Reprinted from: Brain and Behavior. Raju TR, Kutty BM, Sathyaprabha TN and Shanakranarayana Rao BS (eds.
),
National Institute of Mental Health and Neuro Sciences, Bangalore, India. 2004:121-126.
INTRACRANIAL SELF-STIMULATION
Ramkumar K, Raju TR and Shankaranarayana Rao BS
Electrical stimulation of the brain is an
important tool in the study of the neural basis of
behavior. It has, often, the opposite effects of a
lesion to the same site. It can elicit some specific
types of behavior that would have been normally
induced by stimuli naturally sought or attended
to: eating, drinking, copulating, attacks or sleep.
The elicited responses depend on the location of
the electrode in the brain, the parameters of the
current and the test environment in which the
stimulation is administered.
Electrical stimulation of the brain continues to
be a fruitful method for exploring brain-behavior
relationships. The important applications of brain
stimulation are;
a. Study of the electrical self-stimulation
phenomenon
b. Elicitation of complex behaviors by subcortical
stimulation
c. The mapping of “motor” areas in the cerebral
cortex from which limb movements can be
elicited by stimulation
d. The use of electrical brain stimulation as the
conditioned and/or unconditioned stimulus in
classical conditioning studies
e. The elicitation of speech or recall of experience
by brain stimulation in humans and
f. The use of brain stimulation to induce amnesia,
or conversely to facilitate learning
Brain electrical stimulation is a method by
which the functions of some parts of the brain
(limbic system) have been investigated. The
method consists of introducing stimulating
electrodes into specific areas of the brain. Weak
pulses of current produce an immediate increase
in the firing of neurons near the tip of the
electrode. The effects of passing electrical currents
through these electrodes is measured directly from
the observation or recording of some aspects of
behavior.
121
Using the electrical stimulation method, James
Olds and Peter Milner (1954) in McGill University,
observed that some animals seem to behave in a
manner that increased the amount of intracranial
stimulation that they received. Further
investigation demonstrated that rats will press a
lever as rapidly as 2000 times in fifteen minutes to
obtain electrical brain stimulation
(Shankaranarayana Rao et al 1993, 1994), and they
will continue responding at this rate for several
hours. They will ignore other rewards, such as
water or food, and continue working for electrical
stimulation. These very powerful results led to the
adoption of the intracranial self-stimulation method
for investigating the “reward system” in the brain
and remains up to now the principal tool.
Olds and his collaborators have conducted
extensive mapping of the neuronal substrates of
reward in the late 1950s. They reported that the
highest rate of intracranial self-stimulation (ICSS)
was obtained in the septal area, the amygdala and
the anterior hypothalamus. Moderate but
substantial rates of ICSS were observed in related
limbic structures, particularly the hippocampus,
cingulate gyrus, anterior thalamus and posterior
hypothalamus.
General methodology
Subjects: Adult male Wistar rats weighing in
the range of 250-275 grams are used for the
stereotaxic implantation of bipolar electrodes in
substantia nigra-ventral tegmental area (SN-VTA)
bilaterally.
Apparatus: Stereotaxic table, surgical
instruments, stimulator (programmable pulse
generator), Skinner’s box modified for ICSS and
connecting cables.
Electrodes: Either monopolar or bipolar
electrodes can be used for the purpose. Bipolar
electrodes, spaced closely together (0.5-1.0 mm
between tips), are widely used, since the spread of
the stimulation current can be precisely defined,
the maximally stimulated tissue lying between the
electrode tips.
Although platinum is a superior metal for
chronic brain stimulation due to its non-corrosive
property, stainless steel or nichrome wire is
commonly used since it is cheaper and readily
available. The diameter of the wire used is 28G that
should be insulated, except across the tip, with some
inert material (Epoxylite resin).
Surgical procedure: The rat is anesthetized
with sodium pentabarbitone (40mg/kg b.w)
and then positioned in the stereotaxic frame
with rat adaptor. Following injection of
lignocaine anesthesia into the scalp region, the
skull surface was exposed for landmarks. Flat-
skull coordinates adapted from Paxinos and
Watson rat atlas are marked with bregma as
reference point and burr holes are drilled
through the skull. The bipolar electrodes are
implanted chronically into substantia nigra-
ventral tegmental area (SN-VTA).
The stereotaxic coordinates for SN-VTA are: (see
Figure 2)
Antero-posterior (AP) : -3.5 to – 4.5 mm
Medio-lateral (ML) : 1.1 to 1.8 mm
Dorso-ventral (DV) : 8.5 ± 0.2 mm
The indwelled electrodes should be firmly fixed
with acrylic dental cement and one or two
anchoring screws can be fixed for the firm fixation
of the electrodes. Following 5-7 days of post
surgical recovery from surgical trauma the rats can
be tested for the ICSS behavioral response.
