Elevated Plus-maze Model Of Anxiety Research Articles

Behavioral and biochemical characterization of elevated “I-maze” as animal model of anxiety

The elevated I-maze is a modification of the elevated plus-maze model of anxiety in mice. The design of I-maze comprises a straight wooden passage, resembling the English letter “I,” divided equally into three areas; two enclosed areas (close arms) at both ends of the “maze” and an open area in the center of two enclosed areas. The I-maze completely avoids the central platform of elevated plus-maze, removing any ambiguity in time spent on central platform and allowing uninterrupted animal exploration. In this model, diazepam (1 mg/kg) and gabapentin (10 mg/kg) significantly increased the percentage of time spent in the open areas (%TO) and the number of unprotected head dips (uHDIPS), and reduced the number of protected head dips (pHDIPS) and stretch attend postures (SAP) from close to open arm. Similarly, fluoxetine (5 mg/kg) significantly increased %TO and uHDIPS, and significantly decreased SAP from close to open arm, but it did not have any significant effect on pHDIPS. The 5-HT3 receptor antagonist, ondansetron (0.1 mg/kg), did not produce any significant change in all the behaviors, observed, as compared to vehicle-treated control mice. On the other hand, the anxiogenic agent, caffeine (15 mg/kg), did produce a significant decrease in %TO and uHDIPS, and significantly increased pHDIPS and SAP from close to open arm. Mice confined in open area of I-maze bring the relevant biochemical changes associated with anxiety behavior, showing significant increase in the levels of plasma nitrate and plasma corticosterone. These data indicate that a combination of novel design of elevated I-maze and a detailed behavioral analysis provides a sensitive model for the measurement of anxiety.

Anxiolytic effects of Plumeria rubra var. acutifolia (Poiret) L. flower extracts in the elevated plus-maze model of anxiety in mice

Interest in alternative medicine and plant-derived medications that affect the “mind” is growing rapidly since last two decades. The aim of the present study was to investigate the effects of ethanolic extract of flower of Plumeria rubra (PR) along with its fractions in the elevated plus-maze (EPM) model of anxiety. The P. rubra extract or its fractions was administered orally to male Swiss mice, at graded doses, 1h prior to behavioural assessment. The PR extract at the dose of 100mg/kg p.o., significantly increased the time spent in the open arms of the EPM. Further, the anxiolytic properties of hexane, chloroform and butanolic soluble and insoluble fractions at one-fifth of the original dose were also observed in the EPM task. Out of which butanol insoluble fraction showed significant anxiolytic activity comparable to standard anxiolytic drug, diazepam. Further, pretreatment with crude ethanolic extract and butane insoluble fraction showed no significant effects in the horizontal activity, total distance travelled and stereotypy count in the animal activity monitor and had no motor in-coordination side effects in the rotarod test in mice. These observations suggest that the flower extract of P. rubra and its insoluble butanolic fraction might possess significant anxiolytic potential to be pursued further for drug development process.

Anti-anxiety Activity of Cymbopogon citratus (dc.) stapf Leaves Extracts on the Elevated Plus-Maze Model of Anxiety in Mice

Abstract The Methanolic extract of Cymbopogon citratus leaves at the dose of 200 mg/kg increased the percentage of time-spent and the percentage of arm entries in the open arms of the elevated plus-maze (EPM) and decreased the percentage of time-spent in the closed arms of EPM. Moreover, it prolonged the ketamine-induced latency to sleep but had no significant effects on total sleeping time induced by ketamine. Also, the locomotor activity was affected but not to the same extent as observed for diazepam. The anxiolytic effects of methanol extract Cymbopogon citratus leaves may be related to their content of flavonoids. This study validates the traditional use of the plant in management of anxiety.

Behavioral alterations and pro-oxidant effect of a single ketamine administration to mice

A growing body of evidence has pointed to the ionotropic glutamate N-methyl- d-aspartate receptor (NMDA) as an important player in the etiology of psychopathologies, including anxiety and major depression. Clinical findings suggest that ketamine may be used for the treatment of major depression. There is evidence that reactive oxygen species also play an important role in the pathogenesis of many diseases, particularly those which are neurological and psychiatric in nature. This study examined the behavioral and oxidative stress alterations after a single administration of ketamine (5, 10 and 20 mg/kg i.p.) in mice. Ketamine presented a significant anxiogenic effect in the elevated plus-maze model of anxiety, also increasing locomotor activity. In the forced swimming and tail suspension tests, a significant decrease in immobility time after ketamine administration was observed. In addition to the behavioral changes induced by ketamine, this drug also increased lipid peroxidation, nitrite content and catalase activity, while decreased GSH levels in mice prefrontal cortex. In conclusion, our results confirm the antidepressant effects of ketamine, also showing a pro-oxidant effect of this drug.

