SYNOPSIS

For approval of topic of the thesis to be submitted in the partial fulfillment of the
requirement for the degree of Master of Pharmacy in Pharmacology.

Investigations for pharmacodynamic interaction between

agmatine and allopregnanolone in olfactory bulbectomy
induced depression in rats
By

Sandip R. Rahangdale
B. Pharm.

Guide

Co-guide

Prof. C. T. Chopde
M. Pharm., Ph.D., F.I.C.

Dr. R. R. Ugale
M. Pharm., Ph.D.

s
SMT. KISHORITAI BHOYAR COLLEGE OF PHARMACY,
New Kamptee, Nagpur, Maharashtra, 441 002
Rashtrasant Tukadoji Maharaj Nagpur University, Nagpur.
2010-2011

Investigations for pharmacodynamic interaction between

agmatine and allopregnanolone in olfactory bulbectomy
induced depression in rats
Introduction:
Agmatine (l-amino-4-guanidinobutane) is an endogenous amine and a novel
neurotransmitter in the central nervous system (Reis and Regunathan, 2000), synthesized by the
enzymatic decarboxylation of L-arginine by arginine decarboxylase, selective distribution in the
CNS, accumulated by uptake, released by depolarization, and its action is terminated by selective
reuptake or enzymatic degradation by agmatinase (Raasch et al., 2001; Reis and Regunathan,
2000). It shows affinity for 2-adrenergic and imidazoline receptors (Raasch et al., 2001; Reis
and Regunathan, 2000), N-methyl-D-aspartate (NMDA) receptors and all isoforms of nitric
oxide synthase (NOS) in the brain (Reis and Regunathan, 2000), opioid receptors ( and )
(Zomkowski et al., 2005), potassium channels (Budni et al., 2007), serotonin receptors
(Zomkowski et al., 2004).
Exogenous

administration

of

agmatine

produced

several

effects

including

antinociceptive (Onal et al., 2003), neuroprotective (Olmos et al., 1999), anxiolytic (Lavinsky et
al., 2003), anticonvulsant (Bence et al., 2003), mood elevator (Zomkowski et al., 2002; Halaris et
al., 1999) and antidepressant-like effect in the forced swimming test (FST) and tail suspension
test (TST) in mice, through a mechanism that seems to involve an interaction with NMDA
receptors, the L-argininenitric oxide pathway, and/or 2-adrenoreceptors (Zomkowski et al.,
2002). A clinical study has shown an increase in plasma agmatine level in patients with major
depression compared with healthy controls (Halaris et al., 1999). It was also found that brain
agmatine was depleted in depressive patients (Piletz et al., 1994).
Allopregnanolone (3, 5 THP), dehydroepiandrosterone (DHEA) and its sulfated moiety
DHEA(S) in particular are of significance in depression. Both the indirect genomic and nongenomic mediated effects of 3, 5 THP are involved in depression, while DHEA(S) only exerts
non-genomic effect. In patients suffering from major depression, disequilibrium of neurosteroid
concentrations in plasma and cerebrospinal fluid (CSF) from control subjects and normalize after

