Aceite de Lavanda-efectos Neurologicos

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Neurophysiological and behavioural effects of

lavender oil in rats with experimentallyinduced anxiety

Hector W. H. Tsang,a* Samuel C. L. Lo,b Chetwyn C. H. Chan,a

Timothy Y. C. Ho,a Kelvin M. T. Fung,a Alan H. L. Chana and Doreen W. H. Aua

ABSTRACT: This study examined anxiolytic effects of lavender oil on brain serotonin levels and anxiety-related behaviours of rats. The experimental rats were divided into five groups, which respectively received inhalation of saline, 1.25% lavender oil,2.5% lavender oil, chlordiazepoxide (CDP), and 2.5% lavender oil co-administered with CDP. Anxiety was induced in rats usinganimal models including Elevated Plus Maze and Open Field. The levels of serotonin in the pre-frontal cortex and striatum of therats, the anxiolytic effects of lavender oil and its augmentation effect as to co-administration with CDP were evaluated. Theneurophysiological findings showed that groups receiving lavender oils, CDP, and 2.5% lavender oils co-administeredwith CDP had significantly higher level of serotonin in the pre-frontal cortex. However, the anxiolytic behavioural effectsof lavender oil were found to have mixed results. This study provided preliminary evidence that inhalation of lavender oilparalleled effects of CDP in up-regulating synthesis of serotonin in rat pre-frontal cortex, and the co-administration of CDPwith 2.5% lavender oil tended to augment effect of CDP on serotonin in their pre-frontal cortex and striatum. Copyright ©2013 John Wiley & Sons, Ltd.

Keywords: aromatherapy; lavender oil; anxiety; serotonin; tranquilizing drugs

Introduction

Aromatherapy is a commonly used complementary therapy for

anxiety disorders.[1] Its therapeutic effects may result from

physiological effects of inhaled volatile molecules acting onamygdala and hippocampus of the limbic system.[2] Neurochem-

ical responses in the limbic system then lead to the ‘anxiolytic’

effects of stress and anxiety.[3] A systematic review found that

essential oils, including lavender oil, could elicit anxiolytic effects

on rodents[4] and individuals with anxiety symptoms.[5]

Previous studies reported that lavender oil acted like tranqui-

lizing drugs on g-aminobutyric acid in amygdala,[6] and inhibited

acetylcholine release at the neuromuscular junction of mice.[7]

However, the effects of lavender oil on the neurotransmitter

activity that regulates emotion have not been well documented.

The serotonergic system, in particular, is rarely tested in

rodents.[8,9] Besides, several essential oils were reported to have

similar effects as tranquilizers in animal studies, but the possibleaugmentation effects of essential oils on drugs have never been

examined. It therefore remains unclear whether aromatherapy

would have complementary effect with tranquilizing drugs such

as chlordiazepoxide (CDP) and augment the anxiolytic effects in

patients seeking psychiatric treatments.

This study attempted to explore the effect of lavender oils on

the level of serotonin and the corresponding anxiolytic beha-

vioural effects in rats in which three main hypotheses were

tested. First, inhalation of lavender oil would be associated with

increased brain serotonin level of rats in prefrontal cortex and

striatum. Second, inhalation of lavender oil would produce anxi-

olytic effects in rats similar to CDP. Third, inhalation of lavender

oil would augment the effect of CDP.

Materials and Methods

Animals

Adult male Sprague–Dawley albino rats were obtained from theCentralized Animal House of PolyU at 6 weeks old. The rats were

maintained at 22Æ2 C in a regular light–dark cycle (07:00–

19:00 h, light), and provided with free access to food and water.

Each rat was naive to the essential oils and drugs, and used once

only. All rats were randomly allocated to five groups and each

rat was placed individually into the inhalation apparatus for 90

min immediately before the test. Groups 1 to 3 were placed in

the inhalation apparatus containing 0.9% saline, 1.25% w/w lav-

ender oil, and 2.5% w/w lavender oil, respectively. Group 4 was

injected intraperitoneally with CDP (10 mg/kg) 90 min before

each test.[10] Group 5 was injected intraperitoneally with CDP

(10 mg/kg) and placed in the inhalation apparatus containing

2.5% lavender oil for 90 min before the test. The number of rats

per group ranged from five to 10, which was taken with reference

to previous study.[8] All experiments were conducted according to

approved guidelines regarding the care of experimental animals

in PolyU and the Hong Kong Government.

