Effects of Pentobarbital on Intermittently Reinforced Behavior

Author(s): R. J. Herrnstein and W. H. Morse

Source: Science , May 10, 1957, New Series, Vol. 125, No. 3254 (May 10, 1957), pp. 929-
931
Published by: American Association for the Advancement of Science

Stable URL: https://www.jstor.org/stable/1754314

JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide
range of content in a trusted digital archive. We use information technology and tools to increase productivity and
facilitate new forms of scholarship. For more information about JSTOR, please contact support@jstor.org.

Your use of the JSTOR archive indicates your acceptance of the Terms & Conditions of Use, available at
https://about.jstor.org/terms

American Association for the Advancement of Science is collaborating with JSTOR to digitize,
preserve and extend access to Science

This content downloaded from

189.203.104.96 on Wed, 24 Apr 2024 00:34:23 +00:00
All use subject to https://about.jstor.org/terms
advantages.
advantages. WhenWhenthe the
problem
problem
involved
involved
duction
ductionpattern
pattern suggests
suggestsL-galactonate
L-galactonate
as ratia
as marcescens
ratia marcescensto to
form
form
from from
glucuronic
glucuronic
is
is that
thatofof
estimating
estimatinglow concentrations
low concentrations aa possible
possibleintermediate
intermediate andand
reduces
reduces
the the
acid
acid aasubstance
substancetentatively
tentatively
identified
identified
as as
of
of AER
AERcells
cells
(for
(for
example,
example,
0.01 percent) likelihood
0.01 percent)likelihoodthatthatsome
someoften-suggested
often-suggested aa "1,6-ester
"1,6-esterlinked
linkeddihexuronic
dihexuronicacid,"acid,"
in
in aapredominantly
predominantly aer population,
aer population,
the the
schemes
schemes(1) (1)for
forgalacturonic
galacturonic acidacid
catab-
catab-
prompted
promptedanan examination
examination
by us
byofusuronic
of uronic
advantages
advantages of of
thethe
selective
selective
lactatelactate olism
olismare
agar agar areoperative
operative in in
E. carotovora.
E. carotovora. reductase
reductaseactivity
activity in that
in thatmicroorgan-
microorgan-
technique
technique over
over
TTCTTCoverlay
overlay
become become
ap- ap-Glucuronate-grown
Glucuronate-grown intact
intact
cellscells
of Er-
of Er-
ism:
ism: an anenzyme
enzyme preparation
preparation fromfrom
glu- glu-
parent
parent(6). (6). winia
winiacarotovora
carotovora oxidize
oxidize
glucuronate
glucuronate curonate-grown
curonate-grown
and and Serratia
Serratiareduces
reduces
glu- glu-
MAURICE OGUR galacturonate
galacturonate after
after a short
a shortinduction
induction curonate with TPNH or, somewhat
RALPH ST. JOHN phase;
phase;galacturonate-
galacturonate- andandglucuronate-
glucuronate- more slowly, with DPNH. The relation-
S. NAGAI grown
growncells cellsofofAerobacter
Aerobacter cloacae
cloacae
oxi- oxi-
ship of the aforementioned reactions to
Biological Research Laboratory, dize
dize both
bothuronic
uronic acids
acids
rapidly
rapidly
without
without
a thea system described by Isherwood et al.
Southern Illinois University, Carbondale lag lag period;
period;the the last
last
mentioned
mentioned cellscells
are are
(2) in pea mitochondria (which reduces
References and Notes
sequentially
sequentially induced
induced to to
oxidize
oxidize
