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Applied Animal Behaviour Science 114 (2008) 168181

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Anxiety related behavioural disorders and

neurotransmitters in dogs
Jacopo Riva a,*, Gianpietro Bondiolotti b, Manuela Michelazzi a,
Marina Verga a, Corrado Carenzi a
a

Department of Animal Science, Faculty of Veterinary Medicine, University of Milan, via Celoria 10, 20133 Milan, Italy
b
Department of Pharmacology, Chemotherapy and Toxicology, Faculty of Medicine and Surgery,
University of Milan, via Vanvitelli 32, 20133 Milan, Italy
Accepted 30 January 2008
Available online 28 March 2008

Abstract
The aim of this research has been to verify possible correlations among some neurotransmitters and
anxiety related behavioural disorders in the domestic dog.
Twenty dogs affected by anxiety related behavioural disorders and 13 control dogs were studied. The
dogs behaviour has been analysed through a detailed history case. Blood samples have been analysed by
HPLC method to evaluate the plasma levels of dopamine (DA), norepinephrine (NE), serotonin (5-HT), 5hydroxyindolacetic acid (5-HIAA), 3,4-diidroxyfenilacetic acid (DOPAC), L-DOPA and platelets levels of
NE, DA and 5-HT in the two groups.
Plasma levels of DA and 5-HT are significantly higher in the anxious dogs with respect to the control
ones (DA: ng/mg prot. 0.074  0.071 vs. 0.030  0.001, [P < 0.01] and 5HT: ng/mg prot. 24.95  36.64
vs. 5.94  3.20, [P < 0.01]). The plasma levels of the other neurotransmitters are similar in the two groups.
Platelet levels of NE (ng/mg prot. 0.133  0.047 vs. 0.124  0.061) and DA (ng/mg prot. 0.0552  0.018
vs. 0.074  0.039) are also similar in the two groups. A trend to higher 5-HT platelet levels (ng/mg prot.
509  100 vs. 425  117) in the control dogs compared to the anxious ones has been found.
# 2008 Elsevier B.V. All rights reserved.
Keywords: Anxiety; Dogs; Aggression; Serotonin; Neurotransmitters; Dopamine

1. Introduction
Correlations among behavioural problems and neurotransmitters, especially plasma and
platelets concentrations of serotonin (5-HT), dopamine (DA) and norepinephrine (NE), have
* Corresponding author. Tel.: +39 02 50318028; fax: +39 02 50318030.
E-mail address: riva.jacopo@unimi.it (J. Riva).
0168-1591/$ see front matter # 2008 Elsevier B.V. All rights reserved.
doi:10.1016/j.applanim.2008.01.020

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been found in some species, such as rats, rabbits, humans and dogs (Higley et al., 1992, 1996a;
Gray, 1982; Rogeness et al., 1992; Reisner et al., 1996).
5-HT and catecholamines are involved in the regulation of several behavioural systems that play
an important role in the interaction of the organism with its external environment. DA appears to be
involved in the expression of active behavioural patterns, including aggression, sexual behaviour
(Haney et al., 1990; Rogeness et al., 1984) and locomotion (Kiley-Worthington, 1983). On the
contrary behavioural inhibition of affective aggression seems to be regulated by 5-HT (Spoont,
1992). Moreover, in defensive aggression the threshold for agonistic behaviour is lowered by the
release of catecholamines (Higley et al., 1992). Dynamic changes in central 5-HT levels are thought
to be involved in the establishment and maintenance of dominant attitudes and behaviour (Raleigh
et al., 1980, 1985). These interactions have been found to be a common denominator in a very broad
body of literature (Mann, 1995). For example the correlation between 5-HT and aggressive
behaviour has been experimentally obtained inducing shortages of cerebral 5-HT by lesions of the
serotoninergic cells of rafe in laboratory rats: animals were becoming irritable, reacting in an
exaggerated way to light stimulations (Miczek and Donat, 1989). Other studies highlighted this
correlation: mice that respond to isolation by increased inter-male fighting show reduced 5-HT
function (Valzelli and Garattini, 1968). Similar conclusions were reached by researches on
dominance and aggressiveness in non-human primates where low serotonin levels have been
indicated (Higley et al., 1996b). The results are often contradictory; actually some authors have
shown that increased 5HT was correlated to increased aggression (Brunner et al., 1993; Birmaher
et al., 1990). In dogs, a research on CFS (Cerebro-Spinal-Fluid) monoamine metabolites showed
that CSF 5-HIAA concentration (5-hydroxyindolacetic acid is a metabolite of 5-HT and an indirect
measure of central 5-HT concentration) was lower in dogs diagnosed with dominance related
aggression than in the control ones (Reisner et al., 1996) furthermore a research on nuclear imaging
technology showed that 5HT-2a receptors was significantly higher in the cortical regions in dogs
diagnosed with aggressive behaviour, compared to normal dogs (Peremans et al., 2006).
Moreover, dopaminergic and serotoninergic systems play an important role in the
neurotrasmission system involved in stress response (Puglisi-Allegra and Cabib, 1990; Le
Moal and Simon, 1991).
The knowledge of the involvement of NE in aggressive behaviour has been possible thanks to
the availability of brain tissues of animals which had just fought (Welch and Welch, 1965;
Hendley et al., 1973). An increase in NE has been discovered in the brain of aggressive mice
which had been previously held in a state of isolation (Modigh, 1973); increases in noradrenergic
turnover in the brain of cats or rats were shown in association to biting (Salama and Goldberg,
1973; Reis and Fuxe, 1969).
The aim of this research was to find out the possible differences in some neurotransmitters
(DA, NE, 5-HT, 5-HIAA, L-Dopa and DOPAC) in dogs suffering from anxiety related
behavioural disorders compared to control dogs, not suffering from any behavioural disorder.