successful treatment with antidepressants (Romeo et al., 1998; Uzunova et al., 1998; Strohle et
al., 1999, 2000).
The neurosteroid (3, 5 THP) is synthesized from progesterone in the brain (Baulieu,
1981). It alters rapidly (milliseconds to minutes) neuronal excitability through positive allosteric
interaction with steroid recognition site on gamma-aminobutyric acid type A (GABA A) receptorion channel (GRC) (Paul and Purdy, 1992). At lower (nanomolar) concentrations 3, 5 THP
increases the frequency or duration of openings, or both of the GRC (Lambert et al., 1995).
However, at very high (micromolar) concentrations, it exerts a certain intrinsic agonistic activity
at GABAA receptor in the absence of GABA (Puia et al., 1990). It also modulate ligand gated ion
channels including the voltage gated calcium ion channels (Ffrench-Mullen et al., 1994),
nicotinic acetylcholine receptors (Bullock et al., 1997) and 5-HT3 receptors (Wetzel et al., 1998).
Neurosteroids regulate the neuronal function through their concurrent influence on
neuronal excitability and gene expression (Rupprecht and Holsboer, 1999). Exogenous
administration of 3, 5 THP or drugs that increase its endogenous brain content exhibit
antidepressant-like activity in mouse FST (Khisti et al., 2000), behavioral effect such as antistress (Zimmerberg and Blaskey, 1998), anxiolytic (Akwa et al., 1999), anticonvulsant (Frye,
1995), cataleptic (Khisti et al., 1998; Mandhane et al., 1999).
Serotonin (5-HT) system is strongly implicated in the neural regulation of mood.
Dysregulation in 5-HT neurotransmission underlies pathophysiology of depression. Reduced
level of 5-HT and 5-HIAA is reported in CSF of depressed patients (Risch and Nemeroff, 1992).
In addition, enhancement of serotonergic transmission underlies the therapeutic response to
various types of antidepressant treatments (Blier and Montigny, 1994).
The olfactory bulbectomized rat model of depression is a well-characterized animal
model of depression. Bilateral removal of the olfactory bulbs of rats results in a constellation of
behavioral, neurochemical, neuroimmune and neuroendocrine alterations, similar changes
observed in major depressive disorder (Cryan et al., 2002). The most widely studied behavioral
deficit produced by bulbectomy is a response to a novel and brightly lit environment and reduced
gain of body weight (Kelly et al., 1997). It is sensitive almost exclusively to chronic
antidepressant treatment (Cryan et al., 1998). It exhibits ventricular enlargement as well as

altered signal intensity in cortical, caudate, hippocampal and amygdaloid regions as measured by
magnetic resonance imaging (Wrynn et al., 2000).
Considering the fact that i) Agmatine and serotonergic agents produces an antidepressant
effect as well as a well established role of neurosteroids (3, 5 THP) in FST and TST.
ii)Serotonergic agents modulate antidepressant-like effect of 3, 5 THP in mice. iii) Fluoxetine
increases the level of neurosteroids (3, 5 THP) in brain. iv) Agmatine and fluoxetine produce
synergistic effect in sub-effective doses (Zomkowski et al., 2004). Present study sought to
investigate: i) Antidepressant-like effect of agmatine in olfactory bulbectomized rats. ii) Role of
3, 5 THP in antidepressant-like effect of agmatine by altering endogenous neurosteroids using
neusteroidogenic agents or its biosynthesis blocker in olfactory bulbectomized rats.

Objectives:
1) To investigate the antidepressant-like effect of agmatine in olfactory bulbectomized
rats.
2) To investigate the role of 3, 5 THP in antidepressant-like effect of agmatine in
olfactory bulbectomized rats.

Plan of work:
1) The rats will undergo surgery for removal of olfactory bulb, after recovery period of not less
than 14 days.
2) Dose dependent study of agmatine and 3, 5 THP will be carried out by analysing locomotor
activity in open field apparatus and measure body weight. On the basis of this study the final
effective and sub-effective doses will be selected.
3) In another set of experiment the selected sub-effective dose of agmatine will be combined
with sub-effective dose of 3, 5 THP and effect will be analysed in open field test and measure
body weight.
4) In another set of experiment the positive and negative neurosteroid modulators will be
combined with sub-effective and effective doses of agmatine respectively and effects will be
analysed in open field test and measure body weight.