L-gulon- the
L-gulon- methyl ester of galacturonate, but
ate. not galacturonate, to L-galactono-y-lac-
1. C. Raut, Exptl. Cell Research 4, 295 (1953). Cell-free extracts were prepared fromtone) is under investigation, as are ex-
2. W. Laskowski, Heredity 8, 79 (1954).
3. S. Nagai et al., Proc. Natl. Meeting Botan. Soc.
both Erwinia and Aerobacter that had tensions of this general reaction to some
Japan 20, 35 (1955). been grown on either galacturonate of orthe less common uronic acids.
4. M. Ogur and R. St. John, J. Bacteriol. 72, 500 MORTIMER P. STARR*
(1956). glucuronate, by sonic oscillation and cen-
trifugation. The supernatant was dialyzed
5. The tests of replica plating to lactate agar were JOZEF DE LEYt
made by David Pittman. WENDELL W. KILGORE
overnight at 4?C against 0.05M tris-HCl
6. This work was supported in part by grant
N17D from the American Cancer Society. buffer of pH 7.2 and centrifuged at 100,- Department of Bacteriology,
11 February 1957 000 g for 90 minutes; the clear solution University of California, Davis
was used. These preparations contain an
References and Notes
enzyme that catalyzes the reduction of
both galacturonate and glucuronate by 1. S. S. Cohen, J. Biol. Chem. 177, 607 (1949);
G. Buyze, "De Koolhydraatstofwisseling van
TPNH or DPNH. No reduction of
Lactobacillus brevis," dissertation, University
Catabolism of Hexuronic Acids D-mannuronate has ever been observed. of Utrecht (1955); W. J. Payne, J. Bacteriol.
72, 834 (1956).
by Erwinia and Aerobacter The enzyme is provisionally named 2. F. A. Isherwood, Y. T. Chen, L. W. Mapson,
uronic reductase. When a preparation Biochem. J. (London) 56, 1 (1954); L. W.
Only casual information is presently from galacturonate-grown Erwinia is Mapson and F. A. Isherwood, ibid. 64, 13
available on the metabolism of hexuronic (1956).
used, DPNH reacts slightly faster with 3. E. E. B. Smith and G. T. Mills, Biochem. et
acids by microorganisms (1); somewhat galacturonate than it does with glucu- Biophys. Acta 13, 386 (1954); J. J. Burns, P.
greater development of this field is evi- ronate; when TPNH is the reductant, 4. A.Peyser, A. Moltz, Science 124, 1148 (1956).
J. Kraght and M. P. Starr, Arch. Biochem.
dent with plant (2) and animal (3) ma- the reaction with glucuronate is about 5 and Biophys. 42, 271 (1953).
terials. Our interest in this subject stems times faster than it is with galacturonate 5. A. J. Kraght and M. P. Starr, J. Bacteriol. 64,
259 (1952).
from an earlier study of the pectinolytic under the conditions of our experiments. 6. This work was supported in part by research
action of Erwinia soft-rot bacteria in This difference is less distinct with grant RG4544 from the National Institutes of
Health, U.S. Public Health Service.
which it was shown that pectin is broken preparations from cells of Erwinia * Special fellow of the National Institutes of
down to galacturonic acid (4) and that grown on glucuronate, or Aerobacter Health, 1953-54, at the University of Cam-
the uronate is further catabolized to a grown on either uronate. The activity bridge, Cambridge, England, and at the Uni-
versity of Ghent, Ghent, Belgium.
mixture of end-products (5). For the of the uronic reductase is very weak in t Present address: Rijksuniversiteit, Casinoplein