* Correspondence to: Hector W. H. Tsang, Department of Rehabilitation

Sciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong

Kong. E-mail: [email protected]

a Department of Rehabilitation Sciences, The Hong Kong Polytechnic

University, Hung Hom, Kowloon, Hong Kong

b Department of Applied Biology & Chemical Technology, The Hong Kong

Polytechnic University, Hung Hom, Kowloon, Hong Kong

Flavour Fragr. J. 2013 Copyright © 2013 John Wiley & Sons, Ltd.

Research Article

Rec eive d: 25 Octobe r 2012, Revis ed: 16 D ec embe r 2012, Acce pted : 21 D ece mb er 2012 Pub lished o nl ine in W iley Onl ine Library

(wileyonlinelibrary.com) DOI 10.1002/ffj.3148



Chemicals

Lavandula angustifolia (lavender) oil was supplied by DK aroma-

therapy (Hong Kong, China) and prepared by diluting into

double-distilled water (ddH2O) with 1% Tween 20 with mild

stirring. It was then soaked up by cotton set at two sides of the in-

halation box. The inhalation vehicle was 0.9% saline with 1%

Tween 20. CDP was obtained from Sigma-Aldrich (Poole, UK). It

was dissolved in 0.9% saline and 1% Tween. The rat was adminis-tered with CDP at 10 mg/kg via intraperitoneal injection at a

volume of 1 ml/kg according to the rat’s body weight.[11,12] The

lavender oil and drug were freshly prepared before experiments

and applied to the rats 90 min before the tests.

Inhalation Apparatus

The inhalation apparatus[9] consisted of four built chambers

(42Â 30Â 29 cm). The floor was made of acrylic fibre where the

rats were placed individually. The front and back walls were made

of acrylic fibre containing four holes (2 cm in diameter each)

where drug-embedded cotton wool was placed (i.e. 2 ml per unit

of respective substance, lavender oil or vehicle). The top wall

contained 30 small holes for ventilation. There was one perforated

acrylic fibre 3 cm apart from the front and back walls to prevent

rats from biting the drug-embedded cotton wool.

Elevated Plus Maze Test

The elevated plus maze (EPM)[13] consisted of four arms (length:

50 cm, width: 10cm) at right angle to each other, which were

connected to a central square (10 cmÂ 10 cm). The apparatus

was elevated 50 cm above the floor. Two opposite arms had

high walls (closed arms, 40 cm high), and the other two were

opened with a small ridge to provide an additional grip. The

maze was constructed with steel and painted in black. Experi-

ments were performed in a dark room, and illuminated by awhite fluorescent tube. After each test, the maze floor and wall

was thoroughly cleaned with cotton and ethanol. Outcome vari-

ables were the number of entry into open arms and the amount

of time spent within 5 min of test period. Rats which fell down

from the open arm were excluded.

Open Field Test

The apparatus was a round arena made of acrylic fibre with a

diameter of 88 cm and a circular wall of 30 cm high. The floor

and the wall were white. The field consisted of three concentric

circles: an inner circle of 20 cm diameter, a middle circle of

50 cm diameters, and an outer circle. The circles were divided intoroughly equal size areas by radial lines. The outer circle was

divided into eight areas, and the middle circle was divided into

four areas.[14] Experiments were performed in a dark room with

background noise produced from a white-noise generator.[13]

The apparatus was illuminated by an LED giving yellowish light

and positioned 1 m above the centre of the circle. Outcome

variables included immobility time, the number of peripheral

movement, rearing, and defaecations within the 10 min test period.

Serotonin and Protein Quantification

Samples of brain tissues were homogenized in 200 ml 1Â

phosphate buffered saline and centrifuged at 5000 Âg, 4C for

15 min. Supernatant was removed and quantified for serotonin

and protein amount using a serotonin ELISA assay kit (Genway,

San Diego, CA, USA) and the Bradford assay (Bio-Rad Laboratories

Inc, Hercules, CA, USA), respectively, according to the protocols

provided by the manufacturers. The serotonin levels were

measured in nanograms per milligram of protein.

ProcedureExperiments were conducted between 10:00 to 14:00 h. Rats

were housed individually in their cages for at least 1 day prior

to testing for their adaptation to the laboratory environment.

