SEMINAR ON SCREENING

OF NOOTROPICS
(COGNITIVE ENHANCERS)

Presented by:
Souvik dutta
1st M.pharm
Dept. of Pharmacology
gautham college
of pharmacy 1
 CONTENTS
1. Introduction to nootropics
2. Definition of nootropics
3. Definition of Memory
4. Parts of brain and its physiology
5. Constitution of Psyche or mind
6. Indications of nootropics
7. Classification of nootropic agents
8. Common mechanism of action
9. Pharmacological actions of the prototype
10.Screening models
a. invitro models
b. invivo models

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 INTRODUCTION TO NOOTROPICS
 The word Nootropics was coined in 1964
by Dr.Corneliu Giurgea.

 Its derived from greek words

NOOS-mind tropein-bend/turn.

 They are referred to as “smart drugs”,

”memory enhancers”, ”cognitive enhancers”,
“smart nutrients”.

 In the scientific literature they are termed as

nootropics.

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DEFINITION OF:

a) Memory: it’s a unconscious faculty in which mental

impressions are retained and reproduced in mind.
Memory consists of 4 process:
(a) learning (b) retention (C) recall (d) recognition
memory process depends on objects and events.

b) Dementia: acquired global impairment of cognitive

functions in absence of clouding of consciousness or
motor involvement. Memory, capacity to solve problems
of day to day living, social skills, control of emotions are
affected

c) Alzheimer’s disease: A progressive neurodegenerative

disorder which affects older individuals. Atropy of cortical
and subcortical area is associated with deposition of
beta- amyloid protein in form of plaques, and formation
of neurofibrillary tangles. There is marked cholinergic
deficiency in brain.
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DEFINITION OF NOOTROPICS
 These are the drugs that are used to
improve human cognitive abilities.

 Intellect , memory and personality of a

human are called cognitive functions

 Nootropic substances include drugs,

nutrients and herb with cognitive
enhancing effects.

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Typically nootropics work by

 Altering the availability of brain’s

supply of neurochemicals ie,
neurotransmitters, enzymes and
harmones.

 By improving brain’s oxygen supply

 By stimulating nerve growth

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 ANATOMY
 The main parts of brain are:
a) cerebrum b) cerebellum
c) brain stem .
 Cerebrum is the largest part of brain
composed of two similar sized cerebral
hemispheres.
 Each hemisphere is divided into 4 lobes:
occipital, parietal, temporal and frontal.
 The cerebrum (association area) is the
area responsible for learning, memory and
identification activities.

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8
 CONSTITUTION OF
PSYCHE OR MIND
They carry out three functions:
 Cognition : the reception of
environmental stimuli.
 Affect : analysing the information
received and formation of reaction
patterns.
 Conation : the actual behaviourial
response.

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 INDICATIONS OF NOOTROPICS
 Senile dementia of alzheimer type(DAT) and
multi infarct dementia (MID)
 Mental retardation in children, learning
defects, attention deficit disorder.
 Common symptoms of elderly ;dizziness and
memory disturbance.
 Transient ischaemic attack, cerebro vascular
accidents like stroke.
 Sequalae of head injury, ECT, brain surgery.

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 CLASSIFICATION OF NOOTROPICS
 CHOLINERGIC ACTIVATOR
- acetyl –L-carnitine ( ALCAR)
- piracetam
- centrophexine

 SERATONERGICS
- theanine
- typtophan

 DOPAMINERGICS
- L- dopa
- phenylalanine.

 ALZHEIMERS DISEASE
- tacrine
- donepezil
- rivastigmine
- galantamine

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 IMPROVED OXYGEN SUPPLY & BRAIN ENERGY
- lipoic acid
- pyritinol

 MEMORY ENHANCERS
- brahmi
- vasopressin

 MENTAL CONCENTRATION & STAMINA

- caffine
- adrafinil

 NERVE GROWTH STIMULANT & BRAIN CELL PROTECTION

- brahmi
- ginko biloba.