Materials and method:

Animals:
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. Every
possible effort will be made to reduce the suffering of animals in all experimental design.
Drugs:
The drugs and other additives used will be: Agmatine, 3-hydroxy-5-pregnan-20-one
(3, 5 THP), progesterone (neurosteroids precursor), (4-chlorodiazepam (4CD) (diazepam
binding inhibitor) metyrapone (11-hydroxylase inhibitor), indomethacin (3-hydroxysteroid
oxidoreductase inhibitor) (3-HSOR), trilostane (3-hydroxysteroid dehydrogenase inhibitor)
(3-HSD), PK11195 (peripheral benzodiazepine receptors antagonist).
All drugs will be dissolved in 2-hydroxypropyl--cyclodextrin (45% w/v) solution and
further diluted with 0.9% saline except agmatine, injected by intraperitonial (i.p.) route in a
volume of 5ml/kg body weight.
Surgery (bilateral bulbectomy):
Procedure: Surgery will be carried out 1 week after the arrival of rats in the animal
house. Rats will be anaesthetized with a ketamine (75 mg/kg i.p.) and xylazine (5 mg/kg i.p.)
combination prepared in sterile water (1 ml/ kg). Bilateral olfactory bulbectomy will be carried
out as follows: rat will be anaesthetized and skull will be shaved with hair remover. Iodine
solution will be applied as an antiseptic solution to exposed skin. The animal will be fixed in
stereotaxic apparatus for bulbactomy. A skin incision will be made to expose the skull overlying
the olfactory bulbs. Two burr holes will be drilled (each 2 mm diameter) over the right and left
olfactory bulbs, respectively, each 1.5 mm from the midline of the frontal bone and 6.7 mm
anterior to bregma. The bulbs will be removed by suction, care being taken to avoid damage to
the frontal cortex. The burr holes will be then filled with haemostatic sponge and the incision
sutured. Control (sham operated) animals will be treated in a similar manner, however the bulbs

will not be aspirated. Postoperative analgesia will be consisted of buprenorphin (Temgesic) (0.3
mg/kg s.c.) once a day for 3 days (Uzunova et al., 2003).
After surgery, the rats will be placed in a group of 5-6 animals per cage and handled daily
throught recovery period to reduce stress and/or aggressive behavior.

Method:
Apparatus:

Open field test:

Bulbectomized and sham control rats will be subjected to an open field test on the 1, 7,
and 14th day of drug administration. Each rat will be placed individually into the center of the
open field apparatus. The open field apparatus consisted of 80-cm diameter arena with 75-cm
high aluminum walls, divided into 10 cm 2. A 60 W light bulb will be positioned 90 cm above the
base of the arena, and provided the only source of illumination in the testing room. Each animal
will be placed in the center of the open field apparatus, the ambulation scores (the number of
squares crossed) and the number of rearings (number of times animal stood on its hind limbs)
will be measured during a 3-min period. (Ying et al., 2005).

Experimental design:
The rats will undergo surgery for removal of olfactory bulbs and after recovery different
drug treatment will be started at 10-11 a.m. as described above. Effects on locomotor activity and
body weight will be assessed as given below:
Locomotor activity in open field and measurement of body weight of animal:
After recovery period of two week rats will be given treatment of different drugs once a
day for 14 days and accessed for the acute, sub-chronic and chronic effect of different drug
treatment on 1, 7 and 14th day of initiation of treatment respectively. Several positive and
negative modulators will be given at defferent time interval before administration of subeffective and effective dose of agmatine. Effect on locomotor activity will be accessed 30 min
after last dosing of agmatine in open field for ambulations and rearings during a period of 3 min
(as described above) and also measure body weight on same day.

Treatment groups:
1) To study the effect of agmatine:

Bulbectomized animal
A) Control group: Saline (5 ml/kg, i.p.) for 14 days.
B) Treatment group: Agmatine (5, 10 or 20 mg/kg, i.p.) for 14 days.
Sham operated animal
A) Control group: Saline (5 ml/kg, i.p.) for 14 days.
B) Treatment group: Agmatine (5, 10 or 20 mg/kg, i.p.) for 14 days.
Effect on locomotor activity and body weight will be analysed same as described above.

2) To study the effect of 3, 5 THP:

Bulbectomized animal
A) Control group: Vehicle (5 ml/kg, i.p.) for 14 days.
B) Treatment group: 3, 5 THP (0.5, 1 or 2 mg/kg i.p.) for 14 days.
Sham operated animal
A) Control group: Vehicle (5 ml/kg, i.p.) for 14 days.
B) Treatment group: 3, 5 THP (0.5, 1 or 2 mg/kg, i.p.) for 14 days.
Effect on locomotor activity and body weight will be analysed same as described above.