Group 1 served as the vehicle control. Groups 2 and 3 were

the experimental groups, with varying inhaled doses (1.25%

w/w and 2.5% w/w, respectively) of lavender oil. Groups 4 and

5 were positive controls, with administration of CDP alone and

co-administration of CDP with 2.5% lavender oil, respectively.

The animal tests were implemented in sequence: day 1, elevated

plus maze (5 min); day 2, Open field (10 min); and day 3, killing.

The behaviours of animals were individually video-taped and

analysed independently later. Brain samples were obtained by

decapitating the rats followed by removal of prefrontal cortexand striatum before testing. The brain samples were then frozen

in liquid nitrogen and stored at À82C. The serotonin and

protein quantification procedure were used to determine levels

of serotonin in the prefrontal cortex and striatum.

Statistical Analyses

Predictive Analytics Software (PASW) 18 was used for data analyses.

Neurophysiological and behavioural data were presented in bar

charts with meanÆ standarderror (SE). Differences across treatment

groups were analysed by one-way ANOVA followed by a post hoc

Dunnett’s test. Two planned contrasts were used to examine a priori

hypotheses of this study.[15] The first planned contrast was used to

analyse whether there were significant differences in various

outcomes between the two experimental groups (hereafter

referred to as the ‘1.25% lavender oil group’ and the ‘2.5%

lavender oil group’, respectively). The second contrast was used to

examine whether the co-administration of CDP with lavender oils

augmented the effects of CDP (hereafter referred to as the ‘2.5%

lavender oil+ CDP group’ vs. the ‘CDP group’). Differences were

considered statistically significant when p<0.05. The Bonferroni

correction was used to counteract the accumulative effects of type

I errors in multiple comparisons.

Results

Serotonin Levels

The levels of serotonin in the pre-frontal cortex and striatum varied

significantly across treatment groups (Table 1). Post-hoc analyses

found that the 2.5% lavender oil group, the CDP group, and the

2.5% lavender oil + CDP group had significantly higher levels of

serotonin than the vehicle control ( p values< 0.0125, with Bonfer-

roni correction) (Figure 1a). Comparatively, the lavender oil at a

lower dose was not as effective as CDP or lavender oil at higher

dose in up-regulating the serotonin level. Lavender oil at a higher

dose elicited similar levels of serotonin as CDP. By planned

contrast, differences in the levels of serotonin between the

two lavender oil groups (1.25% vs. 2.5%) were not significant

(t = 1.38; p> 0.05). The 2.5% lavender oil + CDP group had

H. W. H. Tsang et al .

Flavour Fragr. J. 2013Copyright © 2013 John Wiley & Sons, Ltd.wileyonlinelibrary.com/journal/ffj



significantly higher level of serotonin than the CDP group

(t = 2.00, p<0.05).

In the striatum, only the 2.5% lavender oil + CDP group exhib-

ited significantly higher levels of serotonin than the vehicle group

( p< 0.0125, with Bonferroni correction). The groups receiving lav-

ender oil or CDP alone demonstrated such regulatory effect on

serotonin, but the effect was not statistically significant (Figure 1b).

The 2.5% lavender oil+ CDP group had significant higher level of

serotonin in striatum than the CDP group (t = 4.03, p< 0.05).

Anxiety-related Behaviours in the Elevated Plus Maze and

Open Field Tests

Levels of anxiety-related behaviours were significantly different

among groups in the EPM and the open-field tests ( p values<0.05)

(Table 2). The two groups of rats respectively receiving CDPalone and co-administration of CDP with 2.5% lavender oil

spent significantly longer time in the open arm compared with

the vehicle control ( p< 0.05 and p<0.0125 with Bonferroni

correction, respectively) (Figure 2). The planned contrast t -test

did not reveal a significant difference between these two

groups (t = 0.62, p> 0.05). The two lavender oil groups spent

longer time in the open arm than did the vehicle control, but

the differences were not statistically significant.

Regarding the open-field test, the 2.5% lavender group

elicited more anxiolytic-related behaviours (increased rearing,

reduced immobility time and defaecation) compared to other

treatment groups and the vehicle control ( p values< 0.02 with

Bonferroni correction) (Table 2). The anxiety-related behavioursvaried in terms of different outcome measures. As to the immo-

bility time, both the 1.25% and 2.5% lavender oil groups and

the CDP group had reduced immobility compared to the vehi-

cle control ( p values< 0.0125 with Bonferroni correction)

(Figure 3a). The co-administration of CDP with 2.5% lavender

oil did not significantly reduce the immobility time of rats

compared to vehicle control ( p>0.05). As to the frequency of

peripheral movement, the peripheral movement of rats signif-

icantly increased in the group receiving CDP alone ( p< 0.0125

with Bonferroni correction) but not in other treatment groups

( p>0.05) (Figure 3b). All treatment groups had increased the

number of rearing compared to the vehicle control, but no

significant differences were found ( p values> 0.05) (Figure 3c).

