Kent1302269033 PDF

THE ROLE OF THE VASOPRESSIN 1B RECEPTOR IN THE REGULATION OF
SENSORIMOTOR GATING
A thesis submitted to Kent State University

in partial fulfillment of the requirements
for the degree of Master of Science
By
Monica Dhakar
May 2011
Thesis written by
Monica Dhakar
M.B.B.S., Byramjee Jeejeebhoy Medical College, 2008
M.S., Kent State University, 2011
Approved by
________________________ Dr. H. Caldwell, Advisor
________________________Dr. R. Dorman, Director, School of Biomedical Sciences
________________________Dr. John Stalvey, Dean, College of Arts and Sciences
ii
TABLE OF CONTENTS
PAGE
LIST OF FIGURES………………………………………………………………vi
LIST OF TABLES……………………………………………………………...viii
ACKNOWLEDGEMENTS………………………………………………………ix
Introduction………………………………………………………………………1
Vasopressin………………………………………………………………..2
Sexual dimorphism in the central vasopressin system…………………….3
Vasopressin and social behavior…………………………………………..3
Vasopressin receptors……………………………………………………..5
The vasopressin 1b receptor………………………………………………6
Vasopressin 1b receptor knockout mice…………………………………..7
Vasopressin, the vasopressin 1b receptor, and neuropsychiatric disorders.8
Sensorimotor gating……………………………………………………...10
Prepulse inhibition of the startle reflex…………………………..11
Cortico-striato-pallido-pontine circuitry…………………………13
Neurotransmitter systems regulating prepulse inhibition of the
startle reflex………………………………………….…………..15
Interactions of vasopressin and the cortico-striato-pallido-pontine
circuitry…………………………………………………………..17
iii
Objective…………………………………………………………………………17
Methods…………………………………………………………………………..20
General Methods…………………………………………………………………20
Animals…………………………………………………………………..20
Prepulse inhibition the startle reflex……………………………………..20
Drugs……………………………………………………………..20
PPI………………………………………………………………..21
Procedure………………………………………………………...21
Study design……………………………………………………...24
Quantitative real time polymerase chain reaction ……………………….26
Procedure………………………………………………………...26
Individual Experiments…………………………………………………………..26
Experiment 1A and 1B…………………………………………………...26
Experiment 2A and 2B…………………………………………………...27
Experiment 3……………………………………………………………..27
Statistics………………………………………………………………….28
Experiment 4A and 4B…………………………………………………..28
Statistics………………………………………………………………….29
Results……………………………………………………………………………30
Experiment 1A…………………………………………………………...31
Experiment 1B…………………………………………………………...31
Experiment 2A…………………………………………………………...37
iv
Experiment 2B…………………………………………………………...43
Experiment 3……………………………………………………………..49
Experiment 4A…………………………………………………………...55
Experiment 4B…………………………………………………………...55
Discussion………………………………………………………………………..64
References………………………………………………………………………..74
Appendix I……………………………………………………………………….88
Appendix II………………………………………………………………………93
v
LIST OF FIGURES
PAGE
Introduction
Fig 1: Role of vasopressin in regulation of social behaviors……………...4
Fig 2: Schematic diagram of PPI………………………………………...12
Fig 3: CSPP circuit………………………………………………………14
Methods
Fig 4: Schematic diagram representing the components of SR-LAB……22
Fig 5: Acoustic startle chamber …………………………………………23
Fig 6: Study design for Experiments 1, 2 and 3………………………….25
Results
Fig7: Experiment 1A Habituation of startle in females………………….32
Fig 8: Experiment 1A Baseline PPI percentage in females ……………..34
Fig 9: Experiment 1A PPI percentage following drug treatment in
females…………………………………………………………….35
Fig 10: Experiment 1B Habituation of startle in males …………………38
Fig 11: Experiment 1B Baseline PPI percentage in males………………40
Fig 12: Experiment 1B PPI percentage following drug treatment in
male………………………………………………….…………..41
Fig 13: Experiment 2A Habituation of startle in females…..…………..44
Fig 14: Experiment 2A Baseline PPI percentage in females…………….46
vi
Fig 15: Experiment 2A PPI percentage following drug treatment in
females…………………………………………………………...47
Fig 16: Experiment 2B Habituation of startle in males………………….50
Fig 17: Experiment 2B Baseline PPI percentage in males………………52
Fig 18: Experiment 2B PPI percentage following drug treatment in
males……………………………………………………………..53
Fig 19: Experiment 3 Habituation of startle in males…………………....56
Fig 20: Experiment 3 Baseline PPI percentage in males………………...58
Fig 21: Experiment 3 PPI percentage following drug treatment in
males…………………………………………………………….59
Fig 22: Experiment 4A Expression of NMDAR1 in males………………62
Fig 23: Experiment 4B Expression of NMDAR2A in males……………..63
Appendix I
Fig 24: PPI percentage in Avpr1b +/− mice following drug treatment….90
Fig 25: PPI percentage following AMP and saline in Avpr1b +/− mice...91
vii
LIST OF TABLES
PAGE
Results
Table 1: Experiment 1A Startle amplitude in females………………...…33
Table 2: Experiment 1A Mean and SEM for PPI in females…………….36
Table 3: Experiment 1B Startle amplitude in males………………..........39
Table 4: Experiment 1B Mean and SEM for PPI in males………………42
Table 5: Experiment 2A Startle amplitude in females…………………...45
Table 6: Experiment 2A Mean and SEM for PPI in females…………….48
Table 7: Experiment 2B Startle amplitude in males……………………..51
Table 8: Experiment 2B Mean and SEM for PPI in males………………54
Table 9: Experiment 3 Startle amplitude in males……………………….57
Table 10: Experiment 3 Mean and SEM for PPI in males……………….60
Appendix II
Table 11: PPI program…………………………………………………...92
viii
ACKNOWLEDGEMENTS
I would like to thank my advisor and mentor Dr. Heather Caldwell for giving me an
opportunity to work in her lab and her invaluable guidance over the course of three years.
Dr. Caldwell has always provided me constant encouragement and endless support in all
my endeavors including research and future career goals. I would also like to thank Dr.
Eric Mintz and Dr. John Johnson for serving on my thesis committee and providing
valuable insights and expert advice on my research project.
I am grateful to my colleagues; Erica Stevenson, Megan Rich and Uju Dike for their
patience in teaching me all the lab techniques that I know today. Without their support
and friendship, this project could not have been completed. I would also like to thank
Shannah Witchey, and all the undergraduate students for their help in conducting some of
the experiments.
I would like to extend my gratitude to my friends; Vivek, Priyanka, Sagar and
Harsh for being the family to me for the last three years. A special thanks to my friend,
Charu; for always being there with me during the ups and downs of the graduate school.
Lastly, a heartfelt thanks to the people, whom I owe everything that I have ever
achieved; Mom, Dad, Neha, and Dhaval! I cannot thank them all enough for everything
that they are to me. I thank each one of them for believing in me and in my dreams.
ix
Without Dhaval’s love and support, I would not be where I am today. I am grateful to
you for being the pillar of strength and encouragement at every step of my life!
x
Introduction:
The expression of normal social behavior is crucial for the development and
survival of many species (Donaldson and Young, 2008). Broadly, social behavior can be
defined as interactions between individuals of the same species and includes affiliation,
aggression, reproduction, and communication (Sokolowski, 2010; Insel and Young,
2000). Successful social interactions require animals to observe the actions of others,
make predictions about their behavior, and respond appropriately (Skuse and Gallagher,
2009). In humans, aberrant social behavior is characteristic of a variety of
neuropsychiatric disorders, such as autism, schizophrenia, obsessive compulsive disorder
(OCD), anxiety, and depression. Across species, the neural substrates and the
mechanisms regulating social behavior are highly conserved (Donaldson and Young,
2008).
A number of hormones and neuropeptides are important to the neural regulation
of social behavior. In particular, two neurohormones, oxytocin (Oxt) and vasopressin
(Avp), have consistently been implicated in the neural regulation of social behavior (as
reviewed in Caldwell et al., 2008; Lee et al., 2009; Ross and Young, 2009).
Evolutionarily, Oxt and Avp and their gene families are largely conserved across species
(Acher et al., 1995). Both peptides are composed of nine amino acids and differ from one
another in only two amino acids, i.e., those in the third and eighth positions. Oxt and Avp
are hydrophilic in nature and are not thought to readily cross the blood brain barrier.
1
2
Further, both exert their actions through membrane bound G protein coupled receptors.
The effectiveness of Oxt and Avp at regulating behavior is attributed to three
characteristics: 1) their localized distribution in the brain 2) their slow and enduring
effects, and 3) their plasticity (Insel and Young, 2000).
Vasopressin
Avp is synthesized as a part of a preprohormone, which is then enzymatically
cleaved to produce the biologically active nonapeptide, a neurophysin, and a
glycopeptide. Once cleaved, Avp is transported along the axons and released (Brownstein
et al., 1980; Acher et al., 2002). Avp is synthesized primarily in the magnocellular
neurons of the supraoptic (SON) and paraventricular nucleus (PVN) of the hypothalamus
that project to the posterior pituitary. Upon appropriate stimulation, Avp is released into
the blood stream from the posterior pituitary to produce systemic effects which include
the regulation of water balance and maintenance of blood pressure by acting on the blood
vessels and the kidneys (Nishimura and Fan, 2003; Bankir, 2001). Centrally, small
quantities of Avp are released from the dendrites of the magnocellular neurons to produce
local effects. In many species, Avp is also made in small populations of parvocellular
neurons within the PVN, the bed nucleus of stria terminalis (BNST), the medial
amygdala (MeA), and the suprachiasmatic nucleus (SCN) (Sofroniew, 1983). These
neurons provide robust projections to a number of brain areas, including, hippocampus,
subiculum, diagonal band of Broca, locus coeruleus, solitary tract nucleus, dorsal motor
nucleus of vagus, olfactory tubercle, lateral septum (LS), lateral habenula (LH), ventral
3
tegmental area (VTA), and spinal cord (Buijs, 1978; De Vries and Buijs, 1983; Millan et
al., 1984).
Sexual dimorphisms in the central vasopressin system
The distribution of Avp immunoreactive (Avp-ir) neurons and fibers shows a
remarkable degree of sexual dimorphism. In most mammals especially rodents, males
have a higher density of Avp-ir neurons and fibers in the MeA, BNST, LH and LS, as
compared to females (De Vries et al., 1983). However, there are exceptions to these
sexual dimorphisms and the distribution varies across species. For example, in primates
such as marmosets, macaques, and humans, there is a similar distribution of Avp-ir
neurons between males and females (Fliers et al., 1986; Caffe et al., 1989; Wang et al.,
1997). In guinea pigs, there is reverse sexual dimorphism with females having higher
Avp-ir fibers in the inferior colliculus, ventral trapezoid body, and dorsal cochlear
nucleus than males (Dubois-Dauphin et al., 1987; Dubois-Dauphin et al., 1989). Since
Avp is involved in the modulation of sex specific behaviors, the distribution of Avp-ir
fibers are also sensitive to circulating gonadal hormones (for full review on the
interactions of gonadal steroids and vasopressin, please see Dhakar et al., in press). For
instance, castration reduces Avp-ergic innervations from BNST and MeA in both males
and female rats and administration of testosterone increases the density of Avp fibers in
these areas (de Vries et al., 1984).
Vasopressin and social behavior

4
VASOPRESSIN
Avpr1 Avpr1
a b
Aggression Social Behaviors

Stress
Social
Affiliation
Memory
Anxiety
Figure 1: Schematic diagram showing the role of vasopressin in the regulation of social
behaviors
5
Centrally released Avp acts as a neuromodulator for a variety of social behaviors
such as, social recognition, social motivation, social memory, aggression, affiliation, and
parental behavior (as reviewed in Caldwell et al., 2008; Donaldson et al., 2008; Lee et al.,
2009) (Figure 1). Some evidence which indicates the importance of Avp in the regulation
of social behaviors comes from studies in Brattleboro rat (BB-Ho), a naturally occurring
Avp mutant, that lacks the ability to synthesize Avp (Bohus and de, 1998), and thus has
diabetes insipidus. BB-Ho rats have deficits in memory, social recognition, social
motivation, and attention (Laycock et al., 1983; Williams et al., 1985; Williams et al.,
1983). Exogenous administration of Avp in these rats improves certain social behaviors
such as social recognition (Engelmann and Landgraf, 1994). Further, in rodents,
intracerebral injection of an Avp analogue facilitates aggression (Ferris et al., 1997;
Caldwell and Albers, 2004), partner preference (Winslow et al., 1993; Cho et al., 1999),
and paternal behavior (Wang et al., 1994). In humans, intranasal administration of Avp
enhances verbal memory and reaction time in males (Born et al., 1998; Beckwith et al.,
1983).
Vasopressin receptors
Avp exerts it effects via three different receptors: 1) the Avp 1a receptor (Avpr1a)
2) the Avp 1b receptor (Avpr1b) and 3) the Avp 2 receptor (Avpr2). While the Avpr1a
and the Avpr1b are found centrally (as well as peripherally), the Avpr2 is only expressed
peripherally (Tribollet et al., 1988; Lolait et al., 1992). Across species, the Avpr1a is
widely distributed in the central nervous system (CNS) in areas known to be important
6
for neural regulation of social behavior (Johnson et al., 1993; Young et al., 2000;
Tribollet et al., 1997). This thesis will focus specifically on the Avpr1b (for a review of
the role of the Avpr in the neural regulation of behavior please see Caldwell et al., 2008;
Bielsky et al., 2004).
The vasopressin 1b receptor
The Avpr1b is a seven transmembrane receptor and belongs to the family of G
protein coupled receptors. It is coupled to Gαq/11 GTP binding proteins which along with
Gβλ activates a downstream 1,4,5-inositol triphosphate (IP3) second messenger pathway
(Michell et al., 1979; Jard et al., 1987). In the rat brain, in situ hybridization and RT-PCR
have localized Avpr1b transcripts to a number of areas that are known to regulate social
behaviors; olfactory bulb, LS, cerebral cortex, hippocampus, PVN, SCN, cerebellum, red
nucleus, and piriform cortical layer II (Lolait et al., 1995; Vaccari et al., 1998; Saito et
al., 1995). However, it should be noted that the probes used in the above mentioned in
situ hybridization studies had some identity to the oxytocin receptor and the Avpr1a. A
subsequent study which used more specific riboprobes, found a more restricted
localization of Avpr1b in the anterior pituitary, and in the CA2 region of the
hippocampus in rat, mouse, and human as detected by in situ hybridization and RT-PCR
(Young et al., 2006). So far, the distribution of Avpr1b has not been studied using
receptor autoradiography, due to the lack of an available pharmacologically specific
radiolabeled ligand, or with immunocytochemistry, due to lack of a specific antibody. Of

7
the species in which the Avpr1b has been sequenced and mapped, there is no evidence of
sexual dimorphism in its distribution.
Evidence for the role of the Avpr1b in the regulation of social behaviors such as
aggression, social recognition and social motivation comes from studies using an oral
Avpr1b antagonist (SSR149415), Avpr1b agonists and genetically engineered Avpr1b
knockout (Avpr1b −/−) mice (Wersinger et al., 2002; Tanoue et al., 2004).The
development of Avpr1b −/− mice, in particular, has been helpful in elucidating the role of
the Avrp1b in the neural regulation of social behaviors. The work described in this thesis
utilized Avpr1b −/− mice to test our central hypothesis.
Vasopressin 1b receptor knockout mice
Avpr1b −/− mice were first generated in 2002 through a targeted disruption of the
Avpr1b gene (Wersinger et al., 2002). Initial characterization of Avpr1b −/− mice found
that the males have deficits in social aggression and social motivation. However,
predatory aggression in these mice seems to be normal, indicating that they do not have a
global deficit in processing and attacking the stimuli but rather have a deficit in
processing the social cues associated with social aggression (Wersinger et al., 2007).
They also have impaired social recognition, despite normal olfactory function (Wersinger
et al., 2004; Wersinger et al., 2002). Avpr1b −/− females have deficits in maternal
aggression and an abnormal Bruce Effect (Wersinger et al., 2008; Wersinger et al., 2007).
(The Bruce Effect is a pheromonally mediated response wherein the female terminates a
pregnancy in response to chemosensory cues from an unfamiliar male). Avpr1b −/−