2. Materials and methods

2.1. Subjects
The research has been carried out on 20 dogs suffering from anxiety related behavioural disorders and 13
control dogs not suffering from behaviour problems. The 20 problem dogs had been presented at the
University of Milan, Faculty of Veterinary Medicine, Behaviour Clinic and diagnosed as suffering from

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anxiety related disorders after taking case. The behaviours of anxious dogs have been described by their
owners during the behaviour visit. The analysis carried out by the therapist has been done through the
description of the owner, the direct observation in the clinic and the results of the questionnaire. In this way
the therapist has correctly carried out different diagnoses, in order to separate the dogs in the two groups
which have been analysed in this study. The control group did not show any disorder problem. The diagnosis
of anxiety related behavioural disorders was done according to the literature models (Overall, 1997; Beaver,
1983). The behavioural problems included: generalised anxiety, separation anxiety, phobias, compulsive
behaviours and anxiety related aggression. The 13 control dogs were randomly chosen and diagnosed as not
suffering from any behavioural problem according to the same history case.
The dogs age range was 8 month to 9 years of age. Breeds represented in anxious dogs included 3
Labrador Retrievers, 2 Doberman Pinschers, 2 German Shepherds, 3 Giant Schnauzers and one of each of
the following breeds: Brittany Spaniel, Akita-inu (Federation Cynologique Internationale standard), English
Cocker Spaniel, Pitbull, English bulldog and Weimaraner. Four mixed-breed dogs were also included.
Breeds represented in the control group included 2 Boxers, 2 Rottweilers and one of each of the following
breeds: Mastiff, Pitbull, German Shepherd, Labrador Retriever. Five mixed-bred dogs were also included.
Sex represented in anxious dogs included 10 intact males, 6 neutered males, 3 intact females and 1 neutered
female. Sex represented in controls dogs included 6 intact males, 3 neutered males, 4 neutered females and
no intact female.
No dog was taking any drug.
2.2. History case recording
A case history has been completed for each subject, recording the following information on the dog and
on the living environment:
(A) individual features: age of adoption, place of origin (rescue kennel, pure breed kennel, private
breeders);
(B) environmental features: kind of family (single, couple, a numerous family, children), house physical
characteristics, number of walks and length of time, active level when walked, walked on leash or not,
sleeping site, food at disposal, presence of other animals;
(C) behavioural features: escaping from physical handling, submission postures toward the owner, not
accepting to be alone, digging holes, destroying objects, showing aggressiveness signals and/or biting
the owners or strangers or other dogs, not allowing the food to be taken away, asking for physical
contact, emotional induced soiling, supine position, coming back in response to recalls, accepting food
or objects or bone to be taken away, continuously following the owner through the house, reacting to
blaming, house-soiling, chasing other animals, excessively barking, pica (which means the ingestion of
non-nutritive, non-food items, such as fabric, plastics, sticks and rocks) (Overall, 1997), coprophagy,
compulsive behaviours, phobia of crackers (phobia is defined as a profound and quickly developed fear
reaction that does not extinguish with gradual exposure to the object or firework or thunderstorm, or
with exposure over time) (Overall, 1997), even if other authors described systematic desensitization
phobia treatment (Mills, 2002).
2.3. Statistical analysis of behaviours
Answers to the single questions in the case history have been scored and analysed through descriptive
statistical analysis and Chi Square tests. The seldom appearing variables have not been considered in the
analysis. The significance was accepted when p < 0.05.
In order to evaluate the interdependence among the different behaviours a principal component analysis
(PCA with Varimax rotation), has been carried out on the most often exhibited behaviours of the dogs in the
two groups (SPSS, 2003). PCA was used in order to point out the relationships among the different variables.
PCA was considered a suitable method to treat this data set, as it presents a number of advantages. First of