Table 1. Treatment group effects on rat serotonin levels at the prefrontal cortex and striatum

5-HT ELISA

quantification

Outcome

measures (units)

Groups F ratio p-value

Vehicle

control

1.25%

Lavender oil

2.5%

Lavender oil

CDP 2.5%

Lavender oil þ CDP

PC: n = 8;

Str: n = 7

PC: n =10;

Str: n = 8

PC: n = 6;

Str: n = 8

PC: n = 8;

Str: n = 8

PC: n = 9;

Str: n = 9

Pre-frontal

cortex

Serotonin 57.55

(2.61)

74.04

(3.26)

90.56

(11.52)

91.72

(5.41)

108.64

(6.50)

F = 11.50 < 0.001*

Striatum Serotonin 106.42

(14.06)

133.17

(12.25)

150.10

(26.76)

114.32

(10.83)

202.30

(14.05)

F = 5.50 0.002*

Values are given as the means and standard errors (in brackets).

Serotonin (5-HT) level is measured in ng/mg protein.

* p< 0.025 with Bonferroni correction.

CPD, chlordiazepoxide; PC, pre-frontal cortex; Str, striatum.

Figure 1. ELISA determination of serotonin in (a) prefrontal cortexand (b)

striatum after 0.9% saline (vehicle control), 1.25% lavender oil inhalation,

2.5% lavender oil inhalation, chlordiazepoxide (CDP, 10 mg/kg, i.p.), and

chlordiazepoxide (CDP, 10 mg/kg, i.p.) co-administered with 2.5% lavender

oil inhalation. Data are expressed as meanÆSE in ng/mg protein. N = 7–10

rats per group. * p<0.05 and ** p<0.0125 (with Bonferroni correction),

respectively, indicates significant differences from vehicle control

Lavender oil on anxiety of rats

Flavour Fragr. J. 2013 Copyright © 2013 John Wiley & Sons, Ltd. wileyonlinelibrary.com/journal/ffj



In terms of the number of defaecations, all treatment groups

had significantly reduced defaecations compared to the

vehicle control ( p values< 0.0125 with Bonferroni correction)

(Figure 3d). By planned contrasts, no significant group differ-

ences were revealed between the two lavender oil groups.

Similarly, no significant group differences were found between

the CDP group and the 2.5% lavender oil+ CDP group.

Discussion

Our results showed that inhalation of 2.5% lavender oil signifi-

cantly increased the synthesis of serotonin in the pre-frontal cortex

but not striatum. This finding aligned well with a previous study on

the anti-stress effect of inhaled lemon vapour on modulating the

serotonergic system in mice.[8] The neurobiological mechanism

that regulates the synthesis of serotonin in the prefrontal cortex

could be considered as a basis for regulating anxiety behaviours.

We also found that co-administration of CDP with 2.5% lavender

oil further enhanced the up-regulation effect of serotonin in the

pre-frontal cortex. It provided evidence that inhalation of lavender

oil possibly augmented the effects of tranquilizing drugs. Results in

the striatum may imply that current doses of inhaled lavender oilor injecting CDP alone might not be suf ficient for promoting

adequate synthesis of serotonin in striatum. However, ourfindings

revealed that once the CDP was co-administrated with 2.5% laven-

der oil, secretion of striatal serotonin increased significantly. It

further provided the neurobiological evidence of the augmenta-

tion effects of lavender oil on tranquilizing drugs.