8
females fail to terminate their pregnancy in presence of unfamiliar male. Given the
discreet localization of the Avpr1b within the CA2 region of the hippocampus in mice,
these data suggest that Avpr1b might be important for the formation and retrieval of
memories about social context (Young et al., 2006). This hypothesis fits well with the
phenotypic data in Avpr1b −/− mice demonstrating that they have deficits in social
recognition tasks. Avpr1b −/− mice also have an attenuated stress response with
suppressed activity of the hypothalamic-pituitary-adrenal (HPA) axis under resting as
well as stress condition (Tanoue et al., 2004). Specifically, Avpr1b −/− mice have a
blunted adrenocorticotrophic hormone (ACTH) and cortisol response to hypoglycemia
and chronic restraint stress (Lolait et al., 2007). Avpr1b −/− mice also have an impaired
stress response following acute immune stress (administration of bacterial
lipopolysaccharide) and alcohol intoxication (Lolait et al., 2007). In summary, these
studies implicate a role for the Avpr1b in the regulation of social behaviors and allow for
the possibility that abnormalities in the Avp system in humans could contribute to
abnormal social behavior.
Vasopressin, the vasopressin 1b receptor, and neuropsychiatric disorders
Alterations in the Avp system have been found in a number of neuropsychiatric
disorders in humans. Compared to controls, patients diagnosed with major depressive
disorder (van et al., 1997), post-traumatic stress disorder (PTSD) (de Kloet et al., 2008),
and OCD (Altemus et al., 1992) all have elevated plasma levels of Avp. In
schizophrenics, baseline levels of plasma Avp are low compared to controls (Elman et al.,
9
2003); however, acute psychosis is associated with increased serum levels of Avp
(Raskind et al., 1987). There is evidence that elevations in plasma Avp correlate with
central Avp, this is consistent with the effects of Avp on the coordination of physiology
and behavior (Dogterom et al., 1978). However, it should be noted that even elevations in
Avp within the CSF may not be reflective of changes in local release.
In depressed patients, Avp mRNA is upregulated within the SON which is
thought to be associated with elevated glucocorticoid levels (Meynen et al., 2006; Inder
et al., 1997; Chakrabarty et al., 2005). Recently, a study found that a single nucleotide
polymorphism (SNP) in the Avpr1b gene seems to be protective against recurrent major
depression in adults (van et al., 2004). In children, variations in the Avpr1b gene have
been associated with the onset of childhood mood disorders, particularly in females
(Dempster et al., 2007). In adolescents and adults with depression and OCD, treatment
with antidepressants such as fluoxetine, a selective serotonin reuptake inhibitor, and
clomipramine, a tricyclic antidepressant, decrease CSF levels of Avp (Altemus et al.,
1994; De et al., 1993). In addition, intranasal administration of Avp or its analogue
ameliorates some of the negative symptoms of schizophrenia and improves memory
(Brambilla et al., 1986; Brambilla et al., 1989; Iager et al., 1986; Korsgaard et al., 1981).
Studies in animal models also confirm a possible role for Avp and the Avpr1b in
psychiatric disorders. SSR149415, which is a specific orally active Avpr1b antagonist,
has been used to study the role of Avpr1b. In rodents, the administration of this agent
reduces aggression and anxiety-like behaviors, suggesting that the Avpr1b is important
10
for regulation of the circuitry that mediates aggression and anxiety (Griebel et al., 2002;
Blanchard et al., 2005; Stemmelin et al., 2005). Administration of SSR149415 in rodents
also produces antidepressant effects in various behavioral tests, such as forced swim in
rats (Griebel et al., 2002; Griebel et al., 2003).
In sum, all the above studies suggest that Avp could potentially contribute to the
pathogenesis of these neuropsychiatric disorders. However, as Avp is a neuromodulator,
it is not likely that it is the cause of neuropsychiatric disorders. Rather, Avp likely
interacts with the neurocircuitry and neurotransmitter systems that underlie
neuropsychiatric disorders, and in turn affects the susceptibility an individual to a certain
psychiatric disorder. Common to many neuropsychiatric disorders, including
schizophrenia, are abnormal thought processing, thought fragmentation, and difficulty
with attention and cognition. This cognitive fragmentation and disordered thinking is
thought to originate in part from deficits in sensorimotor gating.
Sensorimotor gating
Sensorimotor gating is the ability to process and filter incoming sensory and
cognitive information and selectively allocate attention to the specific information. In
normal awake individuals, sensorimotor gating is constantly active to ‘gate’ any excess or
trivial information and selectively process the informational stimuli. Any individual’s
gating abilities are viewed as plastic and are known to be shaped by a combination of
genetic and environmental factors. Sensorimotor gating is also influenced by changes in
the environmental, neurochemical, and hormonal milieu in the body (Swerdlow, 1996).
11
When this ‘gating window’ is pathologically broken, it can lead to abnormalities in
orderly processing of information; resulting in sensory flooding, and ultimately thought
fragmentation and disruption in cognitive processes (Mcghie and Chapman, 1961;
Venables, 1960). This breakdown of the gating window is thought to be responsible for
the cognitive abnormalities which are features of many neuropsychiatric disorders. An
operational measure of sensorimotor gating is called prepulse inhibition of the startle
reflex (PPI).
Prepulse Inhibition of the startle reflex
The startle reflex is a constellation of responses to sudden, relatively intense
stimuli (auditory, visual or tactile), which is usually defensive in nature (Swerdlow et al.,
2000). The primary mammalian acoustic startle circuit is composed of three synapses
from the auditory nerve to the spinal motor neuron (Davis et al., 1982; Koch and
Schnitzler, 1997). Startle demonstrates several forms of plasticity such as habituation,
and fear potentiation, which are regulated by forebrain neural circuitry that is conserved
across species (Brown et al., 1951; Grillon et al., 1994; Geyer and Braff, 1982).
PPI is the profound decrease in the startle magnitude when the startling pulse is
preceded by a prepulse of weaker intensity by a very brief time window (Graham, 1975;
Hoffman and Searle, 1968; Ison et al., 1973) (Figure 2). The ‘gating’ period is
determined to be approximately 30-500 msec and serves to protect the sensory
information within the weak stimulus so that it is processed without being disrupted by
subsequent stronger stimuli (Hoffman and Searle, 1968; Ison et al., 1973; Swerdlow,
12
Figure 2: Schematic diagram showing the phenomenon of prepulse inhibition (PPI). PPI
is the decrease in startle magnitude when a weaker prepulse precedes the startling pulse.
(Modified and adapted from Swerdlow et al., 2000)
13
1996). PPI is a normal phenomenon that occurs virtually in all mammals, including
primates, and does not exhibit habituation or extinction with multiple trials.
PPI is one form of startle plasticity that shows striking similarities across species
from rodents to humans, and is easily quantified (Swerdlow et al., 1999). It is a robust
experimental phenomenon that occurs irrespective of the sensory modality of the startling
stimulus and the prepulse. In humans, PPI is measured by using electromyography of the
orbicularis oculi muscle which is a part of the blink reflex of the startle component (Braff
et al., 1992). In rats and mice, the whole body reflexive flinch to startle stimulus can be
measured by using a stabilimeter chamber (Swerdlow et al., 2000).
Deficits in PPI can be seen in a number of disorders with thought fragmentation
such as schizophrenia (Bolino et al., 1994; Braff et al., 1992; Braff et al., 1978; Braff et
al., 1999; Grillon et al., 1992; Kumari et al., 1999; Weike et al., 2000), OCD (Swerdlow
et al., 1993; Schall et al., 1996), Huntington’s disease (Swerdlow et al., 1995), Tourette
syndrome (Castellanos et al., 1996), nocturnal enuresis, and attention deficit hyperactive
disorder (ADHD) (Ornitz et al., 1992). Deficits in PPI are not unique to a particular
disease but are characteristic of various disorders with pathological disruption of circuitry
regulating it called the cortico-striato-pallido-pontine (CSPP) circuit (Swerdlow et al.,
1992).
Cortico-striato-pallido-pontine circuitry
Studies in rats have identified the neural substrates that regulate PPI. This serial
and parallel neural circuitry is called the CSPP circuit (Figure 3). It connects the limbic
14
Ventral
tegmental area
Hippocampus
mPFC
Amygdala
Nucleus accumbens
Ventral pallidum CSPP Circuit
Pedunculopontine
tegmental nucleus
S R
Cochlear Caudal pontine Motor

nucleus reticular nucleus neuron
Acoustic stimulus
Response
Primary acoustic startle circuit
Figure 3: Schematic diagram showing the main components of the CSPP circuit and
its regulatory control over startle reflex. The acoustic stimulus S produces a motor
response R through the simple pontine circuit. Prepulse effects on R are mediated
via the pedunculopontine tegmental nucleus, which is under the control of serial
and parallel descending projections from the forebrain. (Modified and adapted from
Koch and Schnitzler et al., 1997; Swerdlow et al., 2001) (Abbreviations: mPFC,
medial prefrontal cortex)
15
cortex with the ventral striatum, ventral pallidum, pontine tegmentum and converges with
the primary startle circuit at the level of caudal pons (Koch and Schnitzler, 1997;
Swerdlow and Geyer, 1998; Swerdlow et al., 2001; Koch et al., 1993). Though the main
inhibitory effect of prepulse on startle is exerted at the level of pons, the CSPP circuit
regulates the quality of the inhibitory activity and thus the degree of inhibition of the
response to stimuli when preceded by a prepulse (Swerdlow et al., 2001). The CSPP
circuit is known to be regulated by a variety of neurotransmitter systems in humans and
rodents, including the dopaminergic, serotonergic, and glutamatergic systems.
Neurotransmitter systems regulating prepulse inhibition of the startle reflex
Abnormalities in the dopamine, the glutamate, and the serotonin system have
been consistently implicated in pathogenesis of a number of neuropsychiatric disorders
such as schizophrenia, major depression, panic disorders and OCD (Carlsson et al.,
1997). Since these neurotransmitters are also reported to act on various neural substrates
in the CSPP circuitry, pharmacological manipulation of these neurotransmitter systems
using various psychotomimetics, can be used to produce deficits in PPI in rodents.
The dopamine system has been studied extensively both in humans and rodents.
In healthy human subjects, direct dopamine agonists like bromocriptine (a D2 agonist)
and indirect dopamine agonists like amphetamine (AMP) have been shown to disrupt PPI
(Abduljawad et al., 1998; Abduljawad et al., 1999; Kumari et al., 1998; Hutchison and
Swift, 1999). Similarly, in many preclinical studies in rats and mice, apomorphine
(APO), which is a direct dopamine agonist, and AMP produce robust decreases in PPI
16
(Mansbach et al., 1988; Swerdlow et al., 2000; Dulawa and Geyer, 1996; Ralph et al.,
1999; Ralph et al., 2001). These PPI disrupting effects of dopamine agonists can be
effectively blocked by pretreatment with typical antipsychotics such as haloperidol
(which primarily acts on D2 receptors) suggesting that dopamine and its receptors are
important regulators of PPI (Abduljawad et al., 1998; Abduljawad et al., 1999). In
humans, dopamine hyperactivity has been implicated in the pathogenesis of
schizophrenia and deficits in sensorimotor gating. Few studies report that the PPI deficits
seen in schizophrenic patients can be successfully reversed by administration of typical
and atypical antipsychotics (which have primary actions on dopamine receptors),
suggesting the role of dopamine in the regulation of PPI (Kumari et al., 1999; Kumari et
al., 2000; Weike et al., 2000).
Another important neurotransmitter implicated in the regulation of PPI is
glutamate. Dysregulation of the glutamatergic system is thought to be involved in the
etiology of schizophrenia, OCD, and mania; all disorders with abnormal thought
processing (Chakrabarty et al., 2005; Javitt, 1987). In humans, schizophrenia-like
symptoms, including PPI deficits, can be produced by administering NMDA receptor
antagonists, like ketamine. In rodents and non-human primates, deficits in PPI can be
modeled using other NMDA receptor antagonists, like phencyclidine (PCP) or
dizocilpine (MK-801) (Curzon and Decker, 1998; Mansbach and Geyer, 1989; Furuya et
al., 1999; Linn et al., 1999). Further, the effects of NMDA receptor antagonists on PPI
can be reversed using atypical antipsychotics like clozapine, olanzapine, and quetiapine,
which have actions on multiple receptors (Swerdlow et al., 1996; Swerdlow et al., 1998;
17
Yamada et al., 1999). The reversal of PPI with atypical antipsychotics suggests that the
regulation of PPI by glutamate might not be due to a single receptor interaction, but
might involve many downstream receptors, synapses, and other neurotransmitter systems
such as serotonin (Olney et al., 1999).
Interactions of vasopressin and the cortico-striato-pallido-pontine circuitry
Avp is a neuromodulator that is known to interact with the dopamine, glutamate
and serotonin systems in various brain areas (Skuse and Gallagher, 2009; Syed et al.,
2007). Abnormalities of the Avp system have been implicated in a variety of
neuropsychiatric disorders which are all characterized by a dysfunction of these
neurotransmitters. The Avp mutant, BB-Ho rats have also been found to have
abnormalities in several neurotransmitter systems including dopamine and serotonin
(Feenstra et al., 1990). They also exhibit profound deficits in PPI at baseline, which can
be successfully reversed by using both typical and atypical antipsychotics (Feifel and
Priebe, 2001; Feifel et al., 2007), which suggests that Avp does play some role in
regulating PPI and might interact with more than one neurotransmitter system. Exactly
how Avp interacts with these neurotransmitter systems still needs to be explored further.
Moreover, whether this deficit in PPI is mediated via Avpr1a or Avpr1b has yet to be
investigated. The work described in this thesis set out to determine if the Avpr1b interacts
with the neural circuitry that regulates PPI.
Objective
18
Our central hypothesis is that Avp acting via the Avpr1b contributes to the
regulation of PPI through its interaction with either the glutamatergic or dopaminergic
systems. This hypothesis is based on several pieces of evidence suggesting a role of Avp
in regulation of PPI. First, BB-Ho rats, which are genetically deficient in the production
of Avp, have been shown to have deficits in baseline PPI. Second, these deficits in PPI
can be reversed by infusion of atypical antipsychotics such as clozapine and risperidone
(Feifel et al., 2007). We tested our central hypothesis by treating Avpr1b +/+ and Avpr1b
−/− mice with psychotomimetics and measuring their effects on PPI. The
psychotomimetics used in this study were dopamine agonists (APO and AMP) and
NMDA receptor antagonist (MK-801 and PCP), since these are the drugs most
commonly used to disrupt PPI.
Experiment 1, was performed to examine the effects of APO, AMP, and MK-801
on PPI in Avpr1b −/− and Avpr1b +/+ mice. It was hypothesized that the Avpr1b −/−
mice would be more susceptible to the PPI disruptive effects of psychotics. Experiment 2,
was performed to confirm the results of Experiments 1 by comparing the effects of a
single drug MK-801 to saline. It was hypothesized that Avpr1b −/− mice treated with
MK-801 would have more disrupted PPI as compared to Avpr1b +/+ mice. Experiment 3,
was performed to compare the effects of two different NMDA receptor antagonists MK-
801 and PCP on PPI in Avpr1b −/− and Avpr1b +/+ mice. It was hypothesized that a
stronger disruption of PPI will be seen following treatment with PCP in Avpr1b −/− mice
as compared to MK-801, since PCP augments the effects of glutamate through sigma
19
receptors. Experiment 4 was designed to find out where in the brain the Avp system
interacts with glutamatergic system to affect PPI. Therefore, in Experiment 4,
quantitative real time PCR was performed to quantify the expression of NMDA receptor
mRNA (NMDAR1 and NMDAR2A) in cortex, striatum and hippocampus in male Avpr1b
−/− and Avpr1b +/+ mice. It was hypothesized that developmental lack of Avpr1b in
Avpr1b −/− mice would cause significant changes in expression of NMDA receptor
mRNA as compared to Avpr1b +/+ mice.

