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SYNOPSIS

ASSESSMENT OFANTIPSYCHOTIC POTENTIAL


OF RELAXIN-3 PEPTIDE IN RODENTS.

By
UJWAL T. WANDHARE
Guide:
Mr. N. R. KOTAGALE
INTRODUCTION

Psychosis refers to an abnormal condition of the
mind, and is a generic psychiatric term for a
mental state often described as involving a "loss
of contact with reality. Psychosis may appear as
symptoms of number of mental disorder
including mood, personality disorder. It is also the
defining feature of schizophrenia, the manic
phase of bipolar and psychotic disorder. These
common disorders typically include symptoms of
false beliefs (delusion) and abnormal sensation
(hallucinations).

Schizophrenia is a severe and chronic mental illness
associated with high prevalence. They are usually
classified as positive, negative or cognitive symptoms.
Positive symptoms include hallucination, typically
auditory, delusion which frequently involves
persecution and/or megalomania and severe thought
disorganization. Negative symptoms are a group of
deficits comprising many dimensions such as affect
(flattening), volition (apathy), speech, pleasure
(ahedonia) and social life (withdrawal). Cognitive
symptoms such as deficits in attention and memory are
other prominent features of the illness. Among these
the dopamine and serotonin, glutamate have received
most attention, although other symptoms such as
GABAergic, cholinergic or opoid system have also been
implicated.

Neuropeptides are small protein-like molecules
(peptides) used by neurons to communicate with each
other. Neuropeptides modulate neuronal
communication by acting on cell surface receptors.
Many neuropeptides are co-released with other small-
molecule neurotransmitters. Different neuropeptides
are involved in a wide range of brain functions,
including analgesia, reward, food intake, metabolism,
reproduction, social behaviors, learning and memory.
Relaxin-3 is a two chain, 51 amino acid
neuropeptide that was discovered in 2001. Relaxin
that was observed to relax the pelvic ligament in
guinea pigs almost a century ago. Relaxin-3 is
abundantly expressed within the mammalian brain and
acts as neurotransmitter by activating its cognate G-
protein coupled receptor, RXFP3
Neuroanatomical studies concluded in rat,
mouse and macaque have revealed that relaxin-3
has mainly expressed within neurons of pontine
nucleus incertus while smaller populations are
present in the pontine raphe, periaquiductal gray
and region dorsal to substantia nigra. Relaxin-3
containing neurons in this area innervate a broad
range of target forebrain region rich in RXFP3.
NI relaxin-3 neurons are predominantly
GABAergic and it is likely relaxin-3 signaling
confers complementary inhibitory effects to the
primary transmitter as in cell based studies.
RXFP3 activation is linked to Gi/o and reduces
cAMP accumulation.
Relaxin-3 expression is also negatively regulated by
serotonin/5-HT
1A
receptor signaling

which is strongly
implicated in the aetiology and treatment of anxiety
and depression . Importantly relaxin-3 neurons in NI
express 5HT
1A
serotonin receptors .
The 5HT
1A
receptors are the autoreceptor
(predominantly inhibitory) and regulate the Serotonin
release. 5HT
1A
receptor agonist has also been
suggested to have atypical antipsychotic drug profile.
In view of this background, we speculate that relaxin-3
peptide may possess antipsychotic like activity in
animal model of schizophrenia. This study is designed
with the objective to evaluate the antipsychotic like
activity in animal models of schizophrenia.

To assess the Antipsychotic like effect in of
Relaxin-3 peptide in animal models of
schizophrenia.


The research work has been planned on the following lines:
Cannulation within Nucleus Incertus
Recovery phase/period mock administration of drug.
Distribution of animals into different groups.
Evaluation of following psychopharmacological activity after relaxin-
3 administration
Evaluation animals in Condition Avoidance Response,
Evaluation of animals in Apomorphine induced climbing,
Evaluation of animals in Ketamine/Apomorphine Stereotype
Evaluation of animals in Catalepsy,
Evaluation of animals in Prepulse inhibition,
Biochemical analysis of Prolactin levels.
Separate groups of animals will be used for individual behavior
paradigm.
Involvement of 5HT
1A
receptor in separate group of animals will be
pretreated with 5HT
1A
agonist or antagonist prior to Relaxin-3 and
then subject to above paradigm.