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 PIRACETAM (cholinergic activator)
 This is a GABA derivative that selectively
improves the efficiency of higher telencephalic
activity by:
a) enhancement of learning and memory
b) facilitation of inter hemisphere information
transfer
c) increase tonic cortical control on sub cortical
areas
 piracetam has been claimed to improve ATP/ADP
ratio in telencephalon, stimulate synaptic
transmission and to have an anti thrombotic
effect.
 Side effects are gastric discomfort, excitement,
insomia, dizziness

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 DONEPEZIL ( AD)
 This cerebro selective and reversible anti-
AChE cognitive as well as non cognitive
(activities of daily living ) scores in AD,
elevation of Ach level in cortex, especially
in neurons tat project from basal fore
brain to cerebral cortex and hippocampus

 Its likely to have the greatest impact in

AD among cholinergic agents

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 PYRITINOL
 It’s a vaso active cerebral protective ie, a
drug supposed to counteract the cognitive
and functional defects induced by cerebro
vascular insufficiency
 Its claimed to activate cerebral
metabolism by selectively increasing
glucose transport across BBB, enhance
cerebral cholinergic transmission and
improve regional blood flow in ischaemic
brain area

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 MECHANISM OF ACTION
 Increasing global/ regional blood flow
 Direct support of neuronal metabolism

 Enhancement of neurotransmission.

 Improvement of descrete cerebral

function.
 Improve oxygen supply and brain energy

 Nerve growth stimulation and brain cell

protection.

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 SCREENING MODELS
 INVITRO METHODS:
a) inhibition of acetyl cholinesterase
activity in rat striatum.
b) inhibition of butyryl
cholinesterase activity in human
serum.
c) release of ach and other
transmitters from rat brain slices.

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 IN VIVO METHODS

1. Passive Avoidance
2. Active Avoidance
3. Discriminational learning
4. Conditioned responses
5. Studies in aged monkeys

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 IN- VITRO METHODS
1.Inhibition of acetyl cholinesterase activity
in rat striatum
Purpose and rationale
effects of various cholinesterase inhibitors on the
two major molecular form of acetylcholinesterase
isolated from rat striatum and cerebral cortex.

PROCEDURE
The procedure is divided into three main parts:
I. Preparation and isolation of molecular forms of
Ache
II. Assays for the marker enzymes,
III. Enzyme inhibition studies.

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ASSAY
Blank: 0.8 ml PO4 buffer/DTNB
0.8 ml buffer/Substrate

Control: 0.8 ml PO4 buffer/DTNB/Enzyme

0.8 ml PO4 buffer/Substrate

Drug: 0.8 ml PO4 buffer/DTNB/Drug/Enzyme

0.8 ml PO4 buffer/Substrate

EVALUATION

For IC50 determinations: Substrate concentration is 10

mM diluted 1 : 2 in an assay yielding a final
concentration of 5 mM. DTNB concentration is 0.5 mM
yielding 0.25 mM final concentration 100

% Inhibition = Slope control − Slope

drug /Slope Control × 100
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2.In vitro inhibition of butyrylcholine-esterase
activity in human serum
PURPOSE AND RATIONALE

This assay can be used in conjunction with the

acetylcholine-esterase assay to determine the
enzyme selectivity of various cholinesterase
inhibitors .Butyrylcholine-esterase (BChE), which is
sometimes called pseudocholinesterase,
preferentially hydrolyzes butyrylcholine. This
enzyme is found in the highest amounts in serum,
but its physiological role is not known
Ethopropazine and tetra-isopropyl
pyrophosphoramide are selective inhibitors of
butyrylcholinesterase.

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 Assay

Enzyme activity is measured with spectrophotometer.