Acute effect of Copaifera reticulata Ducke copaiba oil in rats tested in the elevated plus-maze: an ethological analysis

Abstract Objectives Copaiba oil oleoresin exuded from Copaifera reticulata Ducke (CRD) is commonly used in anti-inflammatory, healing and anti-tumoral folk medicines. The purpose of this study was to investigate the putative anxiolytic effect of acute administration of CRD. Methods CRD was administered (100, 400 and 800 mg/kg, p.o.) to male Wistar rats submitted to the elevated plus-maze model of anxiety using an ethopharmacological analysis. Key findings In comparison with control rats, CRD increased the percentage of entries in the open arms over the entire dose range tested (vehicle, 33.6 ± 4.5; CRD 100 mg/kg, 44.67 ± 3.68; CRD 400 mg/kg, 47.2 ± 2.3; CRD 800 mg/kg, 50.7 ± 2.2) and the percentage of time spent in the open arms of the elevated plus-maze at the highest dose (800 mg/kg) (vehicle, 26.4 ± 5.7; CRD 800 mg/kg, 52.0 ± 2.7). A standard anxiolytic, diazepam (3 mg/kg, p.o.), was used as a positive control. In a similar way, diazepam increased the percentage of entries and time spent in the open arms when compared with vehicle (% open entries: vehicle, 45.4 ± 1.3; diazepam, 50.7 ± 1.9; % time spent in open arms: vehicle, 28.2 ± 0.9; diazepam, 38.9 ± 1.2). Regarding ethological measures, CRD at the highest dose (800 mg/kg) reduced peeping out (anxiety-related behaviour) (vehicle, 3.1 ± 0.6; CRD, 0.9 ± 0.2) and increased end-arm activity (vehicle, 0.2 ± 0.2; CRD, 2.0 ± 0.4), indicating an enhanced tendency of the rats to explore actively the potentially dangerous areas of the maze. Diazepam decreased peeping out (vehicle, 3.3 ± 0.3; diazepam, 1.0 ± 0.2) and flat-back approach (vehicle, 0.8 ± 0.2; diazepam, 0.2 ± 0.1) and increased end-arm activity (vehicle, 0.3 ± 0.1; diazepam, 2.5 ± 0.3) and head-dipping (vehicle, 8.2 ± 0.4; diazepam, 12.0 ± 0.5). Conclusions These data showed, for the first time, that acute treatment with CRD copaiba oil produced a dose-dependent anxiolytic-like effect over the dose range tested, on conventional and ethological parameters, without adversely affecting general activity levels.

Screen for genes in periaqueductal grey of male Wistar rats related to reduced exploratory activity in the elevated plus-maze

Aim of a present study was to find genes in the periaqueductal grey (PAG) related to the exploratory behavior in rats. Male Wistar rats were divided according to their exploratory behavior in the elevated plus-maze model of anxiety into two groups: high (non-anxious) and low (anxious) exploratory activity. Differential expression of genes was analyzed using the cDNA representational difference analysis (RDA). Q-RT-PCR was used to confirm most prominent changes and functional annotation of the identified genes was performed to establish pathways related to exploratory behavior of rats. We found different genetic activation related to the exploratory activity of rats. Rats with low exploratory activity showed increase in the intracellular signal transduction and in GABA, vasopressin and adrenergic receptor activities. Functional annotation confirmed significant induction of cAMP system and GTPases in rats with anxious-type behavior. On the other hand, rats with high exploratory activity in the elevated plus-maze (non-anxious type of behavior) had increased activity of genes forming “behavioral fear response” system. These changes were specific to PAG, because they were not found in the cerebellum. In addition, plasma corticosterone levels were significantly higher in rats with non-anxious behavior compared to anxious behavior. Our results show that non-anxious behavior is related to activation of “fear response system” and more intense activation of HPA axis. Possibly it means that this system helps animals to cope with the threatening circumstances. More detailed analysis of this potential “fear response system” is necessary in the further studies for understanding its role in the regulation of emotional behavior.