Materials and Method:-

Subjects:
Male Sprague Dawley rats (200-250 g) will be
used. Food and water will be available ad Libitum. The
experimental procedure will be performed as per the
protocol approved by Institutional animal and ethical
committee according to the guidelines of CPCSEA.

Drugs:
Following drug will be used :
Relaxin-3 agonist- R3 (B)/R3(A11-24)
Relaxin-3 antagonist- R3 (B1-22)R
5HT1A receptor agonist- Lesopitron
5HT1A receptor antagonist- Alprenolol

Behavioral Paradigm:

Conditioned avoidance response (CAR) in rats:
Rats will be trained individually for one week in a two
compartment shuttle box to move from one compartment to
another upon presentation of the buzzer tone (80 dB) for 10 s
(Conditioned stimulus; CS). If the rat failed to respond to CS, the
tone will continue and electric current (0.5 mA) (unconditioned
stimulus, UCS) will simultaneously delivered to the grid floor of the
chamber for a period of 15 s for foot shock. Each animal will be
subjected to a daily session of 10 trials separated by a 20 s interval.
The trial terminate once the rat have move into the other
compartment during CS or UCS period. Crossings made by the rat
during the CS will be recorded as avoidance responses and those
made during UCS period will be recorded as escape responses. Only
those animals characterized by a high level of avoidance responding
(90%) will used for further experiments. The results will be
expressed as avoidance/10 trials.


Apomorphine-induced climbing behavior in Rats:

Apomorphine is dopamine D2 agonist produces psychosis in
rodents. Administration of apomorphine to rat result in a peculiar
climbing behavior characterized initially by rearing and then full
climbing activity. Rats will initially place individually in cylindrical
wire mesh cages (height 13 cm, diameter 14 cm, mesh size 3 mm)
for 60 min to acclimatize to the new environment. Each rat will
place in wire mesh cages 10 min after apomorphine administration
and climbing behavior of individual rat will score at 5 min intervals
for a period of 20 min. The scoring system use will be as: 0 = four
paws on the floor, 1 = one paw on the wall of cage, 2 = two paws on
the wall of cage, 3 = three paws on the wall of cage, and 4 = four
paws on the wall of cage. Climbing scores across each time interval
will be summed and expressed as climbing index, thus providing a
maximum possible Climbing index of 20.



Amphetamine or ketamine induced motor
hyper activity in Rats:

Rat will be transferred from their housing facility to
the testing room in their home cages 15 min before
any experiment. Animals will place in actophotometer
(20 cm 20 cm 10 cm) equipped with six infrared
photo sensors, 2.5 cm apart from each other for 30
min habituation before any testing. Baseline locomotor
activity of each Rat will recorded for 20 min as a total
count of ambulatory, horizontal and vertical activity.


Catalepsy induction in Rats:

The severity of catalepsy in individual rat will
be determine by placing the forepaws of the
animal over a wooden bar 0.4 cm in diameter,
fixed to a height of 3.5 cm above the tabletop.
The time in seconds until rat brings both
forepaws down to the tabletop will recorded,
with a maximum cut-off time of 300 s.

Effects of agmatine on plasma Prolactin
levels in rats:

All present antipsychotics are known to
elevate plasma prolactin levels due to
blockade of DA D2 receptors located on
pituitary gland. Therefore, plasma prolactin
levels will be measure by the manufacturers
protocol.

Prepulse inhibition :

This is one of the excellent animal models
used for assessment of antipsychotic activity
of drug. Prepulse inhibition (PPI) of the
acoustic startle response is used as a measure
of the pre attentive information and
processing. PPI is lowered in schizophrenia
and this impairedment can be mimicked in
experimental animal using the psychomimetic
drug.

Possible outcome:

The present research work may provide
Relaxin-3 may have role in the pathogenesis
or treatment of schizophrenia.

References

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Thank You !!!