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all, it is a statistical method which condenses into a few latent variables the information contained in many
original variables. Secondly, it offers the possibility of using discrete variables as well as continuous
variables, as it is independent from data distribution; furthermore, the use of PCA with binary variables
allows for qualitative considerations (Jolliffe, 1986; Jackson, 1991; Mattiello et al., 1997).
2.4. Blood analyses
Plasma and platelet concentration levels of DA, NE, 5-HT, 5-HIAA, DOPAC and L-Dopa have been
evaluated in the two groups, according to the methodology described below.
Five ml of blood have been taken with addition of 10% in volume of EDTA (disodiumethylenediaminotetraacetate) already diluted to 1%. The blood has been centrifuged to 120  g (15 min at
20 8C); afterwards the plasma has been centrifuged to 2700  g (15 min at 4 8C) (Da Prada and Picotti,
1979; Bondiolotti et al., 1987). The platelet pellet was washed with saline and analysed with the HPLC
method.
Monoamines and their derivatives were detected and measured using liquid chromatography with
electrochemical detection. For the analysis of the catecholamines, platelet pellets were homogenized with
0.3N perchloric acid and plasma deproteinized (1/1 v/v) with 0.6N perchloric acid containing dihydroxybenzylamine as the internal standard. Catecholamines were extracted onto alumina before the injection.
For the analysis of 5-HT and 5-HIAA, a volume of 100 ml of plasma was deproteinized with an equal volume
of 0.6N perchloric acid containing 3-metoxytyramine as internal standard, 50 ml of supernantant was then
injected in the chromatograph. Evaluation of chromatographed products was accomplished by coulometric
detection.
HPLC separation was performed on a reverse-phase analytical column as described elsewhere (Lollis
et al., 1979; Alleva et al., 1998). The protein content of platelets were assayed using a microassay (Lowry
et al., 1951) with bovine serum albumin as a standard.
Platelet levels of DA, NE and 5-HT could be evaluated only on 5 anxious and in 5 control dogs, due to
problems of haemolysis and of difficult separation between plasma and platelets.
2.5. Statistical analysis of blood variables
Plasma and platelet levels have been statistically analysed according to the data distribution, whether
normal or not. Actually plasma DA and 5-HT have been analysed through Mann-Whitney U-test while LDOPA, DOPAC, and 5-HIAA have been analysed through Student T-test.

3. Results
3.1. Individual environmental and behavioural data
3.1.1. Individual features
3.1.1.1. Age of adoption. 77% of control dogs were adopted in the correct period between 60
and 90 days of age (Overall, 1997), while in the anxious group, late adoptions were more frequent
(P < 0.05). In fact 40% of these latter dogs had been adopted after 3 months of age.
3.1.1.2. Place of origin. A statistically significant difference (P < 0.05) has been found
according to the dogs origin: in fact 35% of anxious subjects had come from a kennel or had been
found by chance, being abandoned, vs. 0% in control. This result agrees with the literature
(Beerda et al., 1999), showing that abandoned or kenneled dogs show behavioural problems more
frequently.