Enhanced anxiety, depressive‐like behaviour and impaired recognition memory in mice with reduced expression of the vesicular glutamate transporter 1 (VGLUT1)

Three isoforms of a vesicular glutamate transporter (VGLUT1-3) have been identified. Of these, VGLUT1 is the major isoform in the cerebral cortex and hippocampus where it is selectively located on synaptic vesicles of excitatory glutamatergic terminals. Variations in VGLUT1 expression levels have a major impact on the efficacy of glutamate synaptic transmission. Given evidence linking alterations in glutamate neurotransmission to various neuropsychiatric disorders, we investigated the possible influence of a down-regulation of VGLUT1 transporter on anxiety, depressive-like behaviour and learning. The behavioural phenotype of VGLUT1-heterozygous mice (C57BL/6) was compared to wild-type (WT) littermates. Moreover, VGLUT1-3 expression, hippocampal excitatory terminal ultrastructure and neurochemical phenotype were analysed. VGLUT1-heterozygous mice displayed normal spontaneous locomotor activity, increased anxiety in the light-dark exploration test and depressive-like behaviour in the forced swimming test: no differences were shown in the elevated plus-maze model of anxiety. In the novel object recognition test, VGLUT1(+/-) mice showed normal short-term but impaired long-term memory. Spatial memory in the Morris water maze was unaffected. Western blot analysis confirmed that VGLUT1 heterozygotes expressed half the amount of transporter compared to WT. In addition, a reduction in the reserve pool of synaptic vesicles of hippocampal excitatory terminals and a 35-45% reduction in GABA in the frontal cortex and the hippocampus were observed in the mutant mice. These observations suggest that a VGLUT1-mediated presynaptic alteration of the glutamatergic synapses, in specific brain regions, leads to a behavioural phenotype resembling certain aspects of psychiatric and cognitive disorders.

Behavioural phenotyping of sodium‐myo‐inositol cotransporter heterozygous knockout mice with reduced brain inositol

Inositol plays a key role in dopamine, serotonin, noradrenaline and acetylcholine neurotransmission, and inositol treatment is reported to have beneficial effects in depression and anxiety. Therefore, a reduction in brain intracellular inositol levels could be a cause of some psychiatric disorders, such as depression or anxiety. To determine the behavioural consequences of inositol depletion, we studied the behaviour of sodium-dependent myo-inositol cotransporter-1 heterozygous knockout mice. In heterozygous mice, free inositol levels were reduced by 15% in the frontal cortex and by 25% in the hippocampus, but they did not differ from their wild-type littermates in cholinergic-mediated lithium-pilocarpine seizures, in the apomorphine-induced stereotypic climbing model of dopaminergic system function, in the Porsolt forced-swimming test model of depression, in amphetamine-induced hyperactivity, or in the elevated plus-maze model of anxiety. Reduction of brain inositol by more than 25% may be required to elicit neurobehavioural effects.

Neuropharmacological evaluation of the putative anxiolytic effects ofPassiflora edulis Sims, its sub-fractions and flavonoid constituents

Passiflora edulis Sims together with several other plants of the genus Passiflora have been reported to possess anxiolytic properties. It has been suggested recently that flavonoids may be partly responsible for the neuropharmacological activity of these plants but there are still few data reporting the relation between the constituents of these plants and their activity. This work evaluated the anxiolytic/sedative activity of an aqueous extract of Passiflora edulis Sims and bioguided its fractionation using the elevated plus-maze model of anxiety and other complementary pharmacological tests. The aqueous extract presented an anxiolytic-like activity without any significant effect upon the motor activity whilst the total flavonoid fraction (TFF) presented an anxiolytic-like activity but compromised motor activity. Through fractionation of TFF it was possible to isolate and characterize luteolin-7-O-[2-rhamnosylglucoside] which showed an anxiolytic-like activity without compromising motor activity.

Anxiolytic effects of Salvia reuterana Boiss. on the elevated plus-maze model of anxiety in mice

The anxiolytic and sedative effects of hydroalcoholic extract (HE) of Salvia reuterana Boiss. was evaluated in mice. The HE of Salvia reuterana (100 mg/kg), increased the percentage of time-spent and the percentage of arm entries in the open arms of the elevated plus-maze. Spontaneous locomotor activity count measured in 15 min of the test was significantly decreased in animals pretreated with diazepam and 100 mg/kg of Salvia reuterana extract. Results of the present study provide support for the traditional usage of Salvia reuterana as a sedative and anxiolytic medicinal plant.

