y , the preceding study attempted to experi-

mentally validate the theory that epinephrine is the m a j o r

physiological factor underlyi ng the emotio n f ear and that

nor-epinephrine is the major physiological factor underlying

the emotion anger. However, there are c e r t a i n methodological

considerations which must be discussed in relation to this

experiment.

A result observed in Table I, presented in Chapter III,

revealed that the residual effect was significant at the .01

level. This appears to i n d i c a t e an i n t e r a c t i o n between the

groups and order of presentation. The cause for this inter-

action was not readily explainable, but several factors could

have created this interaction. The order of presentation

could have been affected by the concentration or v o l u m e of

fluid used for injection. When deali ng with small animals

such as raice this variable has been found to be especially

important. There may have been some i ntrai ndividual reactions

to the drug effects. The r a n d o m i z a t i o n of subjects may have

creat ed more w i t h i n subj ect vari abili ty than normally expected

Although there appeared to be some confounding effect, the F

31

value for the treatment effects was so great that any con-

founding would probably not completely erase the treatment

effects.

The function of the placebo, as used here, was to

control any confounding effects caused by the i nj ection

procedure. A result observed from the data was that sub-

jects receiving an i nj ection of a placebo showed a signifi-

cantly greater response of anger directed away from the self

than those receivi ng an injection of epi nephri ne. This is

contrary to the assumption that the mere act of injection

caused the release of epinephrine and that this stimulated

some amount of fear in the animal. This data is documented

by the fact that the treatment condi tions were significantly

different from one another. The placebo condition exceeded

that of the epinephrine treatment but did not exceed that of

the nor-epinephrine condition. This may be related to the

significant within subject variability found in Table II,

Chapter III, or the effectiveness of the nor-epinephrine

treatment.

The i nj ecti ons given in this study were intraperitoneal

This method of administration was chosen for three reasons,

which are as follows: (a) the drugs administered were non-

irritating; (b) the peritoneum of the abdominal cavity

presented a large absorption areaj and (c) the techni que was

simple and could be performed by one person.

32

Thi s v a r i a b l e may have a f f e c t e d the r e s u l t s because a

" s i n g l e - s h o t " dose was used . S ince i t was not p o s s i b l e t o

t i t r a t e t he drug in t h i s type of i n j e c t i o n , an overdose could

be a d m i n i s t e r e d to a s e n s i t i v e an ima l . Most of t h e absorbed

m a t e r i a l e n t e r e d the p o r t a l c i r c u l a t i o n where i t may have

been p a r t i a l l y me tabo l i zed w i t h i n the l i v e r , and t h i s m e t a b o l -

ism could have caused a s i g n i f i c a n t change in the s t r u c t u r e of

t he drug and p o s s i b l y changed i t s p h y s i o l o g i c a l a c t i o n .

Based on t h i s e x p e r i m e n t a l ev idence and t h e assumpt ions

which were made, i t was concluded t h a t e p i n e p h r i n e was a major

p h y s i o l o g i c a l f a c t o r u n d e r l y i n g the emotion f e a r and t h a t n o r -

e p i n e p h r i ne was a raaj or p h y s i o l o g i c a l f a c t o r u n d e r l y i n g t h e

emotion of anger d i r e c t e d away from the s e l f . This c o n c l u -

s i o n can only be made i f t he l i m i t a t i o n s of t h i s s tudy a re

c o n s i d e r e d .

CHAPTER BIBLIOGRAPHY

1. Ax, A, F., "The Physiological Determination Between Fear and Anger in Humans," Psychosomatic Medicine. XV (1953), 433.

2. Funkenstein, D. H., "The Physiology of Fear and Anger," Scientific American, CXCII (1956), 74-80.

3. Hokfelt, B., "Nor-adrenalin and Adrenalin in Mammalian Tissues," Acta Physiology Scandinavia. XCII (1951), 25.

4. Kosraan, M. S. and R. W. Gerard, "The Effect of Adrenalin on a Conditioned avoidance Response," Journal of Comparative Physiology and Psychology. XLVIII (1955), 506-508.

5. Sines, J. 0. and 0. J, Keefe, "The Relationship Between Pharmacologically Modified Overt Response Rate and Heart Rate," Journal of Genetic. Psychology. C (1962), 275-274.

6. Wurtman, R. J . , W. H. Frank, W. H. Morse, and P. B. Dews, "Studies on Behavior. V. Action of 1-epinephrine and Related Compounds," Journal of Pharmacology and Experimental Therapy. CXXVII (1959), 281-287.