past few years we have sought the initial extracts from glucose-grown cells of 11, Ghent, Belgium; Geassocieerde of the Bel-
step in the breakdown of galacturonate. gian National Fonds voor Wetenschappelijk
either species. Onderzoek.
We wish now to report that our investi- The end-products of the reduction of 30 January 1957
gations (6) indicate that the first detect- the uronates by both bacterial species,
able step in the catabolism of D-galac- with either DPNH or TPNH, are non-
turonate and D-glucuronate by cell-free reducing, nonlactonized acids: L-galac-
extracts of Erwinia carotovora and Aero- tonate from galacturonate, and L-gulon-
bacter cloacae is a reduction, with either ate from glucuronate. By boiling for 5 Effects of Pentobarbital on
reduced triphosphopyridine nucleotide minutes with 1N HC1, the acids are
(TPNH) or reduced diphosphopyridine converted into the corresponding lac- Intermittently Reinforced Behavior
nucleotide (DPNH), to the correspond- tones which, on paper chromatograms, The behavioral effects of drugs have
ing hexonic acid, namely, L-galactonate behave in every respect like L-galactono- recently (1-5) been measured in terms
and L-gulonate. y-lactone after galacturonate reduction, of changes in the stable temporal pat-
These reductions are carried out by or L-gulono-y-lactone after glucuronate terns of responding that occur when be-
substrate-induced enzymes. Glucose- reduction. It is highly unlikely that the havior is reinforced intermittently. It
grown intact cells oxidize glucose at reaction proceeds by lactonization of the has been shown that drugs may influence
once in manometric experiments, but act uronic acid followed by reduction to both the average rate and the pattern of
on galacturonate and glucuronate only the corresponding L-hexonolactone; this emission of responses (4). The latter
after an induction period. On the other opinion is based on (i) the failure of effects are of particular interest, since
hand, galacturonate-grown intact cells of extracts of Aerobacter cloacae to reduce these patterns probably represent the
Erwinia carotovora respire galacturonate D-glucurono-y-lactone with DPNH or operation of more subtle behavioral
at once or after a brief induction phase; TPNH, and (ii) the actual accumula- processes than are reflected in the aver-
glucuronate is used very slowly. These tion of the free L-hexonic acids taken age rate of response. In this report, a
galacturonate-grown cells are adapted at together with the observed lack of de- new measure of the temporal pattern of
the same time to the oxidation of lactonizing enzyme activity in the ex-responding is used in order to compare
L-galactonate, but not to D-galactonate, tracts for either of the L-hexono-v-lac- the effects of a drug on temporal pattern-
D- or L-galactose,
L-galactose, mucate,
mucate, D-D- or
or L-Iyxose,
L-Iyxose,
tones.
ing
ingwith
withthe
the
effects
effects
on rate
on of
rate
response.
of response.
D-xylitol,
D-xylitol, L-arabitol,
L-arabitol, L-ascorbate,
L-ascorbate, L-ara-
L-ara-
The recent report by Payne (1) re- One
Oneform
form of of
intermittent
intermittent
reinforce-
reinforce-
binose, or
or D-fucose.
D-fucose. This
This sequential
sequential in-
in-
garding the ability of dried cells of Ser-
ment
mentisisdesignated
designated as a as
fixed-interval
a fixed-interval
10 MAY 1957
929