The present study found that the inhaled lavender oil groups

elicited anxiolytic behaviours in the EPM test and the open field

test, but the effects were not statistically significant in some

behavioural measures. These results are consistent with earlier

conclusion that lavender oil at inhaled dose above 0.5 ml may

have anxiolytic effects that are similar to those of CDP.[10]

However, the neurological basis of the augmentation effect of

inhaled lavender oil on CDPcould only translateinto corresponding

anxiolytic behaviours in the EPM test, but not the open field test. In

EPM test, rats would spend moretime in the openarmsif they were

less anxious.[13] Although the two lavender oil groups did spend

longer time in the open arms than the vehicle control, the

Table 2. Treatment group effects on rat behaviour in the elevated plus maze and open-field tests

Test Outcome measures

(units)

Groups F ratio/Welch’s F a p-value

Vehicle

control

1.25%

Lavender oil

2.5%

Lavender oil CDP

2.5%

Lavender oil

þ CDP

EPM: n = 6;

OF: n = 8

EPM: n = 8;

OF: n = 6

EPM: n = 6;

OF: n = 5

EPM: n =10;

OF: n = 7

EPM: n = 6;

OF: n = 7

Elevated plus

maze

Time spent in

open arms (s)

22.17

(7.27)

54.88

(19.01)

61.17

(12.61)

98.80

(12.96)

120.67

(33.01)

F = 7.46a 0.002*

Open-field Peripheral

movement(mumber of times)

27.63

(4.74)

42.83

(8.76)

67.00

(12.68)

80.14

(15.39)

54.43

(11.42)

F = 3.76 0.014*

Rearing

(number of times)

10.00

(2.10)

22.83

(3.19)

29.00

(8.64)

28.57

(8.25)

26.29

(5.82)

F = 4.40a 0.020*

Immobility

(min)

4.25

(0.65)

0.83

(0.48)

0.40

(0.40)

1.00

(0.49)

2.57

(0.92)

F = 6.34a 0.004**

Defaecation

(number of times)

4.38

(0.50)

2.17

(0.40)

1.00

(0.32)

2.14

(0.51)

1.43

(0.57)

F = 7.48 <0.001**

Results are given as the means and standard errors (in brackets).aWelch’s F was used for the violation of homogeneity assumption.

* p< 0.05

** p< 0.0125 with Bonferroni correction.

CDP, chlordiazepoxide; EPM, elevated plus maze test; OF, open-field test.

Figure 2. Anxiolytic effects of lavender oil in elevated plus maze test.

Data are expressed as meanÆ SE for the time spent in open arms in a

5 min period, monitored 90 min after the administration of 0.9% saline

(vehicle control), 1.25% lavender oil inhalation, 2.5% lavender oil

inhalation, chlordiazepoxide (CDP, 10 mg/kg, i.p.), or chlordiazepoxide

(CDP, 10 mg/kg) co-administered with 2.5% lavender oil inhalation.

N = 6–10 ratsper group. * p<0.05 and ** p<0.0125(with Bonferroni correc-

tion), respectively, indicates significant differences from vehicle control





differences were not significant. Nevertheless, the rats co-adminis-

trated CDP with 2.5% lavender oil spent a longer time in the open

arm than those receiving CDP alone. Such behavioural responses

provided a preliminary observation on the augmented anxiolytic

effects of lavender oil on CDP. As to the openfield test, non-anxious

rats should have shorter immobility time, fewer peripheral move-

ments, fewer defaecations and higher frequency of rearing.[16] While

inhalationof 1.25% or 2.5% lavender oil paralleled theeffects of CDP

in reducing immobility time and defaecations, co-administration of

CDP with 2.5% lavender oil failed to augment such anxiolytic effects.

The opposite results were observed for the frequency of peripheral

movement among rats. The group receiving CDP alone

unexpectedly had the highest frequency of peripheral movements.

Although an increase in the frequency of rearing across treatment

groups was observed, no significant differences were found.

Altogether, this study provided preliminary evidence that

inhalation of 1.25% w/w or 2.5% w/w lavender oil for 90 min

stimulated synthesis of serotonin in the prefrontal cortex

approximated to the effects of CDP. Co-administration of CDP

with 2.5% lavender oil augmented effect of CDP in up-regulating

serotonin levels in prefrontal cortex and striatum. However,

these up-regulatory effects of brain serotonin did not com-

pletely transfer to the reduction of anxiety behaviours in the

EPM and open-field. Factors such as the inhaled dose of laven-

der oil, the experiment period, and the sample size may have

caused the insignificant behavioural change in rats. It is

Figure 3. Anxiolytic effects of lavender oil in the open-field model. Data are expressed as meanÆ SE for (a) immobility time, (b) number of

peripheral movement, (c) rearing, and (d) defaecations in the open-field model for a 10 min period, monitored 90 min after the administration

of 0.9% saline (vehicle control), 1.25% lavender oil inhalation, 2.5% lavender oil inhalation, chlordiazepoxide (CDP, 10 mg/kg, i.p.),

chlordiazepoxide (CDP, 10 mg/kg, i.p.) co-administered with 2.5% lavender oil inhalation, or vehicle (0.9% saline). N = 5–8 rats per

group. ** p<0.0125 (with Bonferroni correction), respectively, indicates significant differences from vehicle control