Electrical stimulation: The electrodes of one side
of the head should be connected to the output
socket of the pulse generator (stimulator) and the
rat placed in the testing chamber (the Skinner’s box
modified for ICSS behavior). The Skinner’s box has
a pedal (lever) on one of the walls of the chamber
that is connected to the micro switch, which has
connection with the pulse generator. Pressing the
lever completes the circuit and delivers a pulse of
current (Figure 1).
Once a current level is found which serves as a
reward, shaping can be used to quickly establish
lever pressing; i.e., by reinforcing first movements,
then locomotion towards the lever, then sniffing
around the lever, then touching and finally
pressing it. Soon the rat learns to predict the
relationship of pedal pressing and the rewarding
stimulation. The current strength and frequency
are monitored by observing the rat’s response to
the stimulus. Thus, the animal self-stimulates
reliably on a continuous reinforcement schedule
resulting in stable self-stimulation behavior.

OSCILLOSCOPE
MODIFIED SKINNER CHAMBER FOR ICSS

TIMER
(STOP WATCH)

Figure 1: Diagrammatic representation of the rat performing ICSS behavior (Shankaranarayana Rao 1996).
122
Histological verification of Electrode
Placement
After the experiments, the position of the
electrode tips can be marked with a small lesion
by passing current through the stimulating
Figure 2. Plate from Paxinos and Watson (1982) stereotaxic atlas of the rat brain. The arrow indicates the placement of
electrodes in the SN-VTA where electrical stimulation should serve as a reinforcer. The vertical mm scale on the right
indicates depth from the surface of the brain.
However, the method of ICSS suffers several
drawbacks:
1. The current would spread over a wider
area of the brain than the targeted neuronal
nucleus.
2. It does not discriminate between different
fibers of passage crossing trough and nearby the
targeted nuclei.
Mesocorticolimbic system a major neural
substrate for ICSS
Research investigating the rewarding properties
of intracranial self-stimulation (ICSS) has identified
dopamine as the neurotransmitter involved in
reward. Regions in the brain that support ICSS,
known as pleasure centers, overlap with known
dopamine systems in the brain. The dopamine
system involved in reward is the mesotelencephalic
dopamine system (referred to as the midbrain
dopamine system). It is constituted of dopaminergic
123
electrodes. Perfuse the brain and section it and stain
the sections with cresyl violet. Under a light
microscope establish the location of the electrode
tips. Reconstruct the position of the tips on plates
taken from a brain atlas.
neurons projecting from the mesencephalon (the
midbrain) into various regions of the telencephalon
(Figure 3). The neurons that compose this system
have their cell bodies in the substantia nigra and
the ventral tegmental area. These are two closely
related nuclei composed of dopamine-containing
neurons, which projects to a number of forebrain
sites. These include regions of the limbic cortex,
prefrontal neocortex, the lateral hypothalamus, the
preoptic area, the olfactory tubercle, the amygdala,
striatum and in particular the nucleus accumbens.
It is dopamine activity in the nucleus accumbens
that is involved with the experience of pleasure.
In bypassing much of the input side of these
neuronal circuit(s), ICSS provides a unique tool in
neuropharmacological research to investigate the
influence of various substances on reward and
reinforcement processes. Intracranial self-
stimulation differs significantly from drug self-
administration in that, in this procedure, the
animal is working to directly stimulate presumed
reinforcement circuits in the brain and the effects
of the drugs are assessed on these reward
thresholds. Drugs of abuse decrease thresholds for
ICSS, and there is a good correspondence between
the ability of drugs to decrease ICSS thresholds and
their abuse potential.
Figure 3. Schematic representation of mesocorticolimbic
dopaminergic system.
ICSS, a Model to Study Neuronal Plasticity
Recently, the electrical self-stimulation
paradigm has proved to be very useful in
delineating the neural substrate involved in
learning and drug abuse. Self-stimulation involves
operant learning, which can induce changes in
the neuronal cytoarchitecture. Accordingly, we
have used this experimental para-digm to study
learning related neuronal plasticity. We have
conducted a series of experiments to determine the
self-stimulation (SS) rewarding experience induced
neuronal plasticity in hippocampal and motor
cortical neurons (Shankaranarayana Rao and Raju,
2001a, Shankaranarayana Rao et al 2001b).