33

C H A P T E R V

S U M M A R Y A N D R E C O M M E N D A T I O N S

Summary

T h i r t y m a l e , b l a c k m i c e w e r e u s e d in this s t u d y . All

subjects were trained to shock avoidance and were injected

w i t h e p i n e p h r i n e , n o r - e p i n e p h r i ne and a p l a c e b o . This s t u d y

was an attempt to test the theory that epinephrine is the

cause of the emotion fear and that n o r - e p i n e p h r i ne is the

cause of the emotion of anger directed away f r o m the self.

The hypotheses that subjects receiving an injection of

epinephrine w o u l d s h o w a significantly greater fear response

t h a n those subj e c t s receiving either an injection of nor-

epinephrine or a p l a c e b o , and t h a t subjects receiving an

injection of n o r - e p i n e p h r i n e w o u l d s h o w a s i g n i f i c a n t l y

greater response of anger directed away from the self than

those subjects receiving either an injection of epinephrine

or a placebo,were tested. A s i g n i f i c a n t d i f f e r e n c e w a s f o u n d

to exist and the hypotheses were accepted.

S t a t i s t i c a l a n a l y s i s of the d a t a r e v e a l e d t h a t all

g r o u p s ( n o n - i nj e c t i o n , e p i n e p h r i n e , n o r - e p i n e p h r i n e , and

placebo) were significantly different from each other. It

was also f o u n d that the order of presentation and groups were

34

35

significant. This indicates that some degree of interaction

was present,but no specific conclusions were made as to the

cause of this interaction.

Eeeoramendatio ns

Based on the results and conclusion of this investiga-

tion, several additional related conditions require further

experimentation and exploration.

1. Future investigations directly related to the

pre ent study should incorporate a design whereby each animal

would serve under one condition and not three.

2. Additional research should be done concerning the

correct concentration and volume of nor-epinephrine to be

i nj ected. This might alleviate the pressure effects which

caused minor paralysis.

3. Future investigations might select a different task,

particularly one which measures anger more directly.

4. Future investigations might select a different

species, one whi ch would manifest the desired physiological

changes wi thout any confoundi ng side effects.

BIBLIOGRAPHY

Books

Barnes, C. D. and L. 6. Eitherington, Drug Dos^e in Laboratory Animals. Berkeley and Los Angeles, University of California, 1954.

Cannon, W. B., Bodily Changes la Fata.. Hunger. Fear and Raae. New York and London, D. Appleton and Company, 1929.

Carr, H. A., A Study of Mental Activity. New York, Longmans, Green, 1925.

James, William, Psycho logy. New York, Harper and Row, 1961.

James, William and C. G. Lange, The Emotions. Baltimore, Williams and Wilkins, 1922.

McDougall, William, Introduction && Swlfll > London, 0 . Appleton and Company, 1908.

Winer, B. J., Statistical Principles in Experimental Design. New York, McGraw-Hill Book Company, Inc., 1962.

Wolff, H. G., Life Distresses and Disease. Baltimore, Williams and Wilkins, 1950.

Articles

Ax, A. F., "The Physiological Differentiation Between Fear and Anger," Psychosomatic Medicine. XV (1953), 433.

Funkenstein, D. H., "The Physiology of Fear and Anger," Scientific A m e r i c a n . CXCII (1955), 74-80.

Funkenstei n, D. H. and L. W. Meade, "Nor-Epi nephri ne-like and Epinephrine-like Substances and the Elevation of Blood Pressure During Acute Stress," Journal of Nervous, and Mental Disorders. CXI v" <1954), 380-397.

36

37

Funkenstein, D. H., S. H. King, and M. Drolette, "The Direction of Anger During a Laboratory Stress-Inducing Situation," Psychosomatic Medicine, XVL (1954), 404-413.

Goldenberg, M., K. L. Pines, E. Baldwin, D. G. Greene and C. E. Boh, "The Hemodynamic Response of Man to Nor-Epinephrine and Epinephrine and Its Relation to the Problem of Hypertension," American Journal of. Medici ne. V (1948), 792.

Hess, W. R. and K. Albert, "Experimental Data on the Role of Hypothalamus in Mechanism of Emotional Behavior," Archives of Neurology and Psychiatry. LXXIII (1955), 127-133.

Hick am, J. B. , W. II. Cargill and A. C. Golden, "Cardio-vascular Reactions to Eraotional Stimuli," Journal of Clinical Invest! gat I on,« XXVI (1947), 1.

Iiokf elt, B., "Nor-adrenalin and Adrenalin in Mammalian Tissues," Acta Physiology Scandinavia. XCII (1951),

25.

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