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3.1.2. Environmental features

3.1.2.1. Kind of family. More problem dogs than the control ones (50% vs. 10%; P < 0.05) live
with a couple as against living in a family with children. In anxiety related behavioural disorders
the family dynamics may play a relevant role, which sometimes might interfere also with the
behaviour treatment plans (Dodman et al., 1996; Dehasse, 1997). Moreover sometimes a dog
may be treated by owners like a human baby.
3.1.2.2. Sleeping site. A statistically significant difference (P < 0.005) has been found as far as
the dog sleeping place is concerned. In fact 45% of anxious dogs sleep on the bed/sofa, 45% in a
dog basket and 10% in other places in the house compared with all the control group dogs
sleeping in the dog basket. This result is important because the dog basket must represent a
relaxing and protective place for the dog.
3.1.2.3. Food at disposal. Dogs in the control group have no food always at disposal; on the
contrary 35% of the anxious dogs have continuously food at disposal (P < 0.05). The food
distribution through a series of regulars and rhythmical acts allows to obtain a clear hierarchy
between the dog and its owner. Actually dogs that are pushed in a rhythm by their owners habits,
may get stressed when this rhythm is broken and their expectations frustrated (Horwitz, 2002).
Beagles living in individual kennels with dry food freely available tend to eat three times a day, at
dawn, at dusk and whenever fresh food is given (Rashotte et al., 1984).
3.1.3. Behaviour
Statistically significant differences have been found on the following dogs behaviours.
3.1.3.1. Escaping from physical handling. More anxious dogs (60%) do not accept to be
handled by the owner compared to the control ones (P < 0.05), where only one subject refused to
be handled. This behaviour could be due to wrong rank in the social group hierarchy (Rowell,
1974), leading to ambiguous and unclear dominance-submission relationship between the dog
and the owner, and consequently raising anxiety in the dog. In fact in a dog pack usually members
which most frequently display aggression are the middle rank, rather than the high rank ones
(Lockwood, 1979).
3.1.3.2. Submission posture towards the owner. 90% of control dogs do not show this
behaviour; on the contrary, in the anxious group 77% of the subjects show it (P < 0.001). Visual
canine communication is conventionally described in terms of the signals performed by the wolf
during dominance/submission interactions within the pack (Goodwin et al., 1997). Submission
posture may be displayed in response to a threatening challenge inducing fear and avoidance.
Not accepting to be alone, digging holes, destruction, excessively barking.
High statistically significant differences have been found between anxious and control dogs on
all these anxiety related behaviours (not accepting to be alone 55% in anxious dogs vs. 0% in control
dogs; digging holes 65% in anxious dogs vs. 0% in control dogs; destruction 75% in anxious dogs
vs. 10% in control dogs; excessively barking 65% in anxious dogs vs. 0% in control dogs)
(P < 0.001). These are the most common distress behaviours seen in dogs showing separation
anxiety (McCrave, 1991; Lund and Jorgensen, 1997; Simpson, 2000), although sometimes
destructiveness may be an element of play or exploratory behaviour in young active animals
(Simpson, 2000). Also vocalisation is common and may be due to outside stimuli, social facilitation
with other dogs, territorial displays or play (Horwitz, 1998). However both destructiveness and