Lavender oil on anxiety of rats

Flavour Fragr. J. 2013 Copyright © 2013 John Wiley & Sons, Ltd. wileyonlinelibrary.com/journal/ffj



noteworthy to point out that we did use the lavender oil at an

amount and adopt the sample size which produced significant

anxiolytic effects following previous protocols.[8,10] Given the

complexity of the interaction between the dose and exposure

to the oil,[17] it seems to be valid to increase the frequency and

the time of exposure to the inhaled lavender oil to optimize its

anxiolytic effects in future studies.

Conclusion

The present findings support the hypothesis that lavender oil has

anxiolytic properties similar to tranquilizing drug and produced

augmentation effect in up-regulating synthesis of serotonin in

pre-frontal cortex and striatum. Whether the neurological basis

of anxiolytic effect of inhaled lavender oil could completely

translate into corresponding behavioural responses remain incon-

clusive. More research to unknot the effect of lavender oil on the

serotonergic system is needed. Nevertheless, the present study

was the first of its kind to provide preliminary evidence on the

up-regulating effects of lavender oil on the levels of serotonin.

Acknowledgement

This material is supported by Internal Grant at The Hong KongPolytechnic University (PolyU). We thank Ms Puiyi Mok andDr Angie Tang of the CAF, and also Dr Danny Gohel and Dr Tony

To at HTI of PolyU for resources and the use of facilities.

References

1. M. Cooke, K. Holzhauser, M. Jones, C. Davis, J. Finucane. J. Clin. Nurs.2007, 16, 1695.

2. H. Cavanagh, J. Wilkinson. Phytother. Res. 2002, 16, 301.3. J. Buckle. In Mosby ’ s Complementary & Alternative Medicine: A

Research-Based Approach, L. W. Freeman (ed.). Mosby Elsevier:Missouri, 2009, 417–435.

4. H. W. H. Tsang, T. Y. C. Ho. Rev. Neurosci. 2010, 21, 141.

5. Y. L. Lee, Y. Wu, H. W. H. Tsang, A. Y. Leung, W. M. Cheung. J. Altern.Complement. Med. 2011, 17 , 101.6. H. Aoshima, K. Hamamoto. Biosci. Biotechnol. Biochem. 1999,

63,743.7. L. Re, S. Barocci, S. Sonnino, A. Mencarelli, C. Vivani, G. Paolucci, A.

Scarpantonio, L. Rinaldi, E. Mosca. Pharmacol. Res. 2000, 42, 177.8. M. Komiya, T. Takeuchi, E. Harada. Behav. Brain Res. 2006,

172 , 240.9. J. Solati, P. Yaghmaei, K. Mohammdadi, Neurophysiology 2012, 44, 49.

10. D. Shaw, J. M. Annett, B. Doherty, J. C. Leslie. Phytomedicine 2007,14, 613.

11. R. N. De Almeida, S. C. Motta, C. De Brito Faturi, B. Catallani, J. R. Leite.Pharmacol. Biochem. Behav. 2004, 77 , 361.

12. D. J. Greenblatt, R. I. Shader, S. M. MacLeod, E. M. Sellers. Clin. Phar-macokinet. 1978, 3, 381.

13. J. N. Crawley. What ’ s Wrong With My Mouse?: Behavioral Phe-notyping of Transgenic and Knockout Mice. Wiley-Liss: New

York, 2000.14. M. J. Anderson, Tasks and Techniques: A Sampling of the Methodolo-

gies for the Investigation of Animal Learning, Behavior, and Cognition.Nova Science Publishers: New York, 2006.

15. A. P. Field, Discovering Statistics Using SPSS: (and Sex and Drugs and Rock ‘ n’ Roll). SAGE Publications: London, 2009.

16. C. Johansson, S. Ahlenius. J. Psychopharmacol. 1989, 3, 32.17. D. Shaw, K. Norwood, J. C. Leslie. Behav. Brain Res.. 2011, 224, 1.



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