Blank: 0.8 ml PO4 buffer/DTNB
0.8 ml buffer/Substrate

Control: 0.8 ml PO4 buffer/DTNB/Enzyme

0.8 ml PO4 buffer/Substrate

Drug: 0.8 ml PO4 buffer/DTNB/Drug/Enzyme

0.8 ml PO4 buffer/Substrate
EVALUATION

For IC50 determinations: Substrate concentration is

10 mM diluted 1 : 2 in assay yielding final concentration
of 5 mM. DTNB concentration is 0.5 mM yielding
0.25 mM final concentration

% Inhibition =slope control - slope drug/

control slope ×100 22

 PASSIVE AVOIDANCE
1. Step down
2. Step through

3. Scopolamine induced amnesia in

rats
4. Up hill avoidance

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 I.PASSIVE AVOIDANCE
1.Scopolamine-induced amnesia in mice
PURPOSE AND RATIONALE
The administration of the antimuscarinic agent scopolamine
to young human volunteers produces transient memory
deficits .Analogously, scopolamine has been shown to
impair memory retention when given to mice shortly before
training in a dark avoidance task .

PROCEDURE
The scopolamine test is performed in groups of 10 male
mice weighing 26–32 g in a one-trial. Five min after i.p.
administration of 3 mg/kg scopolamine hydrobromide, each
mouse is individually placed in the bright part of a two-
chambered apparatus for training. After a brief orientation
period, the mouse enters the second, darker chamber. Once
inside the second chamber, the door is closed which prevents the
mouse from escaping, and a 1 mA, 1-s foot shock is applied
through the grid floor. The mouse is then returned to the home
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cage. Twenty-four hours later, testing is performed by placing
animal again in the bright chamber. The latency in
entering the second darker chamber within a 5 min test
session is measured electronically. Whereas untreated
control animals enter the darker chamber in the second
trial with a latency of about 250 s, treatment with
scopolamine reduces the latency to 50 s. The test compounds
are administered 90 min before training. A prolonged
latency indicates that the animal remembers
that it has been punished and, therefore, does avoid
the darker chamber.

EVALUATION
Using various doses latencies after treatment with test
compounds are expressed as percentage of latencies
in mice treated with scopolamine only.

MODIFICATIONS OF THE METHOD

Amnesia can also be induced by pretreatment with

Benzodiazepines.
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2.Up-hill avoidance
PURPOSE AND RATIONALE
Many animal species exhibit a negative geotaxis, i.e. the
tendency to orient and move towards the top when placed on
a slanted surface. When placed on a tilted platform with
head facing down-hill, rats and mice invariably turn around
and move rapidly up the incline
PROCEDURE
Rats of both sex were used and maintained under standard
conditions. The experimental apparatus is a 50 × 50 cm box
with 35 cm high opaque plastic walls. The box can be
inclined at different angles. The floor consists of 10 mm
diameter stainless steel grid bars placed 13 mm apart. To
deliver the tail-shock, a tail-electrode is constructed,
consisting of a wire clip connected to a constant current
shock source. The animal is first fitted with the tail-electrode
and then placed onto the grid with its nose facing down.
During baseline-trials the animal’s latency to make a 180°
turn and 26
 the first climbing response is measured. Thereafter
the animal is returned to its home cage. During the
experimental trials the latencies are measured and additionally
a tail-shock (1.5 or 2 mA) was administered contingent on
the first climbing response after the 180° turn. Immediately
after the shock the animal is placed in its home cage. Retest
is performed 24 h later.

EVALUATION

The latencies are measured.

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3.Step-down
PURPOSE AND RATIONALE
An animal (mouse or rat) in an open field spends most
of the time close to the walls and in the corners. When
placed on an elevated platform in the center of a rectangular
compartment, it steps down almost immediately
to the floor to explore the enclosure and to approach
the wall.
PROCEDURE
Mice or rats of either sex are used. A rectangular box
(50 × 50 cm) with electrifiable grid floor and 35 cm
fits over the block. The grid floor is connected to a
shock device which delivers scrambled foot shocks.
A typical paradigm consists of three phases:
(1.) Familiarization: The animal is placed on the platform,
released after raising the cylinder, and the latency
to descend is measured. After 10 s of exploration,
it is returned to the home cage. 28
(2.) Learning:
Immediately after the animal has descended from the
platform an unavoidable footshock is applied (Foot-
shock: 50 Hz; 1.5 mA; 1 s) and the animal is returned
to the home cage,
(3.) Retention Test:
24 h after the learning trial the animal is again placed on the
platform and the step-down latency is measured. The test is
finished when the animal steps down or remains on the
platform (cut-off time: 60 s).