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barking become problem behaviours when performed excessively, as shown in anxiety suffering
dogs, mainly in the ones suffering from separation anxiety (Overall, 1997; Houpt et al., 1996).
3.1.3.3. Showing aggressiveness biting owners or strangers. Only the anxious dogs display
aggressive behaviours (85%) towards the owner or towards strangers (P < 0.01). According to
Reisner (2002) anxiety may play an important part in the genesis of aggression, leading to
dangerous attacks also towards human beings. Moreover, aggressive behaviour may appear as a
consequence of affective emotional arousal (Reis, 1974).
3.1.3.4. Phobia of crackers pica compulsive behaviours. Statistically significant differences
have been found between anxious and control dogs also on these anxiety related behaviours
(phobia of cracker 60% in anxious dogs vs. 0% in control dogs; pica 45% in anxious dogs vs. 0%
in control dogs; compulsive behaviours 45% in anxious dogs vs. 0% in control dogs) (P < 0.05
for fear of crackers; P < 0.01 for pica and compulsive behaviours).
Noise phobias may be among the most commonly recognized and exhibited phobic responses
in dogs, though few substantive data exist. As with many fearful, phobic and anxious responses,
noise phobias are characterized by the exhibition of non-specific signs when the dog is presented
with the stimulus or is anticipating the stimulus (Overall, 1997, Overall et al., 2001). Pica may be
exhibited on particular chosen objects in a compulsive way, thus showing a form of compulsive
disorder especially when the dog disregards other activities in favour of stones chewing or
similar activities (Overall, 1997).
The results of the Principal Component Analysis (PCA) performed on the behaviours are
shown in Table 1 and Fig. 1, which include the loadings of the variables explaining the 49.513%
of the total variance on the 1st and 2nd component.
PCA revealed two underlying components. The first component (PC1) shows positive
loadings for the following behaviours: escaping from physical handling, destructiveness, digging
holes and excessively barking and a negative loading for not accepting to be alone (Table 1). The
second component (PC2) shows positive loadings for the following variables: fear of crackers,
pica, compulsive behaviours and a negative loading for the submission posture. Thus the 1st
component identifies the generalised anxiety and separation anxiety problems, while the 2nd
component identifies the anxiety related phobias and compulsive disorders. Each dog has been
plotted on the 2 principal components in Fig. 2. The control dogs are grouped opposite to the
Table 1
Component variance and cumulative variance explained by the 1st and 2nd component
Variables

Components

EH: escaping from physical handling

SP: submission posture
NA: not accepting to be alone
SA: show aggressiveness
DT: destruction
DH: digging holes
EB: excessively barking
FC: phobia of cracker
PC: pica
CB: compulsive behaviours
Variance (%)
Cumulative (%)

0.653
0.281
S0.797
0.292
0.657
0.552
0.776
0.282
0.096
0.036
26.666
26.666

0.299
S0.686
0.068
0.033
0.396
0.294
0.162
0.587
0.790
0.802
22.847
49.513

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Fig. 1. Projection of the loadings of the behavioural variables considered for the first and second principal component.
See Table 1 for the legend.

anxious dogs, particularly for factor 1, thus showing that the two groups are well divided
according to the variables included into the PCA analysis.
3.2. Neurotransmitters
3.2.1. Results in plasma
The plasma concentration values of NE, DA and 5-HT found in this research agree with the
levels shown in the literature by some authors in dogs: in Table 2 the reference average
concentration values of these neurotransmitters in some species, including the dog, are shown, as
well as in Table 3 the average levels of all the neurotransmitters found in this study. In plasma no
reference on dogs have been found as far as 5-HIAA, DOPAC and L-Dopa are concerned.
Statistically significant differences (P < 0.01) have been found between anxious and control
dogs on DA and 5-HT; the anxious dogs showing higher DA and 5-HT plasmatic values
compared to the control dogs (Table 3). However, the levels of the other studied neurotransmitters
do not show any significant difference between the two groups.
3.3. Results in platelets
The platelet levels of 5-HT found in this research also agree with the levels shown in literature
by some authors in dogs (Clagett et al., 1981, 1987; Mezzano et al., 1991). No reference value has

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175

Fig. 2. Plots of scores (anxious and controls dogs) on the first and second principal components. Some subjects in the
control group are overlapping.
Table 2
Plasma concentration values (standard deviation) in different species of norepinephrine (NE), dopamine (DA) and
serotonin (5-HT) according to the literature

Human
Rabbit
Dog
Mice

NE (ng/ml)

DA (ng/ml)

References

5-HT (ng/ml)

References

0.198  0.01
0.392  0.3
0.451  0.046

0.064  0.01
0.216  0.02
0.044  0.004

Da Prada and Picotti (1979)

Da Prada and Picotti (1979)
Roche et al. (2002)

4.7  0.6
6.69  1.70
171  14.6
8.75  0.15

Benedict et al. (1986)

Da Prada and Picotti (1979)
Da Prada and Picotti (1979)
Yamamoto et al. (1986)

Table 3
Blood variables mean plasma values (standard deviation) found in this study
L-DOPA

NE (ng/ml)
Controls
Anxious

0.256  0.087
0.229  0.078a

(ng/ml)
a

1.630  0.587
2.041  0.890a

DOPAC (ng/ml)
a

0.610  0.192
0.767  0.309a

DA (ng/ml)

5-HT (ng/ml)
a

0.030  0.001
0.074  0.071b

5.94  3.20
24.95  36.64b

5-HIAA (ng/ml)
5.59  1.59a
6.68  4.13b

Note: a, b: different superscripts within the same column indicate statistically significant differences (P < 0.01).