3) To study the influence of 3, 5 THP on antidepressant effect of agmatine:

Bulbectomized animal
A) Control group: Vehicle (5 ml/kg, i.p.) 30 min later, agmatine (sub-effective dose, i.p.)
for 14 days.
B) Treatment group: 3, 5 THP (sub-effective dose, i.p.) 30 min later, agmatine (sub-

effective dose, i.p.) for 14 days.

Sham operated animals
A) Control group: Vehicle (5 ml/kg, i.p.) 30 min later, agmatine (sub-effective dose, i.p.)
for 14 days.
B) Treatment group: 3, 5 THP (sub-effective dose, i.p.) 30 min later, agmatine (subeffective dose, i.p.) for 14 days
Effect on locomotor activity and body weight will be analysed same as described above.

4) To study the influence of neurosteroidogenic drugs (positive modulators of

neurosteroids) on antidepressant effect of agmatine:

i) For observing the effect of progesterone:

Bulbectomized animal
A) Control group: Vehicle (5 ml/kg, i.p.) 30 min later, agmatine (sub-effective dose, i.p.)
for 14 days.
B) Treatment group: Progesterone (5 mg/kg, i.p.) 30 min later, agmatine (sub-effective

dose, i.p.) for 14 days.

Sham operated animal
A) Control group: Vehicle (5 ml/kg, i.p.) 30 min later, agmatine (sub-effective dose, i.p.)
for 14 days.
B) Treatment group: Progesterone (5 mg/kg, i.p.) 30 min later, agmatine (sub-effective
dose, i.p.) for 14 days.
Effect on locomotor activity and body weight will be analysed same as described above.

ii) For observing the effect of metyrapone:

Bulbectomized animal
C) Control group: Vehicle (5 ml/kg, i.p.) 30 min later, agmatine (sub-effective dose, i.p.)
for 14 days.
A) Treatment group: Metyrapone (50 mg/kg, i.p.) 30 min later, agmatine (sub-effective

dose, i.p.) for 14 days.

Sham operated animal
A) Control group: Vehicle (5 ml/kg, i.p.) 30 min later, agmatine (sub-effective dose, i.p.)
for 14 days.
B) Treatment group: Metyrapone (50 mg/kg, i.p.) 30 min later, agmatine (sub-effective
dose, i.p.) for 14 days.
Effect on locomotor activity and body weight will be analysed same as described above.

iii) For observing the effect of 4CD:

Bulbectomized animal
A) Control group: Vehicle (5 ml/kg, i.p.) 30 min later, agmatine (sub-effective dose, i.p.)
for 14 days.
B) Treatment group: 4CD (15 mg/kg, i.p.) 30 min later, agmatine (sub-effective dose,

i.p.) for 14 days.

Sham operated animal

A) Control group: Vehicle (5 ml/kg, i.p.) 30 min later, agmatine (sub-effective dose, i.p.)
for 14 days.
B) Treatment group: 4CD (15 mg/kg, i.p.) 30 min later, agmatine (sub-effective dose,
i.p.) for 14 days.
Effect on locomotor activity and body weight will be analysed same as described above.
5) To study the influence of neurosteroid biosynthesis inhibitors (negative modulators of
neurosteroids) on antidepressant effect of agmatine:
i) For observing the effect of indomethacin:

Bulbectomized animal
A) Control group: Vehicle (5 ml/kg, i.p.) 20 min later, agmatine (effective dose, i.p.) for
14 days.
B) Treatment group: Indomethacin (5 mg/kg, i.p.) 20 min later, agmatine (effective

dose, i.p.) for 14 days.