EVALUATION

The time of descent during the learning phase and the

time during the retention test is measured. A prolongation
of the step-down latency is defined as learning.

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4.Step-through
PURPOSE AND RATIONALE
This test uses normal behavior of mice and rats. These
animals avoid bright light and prefer dim illumination.
When placed into a brightly illuminated space connected
to a dark enclosure, they rapidly enter the dark
compartment and remain there.
PROCEDURE
Mice and rats of either sex are used. The test apparatus
consists of a small chamber connected to a larger dark
chamber via a guillotine door. The small chamber is
illuminated with a 7 W/12 V bulb. The test animals are
given an acquisition trial followed by a retention trial
24 h later. In the acquisition trial the animal is placed
in the illuminated compartment at a maximal distance
from the guillotine door, and the latency to enter the
dark compartment is measured.

30
Animals that do not step through the door within a cut-off
time: 90 s (mice) or 180 s (rats) are not used. Immediately
after the animal enters the dark compartment, the door is
shut automatically and an unavoidable footshock (Footshock:
1 mA; 1 s – mice; 1.5 mA; 2 s – rat) is delivered. The animal
is then quickly removed (within 10 s) from the apparatus and
put back into its home cage. The test procedure is repeated
with or without drug. The cut-off time on day 2 is 300 s
(mice) or 600 s (rats), respectively.

EVALUATION

The time to step-through during the learning phase is

measured and the time during the retention test is measured.
In this test a prolongation of the step-through latencies
is specific to the experimental situation. An increase
of the step-through latency is defined as learning.

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 II.ACTIVE AVOIDANCE
1.RUN AWAY AVOIDANCE
PURPOSE AND RATIONALE
A straightforward avoidance situation features a fixed aversive
gradient which can be traversed by the animal. The shock
can be avoided when the safe area is reached within the
time allocated

PROCEDURE
Mice or rats of either sex are used and maintained under
standard conditions and handled for several days before the
experiment. The same box as used in the step-through model
can be used.
The apparatus is uniformly illuminated by an overhead light
source. A loudspeaker, mounted 50 cm above the start-box,
serves for presenting the acoustic conditioned stimulus
(audiogenerator). The footshock is employed .The animal is
allowed to explore the whole apparatus for
5 min.
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 The guillotine door is then closed and the animal
is placed into the light starting area. After 10 s the
acoustic CS is applied and the door is simultaneously
opened. Shock is turned on after 5 s. The CS continuous
until the animal reaches the safe area. It is left there for 50
70 s (inter trial interval, ITI) before returned to the same
area again. The procedure starts again. The training is
continued until the animal attains the criterion of 9
avoidances
in 10 consecutive trials. On the next day the procedure is
repeated until the same learning criterion is reached. The time
needed to reach the safe area is measured.
EVALUATION
The time the animal needs to reach the safe area on
both days is measured. In addition, the number of errors
(not reaching the safe area) is recorded.