Methods:
General Methods
Animals
This line was originally generated at the National Institute of Mental Health
(NIMH) by W. Scott Young, III (Wersinger et al., 2002). These mice were fully
backcrossed into a C57BL/6J background and shipped to Kent State University in 2007
where they have been bred in the vivarium in Cunningham Hall ever since. All the
animals used in the study (Avpr1b −/− and Avpr1b +/+ male and females) were offspring
from heterozygous breeding pairs. After weaning, animals were reared in the same-sex
sibling groups. At the time of weaning (21 days), tails were clipped in order to extract the
DNA, and PCR was performed using the specific primers for Avpr1b for genotyping the
animals (Wersinger et al., 2002). For each Experiment, the animals were adults between
three and six months of age. Food and water were available ad libitum throughout the
studies. All behavior testing was done during the light phase of the 12:12 light dark cycle.
Experiments were conducted in accordance with the protocol approved by the Kent State
University Institutional Animal Care and Use Committee.
Prepulse Inhibition of the Startle Reflex
Drugs
20
21
The following doses of APO, AMP, MK-801 and PCP were used and were
chosen based on the literature (Caldwell et al., 2009; Yee et al., 2004) and pilot
experiments (Appendix I); APO (10mg/kg), AMP (12mg/kg), MK-801 (0.7mg/kg) and
PCP (8mg/kg). All the drugs were dissolved in 0.9% saline. APO and AMP were diluted
to 1.5mg/ml, MK-801 to 10.5mg/ml and PCP to 1.2mg/ml. Saline was given in the same
volume as APO and AMP and served as control.
PPI
Two startle chambers were used for all the experiments (SR-LAB; San Diego
instruments, San Diego, CA, USA) (Figure 4 and 5). The startle chamber consisted of a
small plexiglass cylinder (8 cm diameter, 16cm long) which is mounted on a platform
inside a chamber. The loudspeaker inside the chamber generated the background noise as
well as all the prepulse and pulse alone tones. The startle response was measured from
the whole body reflexive flinch of the animals to acoustic startle stimuli. Vibrations of
the animal were sensed and transduced by the piezoelectric unit (motor sensor) attached
to the bottom of the platform which is then converted into arbitrary units by the
potentiometer and stored in the computer.
Procedure
Each testing session consisted of a 5 min acclimation period followed by 60 trials
for 20 min. During each session, background noise of 74dB was generated by the
loudspeaker. Sessions consisted of NO PULSE, PULSE ALONE and PREPULSE +
PULSE trials. During NO PULSE trails, no tone was presented. The PULSE ALONE
22
Figure 4: Schematic diagrams representing the components of the SR-LAB system.

23
Figure 5: Acoustic startle chamber (SR LAB, San Diego, CA)

24
trial consisted of 120dB pulse of 40ms duration. During the PREPULSE + PULSE trial,
three prepulse tones each 20ms long, of 77dB (3dB above the background), 80dB (6dB
above the background) and 86dB (12dB above the background) were presented 100 msec
prior to the PULSE tone. Each session began and ended with 5 trials of pulse alone tones.
In between the pulse alone trials, each session consisted of 10 blocks of 5 trials, during
which different stimuli were presented in a random order.
Study Design
Prior to drug testing, each animal was tested for baseline PPI without any drug
treatment. Following baseline testing, repeated measures testing with drugs was done
with each animal. APO, AMP, MK-801, PCP and saline (0.9%) were administered
intraperitoneally (i.p.) to animals in a counterbalanced manner, though which drug was
administered depended on the Experiment. A minimum of 1 week between testing was
observed in order to avoid any residual drug effects. PPI percentage was calculated using
the following equation: (1-(startle amplitude of the PREPULSE + PULSE/startle
amplitude of the PULSE ALONE)) × 100. Habituation of startle was calculated using the
average responses from the first 5 PULSE ALONE trials (Pulse 1) and last 5 PULSE
ALONE (Pulse 2) trials. Magnitude of the startle was calculated from the average of all
the PULSE ALONE trials during the entire session excluding the first and last blocks of
PULSE ALONE trials (Figure 6).

25
10 Blocks of 5 Trials
5 Pulse 5 Pulse
Alone Trials NO PULSE Alone Trials
PREPULSE + PULSE (3X)
PULSE 1 PULSE 2
PULSE ALONE
Startle amplitude was Habituation of startle PPI percentage was

calculated from the was measured from calculated from the
average responses to the average responses average response
PULSE ALONE of the first 5 (PULSE from the 10 blocks of
trials. Comparisons 1) and last 5 pulse 5 trials each using
were made between alone (PULSE 2) the equation: (1-
drug treatments as trials and (startle amplitude of
compared to saline. comparisons were the prepulse + pulse
made within drug trial / startle
treatment. amplitude of pulse
alone trial)) × 100.
Comparisons were
made between
genotypes and drug
treatment with
respect to saline.
Figure 6: Study design used to analyze startle amplitude, habituation of startle, and PPI
percentage.
26
Quantitative Real Time Polymerase Chain Reaction
Procedure
Brains were collected from male Avpr1b −/− and Avpr1b +/+ mice and frozen on
dry ice. Tissue punches of 1.00 mm diameter were collected from a 400 µm slab cut on
cryostat, from the striatum, hippocampus and frontal cortex. Samples were homogenized
using homogenizer and RNA was extracted using the RiboPure kit (Ambion, The RNA
Company, Austin, TX). The purity of the sample was established by using the Nanodrop
Spectrophotometer ND-100 (Thermo-Scientific) and 1 µl of extract. Samples having a
ratio of 260/280 between 1.8-2.0 were included in the assay. Purified RNA was reverse
transcribed into cDNA using a ThermoScript RT- PCR system (Invitrogen, Carlsbad, CA,
USA). Specific Taqman primers for NMDAR1 and NMDAR2A were purchased from
Applied Biosystems. Quantitative real time PCR was performed using Stratagene QPCR
System (Agilent Technologies, Foster City, CA, USA). The genes quantified were
NMDAR1 and NMDAR2A within the brain areas mentioned using specific primers.
Triplicates of 20 µl reaction with Taqman Universal master mix and 2 µl of cDNA in
each well were used to perform the assay. GAPDH was used as the control for
normalization.
Individual Experiments
Experiment 1: Assessment of prepulse inhibition of the startle reflex in A) female
Avpr1b +/+ and Avpr1b −/− mice and B) male Avpr1b +/+ and Avpr1b −/− mice
following treatment with APO, AMP, MK-801, and saline

27
In Experiment 1A, 13 Avpr1b +/+ and 10 Avpr1b −/− female mice were used as
subjects. In Experiment 1B, 11 Avpr1b +/+ and 12 Avpr1b −/− male mice were used. The
animals were run in two different cohorts at two different times. Each cohort consisted of
a mix of males and females. However, for analysis purposes, males and females were
considered separately. Each animal was initially tested for baseline PPI measurement
without any drugs. For the drug studies, APO, AMP, MK-801 and saline (0.9%) were
administered i.p. 10 min prior to testing in a counterbalanced manner. Habituation, startle
and PPI were measured as described under statistics section.
Experiment 2: Assessment of the effects of MK-801 and saline on prepulse inhibition
of the startle reflex in A) female Avpr1b +/+ and Avpr1b −/− mice and B) male
Avpr1b +/+ and Avpr1b −/− mice
In Experiment 2A, subjects were 9 Avpr1b +/+ and 7 Avpr1b −/− female mice. In
Experiment 2B, 9Avpr1b +/+ and 10 Avpr1b −/− male mice were used. Males and
females were run at the same time; however analyzed separately. MK-801 and saline
(0.9%) were given i.p. 10 min prior to testing. Data were analyzed as mentioned under
statistics section.
Experiment 3: Assessment of prepulse inhibition of startle reflex in response to MK-
801, PCP, and saline in Avpr1b +/+ and Avpr1b −/− male mice
Only 10 Avpr1b −/− and 7 Avpr1b +/+ male mice were used as study subjects
since a robust effect of MK-801 on PPI was seen only in males in the Experiment 1. MK-
28
801, PCP and saline (0.9%) were administered i.p. in a counterbalanced manner and data
were analyzed as described under statistics section.
Statistics
For Experiments 1, 2, and 3, baseline PPI percentage was analyzed across
genotypes using repeated measure of analysis of variance (ANOVA) comparing the main
effects of genotype, sex and prepulse intensity as well as their interactions. For the drug
studies, PPI percentage was determined using repeated ANOVA and main effects of
genotype, drug treatment, prepulse intensity and their interactions were compared.
Effects in both the sexes were analyzed independently. Animals with PPI percentage
within two standard deviations of the mean were included in statistical analysis. Since we
were interested in habituation across the test session; effect of drugs on habituation was
measured using repeated ANOVA with weight as a covariate. To correct for multiple
comparisons, the α was adjusted to p<0.013. Main effects of the first five PULSE
ALONE trials were compared to last five PULSE ALONE trials as well as those of
genotype and any interactions. Startle was compared using a repeated measures ANOVA.
Main effects of drug treatment, genotype, and their interactions were examined.
Experiment 4: Assessment of gene expression of A) NMDAR1 and B) NMDAR2A in
male Avr1b +/+ and Avpr1b −/− mice
Subjects were 5 Avpr1b −/− and 5 Avpr1b +/+ male mice. Animals were killed by
cervical dislocation and brains removed. Brains were immediately frozen on dry ice and
29
stored at -80C until tissue collection and quantitative real time PCR procedure were
performed as described under quantitative real time PCR methods section.
Statistics
For Experiment 4, the Ct values from the samples were normalized against
GAPDH. The expression values were then calculated using the equation: 2-∆Ct .
Comparisons were then made between genotypes by using t-test.

Results:
Experiment 1A: Assessment of PPI in Avpr1b +/+ and Avprb1 −/− female mice
following treatment with saline, APO, AMP and MK-801.
Habituation of startle
For all the drug treatments, there were no main effects of the drugs, genotype or any
interactions on habituation of startle. There was no habituation to startle between Pulse 1
and Pulse 2 trial sessions within any of the drugs tested (Figure 7).
Startle amplitude
There was a main effect of the drugs on startle amplitude (F3,54 = 16.042, p<0.05), with
APO and AMP decreasing the startle amplitude, whereas MK-801 increased the startle
amplitude. There was no main effect of genotype, or any interactions on the startle (Table
1).
PPI Percentage
Baseline: There were no genotypic differences in the baseline PPI percentage. There were
the expected main effects of prepulse intensity (F2,36 = 25.073, p<0.05), resulting in
increased PPI percentage with increasing prepulse tone (Figure 8).
30
31
Drug treatment: There was a main effect of the drug treatment (F3,54 = 11.636, p<0.05)
with APO, AMP and MK-801 all resulting in disruption of PPI as compared to saline.
There was also a main effect of prepulse intensity (F2,36 = 60.785, p<0.05). However,
there were no genotypic differences in the females on PPI percentage or any other
interactions (Figure 9) and (Table 2).
Experiment 1B: Assessment of PPI in male Avpr1b +/+ and Avprb1 −/− mice
following treatment with saline, APO, AMP and MK-801
There were no main effects of drug treatments, genotype or any interactions on the
habituation of startle across Pulse 1 and Pulse 2 sessions in male Avpr1b +/+ and Avpr1b
−/− mice (Figure 10).
Startle amplitude
There was a main effect of the drugs on startle amplitude (F3,60 = 34.775, p<0.05), with
APO decreasing the startle amplitude whereas AMP and MK-801 increased the startle
amplitude. There was no main effect of genotype, or any interactions on the startle (Table
3).
PPI Percentage
32
Saline APO
300 300
*
Pulse 1 Pulse 1
250 Pulse 2 250 Pulse 2
200
Startle Amplitude
200
Startle Amplitude
150 150
100 100
50 50
0 0
Avpr1b +/+ Avpr1b −/− Avpr1b +/+ Avpr1b −/−
AMP MK-801
300 300
*
Pulse 1 Pulse 1
200
Startle Amplitude
200
Startle Amplitude
150 150
100 100
50 50
0 0
Figure 7: Habituation of startle in female Avpr1b +/+ and Avpr1b −/− mice. There was
no habituation of startle to drugs; Saline (0.9%), APO (10mg/kg), AMP (12mg/kg) and
MK-801 (0.7mg/kg), in Avpr1b +/+ and Avpr1b −/− female mice. Data are expressed as
means + standard error of the mean (Abbreviations: APO, apomorphine; AMP,
amphetamine; MK-801, dizocilpine).
33
Mean ± Standard Error of the Mean (Startle amplitude)
Drugs Avpr1b +/+ Avpr1b −/−
Saline 126.36 ± 18.25 111.83 ± 12.75
APO* 56.91 ± 9.70 47.88 ± 7.95
AMP* 82.50 ± 8.72 93.33 ± 10.01
MK-801* 127.97 ± 27.23 159.25 ± 9.21
Table 1: Startle amplitude in female Avpr1b +/+ and Avpr1b −/− mice. There was a main
effect of the drug treatment (p<0.05), with APO and AMP decreasing the startle
amplitude whereas MK-801 increased the startle amplitude in both the genotypes. There
was no genotypic difference between the Avpr1b +/+ and Avpr1b −/− mice. Startle was
calculated from the averages responses from the first and last pulse alone (120dB) trials.
(*p < 0.05 indicating a main effect of drug compared to saline) (Abbreviations: APO,
apomorphine; AMP, amphetamine; MK-801, dizocilpine).
34
Avpr1b +/+
100
Avpr1b −/−
*
*
80
% Prepulse Inhibition
60
40
20
0
3dB 6dB 12dB
Figure 8: Baseline PPI percentage in female Avpr1b +/+ and Avprb1 −/− mice. There was
no genotypic difference in baseline PPI percentage but there was a main effect of
prepulse intensity with the higher decibel prepulse tone resulting in greater PPI
percentage (* indicates p<0.05). Data are expressed as means + standard error of the
mean.
35
Avpr1b +/+
100 3dB
6dB
12dB
% Prepulse Inhibtion 80
60
40
20
0
SALINE APO AMP MK801
Avpr1b −/−
100 3dB
6dB
12dB
80
% Prepulse Inhibtion
60
40
20
0
Figure 9: PPI percentage in female Avpr1b +/+ and Avpr1b −/− mice following drug
treatment. Treatment with all the three drugs, APO, AMP, and MK-801 resulted in
disruption of PPI as compared to saline (p<0.05). There were however no genotypic
differences between Avpr1b +/+ and Avpr1b −/− female mice. Data are expressed as
means + standard error of the mean. (Abbreviations: APO, apomorphine; AMP,
amphetamine; MK-801, dizocilpine).
36
Mean ± Standard Error of the Mean (PPI percentage)