Sham operated animal
A) Control group: Vehicle (5 ml/kg, i.p.) 20 min later, agmatine (effective dose, i.p.) for
14 days.
B) Treatment group: Indomethacin (5 mg/kg, i.p.) 20 min later, agmatine (effective
dose, i.p.) for 14 days.
Effect on locomotor activity and body weight will be analysed same as described above.

ii) For observing the effect of trilostane:

Bulbectomized animal
A) Control group: Vehicle (5 ml/kg, i.p.) 120 min later, agmatine (effective dose, i.p.) for
14 days.
B) Treatment group: Trilostane (30 mg/kg, i.p.) 120 min later, agmatine (effective

dose, i.p.) for 14 days.

Sham operated animal
A) Control group: Vehicle (5 ml/kg, i.p.) 120 min later, agmatine (effective dose, i.p.) for
14 days.
B) Treatment group: Trilostane (30 mg/kg, i.p.) 120 min later, agmatine (effective
dose, i.p.) for 14 days.
Effect on locomotor activity and body weight will be analysed same as described above.

iii) For observing the effect of PK11195:

Bulbectomized animal
A) Control group: Vehicle (5 ml/kg, i.p.) 30 min later, agmatine (effective dose, i.p.) for
14 days.
B) Treatment group: PK11195 (15 mg/kg, i.p.) 30 min later, agmatine (effective dose,

i.p.) for 14 days.

Sham operated animal
A) Control group: Vehicle (5 ml/kg, i.p.) 30 min later, agmatine (effective dose, i.p.) for
14 days.
B) Treatment group: PK11195 (15 mg/kg, i.p.) 30 min later, agmatine (effective dose,
i.p.) for 14 days.
Effect on locomotor activity and body weight will be analysed same as described above.

Data analysis:
Data will be analysed by two-way analysis of variance (ANOVA). Significant interaction
will be assessed by post hoc Dunnett's or StudentNewmanKeuls test. The criterion for
statistical significance will be P < 0.05.

Possible outcomes:
1) Agmatine may have antidepressant-like effect in olfactory bulbectomized rats.
2) 3, 5 THP may have role in antidepressant-like effect of agmatine in olfactory
bulbectomized rats.

References:
Y. Akwa, R.H. Purdy, G.F. Koob, K.T. Britton, The amygdale mediates the anxiolytic-like effect
of the neurosteroid allopregnanolone in rat, Behav. Brain Res. 106 (1999) 119125.
E.E. Baulieu, Steroid hormones in the brain: Several mechanisms, in: J.A. Fuxe, J.A. Gustafsson,
L. Wetterbreg (Eds.), Steroid Hormone-regulation of the Brain, Pergamon Press, Oxford, (1981)
314.

A.K. Bence, D.R. Worthen, J.P. Stables, P.A. Crooks, An in vivo evaluation of the antiseizure
activity and acute neurotoxicity of agmatine, Pharmacol. Biochem. Behav. 74 (2003) 771775.
P. Blier, C. de Montigny, Current advances and trends in the treatment of depression, Trends
Pharmacol. Sci. 15 (1994) 220226.
J. Budni, V.M. Gadotti, M.P. Kaster, A.R.S. Santos, A.S. Rodrigues, Role of different types of
potassium channels in the antidepressant-like effect of agmatine in the mouse forced swimming
test, Eur. J. Pharmacol. 575 (2007) 8793.
A.E. Bullock, A.L. Clark, S.R. Grady, S.F. Robinson, B.S. Slobe, M.J. Marks, A.C. Collins,
Neurosteroids modulate nicotinic receptor function in mouse striatal and thalamic synaptosomes,
J. Neurochem. 68 (1997) 24122423.
J .F. Cryan, C. McGrath, B.E. Leonard, T.R. Norman, Combining pindolol and paroxetine in an
animal model of chronic antidepressant actioncan early onset of action be detected?, Eur. J.
Pharmacol. 352 (1998) 2328.
J .F. Cryan, A. Markou, I. Lucki, Assessing antidepressant activity in rodents: recent
developments and future needs, Trends Pharmacol. Sci. 23 (2002) 238245.
J.M. ffrench-Mullen, P. Danks, K.T. Spence, Neurosteroids modulate calcium currents in
hippocampal CA1 neurons via a pertussis toxin-sensitive G-protein-coupled mechanism, J.
Neurosci. 14 (1994) 19631977.
C.A. Frye, The neurosteroid 3a, 5a-THP has antiseizure and possible neuroprotective effects in
an animal model of epilepsy, Brain Res. 696 (1995) 113120.
A. Halaris, H. Zhu, Y. Feng, J.E. Piletz, Plasma agmatine and platelet imidazoline receptors in
depression, Ann. N.Y. Acad. Sci. 881 (1999) 445 541.