The electrical self-stimulation has been
considered as one of the intensely rewarding
behavioral experience, perhaps even more
influential than feeding or sexual rewards (Olds
1962). Selfstimulation rewarding experience for 10
days resulted in an long-lasting increase in the
dendritic branching and dendritic length in CA3
hippocampal and layer V motor cortical pyramidal
neurons (Shankaranarayana Rao et al. 1993,
1994). The dendritic growth is associated with
cytoskeletal changes in the hippocampal and
124
cortical neurons. In addition to the dendritic
growth, a significant increase is observed in the
thickness of lacunosum, radiatum and lucidum
laminae in the CA3 region of the hippocampus in
selfstimulation experienced rats
(Shankaranarayana Rao et al. 1993). ICSS also
resulted in a significant increase in numerical
density of dendritic spines in different categories
of both apical and basal dendrites in CA3
hippocampal and layer V motor cortical
pyramidal neurons (Shankaranarayana Rao et al.
1999a) and also an increase in the density of
thorny excrescences in apical dendrites of CA3
neurons of the hippocampus (Shankaranarayana
Rao et al. 1998a). ICSS caused synaptogenesis in
CA3 region, which included moleculare, radiatum
and lucidum layers of the hippocampus and
molecular layer of the motor cortex
(Shankaranarayana Rao et al. 1999b).
In addition, SS results in an increase in the
levels of glutamate, noradrenaline, dopamine and
enhancement of AChE activity in the
hippocampus and the motor cortex
(Shankaranarayana Rao et al 1998b).
Furthermore, the prior SS experience is known
to facilitate the acquisition of operant and spatial
learning tasks in rats (Yoganarasimha et al 1998).
Such facilitation might be due to an increase in
the dendritic arborization associated with
neurochemical changes in the hippocampus.
In summary, the facilitatory effects of ICSS on
operant and spatial learning led us to suggest that
rewarding electrical stimulation could affect
learning and memory by inducing an adequate
activation of the neural system and the pathways
that were able to affect a wide variety of
conditioned responses. Further experiments are
thus necessary to demonstrate a more specific effect
of reinforcing brain stimulation on learning and
memory process.
ICSS, an Animal Model to study Drug Addiction
From the basic neuroscience perspective, study
of the neurobiology of drug addiction offers a novel
opportunity to establish the biological basis of a
complex and clinically relevant behavioral
abnormality. Many prominent aspects of drug
addiction in people can be clearly produced in
laboratory animals, in striking contrast to most of
other forms of neuropsychiatric illness. Thus,
advances made in the study of drug addiction
should provide important insights into the
mechanisms underlying some of the psychiatric
disorders.
In recent conceptualizations of drug
reinforcement, the positive reinforcing properties
of drugs have been thought to play an important
role in drug dependence. It is amply clear that
animals and humans will readily self-administer
drugs in a dependent state and that drugs have
powerful reinforcing properties in that animals will
perform many different tasks to obtain drugs. The
drugs that have positive reinforcing effects
correspond well with the drugs that have high
abuse potential in humans. Electrical self-
stimulation of certain brain areas is rewarding for
animals and humans as demonstrated by the fact
that subjects will readily self-administer the
stimulation (Olds & Milner 1954). The powerful
nature of the reward effect produced by ICSS is
indicated by the behavioral characteristics of the
ICSS response, which include rapid learning and
vigorous execution of the stimulation producing
behavior (Shankaranarayana Rao and Raju, 2001).
ICSS provides a unique tool in
neuropharmacological research to investigate the
influence of various substances on reward and
reinforcement processes.
Drugs of abuse decrease thresholds for ICSS, and
there is a good correspondence between the ability
of drugs to decrease ICSS thresholds and their
abuse potential. ICSS thresholds have been used
to assess changes in systems mediating reward and
reinforcement processes during the course of drug
dependence. Acute administration of
psychostimulant drugs lower ICSS threshold (i.e.,
increases ICSS reward) and withdrawal from
chronic administration of these drugs elevate ICSS
thresholds (i.e., decrease ICSS reward). Similar
results have been observed with precipitated
withdrawal in opiate-dependent rats.
The advantage of the ICSS paradigm as a model
of drug effect on motivation and reward is that by
directly stimulating the putative reward systems,
125
one presumably bypasses the input side of the
system and eliminates the nonspecific effects of
consummatory behaviors, such as feeding, that can
complicate data interpretation. Also, the
behavioral threshold measure provided by ICSS
procedures is easily quantifiable, because ICSS
threshold estimates are very stable over periods of
several months. Another considerable advantage
of the ICSS technique is the high reliability with
which it predicts the abuse liability of drugs
(Shankaranarayana Rao and Raju, 2001a).
ICSS, an Animal Model for Depression
ICSS paradigm provides an operational
measure of anhedonia, a core feature of depression.
Because the anhedonia experienced by depressed
patients suggest that these individuals might exhibit
alterations in reward processes, the ICSS paradigm
has been proposed as a model of depression.