This content downloaded from

189.203.104.96 on Wed, 24 Apr 2024 00:34:23 +00:00
All use subject to https://about.jstor.org/terms
schedule. In this procedure, the first oc- is correlated with t the passage of time, forcement for about 30 daily
currence of the arbitrarily selected re- i be viewed as a tem- mental sessions.
the performance can
sponse after a fixed period of time makes 1. Each pigeon spent 8 hours a day in a
poral discrimination
the reinforcement (for example, food) lerated by the fixed- sound- and light-insulated exp
The behavior ger
accessible to the experimental subject. interval schedule E las been used as a chamber. Reinforcements were deliv-
base line for the stu idy of sodium pento- ered on the 15-minute, fixed-interval
Responses that occur before the fixed
time has elapsed have no explicitly ar- barbital and has ; shown sensitivity in schedule. After each reinforcement, al
ranged consequence. Animals show a terms of changes n both
ii average rate illumination in the chamber was exti
characteristic pattern of response when and temporal pat tern of responding guished for a period of 5 minutes. T
reinforcements are delivered on a fixed- (2, 5). In the pres ent experiment (7), number of key-pecks during this "time
interval schedule (6). For example, if measurements were taken of the changes out" was negligible. Each 15-minute in
the size of the interval is 15 minutes, that occurred in th le fixed-interval per- terval is considered to begin at the te
then there is usually no responding dur- formance when sub; anesthetic dosages of mination of a time-out.
ing the first 5 to 8 minutes of each inter- 1 were administered. Sodium pentobarbital was injected i
sodium pentobarbita
val. After this initial pause, the animal The experimental animals were two tramuscularly during the second or third
begins to respond, at first slowly and It, male, White Car- hour of experimental sessions. Drug ad
food-deprived, adul
then more rapidly; the highest rates oc- 3 response was the ministrations always took place imme
neau pigeons. The
cur just prior to reinforcement. This 'At- pecking of a Plexiiglas disk, and rein- ately after a reinforcement; this left 4
tern of responding is the most stable and forcement was brie f (4 seconds) access to 5 minutes of time-out before the nex
most interesting feature of fixed-interval to grain. Prior todrug administration, interval began. Sessions in which ther
performance. Inasmuch as this highly re- these pigeons weretrained on a 15-min- were injections were separated by at
producible progression of response rates ute, fixed-interval schedule of rein- least one complete experimental session
in which there was no injection. Dosages
of 1, 2, 3, and 4 mg of sodium pentobar-
A. bital were each administered six times
,'~/ \ per pigeon. The various dosages were
Co
w
/ . given in any systematic order.
u)
z
/ '/\ \ The top part of Fig. 1 shows the com-
0
* /\ , ," bined results for both pigeons for all
ui
IL
,/ - injections. The number of responses per
x
.. \ 15-minute interval for 12 consecutive in-
0 ?..?,:.. \ tervals is obtained. Six of the 12 intervals
a: .... \ are prior to injection and six follow.
w
These numbers are then averaged, inter-
I
ao
val by interval, for each dosage of pen-
z
z
tobarbital. Each curve, therefore, pre-
.4
ui
sents average data for 12 administrations
I (six for each pigeon) of each dosage.
POST-INJECTION The graded effect of pentobarbital can
be seen in the first postinjection interval.
In this interval the reduction in average
I -I I I number of responses is a function of the
5 6 1 2
3 4 5 6 size of the dosage. One milligram does
CONSECUTIVE INTERVALS not seem to cause a substantial decrease
in response-output. Beginning with the
15- second postinjection interval, there are
14-
clear instances of increased responding
as a result of administrations of sodium
(0
13-
w
Co
pentobarbital. The magnitude of this
0
z 12- -x....... "excitatory" effect does not appear to be
Co I I- ...-... monotonically related to dosage. There
w
a: I0- x. .. .......I .... ... ..... ..... ..x.--.. . . /.. is some indication that the time at which
LL the maximal increase occurs depends on
9-
Cu, the dosage and that it is later with larger
w
8-
I'
-'/ '^^A---^ doses.
1J z 7- *^~~'~ / ~ The number of responses per interval
aQ: 6-
I'
/ measures the animal's rate of response in
I4-
I successive 15-minute periods. It does not
5- I /.-
'i ? 3 /. ' reflect directly the changes that occur
ot
0
4-
Ii / / in the temporal pattern of responding
2
4 3- within single intervals after drug admin-
I
2- PR E-INJECTION \ /
POST-INJECTION istration. The latter aspect of the drug
I- N/4 effect is presented in the bottom part of
Fig. 1. The data presented in both parts
i[- - I SI I I I I I! - 1 f- -- --1 I ~- of Fig. 1 were obtained from the same
I 2 3 4 5 6 1 2
3 4 5 6 sessions. The ordinate of the bottom part
CONSECUTIVE INTERVALS
of Fig. 1 is a measure of the pattern of
Fig. 1. (Top) Average number of responses per val15-minute
for 12 consecutive
inter)responding characteristic of the fixed-in-
intervals. Each point represents the average of 12Numbers on the curves
determinations. I terval performance. The quarter-life of
) Average quarter-life
refer to dosages in milligrams of sodium pentobarbital. (Bottom) responses is the time taken, in any one
rdinate).
for the same 12 consecutive intervals (see text for interval,
explanation of for
o the first one-fourth of the
930 SCIENCE, VOL. 125

