DETERMINATION OF

ANTIDEPRESSANT
ACTIVITY
SECTION 2B-1
INTRODUCTION
•Determination of the antidepressant activity was determined using the
Swim Stress Test
•Locomotor activity of the mouse was measured by observation
•The plant extract/unknown was screened for its antidepressant
activity and compared to a positive and a negative control
•3 groups per sub section
PORSOLT SWIM TEST
● Widely used classic model for antidepressant activity
● One of the despair-based models which were proposed to have
relatively good predictive validity for monoamine-based
antidepressants
● When rats or mice are forced to swim in narrow cylindrical
apparatus from there is no escape, they adapt a characteristic
immobile posture in which they remain floating in water making
only those movements necessary to keep their head above the
water level– immobility reflects “despair”
Methodology
MATERIALS

• The cylindrical tanks (30 cm height x 20 cm diameters) required

for the mouse forced swim test (FST).
• The dimensions of the tanks should be selected in a way that
the mice will not be able to touch the bottom of the tank, either
water tanks with their feet or their tails, during the swimming test.
• The height of the tank should be high enough to prevent the
mice from escaping from the tank. The diameter of tank and the
depth of water are important parameters that can be adjusted to
change the behavior of mice.

Timer
MATERIALS

Video • Use a video camera supported by a tripod (optional). Since this

test usually involves multiple animals being tested at the same
recording time, live scoring will be very difficult and is not advisable. The
video camera should record in high enough resolution to render

device a quality picture that will be used later for behavioral scoring.

Drying • Before returning the animals to their home cages, it is important

paper and to dry them gently using paper towels and it is helpful to use a
heat lamp (be certain the exposure temperature does not exceed
32°C) to prevent hypothermia.
heat lamp
https://www.ottoenvironmental.com/content/images/thumbs/0003405_porsolt-forced-swim.jpeg
Methodology
Weigh each mouse and compute for the appropriate dose that
would be given. The suggested dosage volumes for test
material administration is 10mL/kg.

Place the camera and the dividers in position. The camera

should be as close as possible to obtain the highest possible
resolution of the mice.

The tanks should be filled with tap water set at the room
temperature (23-25°C) to the determined level, which is
marked on the tank walls.

Hold the animal by its tail, and gently and slowly place in the
water. Once the mice is in the water, slowly release the tail.
Once all mice are in the tanks-start the countdown on the
stopwatch. The usual test length for mice is five minutes in
the FST.
Methodology
Observe for periods and mobility and immobility
taking into account the number of seconds or
minutes the mice have been mobile and immobile.

Remove the mice from the tank and allow to rest

for five minutes under a heat lamp.

Administer the computed dose for each mouse

and let the mice rest for 30 minutes.

Subject mice for another FST for 5 minutes.

Observe for mobility and immobility span. Remove
mice from the tank.
RESULTS AND
DISCUSSION
RESULTS
Table 1. MOBILITY in seconds of mice for the three treatments.
RESULTS
Table 2. IMMOBILITY in seconds of mice for three treatments.
Sertraline: MOA

Under class Selective Serotonin Reuptake Inhibitors

Primary action is the inhibition of serotonin transporters (SERT). Sertraline

potentiates serotonergic activity activity in the central nervous system through
inhibition of neuronal reuptake of serotonin (5-HT)

Flouxetine, Sertraline, Citalopram exist as isomers and are formulated in racemic

forms
Pharmacodynamics of SSRI’s
• The SERT is a glycoprotein with 12 transmembrane regions embedded in the axon
terminal and cell body membrane regions embedded in the axon terminal and cell body
membranes of serotogenic neurons.

• When extracellular serotonin binds to receptors on the transporters, conformational

changes occur in the transporter and serotonin, Na+, and Cl- are moved into cell.

• Binding of intracellular K+ then results in return of transporter to its original conformation

and the release of serotonin inside the cell.

• SSRIs allosterically inhibit the transporter by binding the receptor at a site other than
active binding site for serotonin.
Pharmacodynamics of SSRI’s
• At therapeutic doses, about 80% of the activity of the transporter is inhibited.
Functional polymorphisms exist for SERT that determine the activity of
transporters.

• SSRIs have modest effects on other neurotransmitter.

• Unlike TCAs and SNRIs, there is little evidence that SSRIs have prominent
effects on β- adrenoceptors or norepinephrine transporters, NET.

• Binding to the serotonin transporters is associated with tonic inhibition of the

dopamine system. SSRIs do not bind aggressively to histamine, muscarinic or
other receptors.
Discuss the behavioral theories underlying the Porsolt Swim Test

It is a widely used classic model for antidepressant activity

It is relatively a good predictive validity for monoamine-based antidepressants

It is based on the premise that, when rats or mice are
forced to swim in narrow cylindrical apparatus with no
escape, they adapt a characteristic immobile posture in
which they remain floating in water making only those
movements.

The immobility exhibited reflects the state of “despair” as

they resign themselves to the experimental conditions,
which is taken as an index of depression.
Describe the various theories of depression and
how the different antidepressant agents address
these theories

The Neurotrophic Hypothesis

The Monoamine Hypothesis

Theory of Neuroendocrine Factors

The Neurotrophic Hypothesis
Brain-derived neurotrophic factor (BDNF),
are important in the regulation of
neural plasticity, resilience, and neurogenesis

Evidences suggest that in depression, there is a

decline of this neurotrophic support & that
effective antidepressant therapy is associated with
a rise or resurgence of neurogenesis and synaptic
connectivity in cortical areas
 Stress and pain

Drop in BDNF levels

Loss of
neurotrophic support

Atrophy in the
hippocampus and other
areas such as the medial
frontal cortex and
anterior cingulate
Evidences supporting Neurotrophic theory

Imaging studies: 5-10% loss of hippocampal volume

Studies of direct effects of BDNF on emotional regulation

↑ BDNF levels in animal models with chronic

administration (supported by human studies)

Clinical trials increases BDNF levels and probably

hippocampal volume
The Monoamine Hypothesis
Depression is associated with changes in serotonin or
norepinephrine signalling in the brain (or both)

There is deficiency in function and/or amount of

serotonin (5HT), norepinephrine (NE), and
dopamine (DA) in the cortical and limbic areas

Goal of treatment is to replace or increase deficient NTA’s

Evidences supporting
Monoamine Theory

•Effect of reserpine treatment

•Effect of tryptophan-free diet
•Effects of NE synthesis inhibitor
•Genetic studies
Theory of Neuroendocrine Factors

Several hormonal abnormalities are assoc. w/ MDD:

Abnormalities in HPA

Elevated cortisol levels, nonsuppression of ACTH

release in dexamethasone suppresion test

Chronically elevated levels of CRH

What are these
neuroendocrine factors?
↑ cortisol

negative feedback

↓ CRH

no β-endorphins (feel good

hormones)

Thyroid Hypofunction (negative

feedback upregulates catecholamine)
 What are the limitations of the Porsolt Swim Test?

• The end result of the test is not an equivalent of the entire spectrum of human
depressive states

• The observable outcome of this test is one-dimensional, it cannot determine the

mechanisms by which that compound acts

• Tested drugs may affect overall activity levels of the animals, which could lead
to unreliable results