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2.Shuttle box avoidance (two-way
shuttle box)
PURPOSE AND RATIONALE
Compared to runway avoidance, shuttle box avoidance
(two-way-shuttle-box) is a more difficult task. Since
the animal is not handled between trials, the shuttle-
box can be easily automated
PROCEDURE
Rats of both sex are used and maintained under standard
conditions. The apparatus used consists of a rectangular
box 50 × 15 cm with 40 cm high metal walls, and an
electrifiable grid floor. The box is divided by a wall
with a manually or solenoid-operated guillotine door
(10 × 10 cm). into two 25 × 15 cm compartments. Each
compartment can be illuminated by a 20 W bulb mounted
in the hinged Plexiglas lids.
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A fixed resistance shock source with an automatic switch (0.5
s on 1.5 s off) is used. Simple programming equipment
provides for automatic delivery of the conditioned stimulus
(CS) and the unconditioned stimulus (US). The apparatus is
placed in a dimly lit room with a masking noise background
(white noise) of 60 dB. The animal is allowed to explore the
apparatus for 5 min with the connecting door open and the
compartment lights switched off. The guillotine door is then
closed. After 20 s the light is switched on in the compartment
containing the animal, and the door
is opened. A tone (CS) is presented and 5 s later the
floor shock is applied in the illuminated compartment
and continued until the animal escapes to the dark side
of the compartment, the connecting door is closed and
the shock discontinued. After a variable intertrial interval
(ITI; 30–90 s) the light is switched on in the previous
dark compartment, the door is opened and the
animal is required to cross to the other side.

35
 The training is continued until the animal reaches the
criterion of 9 avoidances in 10 consecutive trials.
Retention is tested at different intervals after the
original training by retraining the animal to the same
criterion again.

EVALUATION

The time the animal needs to reach the safe area on

both days is measured.

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Active avoidance – Shuttle box avoidance

37
3.Jumping avoidance (one-way shuttle
box)
PURPOSE AND RATIONALE
Since a high degree of automation and minimum handling
are additional requirements for this model, the
obvious solution is a simplified one-way avoidance,
allowing for the spontaneous or forced return of the
animal to the start. In order to enhance the start-goal
distinction a vertical gradient is introduced which requires
the animal to perform a discrete response of an
all-or-none character, such as the jump, which clearly
differs from the more continuous translational movements
required in the usual avoidance tasks
PROCEDURE
Rats of both sex are used and maintained under standard
conditions. The apparatus used consists of a rectangular
box 40 × 25 cm with 40 cm high metal walls, an electrifiable grid
floor and a Plexiglas ceiling. A 12 × 12 × 25 cm opaque plastic
pedestrial, mounted onto one of the narrow walls of the box provides
the 38
isolated goal area
Flush with the horizontal surface of the pedestal moves a
vertical barrier, which can either be retracted to the rear wall
of the apparatus to expose the goal area or pushed forward
to block access to the goal completely. The animal is placed
into the apparatus for 5 min with the goal area exposed
(barrier retracted). The barrier is then moved forwards and
the goal is blocked for 2 s. The first trial starts by exposing
the goal area and applying an acoustic CS (1 000 Hz,
85 dB). Electric shocks – US (1.0 mA; 50 Hz; 0.5 s) –
are applied 5 s later (once per 2 s), and continued together
with the CS until the animal jumps onto the platform.
After 30 s the barrier pushes the animal off the
platform onto the grid floor. The sequence is repeated
until the criterion of 10 consecutive avoidances is
reached. Retention is tested on the next day until the
animal reaches criterion.
EVALUATION
The time the animal needs to reach the safe area on
both days is measured. In addition, the number of errors
(not reaching the safe area) is recorded.
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 III.DISRIMINATIONAL LEARNING
1. SPATIAL HABITUAL LEARNING
PURPOSE AND RATIONALE

The open-field test utilizes the natural tendency of rodents

to explore novel environments in order to open
up new nutrition, reproduction and lodging resources
The rate of exploratory behaviors exhibited in an unfamiliar
environment is limited through the inherent
necessity to avoid potential dangers. The observed behavior
therefore is always a compromise between these
conflicting interests and is regulated in part by the
momentary physiological needs.Spatial habituation learning is
defined as a decrement in reactivity to a novel environment
after repeated exposure to that now familiar
environment. This reduction in exploratory behaviors
during re-exposures is interpreted in terms of remembering
or recognition of the specific physical characteristics
of the environment. The test can be used to
examine short-term spatial memory and/or long-term spatial
memory 40
PROCEDURE