This content downloaded from

189.203.104.96 on Wed, 24 Apr 2024 00:34:23 +00:00
All use subject to https://about.jstor.org/terms
 References and Notes
total
totalnumber
numberof of
responses
responses
in that
in interval
that interval coupled
coupledwith
withmetabolic
metabolicreactions,
reactions,
with-
with-
to
to be
beemitted.
emitted. ForFor
example,
example,
if theifre-
the re-
1. J. V. Brady, Science 123, 1033 (1956). out reference to the direction of the re-
sponse
sponserate
rate
in in
some
some
interval
interval
were were
con- con-2. P. B. Dews, J. Pharmacol. Exptl. Therap. 113, sulting net flux. However, in the absence
393 (1955).
stant,
stant,then
thenthethe
quarter-life
quarter-life
wouldwould
be sim-be sim-
3. D. S. Blough, Ann. N.Y. Acad. Sci. 65, 334 of a precise knowledge of the mecha-
ply
plyone-fourth
one-fourthof the
of the
duration
duration
of theof the (1956). nisms involved or of a possibility of
4. W. H. Morse and R. J. Herrnstein, Ann. N.Y.
interval.
interval.That
Thatis to
is say,
to say,
whenwhen
the rate
theis rate is Acad. Sci. 65, 303 (1956); M. Sidman, Ann. measuring "passive" permeability coeffi-
constant,
constant,one-fourth
one-fourth
of the
of responses
the responses
will will N.Y. Acad. Sci. 65, 282 (1956). cients in the undisturbed living system,
occur in one-fourth of the time. If the 5. R. J. Herrnstein and W. H. Morse, Science such a definition does not prove experi-
124, 367 (1956).
responding within an interval has posi-6. C. B. Ferster and B. F. Skinner, Schedules of mentally meaningful. Numerous at-
tive acceleration, then the quarter-life Reinforcement (Appleton-Century-Crofts, in tempts have been made to arrive at a
press).
will be greater than one-fourth of the in-
7. This research was carried on in the Psychologi-
value for the thus defined "passive" term
terval. On the other hand, if the rate cal Laboratories, Harvard University, with the through the use of metabolic inhibitors.
should show a decline within an inter- support of the William F. Milton Fund, ONR That it is, however, impossible in prin-
contract N5 ori-76, and a grant from the Na-
val, then the quarter-life will fall below
tional Science Foundation. ciple to make an unequivocal distinction
one-fourth. *Present address: Walter Reed Army Institute between "active" and "passive" or even
of Research, Walter Reed Army Medical Cen-
The quarter-life prior to injection ter, Washington, D.C. "metabolically dependent" and "meta-
tends to lie between 11 and 12 minutes. bolically independent" components of
25 February 1957
This means that about four-fifths of the flux solely on the basis of experiments
15-minute fixed interval has elapsed with metabolic inhibitors can be seen
when the first one-fourth of the responses from the following considerations.
in the interval have been emitted. The According to the most general defini-
high value of the quarter-life expresses On the Distinction between the tion, the isotopically measured flux in
the rate increase that is characteristic of either direction would be expressed as a
Effects of Agents on Active and
fixed-interval performance. Passive Transport of Ions sum of a "passive" and an "active"
In the first interval following injec- term. The passive term should in prin-
tion, the average quarter-life falls in- Several recent studies (1-3) designed ciple be given by the product of a
creasingly further below the base-line to distinguish between the effects of permeability coefficient, which is deter-
value, for 2, 3, and 4 mg of pentobarbi- pharmacologic and other agents on the mined by the properties of the cell mem-
active and passive components of ionicbrane and the activity of the ion on the
tal, respectively. This change reflects the
loss of the characteristic rate increase flux have brought out the difficulties ofside from which the flux originates. In
within the fixed interval. In the case of posing this question within the frame-the absence of evidence to the contrary
3 and 4 mg of pentobarbital, the quarter- work of current definitions of active -and such evidence is unobtainable
life has fallen below 334 minutes. This transport. A brief analysis of the as- without a clear-cut identification of the
indicates that the responding in these sumptions on which an unambiguous "active" term-the permeability coeffi-
instances shows negative, rather than the distinction of this kind can be founded
cient must be assumed to depend on the
customary positive, acceleration. seems, therefore, very desirable. state of metabolism as well as on ionic