Substantial evidence suggests that ICSS thresholds
are reliable measures of reward that reflect the
whole continuum from hedonia to anhedonia.
Because it appears that ICSS acts directly on some
of the same neuronal substrates that mediate the
rewarding effects of natural reinforcers, it is
considered to be a valuable tool for the investigation
of brain-reward systems (Shankaranarayana Rao
and Raju, 2001a).
The study of neural substrates of ICSS behavior
following experiential or pharmacological
manipulations promises to promote our
understanding of reward mechanisms that seem
to be altered in several psychiatric disorders,
including depression and schizophrenia. Two
manipulations have been used to produce an
anhedonic state in animals, as operationally
defined by decreases in ICSS response rates or
elevations of thresholds, (i) exposure to
uncontrollable stress and (ii) withdrawal from
long-term exposure to psychomotor stimulants.
Tricyclics appear to be effective in reversing the
effects of withdrawal from amphetamine on ICSS.
Although many more studies are needed, the
evidence to date suggests that ICSS model has good
construct and etiological validity and exhibits
pharmacological isomorphism as a model for the
drug induced anhedonia. Nevertheless, the precise
relationship between drug-induced and non-
drug-induced depression in humans is not
known. Thus, the possible etiological validity of
the ICSS paradigm as a model of non-drug-
induced depression remains unclear. Future
clinical and preclinical research needs to address
this issue further.
References:
1. Olds, J. and Milner, P. (1954) Positive reinforcement
produced by electrical stimulation of septal area and
other regions of the rat brain. J. Comp. Physiol. Psychol.,
47:419-429.
2. Paxinos G and Watson C (1982) The rat brain in
stereotaxic coordinates. Academic Press. Australia.
3. Shankaranarayana Rao BS (1996) Neuronal plasticity
induced by self-stimulation rewarding behavioural
experience: Morphological, biochemical and immuno-
cytochemical evaluation. Ph.D. Thesis, NIMHANS
Deemed University, Bangalore. India.
4. Shankaranarayana Rao BS and Raju TR (2001a)
Intracranial self-stimulation : An animal model to
study drug addiction, depression and neuronal
plasticity – A Review. Proc Indian Natnl Sci Acad B67:
155-188.
5. Shankaranarayana Rao BS, Desiraju T and Raju TR
(1993) Neuronal plasticity induced by self-
stimulation rewarding experience in rats - a study
on alterations in dendritic branching in pyramidal
neurons of hippocampus and motor cortex. Brain Res
627:216-224.
6. Shankaranarayana Rao BS, Desiraju T, Meti BL and
Raju TR (1994) Plasticity of hippocampal and motor
cortical pyramidal neurons induced by self-
stimulation experience. Indian J Physiol Pharmacol
38:23-28.
126
7. Shankaranarayana Rao BS, Meti BL and Raju TR
(2001b) Neuronal plasticity - A unique property for
Neurorehabilitation. In : Neurorehabilitation :
Principles and Practice, (Taly AB, Nair KPS and Murali
T eds) 2nd edition, Ahuja Book Company, New Delhi,
India, pp.27-37.
8. Shankaranarayana Rao BS, Raju TR and Meti BL
(1998a) Alterations in the density of excrescences in
CA3 neurons of hippocampus in rats subjected to self-
stimulation experience. Brain Res 804:320-324.
9. Shankaranarayana Rao BS, Raju TR and Meti BL
(1998b) Self-stimulation of lateral hypothalamus and
ventral tegmentum increases the levels of
noradrenaline, dopamine, glutamate and AChE
activity, but not 5-hydroxytryptamine and GABA levels
in hippocampus and motor cortex. Neurochem Res
23:1053-1059.
10. Shankaranarayana Rao BS, Raju TR and Meti BL
(1998e) Long-lasting structural changes in CA3
hippocampal and layer V motor cortical pyramidal
neurons associated with self-stimulation - a
quantitative Golgi study. Brain Res Bull 47:95-101.
11. Shankaranarayana Rao BS, Raju TR and Meti BL
(1999a) Self-stimulation rewarding experience induced
alterations in dendritic spine density in CA3
hippocampal and layer V motor cortical pyramidal
neurons. Neuroscience 89:1067-1077.
12. Shankaranarayana Rao BS, Raju TR and Meti BL
(1999b) Increased numerical density of synapses in
CA3 region of hippocampus and molecular layer of
motor cortex after self-stimulation rewarding
experience. Neuroscience 91:799-803.
13. Yoganarasimha D, Shankaranarayana Rao BS, Raju
TR and Meti BL (1998) Facilitation of acquisition and
performance of operant and spatial learning tasks in
self-stimulation experienced rats. Behav Neurosci
112:725-729.