The open-field apparatus is a rectangular chamber (rats:

60 × 60 × 40 cm, mice: 26 × 26 × 40) made of painted
wood or grey PVC. A 25 W red or green light bulb is
placed either directly above or beneath the maze to
achieve an illumination density at the centre of approximately
0.3 lx. Masking noise is provided by a broad
spectrum noise generator (60 dB). Prior to each trial,
the apparatus is swept out with water containing 0.1%
acetic acid. Housing room and the testing location are
separated and animals are transported to the testing
room 30 min before testing. The digitized image of
the path taken by each animal is stored and analyzed
post hoc with a semi-automated analysis system.
The rodent is placed on the center or in a corner of the open-
field for 5–10 minute sessions (mice: up to 20 min, because of
the high basal activity level). The animals are re-exposed to
the open-field 24 and 96 h after the initial trial

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EVALUATION

The exploratory behaviours registered are:

(1) Rearings or vertical activity: the number of
times an animal was standing on its hind legs
with forelegs in the air or against the wall.
(2) The duration of single rearings as
a measure of non-selective attention
(3) Locomotion or horizontal activity: the distance
in centimeters an animal moved.

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3.Visual discrimination
PURPOSE AND RATIONALE

Vision is better than any other sensory system for the

analysis of spatial relationships in the environment of
the animal. From the retina to the cerebral cortex, the
organization of the visual system ensures processing
of visual information according to simple principles,
i.e. by fitting the distribution of light over the receptive
surface to elementary geometrical concepts and by comparing
these patterns with images stored in the memory.
Experimental studies of pattern discrimination
must take into account the visual capability of the given
species and present the discriminanda under conditions
compatible with light sensitivity and acuity of the eye.
The constructions of the apparatus should ensure that
the discriminanda are viewed from one optimum distance
and for a sufficient period of time.

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PROCEDURE
Rats and mice of both sexes are used and maintained under
standard conditions. The apparatus consists of a square 10 ×
10 cm start area separated by a Plexiglas sliding door from
the choice area, which is connected by swing doors to the goal
compartment. The grid floor in the starting and the choice
areas is electrifiable. The stimulus cards can be attached to
the swing doors. The patterns are black on a white
background and have different forms. The apparatus is
illuminated by a dim light. The animal is placed into the
apparatus with all doors open and allowed to explore it. Then
it is placed in the start and after 5 s released by
raising the Plexiglas door. After another 5 s, electric shocks
are applied until the animal escapes through either of the
open doors to the safe goal compartment where it is left for
some seconds. As soon as this preliminary step is mastered,
the stimulus cards are inserted, the negative door is locked
and the grid section in front of this door is electrified. The
animal is trained to a criterion.

44
 On the next day the animal is retrained to the same
criterion and retention is expressed in savings. Another
parameter which can be used to evaluate the savings is the
cumulative number of errors until the criterion is reached .

EVALUATION

The number of correct answers as well as the

number of trials until the criterion is reached are
counted.

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3.Spatial learning in the water maze
PURPOSE AND RATIONALE

A task was developed where rats learn to swim in a

water tank to find an escape platform hidden under
the water .As there are no proximal cues
to mark the position of the platform, the ability to locate
it efficiently will depend on the use of a configuration
of the cues outside the tank. Learning is reflected
on the shorter latencies to escape and the decrease on
the length of the path to find the platform. Although
rodents can find the platform by using non-spatial strategies,
the use of a spatial strategy is the most efficient
way to escape and young animals develop the spatial
strategy after a small number of trials.

46
PROCEDURE
Different strains of rats are generally used. The
apparatus is a circular water tank filled to a depth
of 20 cm with 25 °C water . Four points equally
distributed along the perimeter of the tank serve
as starting locations. The tank is divided in four
equal quadrants and a small platform (19 cm
height) is located in the centre of one of the
quadrants. The platform remains in the same
position during the training days. The rat is
released into the water and allowed 60–90 s. to
find the platform. Animals usually receive 2–4
trials per day for 4–5 days until they escape onto
the platform. Well-trained rats escape in less
than 10 s.