J .P. Kelly, A.S. Wrynn, B.E. Leonard, The olfactory bulbectomized rat as a model of depression:
an update, Pharmacol. Ther. 74 (1997) 299316.

R.T. Khisti, C.T. Chopde, and S.P. Jain, Antidepressant-like effect of the neurosteroid 3ahydroxy-5a-pregnan-20-one in mice forced swim test, Pharmacol. Biochem. Behav. 67(2000)
137-143.
R.T. Khisti, S.N. Mandhane, C.T. Chopde, The neurosteroid 3a-hydroxy-5a-pregnan-20-one
induces catalepsy in mice, Neurosci. Lett. 251 (1998) 8588.
M. Krass, G. Wegener, E. Vasar, V. Volke, Antidepressant-like effect of agmatine is not
mediated by serotonin, Behav. Brain Res. 188 (2008) 324328.
J.J. Lambert, D. Belelli, C. Hill-Venning, J.A. Peters, Neurosteroids and GABA receptor
function, Trends Pharmacol. Sci. 16 (1995) 295303.
D. Lavinsky, N.S. Arteni, C.A. Netto, Agmatine induces anxiolysis in the elevated plus maze task
in adult rats, Behav. Brain Res. 141 (2003) 19 24.
S.N. Mandhane, R.T. Khisti, C.T. Chopde, Adenosinergic modulation of 3a-hydroxy-5a-pregnan20-one induced catalepsy in mice, Psychopharmacology (Berl.) 144 (1999) 398404.
B.S. McEwen, Non-genomic and genomic effects of steroids on neural activity, Trends
Pharmacol. Sci. 12 (1991) 141-147.
G. Olmos, N. DeGregorio-Rocasolano, M. Paz-Regalado, T. Gasull, M. Assumpcio-Boronat, R.
Trullas, A. Villarroel, J. Lerma, J.A. Garcia-Sevilla, Protection by imidazoline drugs and
agmatine of glutamate-induced neurotoxicity in cultured cerebellar granule cells through
blockade of NMDA receptor, Br. J. Pharmacol. 127 (1999) 1317 1326.

A. Onal, Y. Delen, S. Ulker, N. Soykan, Agmatine attenuates neuropathic pain in rats: possible
mediation of nitric oxide and noradrenergic activity in the brainstem and cerebellum, Life Sci. 73
(2003) 413428.
S.M. Paul, R.H. Purdy, Neuroactive steroids, FASEB J. 6 (1992) 23112322.
J.E. Piletz, A. Halaris, P.R. Ernsberger, Psychopharmacology of imidazoline and alpha 2adrenergic receptors: implications for depression, Crit. Rev. Neurobiol. 9 (1994) 29 66.
G. Puia, M.R. Santi, S. Vicini, D.B. Pritchett, R.H. Purdy, S.M. Paul, P.H. Seeburg, E. Costa,
Neurosteroids act at recombinant human GABAA receptors, Neuron 4 (1990) 759765.
W. Raasch, U. Schafer, J. Chun, P. Dominiak, Biological significance of agmatine, an
endogenous ligand at imidazoline binding sites, Br. J. Pharmacol. 133 (2001) 755780.
D.J. Reis, S. Regunathan, Is agmatine a novel neurotransmitter in brain? Trends Pharmacol. Sci.
21 (2000) 187193.
S.C. Risch, C.B. Nemeroff, Neurochemical alterations of serotonergic neuronal systems in
depression, J. Clin. Psychiatry 53 (Suppl) (1992) 37.
E. Romeo, A. Strohle, G. Spalletta, F. Michele, B. Hermann, F. Holsboer, A. Pasini, R.
Rupprecht, Effect of antidepressant treatment on neuroactive steroid in major depression, Am. J.
Psychiatry. 155 (1998) 910-913.
R.