Drugs and Prepulse
Avpr1b +/+ Avpr1b −/−
Levels
Saline 77 63.40 ± 4.56 66.61 ± 3.53
Saline 80 73.42 ± 3.19 74.14 ± 3.52
Saline 86 78.68 ± 2.21 82.47 ± 1.74
APO 77 48.91 ± 4.02 38.60 ± 9.87
APO 80 57.35 ± 3.62 52.87 ± 6.67
APO 86 58.46 ± 3.02 55.39 ± 10.41
AMP 77 46.51 ± 4.76 38.92 ± 7.58
AMP 80 55.60 ± 5.01 55.75 ± 2.71
AMP 86 63.22 ± 4.14 62.53 ± 3.51
MK-801 77 31.49 ± 9.29 40.22 ± 4.28
MK-801 80 47.78 ± 5.99 42.98 ± 6.62
MK-801 86 63.31 ± 5.62 68.43 ± 4.37
Table 2: Mean and standard error of the mean for drug treatment in female Avpr1b +/+
and Avpr1b −/− mice. (Abbreviations: APO, apomorphine; AMP, amphetamine; MK-
801, dizocilpine)
37
Baseline: There was a main effect of the prepulse intensity (F2,40 = 38.869, p<0.05), with
greater PPI percentage with higher intensity tone. However there were no genotypic
differences in baseline PPI percentage in Avpr1b +/+ and Avpr1b −/− males (Figure 11).
Drug treatment: There was a main effect of drug treatment (F3,60 = 28.954, p<0.05) and
prepulse intensity (F2,40 = 121.057, p<0.05). APO, AMP and MK-801 disrupted the PPI
as compared to saline in both Avpr1b +/+ and Avpr1b −/− male mice but there was no
main effect of genotype. However, there was a genotype × drug interaction (F3,60 = 6.018,
p<0.05). Post hoc tests with repeated ANOVA revealed that treatment with MK-801
resulted in more disruption of PPI in Avpr1b −/− mice as compared to Avpr1b +/+ mice
(p<0.05). There were no other interactions of genotype with any other drug treatment
(Figure 12) and (Table 4).
Experiment 2A: Assessment of prepulse inhibition of startle reflex in female
Avpr1b +/+ and Avpr1b −/− mice following administration of MK-801
Habituation of Startle
There was no habituation to startle reflex in female Avpr1b +/+ and Avpr1b −/− mice
with saline or MK-801 (Figure 13). There was no main effect of genotype, drug or any
other interaction.
Startle amplitude
38
Saline APO
300 300
* *
Pulse 1 Pulse 1
200
Startle Amplitude
200
Startle Amplitude
150 150
100 100
50 50
0 0
AMP MK-801
300 300
* *
Pulse 1 Pulse 1
200
Startle Amplitude
200
Startle Amplitude
150 150
100 100
50 50
0 0
Figure 10: Habituation of startle in male Avpr1b +/+ and Avpr1b −/− mice. There were
no main effects of the drugs; Saline (0.9%), APO (10mg/kg), AMP (12mg/kg) and MK-
801 (0.7mg/kg) on habituation of startle on Avpr1b +/+ and Avpr1b −/− mice. Data are
expressed as mean + standard error of the mean. (Abbreviations: APO, apomorphine;
AMP, amphetamine; MK-801, dizocilpine)
39
Saline 137.12 ± 18.95 116.91 ± 13.95
APO* 76.19 ± 11.65 96.99 ± 11.38
AMP* 142.35 ± 13.80 117.23 ± 14.56
MK-801* 243.21 ± 29.89 232.95 ± 32.07
Table 3: Startle amplitude in male Avpr1b +/+ and Avpr1b −/− mice. There was a main
effect of the drug treatment on startle amplitude (p<0.05), with APO decreasing the
startle amplitude whereas AMP and MK-801 increased the startle amplitude in both
genotypes. There was no genotypic difference between the Avpr1b +/+ and Avpr1b −/−
mice. Startle was calculated from the averages responses from the first and last pulse
alone (120dB) trials. (*p < 0.05 indicating a main effect of drug compared to saline)
(Abbreviations: APO, apomorphine; AMP, amphetamine; MK-801, dizocilpine).
40
Avpr1b +/+
100
Avpr1b −/−
*
*
80
60
40
20
0
3dB 6dB 12dB
Figure 11: Baseline PPI percentage in male Avpr1b +/+ and Avprb1 −/− mice. There was
no genotypic difference in baseline PPI percentage but there was a main effect of
prepulse intensity with the higher decibel prepulse tone resulting in greater PPI
percentage (* indicates p<0.05). Data are expressed as mean + standard error of the
mean.
41
Avpr1b +/+
100 3dB
6dB
12dB
80
60
40
20
0
Avpr1b −/−
100 3dB
6dB
12dB
80
*
60
*
40
20
0
Figure 12: PPI percentage following drug treatment in males. Treatment with all the three
drugs, APO, AMP, and MK-801 resulted in disruption of PPI as compared to saline
(p<0.05). There was no main effect of the genotype; however there was a genotype ×
drug interaction (p<0.05). Specifically, treatment with MK-801 resulted in greater
disruption of PPI in male Avpr1b −/− mice as compared to Avpr1b +/+ mice (* indicates
p<0.05). Data are expressed as mean + standard error of the mean. (Abbreviations: APO,
apomorphine; AMP, amphetamine; MK-801, dizocilpine)
42
Mean ± Standard Error of the Mean (PPI percentage)

Drugs and Prepulse
Levels
Saline 77 67.33 ± 3.62 57.16 ± 3.89
Saline 80 73.25 ± 3.63 71.90 ± 2.08
Saline 86 78.06 ± 2.47 80.03 ± 1.79
APO 77 51.56 ± 6.30 50.17 ± 5.53
APO 80 66.23 ± 4.03 62.22 ± 4.41
APO 86 65.91 ± 4.11 65.54 ± 4.66
AMP 77 30.11 ± 2.26 39.46 ± 3.98
AMP 80 46.88 ± 5.10 54.62 ± 4.83
AMP 86 54.28 ± 3.62 62.29 ± 3.29
MK-801 77 45.37 ± 3.86 34.62 ± 5.09
MK-801 80 57.80 ± 6.35 42.88 ± 6.20
MK-801 86 73.44 ± 3.55 54.57 ± 7.72
Table 4: Mean and standard error of the mean for drug treatments on male Avpr1b +/+
and Avpr1b −/− mice. (Abbreviations: APO, apomorphine; AMP, amphetamine; MK-
801, dizocilpine)
43
There was a main effect of the drug on startle amplitude (F1,14 = 13.842, p<0.05), with
MK-801 increasing the startle amplitude. There was no main effect of genotype, or any
interactions on the startle amplitude (Table 5).
PPI Percentage
Baseline: There was no genotypic difference between Avpr1b +/+ and Avpr1b −/− female
mice in baseline PPI percentage. However, there was a main effect of prepulse intensity
(F2,26 = 14.393, p< 0.05), with higher prepulse tone resulting in greater PPI percentage
(Figure 14).
Drug treatment: There was a main effect of the drug treatment (F1,14 = 13.095, p<0.05)
and prepulse intensity (F2,28 = 34.016, p<0.05). There was no genotypic effect or any
interactions in Avpr1b +/+ and Avpr1b −/− female mice (Figure 15) and (Table 6).
Experiment 2B: Assessment of prepulse inhibition of startle reflex in Avpr1b
+/+ and Avpr1b −/− male mice following administration of MK-801
Habituation of startle reflex
Male Avpr1b +/+ and Avpr1b −/− mice did not show habituation to startle reflex across
the test sessions with either saline or MK-801 (Figure 16).
Startle amplitude
There was a main effect of the drug (MK-801) on startle amplitude, (F1,15 = 5.587,
p<0.05), with MK-801 increasing the amplitude of the startle reflex. There was no
44
Saline MK-801
300 300
* *
Pulse 1 Pulse 1
200 200
Startle Amplitude
Startle Amplitude
150 150
100 100
50 50
0 0
Figure 13: Habituation of startle in Avpr1b +/+ and Avpr1b −/− female mice. There was
no habituation of startle between PULSE 1 and PULSE 2 trials with saline (0.9%) and
MK-801 (0.7mg/kg) in both genotypes. Data are expressed as mean + standard error of
the mean. (Abbreviations: MK-801, dizocilpine)
45
Saline 49.94 ± 6.60 56.84 ± 11.46
MK-801* 95.06 ± 22.44 93.43 ± 18.18
Table 5: Startle amplitude in female mice following drug treatment. There was a main
effect of the drug on startle amplitude (p<0.05), with MK-801 increasing the startle
amplitude in both the genotypes. There was no genotypic difference between the Avpr1b
+/+ and Avpr1b −/− mice. Startle was calculated from the averages responses from the
first and last pulse alone (120dB) trials. (*p < 0.05 indicating a main effect of drug
compared to saline) (Abbreviations: MK-801, dizocilpine)
46
Avpr1b +/+
100
Avpr1b −/−
*
*
80
60
40
20
0
3dB 6dB 12dB
Prepulse Intensities
dBA above background
Figure 14: Baseline PPI percentage in females. There was no genotypic difference
between Avpr1b +/+ and Avpr1b −/− females, but an effect of prepulse intensity was
present (p<0.05), with higher prepulse tone resulting in greater PPI percentage. (*
indicates p<0.05). Data are expressed as mean + standard error of the mean.
47
Avpr1b +/+
100
3dB
6dB
12dB
% Prepulse Inhibtion 80
60
40
20
0
SALINE MK801
Avpr1b −/−
100 3dB
6dB
12dB
80
60
40
20
0
SALINE MK801
Figure 15: PPI percentage following drug treatment in females. There was a main effect
of the drug (p<0.05) with MK-801 disrupting the PPI in both Avpr1b +/+ and Avpr1b
−/− female mice. However, there was no genotypic difference in PPI between Avpr1b +/+
and Avpr1b −/− females. Data are expressed as mean + standard error of the mean.
(Abbreviations: MK-801, dizocilpine)
48
Mean ± Standard Error of the Mean (PPI Percentage)

Drugs and Prepulse
Levels
Saline 77 64.24 ± 6.57 67.29 ± 4.05
Saline 80 71.46 ± 4.33 72.62 ± 3.25
Saline 86 78.86 ± 2.49 78.43 ± 2.98
MK-801 77 34.72 ± 7.63 46.68 ± 6.31
MK-801 80 51.19 ± 7.54 58.20 ± 6.63
MK-801 86 60.58 ± 4.53 65.38 ± 6.63
Table 6: Mean and standard error of the mean for drug treatment in female Avpr1b +/+
and Avpr1b −/− mice. (Abbreviations: MK-801, dizocilpine)
49
genotypic difference between Avpr1b +/+ and Avpr1b −/− male mice in startle amplitude
(Table 7).
PPI Percentage
Baseline: As expected there was a main effect of the prepulse intensity (F2,32 = 17.968,
p<0.05), but there was no genotypic difference (Figure 17).
Drug Treatment: There was a main effect of the drug treatment (F1,17 = 20.883, p<0.05)
with MK-801 disrupting the PPI as compared to saline in both Avpr1b +/+ and Avpr1b
−/− male mice. There was also a main effect of the prepulse intensity (F2, 34 = 71.671,
p<0.05), but there was no main effect of the genotype or any other interactions (Figure
18) and (Table 8).
Experiment 3: Assessment of prepulse inhibition of startle reflex in male Avpr1b +/+
and Avpr1b −/− mice following administration of MK-801, PCP and saline
There was no habituation to startle reflex across the test trials PULSE 1 and PULSE 2 in
both Avpr1b +/+ and Avpr1b −/− males (Figure 19).
Startle amplitude
There was a main effect of the drug on startle amplitude (F2,28 = 10.803, p<0.05), with
MK-801 increasing the startle amplitude and PCP decreasing the startle amplitude.
50
Saline MK-801
300 300
* *
Pulse 1 Pulse 1
200 200
Startle Amplitude
Startle Amplitude
150 150
100 100
50 50
0 0
Figure 16: Habituation of startle reflex in males. There was no main effect of saline
(0.9%) and MK-801 (0.7mg/kg) on habituation of startle across test trials PULSE 1 and
PULSE 2 on male Avpr1b +/+ and Avpr1b −/− mice. Data are expressed as mean +
standard error of the mean. (Abbreviations: MK-801, dizocilpine)
51
Mean ± Standard Error of the Mean (Startle Amplitude)
Saline 58.00 ± 4.02 73.01 ± 8.09
MK-801* 110.33 ± 21.32 109.30 ± 20.55
Table 7: Effects of drugs on startle amplitude on males. Treatment with MK-801 resulted
in an increase in the startle amplitude as compared to saline in both Avpr1b +/+ and
Avpr1b −/− male mice (p<0.05). There was no genotypic difference between the Avpr1b
+/+ and Avpr1b −/− male mice. Startle was calculated from the averages responses from
the first and last pulse alone (120dB) trials. (*p < 0.05 indicating a main effect of drug
compared to saline) (Abbreviations: MK-801, dizocilpine)
52
Avpr1b +/+
100 *
Avpr1b −/−
*
80
60
40
20
0
3dB 6dB 12dB
Figure 17: Baseline PPI percentage in males. There was a main effect of the prepulse
intensity (p<0.05); with increased PPI percentage with higher decibel prepulse tone.
However, there was no effect of the genotype on baseline PPI percentage in male Avpr1b
+/+ and Avpr1b −/− mice. (* indicates p<0.05). Data are expressed as mean + standard
error of the mean.
53
Avpr1b +/+
100 3dB
6dB
12dB
80
60
40
20
0
SALINE MK801
Avpr1b −/−
100 3dB
6dB
12dB
80
60
40
20
0
SALINE MK801
Figure 18: PPI percentage following drug treatment in males. There was a main effect of
the drug (p<0.05), with MK-801 disrupting the PPI in both Avpr1b +/+ and Avpr1b
−/− male mice. However, there was no genotypic difference in PPI percentage between
Avpr1b +/+ and Avpr1b −/− males. Data are expressed as mean + standard error of the
mean. (Abbreviations: MK-801, dizocilpine)
54
Mean ± Standard Error of the Mean (PPI Percentage)
Saline 77 72.11 ± 5.88 58.29 ± 7.90
Saline 80 77.75 ± 3.78 64.66 ± 8.37
Saline 86 86.59 ± 1.58 77.40 ± 4.11
MK-801 77 48.53 ± 4.47 30.29 ± 6.78
MK-801 80 63.12 ± 5.46 51.21 ± 6.07
MK-801 86 72.03 ± 4.19 56.27 ± 6.40
Table 8: Mean and standard error of the mean of PPI percentage following drug treatment
in Avpr1b +/+ and Avpr1b −/− males. (Abbreviations: MK-801, dizocilpine)
55
However there was no main effect of genotype or any other interactions on Avpr1b+/+
and Avpr1b −/− mice (Table 9).
PPI Percentage
Baseline: There was a main effect of prepulse intensity (F2,28 = 24.079, p<0.05), with
greater PPI percentage with higher decibel prepulse tone. There was no main effect of the
genotype on baseline PPI percentage (Figure 20).
Drug Treatment: There was a main effect of drug treatment (F2,28 = 4.659, p<0.05) with
both MK-801 and PCP disrupting the PPI in Avpr1b +/+ and Avpr1b −/− mice as
compared to saline. There was also a main effect of the prepulse intensity (F2,28 =
105.553, p<0.05). However there was no genotypic effect or any other interactions
(Figure 21) (Table 10).
Experiment 4A: Gene expression of NMDAR1 in Avpr1b +/+ and Avpr1b −/− male
mice
In this experiment, expression of NMDAR1 was examined in cortex, striatum and
hippocampus in Avpr1b +/+ and Avpr1b −/− male mice. There were no genotypic
differences in mRNA levels of NMDAR1 receptors in cortex (p=0.365), striatum
(p=0.083) or hippocampus (p=0.137) (Figure 22).
Experiment 4B: Gene expression of NMDAR2A in Avpr1b +/+ and Avpr1b −/− male
mice
56
Saline
300
*
Pulse 1
250 Pulse 2
200
Startle Amplitude 150
100
50
0
MK-801 PCP
300 300
* *
Pulse 1 Pulse 1
200 200
Startle Amplitude
Startle Amplitude
150 150
100 100
50 50
0 0
Figure 19: Habituation of startle in males. There was no habituation of startle between
PULSE 1 and PULSE 2 in both Avpr1b +/+ and Avpr1b −/− males on treatment with
saline (0.9%), MK-801 (0.7mg/kg) and PCP (1.2mg/kg). Data are expressed as mean +
standard error of the mean. (Abbreviations: MK-801, dizocilpine; PCP, phencyclidine)
57
Saline 90.83 ± 7.11 68.51 ± 8.31
MK-801* 103.83 ± 16.36 143.71 ± 17.83
PCP* 85.91 ± 13.20 96.14 ± 11.53
Table 9: Effects of drugs on startle amplitude on males. There was a main effect of the
drugs on startle amplitude (p<0.05) with MK-801 increasing the startle amplitude and
PCP decreasing the startle amplitude. There was no genotypic difference between the
Avpr1b +/+ and Avpr1b −/− mice. Startle was calculated from the averages responses
from the first and last pulse alone (120dB) trials. (*p < 0.05 indicating a main effect of
drug compared to saline) (Abbreviations: MK-801, dizocilpine; PCP, phencyclidine)
58
Avpr1b +/+
100
Avpr1b −/−
*
80 *
60
40
20
0
3dB 6dB 12dB
Figure 20: Baseline PPI percentage in males. There was no genotypic difference between
male Avpr1b +/+ and Avpr1b −/− mice in baseline PPI percentage. There was the
expected main effect of prepulse intensity with greater PPI percentage with higher
decibel prepulse tone (* indicates p<0.05). Data are expressed as mean and standard error
of the mean.
59
Avpr1b +/+
100 3dB
6dB
12dB
80
60
40
20
0
SALINE MK-801 PCP
Avpr1b −/−
100 3dB
6dB
12dB
80
60
40
20
0
SALINE MK-801 PCP
Figure 21: PPI percentage following drug treatment in males. Treatment with MK-80 and
PCP resulted in disruption of PPI as compared to saline in both Avpr1b +/+ and Avpr1b
−/− mice (p<0.05). However there was no genotypic difference in PPI percentage. Data
are expressed as mean + standard error of the mean. (Abbreviations: MK-801,
dizocilpine; PCP, phencyclidine)
60
Mean ± Standard Error of the Mean

Drugs and Prepulse
Levels
Saline 77 57.89 ± 6.61 51.75 ± 6.45
Saline 80 70.53 ± 4.36 63.39 ± 4.59
Saline 86 78.39 ± 2.50 73.78 ± 5.38
MK-801 77 51.97 ± 6.27 35.92 ± 9.39
MK-801 80 63.81 ± 6.03 47.16 ± 7.83
MK-801 86 69.41 ± 7.37 60.94 ± 7.48
PCP 77 41.95 ± 6.00 39.72 ± 8.23
PCP 80 50.63 ± 6.00 53.48 ± 6.96
PCP 86 67.80 ± 4.93 62.27 ± 7.62
Table 10: Mean and stand error of the mean for drug treatment on PPI percentage in male
Avpr1b +/+ and Avpr1b −/− mice. (Abbreviations: MK-801, dizocilpine; PCP,
phencyclidine)
61
In this experiment, expression of NMDAR2A was evaluated in cortex, striatum and
hippocampus of Avpr1b +/+ and Avpr1b −/− male mice. Avpr1b −/− mice had
significantly higher mRNA levels of NMDAR2A receptors as compared to Avpr1b +/+
mice in the hippocampus (p= 0.003). No differences in NMDAR2A levels were found in
cortex (p=0.897) and striatum (p=0.527) (Figure 23).