The time course of the drug effect on The ability to separate the effects of activity. Any metabolic inhibition must
an agent on the two components of flux
the quarter-life is seen in the consecutive therefore be assumed to alter the per-
hinges, of course, on an experimentally
postinjection intervals. By the sixth post- meability coefficient by an unknown
injection interval, the quarter-life has unequivocal distinction between the ac- amount and in an unknown direction.
almost returned to the base-line value. tive or passive components themselves. Thus one may not regard its measured
The effect of sodium pentobarbital,However, as such a satisfactory quantita- value even as a meaningful extremum,
measured by the quarter-life, is a change tive distinction does not exist at present,making a distinction between the "ac-
in the characteristic pattern of responsesand one is limited to a distinction based
tive" and "passive" terms on this basis
associated with the fixed-interval sched- on the thermodynamic definition of impossible.
ac-
ule. The magnitude of the change varies tive transport as transport against an elec-More difficult to foresee is the impos-
trochemical potential gradient. This defi-
directly with the size of the dose and dis- sibility of distinguishing by this approach
sipates gradually in time. nition was given by Rosenberg (4). The between the components of flux which
The drug effect appears to be analyz- principal objection to this formulation are and those which are not dependent
able into two components: a depressive is that it defines active transport in on metabolism. We may represent the
terms of a net-that is, a necessarily uni-
effect and a loss of the positive accelera- total (measured) flux by an equation
tion in responding within the 15-minute directional-active flux. It specifically such as
interval. The increase in responding,excludes metabolically linked transport
shown in the top part of Fig. 1, is prob- (i) that is codirectional with and in ex- fi = P?(m)ao + fi*(m) (1)
ably a consequence of the change in the cess of that expected from the electro-
temporal pattern of responding within chemical potential gradient and (ii) where
wherefifiis is thethe
total
total
inward
inward
flux, flux,
po thepo the
that is opposing but incompletely com-
intervals. As the depressive effect disap- value
valueofofthe the permeability
permeability coefficient
coefficient
in in
pears, the absence of positive accelera- pensating the flux resulting from the the
the complete
complete absence
absence
of metabolism
of metabolism but but
tion produces responding that occursgradient. One would intuitively wishwith with
to allallother
other independent
independent variables
variables
re- re-
include both of these. The definition con-
throughout, rather than at the end of, the turning
turningtheirtheir values,
values,
Jt (m)
Jt an
(m) unknown
an unknown
15-minute interval, thus increasing the tains nothing to justify an interpretation function
functiondescribing
describing its dependence
its dependence on on
over-all output of responses. The fact of the net flux as a difference between the
the rates
ratesofof metabolic
metabolic reactions,
reactions,
ao theao the
that the depressive effect (Fig. 1, top) two "active" components in the two di- activity
activityofof thethe
ionion
on the
on outside,
the outside,and and
disappears more rapidly than the loss rections,
of as has been suggested. In addi-fi
fi (m)
(m)thetheunknown
unknown active
active
transport
transport
discrimination (Fig. 1, bottom) prob-tion, it encompasses transport processes term.
term.EvenEven if if
perfect
perfect
metabolic
metabolicinhibi-inhibi-
ably accounts for at least some of the that require a supply of energy but do tion
tionwere
wereachieved,
achieved,there
there
is noismeans
no means
of of
increase in responding. not derive it from metabolic reactions- ruling
rulingoutout anyanyeffect
effect
of the
of inhibitor
the inhibitor
on on
R.
R. J.
J. HERRNSTEIN*
HERRNSTEIN* for example, those leading to a Donnan the
the membranes,
membranes, which
which
itself
itself
is indepen-
is indepen-
W. H. MORSE equilibrium. dent
dentof ofmetabolism,
metabolism, so that
so that
again again
the the
Psychological Laboratories, Harvard It may seem that one could define measured measured
ac- value
valuepotpot
maymay
in noinwayno reflect
way reflect
University, Cambridge, Massachusetts tive transport as transport specifically even an extreme value of po. In other
10 MAY 1957 931

This content downloaded from

189.203.104.96 on Wed, 24 Apr 2024 00:34:23 +00:00
All use subject to https://about.jstor.org/terms