Rupprecht,

F.

Holsboer,

Neuroactive

steroids:

mechanisms

of

action

and

neuropsychopharmacological perspectives, Trends Neurosci. 22 (1999) 410416.

A. Strohle, A. pasini, E. Romeo, B. Hermann, G. Spalletta, F. Michele, F. Holsboer, R.
Rupprecht, Fluoxetine decreases conctrations of 3alpha, 5alpha-tetrahydrodeoxycorticosterone
(THDOC) in major depression, J. Psychiatry Res. 34 (2000) 183-186.

R.R. Ugale, A.N. Sharma, D.M. Kokare, K. Hirani, N.K. Subhedar, C.T. Chopde, Neurosteroid
allopregnanolone mediates anxiolytic effect of etifoxine in rats, Brain Res. 1184 (2007) 193-201.
V. Uzunova, Y. Sheline, J.M. Davis, A. Rasmusson, D.P. Uzunov, E. Costa, A. Guidotti, Increase
in the cerebrospinal fluid content of neurosteroids in patients with unipolar major depression
who are receiving fluoxetine or fluvoxamine, Proc. Natl. Acad. Sci. U.S.A. 95 (1998) 3239
3244.
V. Uzunova, M. Ceci, C. Kohler, D.P. Uzunov, A.S. Wrynn, Region-specific dysregulation of
allopregnanolone brain content in the olfactory bulbectomized rat model of depression, Brain
Res. 976 (2003) 18.
C.H. Wetzel, B. Hermann, C. Behl, E. Pestel, G. Rammes, W. Zieglgansberger, F. Holsboer, R.
Rupprecht, Functional antagonism of gonadal steroids at the 5-hydroxytryptamine type 3
receptor, Mol. Endocrinol. 12 (1998) 14411451.
A .S. Wrynn, C.P. MacSweeney, F. Franconi, L. Lemaire, D. Pouliquen, S. Herlidou, B.E.
Leonard, J.-M. Gandon, J.D. de Certaines, An in-vivo magnetic resonance study of the olfactory
neurosteroids in patients with unipolar major depression who are bulbectomized rat model of
depression, Brain. Res. 879 (2000) 193199.
X. Ying, K. Bao-Shan, Y. Hai-Yan, L. Yan-Hua, M. Xing, Z. Yong-He, L. Xue-Jun,
Antidepressant effects of curcumin in the forced swim test and olfactory bulbectomy models of
depression in rats, Pharmacol. Biochem. Behav. 82 (2005) 200 206.
B. Zimmerberg, L.G. Blaskey, Prenatal stress effects are partially ameliorated by prenatal
administration of the neurosteroid allopregnanolone, Pharmacol. Biochem. Behav. 59 (1998)
819827.

A.D.E. Zomkowski, L. Hammes, J. Lin, J.B. Calixto, A.R.S. Santos, A.L.S. Rodrigues, Agmatine
produces antidepressant-like effects in two models of depression in mice, NeuroReport 13 (2002)
387391.
A.D.E. Zomkowskia, A.O. Rosa, J. Lin, A.R.S. Santos, J.B. Calixtoc, A.L.S. Rodrigues,
Evidence for serotonin receptor subtypes involvement in agmatine antidepressant like-effect in
the mouse forced swimming test, Brain Res. 1023 (2004) 253 263.
A.D.E. Zomkowski, A.R.S. Santos, A.L.S. Rodrigues, Evidence for the involvement of the
opioid system in the agmatine antidepressant-like effect in the forced swimming test, Neurosci.
Lett. 381 (2005) 279283.
Signature of Guide
Prof. C. T. Chopde

Date:
Place:

Signature of Co-guide
Dr. R. R. Ugale

Signature of Candidate
S. R. Rahangdale