62
200 p=0.14
Avpr1b +/+
Avpr1b −/−
150
Relative Expression
p=0.08
100
50
0
Cortex Striatum Hippocampus
Figure 22: Expression of NMDAR1 in male Avpr1b +/+ and Avpr1b −/− mice. There
were no genotypic differences in mRNA levels of NMDAR1 in cortex, striatum and
hippocampus. Data are expressed as mean + standard error of the mean.
63
**
200
Avpr1b +/+
Avpr1b −/−
150
Relative Expression
100
50
0
Cortex Striatum Hippocampus
Figure 23: Expression of NMDAR2A in male Avpr1b +/+ and Avpr1b −/− mice. Avpr1b
−/− males had significantly higher mRNA levels of NMDAR2A expression in the
hippocampus as compared to Avpr1b +/+ mice (** indicates p<0.01). There were no
genotypic differences in NMDAR2A expression in cortex and striatum. Data are
expressed as mean + standard error the mean.
Discussion
The work described in this thesis has helped elucidate the role of the Avpr1b in
the neural circuitry that underlies sensorimotor gating. In Experiments 1A and 2A, we
examined the effect of various PPI-disrupting drugs on PPI and habituation in female
Avpr1b +/+ and Avpr1b −/− mice. It was found that treatment with APO, AMP, MK-801,
all resulted in disruption of PPI in both Avpr1b +/+ and Avpr1b −/− females. However,
no genotype or genotype × drug differences were observed between Avpr1b +/+ and
Avpr1b −/− females. Our results are consistent with many other studies where no
genotypic differences have been found in PPI in females despite its presence in males.
Further, given the lack of effect in females, we did not include them in Experiment 3.
In Experiment 1B, we found that treatment with MK-801, but not APO and AMP,
resulted in greater disruption of PPI in male Avpr1b −/− mice as compared to male
Avpr1b +/+ mice, indicating that male Avpr1b −/− mice are more susceptible to the PPI-
disrupting effects of MK-801. As MK-801 is a selective NMDA receptor antagonist
(Wong et al., 1986), these data suggest that male Avpr1b −/− mice have an impairment in
their glutamatergic system.
Experiment 2B was performed to verify the results of Experiment 1B, though,
only two drugs were used, MK-801 and saline. We found that in male Avpr1b +/+ and
Avpr1b −/−mice, there were no genotypic differences in PPI percentage following
64
65
treatment with MK-801. Although the differences in PPI percentage between the
genotypes were not statistically significant, they showed a similar trend as in Experiment
1B; with male Avpr1b −/− mice having an inclination for greater PPI disruption by MK-
801 as compared to male Avpr1b +/+ mice (p=0.19).
In Experiment 3, we wanted to repeat aspects of Experiment 1 and 2 and add
another NMDA antagonist, PCP, which has a slightly different pharmacology than MK-
801. PCP is the most commonly used NMDA receptor antagonist to study PPI (Geyer et
al., 2001 and the references within). However, PCP binds to receptors other than NMDA
receptors, such as sigma opioid receptors (Contreras et al., 1988; Largent et al., 1986;
Sonders et al., 1988),which are known to further facilitate glutamate transmission (Su and
Hayashi, 2003). It was found that PCP produced a stronger disruption of PPI than MK-
801 in both Avpr1b +/+ and Avpr1b −/− mice. However, contrary to our hypothesis, there
were no drug specific genotypic differences.
Based on the finding from Experiment 1B, we decided to explore possible
changes in the glutamatergic system in male Avpr1b −/− mice in Experiment 4, using
quantitative real time PCR to quantify mRNA levels of the NMDA receptors subunits
NMDAR1 and NMDAR2A. Interestingly, we found that the NMDAR2A subunit is
increased in the hippocampus of Avpr1b −/− mice as compared to Avpr1b +/+ mice.
The lack of differences in PPI between Avpr1b +/+ and Avpr1b −/− females in
Experiment 1A and 2A was not surprising since there are other studies which have found
a differential response of females to these drugs as compared to males. The lack of a

66
genotypic effect in females may be due to the variability in the hormonal milieu
throughout the estrous cycle. This idea is supported by one study in humans, which found
that there are sex differences in PPI, with women having lower PPI than males
(Swerdlow et al., 1993). Further, there is evidence that cyclic changes in gonadal steroids,
in particular estrogens, can affect PPI across the menstrual cycle (Swerdlow et al., 1997).
These data from humans are consistent with data in female rats, that show that PPI varies
across the estrous cycle (Aurebach et al., 1997, (Koch, 1998). In a number of other
studies, estrogens have been reported to influence monoamines in brain regions which are
important for regulation of PPI (Becker, 1990; Joyce and van, 1984; Thompson and
Moss, 1997; Van and Joyce, 1986). These hormonal effects of estrogens on PPI in
females make it difficult to interpret the results seen in our study. It would be interesting
to investigate PPI in ovarectomized female mice treated with estradiol.
The data in Experiment 2B and 3 did not support our findings from Experiment
1B, where a genotypic difference was found in Avpr1b males following treatment with
MK-801. The lack of genotypic difference in PPI percentage with MK-801 and PCP seen
in Experiment 2B and 3, could be due to a number of reasons. First, in Experiment 2B, by
using just two drugs instead of four as in Experiment 1B, we might have eliminated the
possible drug interactions which could have resulted in a significant difference. Second, a
careful analysis of the PPI data in Experiment 2B revealed that saline treatment resulted
in lower PPI in Avpr1b −/− mice as compared to Avpr1b +/+ mice. Since the control
itself resulted in differences between Avpr1b +/+ and Avpr1b −/− males, it is possible
that the disruption of PPI that was due to MK-801 did not appear as robust. This
67
genotypic difference in PPI following saline treatment between Avpr1b +/+ and Avpr1b
−/− mice has not been observed in any previous experiments and we are unclear as to
why this is happening. We are hopeful that a repeat of this study will shed some light
onto these aspects. In Experiment 3, we are fairly confident that our lack of effect in
Avpr1b −/− mice is due to a decrease in the potency and efficacy of drugs in disrupting
PPI. The drugs used in this Experiment were at least a year old and it is likely that there
was a decrease in their effects due to their breakdown over time. This hypothesis is
supported by the fact that the disruptions of PPI by MK-801 and PCP were not as
profound as that observed in previous Experiments. Another reason that there were not
drug-specific genotypic differences in PPI may be due to the smaller numbers of animals
that were tested. It is likely that a repeat of this study with a larger sample size and using
freshly prepared drugs would reproduce the results of Experiment 1B.
With regards to startle magnitude, in Experiment 1B, the genotypic difference in
PPI percentage does not seem to be due to changes in the startle magnitude. So far, the
relationship between startle and PPI is not well understood. One study compared the
prepulse and startle magnitude in 12 different inbreed mouse strains and found no
association between the two; while another study with 20 different strains found a
positive correlation between the two (Paylor and Crawley, 1997; Logue et al., 1997). Our
present findings suggest that the Avpr1b plays a role in regulation of PPI by interacting
with the glutamatergic system.

68
Data suggesting a link between glutamate and PPI has been established by a
number of studies. Across species, treatment with PCP and MK-801 has been shown to
disrupt PPI through antagonism of NMDA receptors (Caldwell et al., 2009; Jerram et al.,
1996; Yee et al., 2004). The lack of genotypic difference in PPI following treatment with
AMP and APO in Experiment 1B suggests that the dopaminergic system is not involved
in the regulation of PPI in Avpr1b −/− male mice. However, it is naïve to think that an
altered glutamatergic system is solely responsible for the deficient sensorimotor gating
observed in Avpr1b −/− mice following treatment with MK-801. In a number of studies,
it has been shown that there is a great degree of crosstalk between the various
neurotransmitters- glutamate, dopamine and serotonin (Carlsson et al., 2000; Carlsson et
al., 1999). It has been reported that drugs that block NMDA receptors not only increase
the release of glutamate, but also dopamine and serotonin (margos-Bosch et al., 2006;
Martin et al., 1998; Mathe et al., 1999; Moghaddam et al., 1997; Schmidt and Fadayel,
1996). Further in rats and mice, PPI deficits induced by PCP and MK-801, which are
specific NMDA antagonists, can be effectively reversed by pretreatment with atypical
antipsychotics such as clozapine and olanzapine, which act on many different receptors
(Bubenikova et al., 2005). Additionally, there are few reports which show that the release
of serotonin and dopamine, due to administration of MK-801 or PCP, can be effectively
suppressed by atypical neuroleptics (Lopez-Gil et al., 2007). Hence it is possible that
altered functioning of glutamate receptors in Avpr1b −/− mice may lead to deficits in PPI
through some indirect mechanism involving other neurotransmitters, such as by

69
increasing the levels of dopamine. Further studies will be required to explore the
relationship of different neurotransmitters and their effects on PPI.
Our findings from Experiment 4 support our hypothesis that the glutamatergic
system is altered in male Avpr1b −/− mice. The increases in the NMDA receptor subunit
NMDAR2A within the hippocampus are consistent with Avpr1b localization in the brain,
with the highest density of Avpr1b in the CA2 region of hippocampus (Young et al.,
2006). Thus, the developmental lack of Avpr1b could alter the PPI neurocircuitry by
changing the levels of expression of NMDA receptor subunits in this area. Although the
levels of NMDAR1 did not differ significantly between Avpr1b +/+ and Avpr1b −/−
mice, the level of NMDAR1 was decreased in the striatum in Avpr1b −/− mice as
compared to Avpr1b +/+ (P=0.08). We are currently quantifying these same subunits
(NMDAR1 and NMDAR2A) in Avpr1b +/+ and Avpr1b −/− mice by western blot
analysis to determine whether these observed changes in mRNA result in changes in
protein levels as well.
Currently, hypo-functioning of the NMDA receptor is believed to partly
contribute to the PPI deficits seen in patients with schizophrenia or mood disorders.
However, in individuals with psychiatric disorders characterized by PPI deficits, the data
regarding NMDA transcripts within the brain are highly inconsistent. In one study by
Gao and colleagues (2000), they reported an increase in NMDAR2A and decrease in
NMDAR1 transcripts in the hippocampus of schizophrenic patients as compared to
healthy controls, with no changes in protein levels. Conversely, other studies of NMDA
70
receptors in individuals diagnosed with either bipolar disorder or schizophrenia have
shown decreases in NMDAR1 and NMDAR2A transcripts in hippocampus
(McCullumsmith et al., 2007). As NMDA receptors are tetrameric and composed of two
obligatory NMDAR1 subunits and either two facultative NMDAR2 (A-D) or NMDAR3
(A-B) subunits (Moriyoshi et al., 1991; Sugihara et al., 1992; Paoletti and Neyton, 2007),
it is likely that absolute numbers may not be particularly telling. Rather, the ratios of
subunits within the receptors are more likely to be altered in these diseased states which
in turn would result in altered functioning. Therefore, further studies are required to
confirm these results and explore their functional significance.
While the role of glutamate in PPI is well known, the interaction of Avp and
glutamate is still under investigation. In a couple of studies, it has been found that MK-
801 abolishes Avp-induced increases in memory, suggesting that Avp does interact with
NMDA receptors in the process of memory formation (Artemowicz and Wisniewski,
1998; Wisniewski et al., 1996). A recent study also reported that Avp increases
glutamate release in the astrocytes of the hippocampus and cortex (Syed et al., 2007).
This effect on hippocampal astrocytes was shown to be specifically mediated via Avpr1b
receptors. Given all these interactions between Avp and glutamate, it would be interesting
to examine the levels of glutamate within the hippocampus of Avpr1b −/− mice by
microdialysis. It can be speculated that the similar to the mRNA levels of NMDA
receptors, the amount of glutamate would also be different between Avpr1b +/+ and
Avpr1b −/− mice.

71
There is another line of Avpr1b −/− mice that were engineered by another
research group. Contrary to the work from this thesis, in this line there are reported
baseline differences in PPI between Avpr1b +/+ and Avpr1b −/− mice (Egashira et al.,
2005). There are several reasons why these inconsistencies may exist. First, the mice used
in the above study were hybrids between 129Sv and C57BL/6J from F3 to F5
generations, whereas the mice used our study were fully backcrossed into the C57BL/6J
background. This is significant because there are reports which indicate that PPI differs
between the different inbred mouse strains (Logue et al., 1997). Thus, it is possible that
the differences in the background could have resulted in differences in the results seen
with PPI. A second difference between the two studies is the technique of creating the
Avpr1b −/− mice (Tanoue et al., 2004; Wersinger et al., 2002). It can be speculated that
the differences in generation of knockout could have differently altered other systems as
well which resulted in discrepancy in the results.
Nevertheless, the study with two different Avpr1b −/− mice as well as BB-Ho rats
suggests that Avp and Avpr1b do play a role in the regulation of sensorimotor gating.
Interestingly, PPI deficits in Avpr1b −/− mice and BB-Ho rats are effectively reversed by
atypical antipsychotics (Feifel et al., 2004; Egashira et al., 2005). These deficits might be
sensitive to some particular property of these atypical neuroleptics. It would be
interesting to see if we can reverse the deficits with atypical antipsychotics.
One of caveat of our study is that Avpr1b −/− mice completely lack the Avpr1b
since the time of conception. This developmental lack of Avpr1b could have resulted in
72
some compensatory changes in these animals. It is also possible that all the results seen
are due to some compensatory changes due to developmental lack of Avpr1b and not
primarily due to its absence. The same reason also makes it difficult to extrapolate the
results for humans in the pathogenesis of neuropsychiatric disorders. Hence, while our
studies strongly suggest the role of Avpr1b in regulating sensorimotor gating; its
translational implications for humans need to be explored further.
Overall conclusions
In summary, our studies have provided evidence for the role of Avpr1b in the
regulation of sensorimotor gating. However, lack of Avpr1b by itself is not sufficient to
cause deficits in sensorimotor gating. We have demonstrated that the glutamatergic
system is altered in Avpr1b −/− mice, which might be contributing to the PPI deficits
found in Experiment 1B. Thus, we hypothesize that under altered glutamatergic signaling
following treatment with psychotomimetics, the lack of the Avpr1b increases the
susceptibility to develop deficits in sensorimotor gating. However, since glutamate and
Avp are both known to interact with other neurotransmitters systems, it is likely these
may also be altered in these mice. Therefore, further research will be required to explore
how and where Avpr1b is interacting with these neurotransmitter systems to regulate PPI.
In order to further explore the relationship of glutamate and Avp, it would be
interesting to examine the changes in NMDA receptors after acute and chronic
administration of MK-801 and PCP in Avpr1b +/+ and Avpr1b −/− mice. We could also
examine other aspects of glutamatergic system such as measuring the glutamate levels in
73
specific brain regions at baseline and following administration of psychotics to look for
changes.
In conclusion, our study further strengthened the evidence for the role of central
Avp system in pathogenesis of neuropsychiatric disorders. With more evidence emerging
implicating Avp in regulation of social behavior and its alteration in many diseases states,
further research could open new avenues for development of new treatment options.
Reference List
Abduljawad,K.A., Langley,R.W., Bradshaw,C.M., and Szabadi,E. (1998). Effects of

bromocriptine and haloperidol on prepulse inhibition of the acoustic startle response in
man. J. Psychopharmacol. 12, 239-245.
Abduljawad,K.A., Langley,R.W., Bradshaw,C.M., and Szabadi,E. (1999). Effects of

bromocriptine and haloperidol on prepulse inhibition: comparison of the acoustic startle
eyeblink response and the N1/P2 auditory-evoked response in man. J. Psychopharmacol.
13, 3-9.
Acher,R., Chauvet,J., and Chauvet,M.T. (1995). Man and the chimaera. Selective versus
neutral oxytocin evolution. Adv. Exp. Med. Biol. 395, 615-627.
Altemus,M., Pigott,T., Kalogeras,K.T., Demitrack,M., Dubbert,B., Murphy,D.L., and