47
EVALUATION

The latency to reach the escape platform is measured

during the training days. A free-swim trial is generally
performed after the training days where the escape platform
is removed and the animal is allowed to swim for
30 s. With the help of a video system, the latency to
reach the previous position of the platform, the number
of annulus crossings as well as the time the rat spent in
the training quadrant are measured. Well-trained rats
show short latencies, a large number of annulus crossings
and bias to the quadrant where the escape platform
was located during the training sessions.

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 IV.STUDIES IN AGED MONKEYS
1.Long-term potentiation in hippocampal
slices
PURPOSE AND RATIONALE

Long-term potentiation (LTP) in the hippocampus is

the most dramatic example of activity-dependent
synaptic plasticity that has yet been identified in
the mammalian brain.. The fact that it occurs in the
hippocampus has done much to stimulate interest in LTP as a
synaptic model of memory, since the importance of the
hippocampus for memory processing has been evident ever
since the discovery that its bilateral removal in man causes
a profound impairment in the ability to lay down new
memories.
The particular popularity of the slice preparation
prepared from the rodent hippocampus rests on its
lamellar and laminar organization.
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PROCEDURE

Transverse slices, 400 mm thick, are cut from the

hippocampus of male albino guinea pigs weighing
250–300 g and prepared for electrophysiological recordings.
slices are incubated for 90–120 min in the recording chamber
to allow equilibration with artificial cerebrospinal
fluid. They are submerged, placed on a nylon mesh
and perfused at a flow rate of 2–2.5 ml/min with oxygenated
(95% O2/5% CO2) cerebrospinal fluid having the following
composition (in mM): NaCl 124, KCl 3.3, CaCl2 2.5, KH2PO4
1.25, MgSO4 2, NaHCO3 25,7, glucose 10. The recording
chamber is maintained at 33 ±2 °C. The extra cellular
population spike is obtained
using glass microelectrodes filled with 2 M
NaCl, The electrodes are placed into the stratum pyramidal
of CA1 or CA3. The signal is amplified and stored on
magnetic discs for later analysis. The evoked responses are
averaged and analyzed off-line using a personal computer.

50
The magnitude of the population spike is evaluated
by taking the voltage difference between the negative
peak and the following positive peak. Either mossy
fibers in the hilus fasciae or commissural/associational
fibers in the stratum radiatum are activated via bipolar,
sharpened silver wire electrodes insulated except
for the tips. Constant current pulses (100 ms) are delivered
with a frequency of 0.2 Hz, only during the
test intervals. The stimulation intensity is adjusted to
elicit the population spike of about 40% and 80% of
its maximal amplitude in CA1 and CA3, respectively.
After the baseline is recorded for 10–20 min, LTP is
induced by repetitive stimulation of 100 pulses at 20 Hz
for 5 s in CA1 and at 50 Hz for 2 s in CA3 at the same
strength as for the test pulses. Responses by test pulses
are recorded 0, 10, 20 and 30 min after repetitive
stimulation.
The test drugs are dissolved in the artificial cerebrospinal
fluid and applied extracellularly at various
concentrations by switching perfusion reservoirs.
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 EVALUATION

The time course of LTP is registered for CA1 and CA3.

The mean percent increase in the amplitude of the
population spike from baseline responses after drug
application is compared with controls.

52
 REFERENCE
 Tripathi .K.D.; CNS stimulants and cognition
enhancers. In: Essentials of medical
pharmacology ;Edn no 5, Publisher: Jaypee
brothers EMCA house , New Delhi.
pp: 435 – 442.
H.Gerhard Vogel and
Wolfgang.H.Vogel.Psychotropic and neurotropic
activity,In; Drug Discovery and Evaluation, 5th
edition,Springer-Verlag Berlein Heidelberg,Germany,
pp-496-548.

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