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Table 4
Platelet concentration values (standard deviation) of norepinephrine (NA), dopamine (DA) and serotonin (5-HT) in
different species according to the literature
NE (ng/mg)

DA (ng/mg)

References

5-HT (ng/mg)

References

Human

0.241  0.057

0.012  0.001

Picotti et al. (1984)

Mice
Guinea pig
Rabbit
Dog

0.181  0.043
0.263  0.023
0.698  0.13

0.041  0.009
0.035  0.009
0.186  0.127

Picotti et al. (1984)

Picotti et al. (1984)
Picotti et al. (1984)

139  61.6
227  55
211

Maurer-Spurej et al. (2004)

Sagud et al. (2007)
Da Prada and Picotti (1979)

9680
956  126
796  103
626  196

Da Prada and Picotti (1979)

Clagett et al. (1981)
Clagett et al. (1987)
Mezzano et al. (1991)

Table 5
Platelets amine concentration mean values (standard deviation) found in this study

Controls
Anxious

NE (ng/mg)

DA (ng/mg)

5-HT (ng/mg)

0.133  0.047
0.124  0.061

0.0552  0.018
0.074  0.039

509  100
425  117

been found in the literature on DA and NE platelet values in dogs: in Table 4 the reference levels
of DA-NE in some species, and 5-HT in the dog are shown, for comparison with the average
concentration values of platelet DA, NE and 5-HT found in this study (Table 5). The platelet DA
and NE values are similar in the anxious compared to the control dogs, while a trend to higher
values of 5-HT has been found in control dogs.
Peripheral measures of 5-HT activity may reflect central 5-HT functioning despite the fact that
the major portion of 5-HT in the periphery is derived from the gut (Coccaro et al., 1997).
4. Discussion
5-HT seem to inhibit affective aggression, while DA seems to enhance it (Spoont, 1992;
Dodman and Shuster, 1998). Increased DA and indices of DA synthesis and turnover in the whole
brain of laboratory mice that had just fought have been shown (Modigh, 1973), especially in
nucleus accumbens, striatum, frontal cortex and hypothalamus (Haney et al., 1990; Tizabi et al.,
1979; Hutchins et al., 1975). Moreover, plasma DA levels increase together with the sympathetic
activation (Van Loon, 1983). In children showing high affective aggressiveness and
impulsiveness a reduction of the serotonergic function in the brain and lower concentrations
of 5-HIAA in the cerebrospinal fluid (CSF) have been found (Tuinier et al., 1995). Serotoninergic
mechanisms have been shown to operate in the acquisition and maintenance of dominance in
primates. Engaging in and winning dominance related struggles in some monkeys, generate high
blood levels of serotonin (Raleigh et al., 1991a). In our research, DA and 5-HT higher plasma
levels in anxious dogs compared to the control ones suggest that also in dogs a relationship exists
between these two neurotransmitters and more frequent exhibition of anxiety related behaviours.
However a question remains to be solved, that is the possibility that plasma levels of the
evaluated neurotransmitters may reflect what is happening at the brain level. According to some
authors, this is true (Da Prada et al., 1988; Coccaro et al., 1997). Yan et al. (1993) found a
statistically significant relationship between serotoninergic measures (including 5-HIAA) in CSF