Gold,P.W. (1992). Abnormalities in the regulation of vasopressin and corticotropin
releasing factor secretion in obsessive-compulsive disorder. Arch. Gen. Psychiatry 49, 9-
20.
Altemus,M., Swedo,S.E., Leonard,H.L., Richter,D., Rubinow,D.R., Potter,W.Z., and

Rapoport,J.L. (1994). Changes in cerebrospinal fluid neurochemistry during treatment of
obsessive-compulsive disorder with clomipramine. Arch. Gen. Psychiatry 51, 794-803.
Artemowicz,B. and Wisniewski,K. (1998). Role of NMDA receptor in the effects of

arginine-vasopressin on memory processes. Pol. J. Pharmacol. 50, 5-14.
Bankir,L. (2001). Antidiuretic action of vasopressin: quantitative aspects and interaction

between V1a and V2 receptor-mediated effects. Cardiovasc. Res. 51, 372-390.
Becker,J.B. (1990). Estrogen rapidly potentiates amphetamine-induced striatal dopamine

release and rotational behavior during microdialysis. Neurosci. Lett. 118, 169-171.
Beckwith,B.E., Couk,D.I., and Till,T.S. (1983). Vasopressin analog influences the

performance of males on a reaction time task. Peptides 4, 707-709.
Bielsky,I.F., Hu,S.B., Szegda,K.L., Westphal,H., and Young,L.J. (2004). Profound

impairment in social recognition and reduction in anxiety-like behavior in vasopressin
V1a receptor knockout mice. Neuropsychopharmacology 29, 483-493.
74
75
Blanchard,R.J., Griebel,G., Farrokhi,C., Markham,C., Yang,M., and Blanchard,D.C.

(2005). AVP V1b selective antagonist SSR149415 blocks aggressive behaviors in
hamsters. Pharmacol. Biochem. Behav. 80, 189-194.
Bohus,B. and de,W.D. (1998). The vasopressin deficient Brattleboro rats: a natural
knockout model used in the search for CNS effects of vasopressin. Prog. Brain Res. 119,
555-573.
Bolino,F., Di,M., V, Di,C.L., Manna,V., Daneluzzo,E., and Casacchia,M. (1994).

Sensorimotor gating and habituation evoked by electro-cutaneous stimulation in
schizophrenia. Biol. Psychiatry 36, 670-679.
Born,J., Pietrowsky,R., and Fehm,H.L. (1998). Neuropsychological effects of vasopressin

in healthy humans. Prog. Brain Res. 119, 619-643.
Braff,D., Stone,C., Callaway,E., Geyer,M., Glick,I., and Bali,L. (1978). Prestimulus

effects on human startle reflex in normals and schizophrenics. Psychophysiology 15, 339-
343.
Braff,D.L., Grillon,C., and Geyer,M.A. (1992). Gating and habituation of the startle
reflex in schizophrenic patients. Arch. Gen. Psychiatry 49, 206-215.
Braff,D.L., Swerdlow,N.R., and Geyer,M.A. (1999). Symptom correlates of prepulse

inhibition deficits in male schizophrenic patients. Am. J. Psychiatry 156, 596-602.
Brambilla,F., Aguglia,E., Massironi,R., Maggioni,M., Grillo,W., Castiglioni,R.,

Catalano,M., and Drago,F. (1986). Neuropeptide therapies in chronic schizophrenia:
TRH and vasopressin administration. Neuropsychobiology 15, 114-121.
Brambilla,F., Bondiolotti,G.P., Maggioni,M., Sciascia,A., Grillo,W., Sanna,F., Latina,A.,

and Picotti,G.B. (1989). Vasopressin (DDAVP) therapy in chronic schizophrenia: effects
on negative symptoms and memory. Neuropsychobiology 20, 113-119.
Brown,J.S., Kalish,H.I., and Farber,I.E. (1951). Conditioned fear as revealed by

magnitude of startle response to an auditory stimulus. J. Exp. Psychol. 41, 317-328.
Bubenikova,V., Votava,M., Horacek,J., Palenicek,T., and Dockery,C. (2005). The effect

of zotepine, risperidone, clozapine and olanzapine on MK-801-disrupted sensorimotor
gating. Pharmacol. Biochem. Behav. 80, 591-596.
Buijs,R.M. (1978). Intra- and extrahypothalamic vasopressin and oxytocin pathways in

the rat. Pathways to the limbic system, medulla oblongata and spinal cord. Cell Tissue
Res. 192, 423-435.
76
Caffe,A.R., Van Ryen,P.C., Van der Woude,T.P., and Van Leeuwen,F.W. (1989).
Vasopressin and oxytocin systems in the brain and upper spinal cord of Macaca
fascicularis. J. Comp Neurol. 287, 302-325.
Caldwell,H.K. and Albers,H.E. (2004). Effect of photoperiod on vasopressin-induced

aggression in Syrian hamsters. Horm. Behav. 46, 444-449.
Caldwell,H.K., Lee,H.J., Macbeth,A.H., and Young,W.S., III (2008). Vasopressin:

behavioral roles of an "original" neuropeptide. Prog. Neurobiol. 84, 1-24.
Caldwell,H.K., Stephens,S.L., and Young,W.S., III (2009). Oxytocin as a natural

antipsychotic: a study using oxytocin knockout mice. Mol. Psychiatry 14, 190-196.
Carlsson,A., Hansson,L.O., Waters,N., and Carlsson,M.L. (1997). Neurotransmitter

aberrations in schizophrenia: new perspectives and therapeutic implications. Life Sci. 61,
75-94.
Carlsson,A., Waters,N., and Carlsson,M.L. (1999). Neurotransmitter interactions in

schizophrenia-therapeutic implications. Eur. Arch. Psychiatry Clin. Neurosci. 249 Suppl
4, 37-43.
Carlsson,A., Waters,N., Waters,S., and Carlsson,M.L. (2000). Network interactions in

schizophrenia - therapeutic implications. Brain Res. Brain Res. Rev. 31, 342-349.
Castellanos,F.X., Fine,E.J., Kaysen,D., Marsh,W.L., Rapoport,J.L., and Hallett,M.

(1996). Sensorimotor gating in boys with Tourette's syndrome and ADHD: preliminary
results. Biol. Psychiatry 39, 33-41.
Chakrabarty,K., Bhattacharyya,S., Christopher,R., and Khanna,S. (2005). Glutamatergic

dysfunction in OCD. Neuropsychopharmacology 30, 1735-1740.
Cho,M.M., DeVries,A.C., Williams,J.R., and Carter,C.S. (1999). The effects of oxytocin

and vasopressin on partner preferences in male and female prairie voles (Microtus
ochrogaster). Behav. Neurosci. 113, 1071-1079.
Contreras,P.C., Contreras,M.L., O'Donohue,T.L., and Lair,C.C. (1988). Biochemical and

behavioral effects of sigma and PCP ligands. Synapse 2, 240-243.
Curzon,P. and Decker,M.W. (1998). Effects of phencyclidine (PCP) and (+)MK-801 on

sensorimotor gating in CD-1 mice. Prog. Neuropsychopharmacol. Biol. Psychiatry 22,
129-146.
Davis,M., Gendelman,D.S., Tischler,M.D., and Gendelman,P.M. (1982). A primary

acoustic startle circuit: lesion and stimulation studies. J. Neurosci. 2, 791-805.
77
de Kloet,C.S., Vermetten,E., Geuze,E., Wiegant,V.M., and Westenberg,H.G. (2008).

Elevated plasma arginine vasopressin levels in veterans with posttraumatic stress
disorder. J. Psychiatr. Res. 42, 192-198.
De Vries,G.J., Best,W., and Sluiter,A.A. (1983). The influence of androgens on the

development of a sex difference in the vasopressinergic innervation of the rat lateral
septum. Brain Res. 284, 377-380.
De Vries,G.J. and Buijs,R.M. (1983). The origin of the vasopressinergic and

oxytocinergic innervation of the rat brain with special reference to the lateral septum.
Brain Res. 273, 307-317.
de Vries,G.J., Buijs,R.M., and Sluiter,A.A. (1984). Gonadal hormone actions on the

morphology of the vasopressinergic innervation of the adult rat brain. Brain Res. 298,
141-145.
De,B., Gold,P.W., Geracioti,T.D., Jr., Listwak,S.J., and Kling,M.A. (1993). Association

of fluoxetine treatment with reductions in CSF concentrations of corticotropin-releasing
hormone and arginine vasopressin in patients with major depression. Am. J. Psychiatry
150, 656-657.
Dempster,E.L., Burcescu,I., Wigg,K., Kiss,E., Baji,I., Gadoros,J., Tamas,Z.,

Kennedy,J.L., Vetro,A., Kovacs,M., and Barr,C.L. (2007). Evidence of an association
between the vasopressin V1b receptor gene (AVPR1B) and childhood-onset mood
disorders. Arch. Gen. Psychiatry 64, 1189-1195.
Dhakar M.B., Stevenson E.L., and Caldwell H.K. (in press). The interplay between
oxytocin, vasopressin and gonadal steroids. In Oxytocin, Vasopressin and Related
Peptides in the Regulation of Behavior, Elena Choleris, Donald Pfaff, and Martin
Cavaliers, eds. Cambridge University Press).
Dogterom,J., van Wimersma Greidanus,T.B., and De,W.D. (1978). Vasopressin in

cerebrospinal fluid and plasma of man, dog, and rat. Am. J. Physiol 234, E463-E467.
Donaldson,Z.R. and Young,L.J. (2008). Oxytocin, vasopressin, and the neurogenetics of

sociality. Science 322, 900-904.
Dubois-Dauphin,M., Tribollet,E., and Dreifuss,J.J. (1987). A sexually dimorphic

vasopressin innervation of auditory pathways in the guinea pig brain. Brain Res. 437,
151-156.
Dubois-Dauphin,M., Tribollet,E., and Dreifuss,J.J. (1989). Distribution of

neurohypophysial peptides in the guinea pig brain. I. An immunocytochemical study of
the vasopressin-related glycopeptide. Brain Res. 496, 45-65.
78
Dulawa,S.C. and Geyer,M.A. (1996). Psychopharmacology of prepulse inhibition in

mice. Chin J. Physiol 39, 139-146.
Egashira,N., Tanoue,A., Higashihara,F., Fuchigami,H., Sano,K., Mishima,K., Fukue,Y.,

Nagai,H., Takano,Y., Tsujimoto,G., Stemmelin,J., Griebel,G., Iwasaki,K., Ikeda,T.,
Nishimura,R., and Fujiwara,M. (2005). Disruption of the prepulse inhibition of the startle
reflex in vasopressin V1b receptor knockout mice: reversal by antipsychotic drugs.
Neuropsychopharmacology 30, 1996-2005.
Elman,I., Lukas,S., Shoaf,S.E., Rott,D., Adler,C., and Breier,A. (2003). Effects of acute
metabolic stress on the peripheral vasopressinergic system in schizophrenia. J.
Psychopharmacol. 17, 317-323.
Engelmann,M. and Landgraf,R. (1994). Microdialysis administration of vasopressin into

the septum improves social recognition in Brattleboro rats. Physiol Behav. 55, 145-149.
Feenstra,M.G., Snijdewint,F.G., Van,G.H., and Boer,G.J. (1990). Widespread alterations

in central noradrenaline, dopamine, and serotonin systems in the Brattleboro rat not
related to the local absence of vasopressin. Neurochem. Res. 15, 283-288.
Feifel,D., Melendez,G., Priebe,K., and Shilling,P.D. (2007). The effects of chronic

administration of established and putative antipsychotics on natural prepulse inhibition
deficits in Brattleboro rats. Behav. Brain Res. 181, 278-286.
Feifel,D., Melendez,G., and Shilling,P.D. (2004). Reversal of sensorimotor gating

deficits in Brattleboro rats by acute administration of clozapine and a neurotensin agonist,
but not haloperidol: a potential predictive model for novel antipsychotic effects.
Feifel,D. and Priebe,K. (2001). Vasopressin-deficient rats exhibit sensorimotor gating

deficits that are reversed by subchronic haloperidol. Biol. Psychiatry 50, 425-433.
Ferris,C.F., Melloni,R.H., Jr., Koppel,G., Perry,K.W., Fuller,R.W., and Delville,Y.

(1997). Vasopressin/serotonin interactions in the anterior hypothalamus control
aggressive behavior in golden hamsters. J. Neurosci. 17, 4331-4340.
Fliers,E., Guldenaar,S.E., van de,W.N., and Swaab,D.F. (1986). Extrahypothalamic

vasopressin and oxytocin in the human brain; presence of vasopressin cells in the bed
nucleus of the stria terminalis. Brain Res. 375, 363-367.
Furuya,Y., Kagaya,T., Ogura,H., and Nishizawa,Y. (1999). Competitive NMDA receptor

antagonists disrupt prepulse inhibition without reduction of startle amplitude in a
dopamine receptor-independent manner in mice. Eur. J. Pharmacol. 364, 133-140.
79
Gao,X.M., Sakai,K., Roberts,R.C., Conley,R.R., Dean,B., and Tamminga,C.A. (2000).

Ionotropic glutamate receptors and expression of N-methyl-D-aspartate receptor subunits
in subregions of human hippocampus: effects of schizophrenia. Am. J. Psychiatry 157,
1141-1149.
Geyer,M.A. and Braff,D.L. (1982). Habituation of the Blink reflex in normals and
schizophrenic patients. Psychophysiology 19, 1-6.
Geyer,M.A., Krebs-Thomson,K., Braff,D.L., and Swerdlow,N.R. (2001).

Pharmacological studies of prepulse inhibition models of sensorimotor gating deficits in
schizophrenia: a decade in review. Psychopharmacology (Berl) 156, 117-154.
Graham,F.K. (1975). Presidential Address, 1974. The more or less startling effects of
weak prestimulation. Psychophysiology 12, 238-248.
Griebel,G., Simiand,J., Serradeil-Le,G.C., Wagnon,J., Pascal,M., Scatton,B.,

Maffrand,J.P., and Soubrie,P. (2002). Anxiolytic- and antidepressant-like effects of the
non-peptide vasopressin V1b receptor antagonist, SSR149415, suggest an innovative
approach for the treatment of stress-related disorders. Proc. Natl. Acad. Sci. U. S. A 99,
6370-6375.
Griebel,G., Simiand,J., Stemmelin,J., Gal,C.S., and Steinberg,R. (2003). The vasopressin

V1b receptor as a therapeutic target in stress-related disorders. Curr. Drug Targets. CNS.
Neurol. Disord. 2, 191-200.
Grillon,C., Ameli,R., Charney,D.S., Krystal,J., and Braff,D. (1992). Startle gating deficits
occur across prepulse intensities in schizophrenic patients. Biol. Psychiatry 32, 939-943.
Grillon,C., Falls,W.A., Ameli,R., and Davis,M. (1994). Safety signals and human
anxiety: a fear-potentiated startle study. Anxiety. 1, 13-21.
Hoffman,H.S. and Searle,J.L. (1968). Acoustic and temporal factors in the evocation of
startle. J. Acoust. Soc. Am. 43, 269-282.
Hutchison,K.E. and Swift,R. (1999). Effect of d-amphetamine on prepulse inhibition of

the startle reflex in humans. Psychopharmacology (Berl) 143, 394-400.
Iager,A.C., Kirch,D.G., Bigelow,L.B., and Karson,C.N. (1986). Treatment of

schizophrenia with a vasopressin analogue. Am. J. Psychiatry 143, 375-377.
Inder,W.J., Donald,R.A., Prickett,T.C., Frampton,C.M., Sullivan,P.F., Mulder,R.T., and

Joyce,P.R. (1997). Arginine vasopressin is associated with hypercortisolemia and suicide
attempts in depression. Biol. Psychiatry 42, 744-747.
80
Insel,T.R. and Young,L.J. (2000). Neuropeptides and the evolution of social behavior.
Curr. Opin. Neurobiol. 10, 784-789.
Ison,J.R., McAdam,D.W., and Hammond,G.R. (1973). Latency and amplitude changes in

the acoustic startle reflex of the rat produced by variation in auditory prestimulation.
Physiol Behav. 10, 1035-1039.
Jard,S., Barberis,C., Audigier,S., and Tribollet,E. (1987). Neurohypophyseal hormone

receptor systems in brain and periphery. Prog. Brain Res. 72, 173-187.
Javitt,D.C. (1987). Negative schizophrenic symptomatology and the PCP (phencyclidine)

model of schizophrenia. Hillside. J. Clin. Psychiatry 9, 12-35.
Jerram,A.H., Smith,P.F., and Darlington,C.L. (1996). A dose-response analysis of the

behavioral effects of (+)MK-801 in guinea pig: comparison with CPP. Pharmacol.
Biochem. Behav. 53, 799-807.
Johnson,A.E., Audigier,S., Rossi,F., Jard,S., Tribollet,E., and Barberis,C. (1993).

Localization and characterization of vasopressin binding sites in the rat brain using an
iodinated linear AVP antagonist. Brain Res. 622, 9-16.
Joyce,J.N. and van,H.C. (1984). Behaviors induced by intrastriatal dopamine vary

independently across the estrous cycle. Pharmacol. Biochem. Behav. 20, 551-557.
Koch,M. (1998). Sensorimotor gating changes across the estrous cycle in female rats.
Physiol Behav. 64, 625-628.
Koch,M., Kungel,M., and Herbert,H. (1993). Cholinergic neurons in the

pedunculopontine tegmental nucleus are involved in the mediation of prepulse inhibition
of the acoustic startle response in the rat. Exp. Brain Res. 97, 71-82.
Koch,M. and Schnitzler,H.U. (1997). The acoustic startle response in rats--circuits

mediating evocation, inhibition and potentiation. Behav. Brain Res. 89, 35-49.
Korsgaard,S., Casey,D.E., mgaard Pedersen,N.E., Jorgensen,A., and Gerlach,J. (1981).

Vasopressin in anergic schizophrenia. A cross-over study with lysine-8-vasopressin and
placebo. Psychopharmacology (Berl) 74, 379-382.
Kumari,V., Mulligan,O.F., Cotter,P.A., Poon,L., Toone,B.K., Checkley,S.A., and

Gray,J.A. (1998). Effects of single oral administrations of haloperidol and d-
amphetamine on prepulse inhibition of the acoustic startle reflex in healthy male
volunteers. Behav. Pharmacol. 9, 567-576.
81
Kumari,V., Soni,W., Mathew,V.M., and Sharma,T. (2000). Prepulse inhibition of the

startle response in men with schizophrenia: effects of age of onset of illness, symptoms,
and medication. Arch. Gen. Psychiatry 57, 609-614.
Kumari,V., Soni,W., and Sharma,T. (1999). Normalization of information processing

deficits in schizophrenia with clozapine. Am. J. Psychiatry 156, 1046-1051.
Largent,B.L., Gundlach,A.L., and Snyder,S.H. (1986). Pharmacological and

autoradiographic discrimination of sigma and phencyclidine receptor binding sites in
brain with (+)-[3H]SKF 10,047, (+)-[3H]-3-[3-hydroxyphenyl]-N-(1-propyl)piperidine
and [3H]-1-[1-(2-thienyl)cyclohexyl]piperidine. J. Pharmacol. Exp. Ther. 238, 739-748.
Laycock,J.F., Gartside,I.B., and Chapman,J.T. (1983). A comparison of the learning

abilities of Brattleboro rats with hereditary diabetes insipidus and Long-Evans rats using
positively reinforced operant conditioning. Prog. Brain Res. 60, 183-187.
Lee,H.J., Macbeth,A.H., Pagani,J.H., and Young,W.S., III (2009). Oxytocin: the great
facilitator of life. Prog. Neurobiol. 88, 127-151.
Linn,G.S., O'Keeffe,R.T., Schroeder,C.E., Lifshitz,K., and Javitt,D.C. (1999). Behavioral

effects of chronic phencyclidine in monkeys. Neuroreport 10, 2789-2793.
Logue,S.F., Owen,E.H., Rasmussen,D.L., and Wehner,J.M. (1997). Assessment of

locomotor activity, acoustic and tactile startle, and prepulse inhibition of startle in inbred
mouse strains and F1 hybrids: implications of genetic background for single gene and
quantitative trait loci analyses. Neuroscience 80, 1075-1086.
Lolait,S.J., O'Carroll,A.M., Mahan,L.C., Felder,C.C., Button,D.C., Young,W.S., III,

Mezey,E., and Brownstein,M.J. (1995). Extrapituitary expression of the rat V1b
vasopressin receptor gene. Proc. Natl. Acad. Sci. U. S. A 92, 6783-6787.
Lolait,S.J., O'Carroll,A.M., McBride,O.W., Konig,M., Morel,A., and Brownstein,M.J.

(1992). Cloning and characterization of a vasopressin V2 receptor and possible link to
nephrogenic diabetes insipidus. Nature 357, 336-339.
Lolait,S.J., Stewart,L.Q., Roper,J.A., Harrison,G., Jessop,D.S., Young,W.S., III, and

O'Carroll,A.M. (2007). Attenuated stress response to acute lipopolysaccharide challenge
and ethanol administration in vasopressin V1b receptor knockout mice. J.
Neuroendocrinol. 19, 543-551.
Lopez-Gil,X., Babot,Z., margos-Bosch,M., Sunol,C., Artigas,F., and Adell,A. (2007).

Clozapine and haloperidol differently suppress the MK-801-increased glutamatergic and
serotonergic transmission in the medial prefrontal cortex of the rat.
82
Mansbach,R.S. and Geyer,M.A. (1989). Effects of phencyclidine and phencyclidine

biologs on sensorimotor gating in the rat. Neuropsychopharmacology 2, 299-308.
Mansbach,R.S., Geyer,M.A., and Braff,D.L. (1988). Dopaminergic stimulation disrupts

sensorimotor gating in the rat. Psychopharmacology (Berl) 94, 507-514.
margos-Bosch,M., Lopez-Gil,X., Artigas,F., and Adell,A. (2006). Clozapine and

olanzapine, but not haloperidol, suppress serotonin efflux in the medial prefrontal cortex
elicited by phencyclidine and ketamine. Int. J. Neuropsychopharmacol. 9, 565-573.
Martin,P., Carlsson,M.L., and Hjorth,S. (1998). Systemic PCP treatment elevates brain
extracellular 5-HT: a microdialysis study in awake rats. Neuroreport 9, 2985-2988.
Mathe,J.M., Nomikos,G.G., Blakeman,K.H., and Svensson,T.H. (1999). Differential

actions of dizocilpine (MK-801) on the mesolimbic and mesocortical dopamine systems:
role of neuronal activity. Neuropharmacology 38, 121-128.
McCullumsmith,R.E., Kristiansen,L.V., Beneyto,M., Scarr,E., Dean,B., and Meador-

Woodruff,J.H. (2007). Decreased NR1, NR2A, and SAP102 transcript expression in the
hippocampus in bipolar disorder. Brain Res. 1127, 108-118.
Mcghie,A. and Chapman,J. (1961). Disorders of attention and perception in early

schizophrenia. Br. J. Med. Psychol. 34, 103-116.
Meynen,G., Unmehopa,U.A., van Heerikhuize,J.J., Hofman,M.A., Swaab,D.F., and

Hoogendijk,W.J. (2006). Increased arginine vasopressin mRNA expression in the human
hypothalamus in depression: A preliminary report. Biol. Psychiatry 60, 892-895.
Michell,R.H., Kirk,C.J., and Billah,M.M. (1979). Hormonal stimulation of

phosphatidylinositol breakdown with particular reference to the hepatic effects of
vasopressin. Biochem. Soc. Trans. 7, 861-865.
Millan,M.J., Millan,M.H., Czlonkowski,A., and Herz,A. (1984). Vasopressin and

oxytocin in the rat spinal cord: distribution and origins in comparison to [Met]enkephalin,
dynorphin and related opioids and their irresponsiveness to stimuli modulating
neurohypophyseal secretion. Neuroscience 13, 179-187.
Moghaddam,B., Adams,B., Verma,A., and Daly,D. (1997). Activation of glutamatergic

neurotransmission by ketamine: a novel step in the pathway from NMDA receptor
blockade to dopaminergic and cognitive disruptions associated with the prefrontal cortex.
J. Neurosci. 17, 2921-2927.
Moriyoshi,K., Masu,M., Ishii,T., Shigemoto,R., Mizuno,N., and Nakanishi,S. (1991).

Molecular cloning and characterization of the rat NMDA receptor. Nature 354, 31-37.
83
Nishimura,H. and Fan,Z. (2003). Regulation of water movement across vertebrate renal
tubules. Comp Biochem. Physiol A Mol. Integr. Physiol 136, 479-498.
Olney,J.W., Newcomer,J.W., and Farber,N.B. (1999). NMDA receptor hypofunction

model of schizophrenia. J. Psychiatr. Res. 33, 523-533.
Ornitz,E.M., Hanna,G.L., and de,T.J. (1992). Prestimulation-induced startle modulation

in attention-deficit hyperactivity disorder and nocturnal enuresis. Psychophysiology 29,
437-451.
Paoletti,P. and Neyton,J. (2007). NMDA receptor subunits: function and pharmacology.
Curr. Opin. Pharmacol. 7, 39-47.
Paylor,R. and Crawley,J.N. (1997). Inbred strain differences in prepulse inhibition of the
mouse startle response. Psychopharmacology (Berl) 132, 169-180.
Ralph,R.J., Paulus,M.P., and Geyer,M.A. (2001). Strain-specific effects of amphetamine

on prepulse inhibition and patterns of locomotor behavior in mice. J. Pharmacol. Exp.
Ther. 298, 148-155.
Ralph,R.J., Varty,G.B., Kelly,M.A., Wang,Y.M., Caron,M.G., Rubinstein,M.,

Grandy,D.K., Low,M.J., and Geyer,M.A. (1999). The dopamine D2, but not D3 or D4,
receptor subtype is essential for the disruption of prepulse inhibition produced by
amphetamine in mice. J. Neurosci. 19, 4627-4633.
Raskind,M.A., Courtney,N., Murburg,M.M., Backus,F.I., Bokan,J.A., Ries,R.K.,

Dorsa,D.M., and Weitzman,R.E. (1987). Antipsychotic drugs and plasma vasopressin in
normals and acute schizophrenic patients. Biol. Psychiatry 22, 453-462.
Ross,H.E. and Young,L.J. (2009). Oxytocin and the neural mechanisms regulating social
cognition and affiliative behavior. Front Neuroendocrinol. 30, 534-547.
Saito,M., Sugimoto,T., Tahara,A., and Kawashima,H. (1995). Molecular cloning and

characterization of rat V1b vasopressin receptor: evidence for its expression in extra-
pituitary tissues. Biochem. Biophys. Res. Commun. 212, 751-757.
Schall,U., Schon,A., Zerbin,D., Eggers,C., and Oades,R.D. (1996). Event-related

potentials during an auditory discrimination with prepulse inhibition in patients with
schizophrenia, obsessive-compulsive disorder and healthy subjects. Int. J. Neurosci. 84,
15-33.
Schmidt,C.J. and Fadayel,G.M. (1996). Regional effects of MK-801 on dopamine

release: effects of competitive NMDA or 5-HT2A receptor blockade. J. Pharmacol. Exp.
Ther. 277, 1541-1549.
84
Skuse,D.H. and Gallagher,L. (2009). Dopaminergic-neuropeptide interactions in the

social brain. Trends Cogn Sci. 13, 27-35.
Sofroniew,M.V. (1983). Morphology of vasopressin and oxytocin neurones and their

central and vascular projections. Prog. Brain Res. 60, 101-114.
Sokolowski,M.B. (2010). Social interactions in "simple" model systems. Neuron 65, 780-
794.
Sonders,M.S., Keana,J.F., and Weber,E. (1988). Phencyclidine and psychotomimetic

sigma opiates: recent insights into their biochemical and physiological sites of action.
Trends Neurosci. 11, 37-40.
Stemmelin,J., Lukovic,L., Salome,N., and Griebel,G. (2005). Evidence that the lateral
septum is involved in the antidepressant-like effects of the vasopressin V1b receptor
antagonist, SSR149415. Neuropsychopharmacology 30, 35-42.
Su,T.P. and Hayashi,T. (2003). Understanding the molecular mechanism of sigma-1

receptors: towards a hypothesis that sigma-1 receptors are intracellular amplifiers for
signal transduction. Curr. Med. Chem. 10, 2073-2080.
Sugihara,H., Moriyoshi,K., Ishii,T., Masu,M., and Nakanishi,S. (1992). Structures and

properties of seven isoforms of the NMDA receptor generated by alternative splicing.
Biochem. Biophys. Res. Commun. 185, 826-832.
Swerdlow,N.R., Bakshi,V., and Geyer,M.A. (1996). Seroquel restores sensorimotor

gating in phencyclidine-treated rats. J. Pharmacol. Exp. Ther. 279, 1290-1299.
Swerdlow,N.R., Bakshi,V., Waikar,M., Taaid,N., and Geyer,M.A. (1998). Seroquel,

clozapine and chlorpromazine restore sensorimotor gating in ketamine-treated rats.
Psychopharmacology (Berl) 140, 75-80.
Swerdlow,N.R., Benbow,C.H., Zisook,S., Geyer,M.A., and Braff,D.L. (1993). A

preliminary assessment of sensorimotor gating in patients with obsessive compulsive
disorder. Biol. Psychiatry 33, 298-301.
Swerdlow,N.R., Braff,D.L., and Geyer,M.A. (1999). Cross-species studies of

sensorimotor gating of the startle reflex. Ann. N. Y. Acad. Sci. 877, 202-216.
Swerdlow,N.R. and Geyer,M.A. (1998). Using an animal model of deficient sensorimotor

gating to study the pathophysiology and new treatments of schizophrenia. Schizophr.
Bull. 24, 285-301.
85
Swerdlow,N.R., Geyer,M.A., and Braff,D.L. (2001). Neural circuit regulation of prepulse

inhibition of startle in the rat: current knowledge and future challenges.
Psychopharmacology (Berl) 156, 194-215.
Swerdlow,N.R., Hartman,P.L., and Auerbach,P.P. (1997). Changes in sensorimotor

inhibition across the menstrual cycle: implications for neuropsychiatric disorders. Biol.
Psychiatry 41, 452-460.
Swerdlow,N.R., Martinez,Z.A., Hanlon,F.M., Platten,A., Farid,M., Auerbach,P.,

Braff,D.L., and Geyer,M.A. (2000). Toward understanding the biology of a complex
phenotype: rat strain and substrain differences in the sensorimotor gating-disruptive
effects of dopamine agonists. J. Neurosci. 20, 4325-4336.
Swerdlow,N.R., Paulsen,J., Braff,D.L., Butters,N., Geyer,M.A., and Swenson,M.R.