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177

and blood in sub-human primates, indicating that measurements in blood may be used as a
peripheral indicator of central 5-HT functioning (Stahl et al., 1982). No indications are up to date
at disposal on dogs in the literature.
Blood concentrations of catecholamines in many species such as humans beings, cats, mice
and rabbits (Da Prada and Picotti, 1979; Picotti et al., 1984) are higher in the platelets compared
to in plasma. Some factors affect the distribution of 5-HT and DA in platelets and plasma
(Pletscher, 1968). In vitro findings on rats platelets show that 5-HT and DA compete for the same
active transport mechanism (carrier) but 5-HT has a higher affinity than DA for this transport
process (Gordon and Olverman, 1978). In vivo this competition is likely to occur, since it has
been shown both in humans and in rabbits that the concentrations measured in plasma were
higher for 5-HT than for DA (Da Prada and Picotti, 1979). Moreover, the respective neuronal and
platelet proteins have been shown to be encoded, both for 5-HT and for DA, by the same genes
(Cook et al., 1994; Ramamoorthy et al., 1993). These factors may explain why different
concentrations of DA and 5-HT are found between platelets and plasma.
As far as the relationships between blood and platelet neurotransmitters concentrations are
concerned, our study suggests that also in dogs anxiety related behavioural disorders may be
enhanced by high DA and 5-HT levels in plasma, and inhibited by low 5-HT concentrations in
platelets. According to Dodman and Shuster (1998) the threshold for aggression may be raised also
by dopaminergic antagonists, being aggressive behaviour increased by the release of
catecholamines (Eichelman, 1987). The important role of catecholamines in the flight or fight
stress response, where the limbic system is involved (Henry and Stephens, 1977) is well known.
Moreover, DA may be involved also in compulsive behavioural disorders. The treatment with
dopamine antagonists has reduced some compulsive behaviours in cats and humans (Cools and Van
Rossum, 1970; Stein and Hollander, 1992) as well the Tourette-like self-injurious behaviour in
horses (Dodman and Shuster, 1998) and dopamine agonist-induced stereotypic grooming and selfmutilation in rats (Hartgraves and Randall, 1986). High blood levels of 5-HT have been revealed in
relation with affective aggressive behaviour for example in vervet monkeys, showing the
involvement of serotonin dynamics in this stress response (Raleigh et al., 1980, 1985, 1991a,b).
The involvement of the 5-HT with aggressive behaviour has been experimentally proven,
inducing shortages of brain 5-HT (Garattini et al., 1967; Salama and Goldberg, 1973). A few
researches on aggressive behaviour together with reduced brain 5-HT concentration have
highlighted this correlation in mice after isolation, revealing an increase of fights between males
in response to the treatment (Valzelli and Bernasconi, 1979). Reisner et al. (1996) studied the
relationship between cerebrospinal fluid (CSF) monoamine metabolite levels in dominantaggressive and non-aggressive dogs. Post-mortem CSF 5-HIAA levels were measured by HPLC:
concentrations of 5-HIAA (202.0 pmol/ml) were lower in the aggressive group than in the control
one (298.0 pmol/ml). This study suggests that reduced serotonergic function is associated with
aggressive behaviour and impaired impulse control in dogs and that serotonin may modulate
aggressive behaviour in mammals.
As far as the relationship between platelet 5-HT and disturbed behaviours is concerned, 5-HT
platelet content was reported to be lower in human adult depressed patients who attempted
suicide than in depressed patients who did not (Mann et al., 1992a,b). Platelet serotonin content
was positively correlated also with a lifetime history of aggression and current hostility in
humans (Pliszka et al., 1988). According to Dodman and Shuster (1998), anxiety related
aggressive behaviour may be decreased enhancing central serotoninergic activity. We also have
found a trend to lower platelet 5-HT levels in anxious dogs compared to the control ones, thus
suggesting too an involvement of serotoninergic function in anxiety related behaviours in dogs.

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5. Conclusion
This study shows that 5-HT plasma and DA are higher in the anxious dogs than in the control
ones, while NE plasma levels are similar in the two groups. Another important result of this study
is related to the platelet concentrations of 5-HT, showing a trend to lower concentration levels in
anxious dogs compared to the control ones.
This result could demonstrate that also in dogs the serotoninergic and dopaminergic systems
play an important role in determining affective reactions such as the exhibition of anxiety related
behaviour problems. This hypothesis raised by the present research needs to be confirmed on a
larger sample of dogs possibly comparing the platelet 5-HT levels to the 5-HT levels at the CNS
level.
Acknowledgments
We wish to thank Dr. Michela Minero for the support in data statistical analysis, Dr.
Alessandra Trotti and Prof. Elena Zenoni for the English language final check.
Tanks to The European Social Fund, Ministero del Lavoro e delle Politiche Sociali, Regione
Lombardia and Ingenio to partially funding this work.
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