(1995). Impaired prepulse inhibition of acoustic and tactile startle response in patients
with Huntington's disease. J. Neurol. Neurosurg. Psychiatry 58, 192-200.
Syed,N., Martens,C.A., and Hsu,W.H. (2007). Arginine vasopressin increases glutamate

release and intracellular Ca2+ concentration in hippocampal and cortical astrocytes
through two distinct receptors. J. Neurochem. 103, 229-237.
Tanoue,A., Ito,S., Honda,K., Oshikawa,S., Kitagawa,Y., Koshimizu,T.A., Mori,T., and

Tsujimoto,G. (2004). The vasopressin V1b receptor critically regulates hypothalamic-
pituitary-adrenal axis activity under both stress and resting conditions. J. Clin. Invest 113,
302-309.
Thompson,T.L. and Moss,R.L. (1997). Modulation of mesolimbic dopaminergic activity

over the rat estrous cycle. Neurosci. Lett. 229, 145-148.
Tribollet,E., Barberis,C., and Arsenijevic,Y. (1997). Distribution of vasopressin and

oxytocin receptors in the rat spinal cord: sex-related differences and effect of castration in
pudendal motor nuclei. Neuroscience 78, 499-509.
Tribollet,E., Barberis,C., Dreifuss,J.J., and Jard,S. (1988). Autoradiographic localization

of vasopressin and oxytocin binding sites in rat kidney. Kidney Int. 33, 959-965.
Vaccari,C., Lolait,S.J., and Ostrowski,N.L. (1998). Comparative distribution of

vasopressin V1b and oxytocin receptor messenger ribonucleic acids in brain.
Endocrinology 139, 5015-5033.
Van,H.C. and Joyce,J.N. (1986). Effects of estrogen on the basal ganglia. Neurosci.
Biobehav. Rev. 10, 1-14.
86
van,L.L., Goekoop,J.G., van Kempen,G.M., Frankhuijzen-Sierevogel,A.C.,

Wiegant,V.M., van,d., V, and De,W.D. (1997). Plasma levels of arginine vasopressin
elevated in patients with major depression. Neuropsychopharmacology 17, 284-292.
van,W.D., Del-Favero,J., Aulchenko,Y., Oswald,P., Souery,D., Forsgren,T., Sluijs,S.,

Bel-Kacem,S., Adolfsson,R., Mendlewicz,J., Van,D.C., Deboutte,D., Van,B.C., and
Claes,S. (2004). A major SNP haplotype of the arginine vasopressin 1B receptor protects
against recurrent major depression. Mol. Psychiatry 9, 287-292.
Venables,P.H. (1960). The effect of auditory and visual stimulation on the skin potential
response of schizophrenics. Brain 83, 77-92.
Wang,Z., Ferris,C.F., and De Vries,G.J. (1994). Role of septal vasopressin innervation in

paternal behavior in prairie voles (Microtus ochrogaster). Proc. Natl. Acad. Sci. U. S. A
91, 400-404.
Wang,Z., Toloczko,D., Young,L.J., Moody,K., Newman,J.D., and Insel,T.R. (1997).

Vasopressin in the forebrain of common marmosets (Callithrix jacchus): studies with in
situ hybridization, immunocytochemistry and receptor autoradiography. Brain Res. 768,
147-156.
Weike,A.I., Bauer,U., and Hamm,A.O. (2000). Effective neuroleptic medication removes

prepulse inhibition deficits in schizophrenia patients. Biol. Psychiatry 47, 61-70.
Wersinger,S.R., Caldwell,H.K., Christiansen,M., and Young,W.S., III (2007). Disruption

of the vasopressin 1b receptor gene impairs the attack component of aggressive behavior
in mice. Genes Brain Behav. 6, 653-660.
Wersinger,S.R., Ginns,E.I., O'Carroll,A.M., Lolait,S.J., and Young,W.S., III (2002).

Vasopressin V1b receptor knockout reduces aggressive behavior in male mice. Mol.
Psychiatry 7, 975-984.
Wersinger,S.R., Kelliher,K.R., Zufall,F., Lolait,S.J., O'Carroll,A.M., and Young,W.S., III

(2004). Social motivation is reduced in vasopressin 1b receptor null mice despite normal
performance in an olfactory discrimination task. Horm. Behav. 46, 638-645.
Wersinger,S.R., Temple,J.L., Caldwell,H.K., and Young,W.S., III (2008). Inactivation of

the oxytocin and the vasopressin (Avp) 1b receptor genes, but not the Avp 1a receptor
gene, differentially impairs the Bruce effect in laboratory mice (Mus musculus).
Endocrinology 149, 116-121.
Williams,A.R., Carey,R.J., and Miller,M. (1985). Altered emotionality of the

vasopressin-deficient Brattleboro rat. Peptides 6 Suppl 1, 69-76.
87
Williams,A.R., Carey,R.J., and Miller,M. (1983). Behavioral differences between

vasopressin-deficient (Brattleboro) and normal Long-Evans rats. Peptides 4, 711-716.
Winslow,J.T., Hastings,N., Carter,C.S., Harbaugh,C.R., and Insel,T.R. (1993). A role for

central vasopressin in pair bonding in monogamous prairie voles. Nature 365, 545-548.
Wisniewski,K., Artemowicz,B., and Lutostanska,A. (1996). The estimation of

interactions between arginine-vasopressin (AVP) and NMDA receptors in memory and
learning processes. Acta Physiol Hung. 84, 477-479.
Wong,E.H., Kemp,J.A., Priestley,T., Knight,A.R., Woodruff,G.N., and Iversen,L.L.

(1986). The anticonvulsant MK-801 is a potent N-methyl-D-aspartate antagonist. Proc.
Natl. Acad. Sci. U. S. A 83, 7104-7108.
Yamada,S., Harano,M., Annoh,N., Nakamura,K., and Tanaka,M. (1999). Involvement of

serotonin 2A receptors in phencyclidine-induced disruption of prepulse inhibition of the
acoustic startle in rats. Biol. Psychiatry 46, 832-838.
Yee,B.K., Chang,D.L., and Feldon,J. (2004). The Effects of dizocilpine and

phencyclidine on prepulse inhibition of the acoustic startle reflex and on prepulse-elicited
reactivity in C57BL6 mice. Neuropsychopharmacology 29, 1865-1877.
Young,L.J., Wang,Z., Cooper,T.T., and Albers,H.E. (2000). Vasopressin (V1a) receptor

binding, mRNA expression and transcriptional regulation by androgen in the Syrian
hamster brain. J. Neuroendocrinol. 12, 1179-1185.
Young,W.S., Li,J., Wersinger,S.R., and Palkovits,M. (2006). The vasopressin 1b receptor

is prominent in the hippocampal area CA2 where it is unaffected by restraint stress or
adrenalectomy. Neuroscience 143, 1031-1039.
APPENDIX I
PILOT STUDY
Objective: To determine the optimum dose of drug to disrupt the PPI in both male and
female mice
Methods: 6 female and 6 male heterozygous (Avpr1b +/−) animals of ages 3-6 months
were used for the study. All the testing was conducted in the light phase of the 12:12 light
dark cycle. APO, AMP and MK-801 were in the following doses; APO (10mg/kg), AMP
(10mg/kg) and (12mg/kg), MK-801 (0.7mg/kg). All the drugs were dissolved in 0.9%
saline. Saline was given in the same volume as APO and AMP and served as control.
Animals were tested as per the protocol mentioned in the methods section. PPI
percentage was calculated using the following equation: (1−(startle amplitude of the
PREPULSE + PULSE/startle amplitude of the PULSE ALONE)) × 100. PPI percentage
was determined using repeated ANOVA and main effects of drug treatment, prepulse
intensity and their interactions were compared. Data from both males and females were
collapsed and analyzed together.
Results:
PPI percentage: There was a main effect of the drug treatment with APO (10mg/kg) (F1,9
= 6.005, P<0.05) and MK-801(0.7mg/kg) (F1,9= 16.384, P<0.05) all resulting in
88
89
disruption of PPI as compared to saline. There was also a main effect of prepulse
intensity (F2,18 = 58.048, P<0.001) with higher decibel tone resulting in increased PPI
percentage. However, treatment with AMP (10mg/kg) did not result in significant
disruption of PPI. Hence the experiment was repeated with an increased dose of AMP-
12mg/kg. Treatment with AMP (12mg/kg) resulted in significant disruption of PPI (F1,9=
11.220, P<0.05) as compared to saline (P<0.05).
Conclusions:
APO (10mg/kg), AMP (12mg/kg) and MK-801 (0.7mg/kg) resulted in significant
disruption of PPI as compared to saline in Avpr1b +/− mice. The above mentioned doses
were selected for conducting experiments 1, 2, and 3.

90
100 3dB
6dB
12dB
a b
80
60 3DB
6DB
12 DB
Col 8
40
20
0
0.9% 10mg/kg 10mg/kg 0.7mg/kg
Figure 24: PPI percentage in Avpr1b +/− mice following drug treatment: Treatment with
APO (a), and MK-801 (b) resulted in disruption of PPI as compared to saline (P<0.05).
There was also a main effect of prepulse intensity, with higher decibel tone resulting in
increased PPI percentage. Treatment with AMP at 10mg/kg did not result in significant
disruption of PPI as compared to saline. Data are expressed as means and standard error
of means. (Abbreviations: APO, apomorphine; AMP, amphetamine; MK-801,
dizocilpine).
91
100 * 3dB
6dB
12dB
80
60 3DB
6DB
12 DB
Col 8
40
20
0
SALINE AMP
0.9% 12mg/kg
Figure 25: PPI percentage in Avpr1b +/− mice following treatment with AMP: Treatment
with AMP (12mg/kg) resulted in disruption of PPI as compared to saline (P<0.05). Data
are expressed as means and standard error of means. (Abbreviations: AMP,
amphetamine)
APPENDIX II
PPI Program used in Experiment 1, 2, and 3
Trial Decibel
1 pulse120 Parameters
2 pulse120
PULSE 1 3 pulse120 5 min acclimation
4 pulse120 Background 74dB
5 pulse120
6 pulse120 Pulse 120- 40 msec duration
7 PPI77
Block 1 8 PPI80 After variable ITI- 12-30 sec
9 PPI86
10 nopulse Prepulse 77- 20 msec duration,
11 PPI80 100 msec wait and pulse 120
12 pulse120
Block 2 Prepulse 80- 20 msec duration,
13 nopulse
100 msec wait and pulse 120
14 PPI77
15 PPI86
Prepulse 86- 20 msec duration,
16 PPI77 100 msec wait and pulse 120
17 Nopulse
Block 3 18 PPI86
19 pulse120
20 PPI80
21 Nopulse
22 PPI86
Block 4 23 PPI80
24 PPI77
25 pulse120
26 PPI86
27 PPI80
Block 5 28 pulse120
29 nopulse
30 PPI77
31 pulse120
32 PPI86
Block 6 33 PPI77
92
93
34 nopulse
35 PPI80
36 PPI77
37 pulse120
Block 7 38 nopulse
39 PPI86
40 PPI80
41 pulse120
42 PPI80
Block 8 43 PPI86
44 nopulse
45 PPI77
46 PPI80
47 nopulse
Block 9 48 PPI86
49 PPI77
50 pulse120
51 nopulse
52 PPI86
Block 10 53 PPI77
54 PPI80
55 pulse120
56 pulse120
57 pulse120
PULSE 2 58 pulse120
59 pulse120
60 pulse120

Kent1302269033 PDF

Загружено:

Сведения о документе

Оригинальное название

Авторское право

Доступные форматы

Поделиться этим документом

Поделиться или встроить документ

Параметры публикации

Этот документ был вам полезен?

Это неприемлемый материал?

Авторское право:

Доступные форматы

Kent1302269033 PDF

Загружено:

Авторское право:

Доступные форматы

THE ROLE OF THE VASOPRESSIN 1B RECEPTOR IN THE REGULATION OF

A thesis submitted to Kent State University

________________________ Dr. H. Caldwell, Advisor

________________________Dr. R. Dorman, Director, School of Biomedical Sciences

________________________Dr. John Stalvey, Dean, College of Arts and Sciences

Sexual dimorphism in the central vasopressin system…………………….3

Vasopressin and social behavior…………………………………………..3

The vasopressin 1b receptor………………………………………………6

Vasopressin 1b receptor knockout mice…………………………………..7

Vasopressin, the vasopressin 1b receptor, and neuropsychiatric disorders.8

Prepulse inhibition of the startle reflex…………………………..11

Neurotransmitter systems regulating prepulse inhibition of the

Interactions of vasopressin and the cortico-striato-pallido-pontine

Prepulse inhibition the startle reflex……………………………………..20

Quantitative real time polymerase chain reaction ……………………….26

Experiment 1A and 1B…………………………………………………...26

Experiment 2A and 2B…………………………………………………...27

Experiment 4A and 4B…………………………………………………..28

Fig 1: Role of vasopressin in regulation of social behaviors……………...4

Fig 2: Schematic diagram of PPI………………………………………...12

Fig 3: CSPP circuit………………………………………………………14

Fig 4: Schematic diagram representing the components of SR-LAB……22

Fig 5: Acoustic startle chamber …………………………………………23

Fig 6: Study design for Experiments 1, 2 and 3………………………….25

Fig7: Experiment 1A Habituation of startle in females………………….32

Fig 8: Experiment 1A Baseline PPI percentage in females ……………..34

Fig 9: Experiment 1A PPI percentage following drug treatment in

Fig 10: Experiment 1B Habituation of startle in males …………………38

Fig 11: Experiment 1B Baseline PPI percentage in males………………40

Fig 12: Experiment 1B PPI percentage following drug treatment in

Fig 13: Experiment 2A Habituation of startle in females…..…………..44

Fig 14: Experiment 2A Baseline PPI percentage in females…………….46

Fig 16: Experiment 2B Habituation of startle in males………………….50

Fig 17: Experiment 2B Baseline PPI percentage in males………………52

Fig 18: Experiment 2B PPI percentage following drug treatment in

Fig 19: Experiment 3 Habituation of startle in males…………………....56

Fig 20: Experiment 3 Baseline PPI percentage in males………………...58

Fig 21: Experiment 3 PPI percentage following drug treatment in

Fig 22: Experiment 4A Expression of NMDAR1 in males………………62

Fig 23: Experiment 4B Expression of NMDAR2A in males……………..63

Table 1: Experiment 1A Startle amplitude in females………………...…33

Table 2: Experiment 1A Mean and SEM for PPI in females…………….36

Table 3: Experiment 1B Startle amplitude in males………………..........39

Table 4: Experiment 1B Mean and SEM for PPI in males………………42

Table 5: Experiment 2A Startle amplitude in females…………………...45

Table 6: Experiment 2A Mean and SEM for PPI in females…………….48

Table 7: Experiment 2B Startle amplitude in males……………………..51

Table 8: Experiment 2B Mean and SEM for PPI in males………………54

Table 9: Experiment 3 Startle amplitude in males……………………….57

Table 10: Experiment 3 Mean and SEM for PPI in males……………….60

Table 11: PPI program…………………………………………………...92

valuable insights and expert advice on my research project.

I would like to extend my gratitude to my friends; Vivek, Priyanka, Sagar and

aggression, reproduction, and communication (Sokolowski, 2010; Insel and Young,

2009). In humans, aberrant social behavior is characteristic of a variety of

neuropsychiatric disorders, such as autism, schizophrenia, obsessive compulsive disorder

A number of hormones and neuropeptides are important to the neural regulation

of social behavior. In particular, two neurohormones, oxytocin (Oxt) and vasopressin

The effectiveness of Oxt and Avp at regulating behavior is attributed to three

effects, and 3) their plasticity (Insel and Young, 2000).

Avp is synthesized as a part of a preprohormone, which is then enzymatically

cleaved to produce the biologically active nonapeptide, a neurophysin, and a

neurons provide robust projections to a number of brain areas, including, hippocampus,