Anxiolytic effects of Echium amoenum on the elevated plus-maze model of anxiety in mice

The ethanolic extract of Echium amoenum flowers at the dose of 50 mg/kg increased the percentage of time-spent and the percentage of arm entries in the open arms of the elevated plus-maze (EPM) and decreased the percentage of time-spent in the closed arms of EPM. Moreover, it prolonged the ketamine-induced latency to sleep but had no significant effects on total sleeping time induced by ketamine. Also, the locomotor activity was affected but not to the same extent as observed for diazepam. These results suggested that the extract of E. amoenum seems to possess anxiolytic effect with lower sedative activity than that of diazepam.

Anxiolytic effects of Stachys lavandulifolia Vahl on the elevated plus-maze model of anxiety in mice

Interest in alternative medicine and plant-derived medications that affect the “mind” is growing. The aim of the present study was to investigate the effects of a hydroalcoholic extract and essential oil of Stachys lavanduifolia Vahl on the elevated plus-maze (EPM) model of anxiety. The Stachys lavandulifolia extract or its essential oil was administered intraperitoneally to male TO mice, at various doses, 30 min before the behavioral evaluation. The extract of Stachys lavandulifolia at the dose of 100 mg/kg increased the percentage of time spent and the percentage of arm entries in the open arms of the EPM and decreased the percentage of time spent and the percentage of arm entries in the closed arms of the EPM. The plant extract at doses lower than 100 mg/kg had no significant effects on any of the parameters measured on the EPM. This dose of the plant extract prolonged the ketamine-induced sleeping time, and decreased the locomotor activity in mice. These results suggested that the extract of Stachys lavandulifolia possessed anxiolytic effect with relatively lower sedative activity than diazepam. The essential oil of Stachys lavandulifolia, however, at doses of up to 100 mg/kg did not have any significant effects on the mice behaviour on the EPM.

P.1.193 Thyroid auto-antibodies in bipolar affective disorder: Is there a link with lithium treatment?

The present studies were designed to evaluate the competitive binding properties and functional effects of a novel nonpeptide CRF1 receptor antagonist, R121919. R121919 administered in doses of 0.63 to 20 mg/kg p.o. 60 min pretest in Wistar rats dose dependently attenuated the swim stress-induced anxiogenic-like behavior in the elevated plus-maze model of anxiety. Moreover, receptor autoradiography revealed that R121919 dose-dependently occupied brain CRF1 receptors in subjects tested in the plus-maze experiment. Orally administered doses of up to 20 mg/kg R121919 also blunted basal and swim stress-induced pituitary-adrenocortical activation, produced additional anxiolytic-like behavioral actions in the defensive withdrawal and defensive burying paradigms, and functionally antagonized the locomotor stimulatory properties of exogenously administered CRF. Taken together, these results suggest that the anxiolytic-like efficacy of R121919 in attenuating the stress-, novelty-, shock-, and CRF-induced increases in behavioral arousal is correlated with competitive blockade of central CRF1 receptors.

Rats with low exploratory activity in the elevated plus-maze have the increased expression of limbic system-associated membrane protein gene in the periaqueductal grey

The aim of a present study was to analyse the gene expression profiles in the periaqueductal grey (PAG) of rats related to their exploratory activity in the elevated plus-maze model of anxiety. Animals were divided into the groups according to their exploratory activity in the plus-maze as follows: rats with low activity (‘anxious’), moderate activity (‘intermediate’) and high activity (‘non-anxious’). Control animals were not exposed to the elevated plus-maze. The differential expression of genes was analysed using the cDNA representational difference analysis (RDA) in combination with the sequencing and database search. Reverse transcription-polymerase chain reaction with specific primers was applied to confirm the differences found by the RDA. We established that animals displaying the different exploratory activity have also the different gene expression profiles in the PAG. Among the identified genes, we were able to confirm the increased expression of limbic system-associated membrane protein (LSAMP) in animals having the reduced exploratory activity in the elevated plus-maze. ‘Anxious’ group of rats had 1.6-fold higher expression of LSAMP gene compared to ‘non-anxious’ animals. By contrast, ‘home-cage’ control rats and ‘intermediate’ group did not differ significantly by their LSAMP gene expression level. In conclusion, it is likely that LSAMP plays a role in the regulation of exploratory behaviour of rats in the novel aversive environment.

Rats with low exploratory activity in the elevated plus-maze have the increased expression of limbic system-associated membrane protein gene in the periaqueductal grey

The aim of a present study was to analyse the gene expression profiles in the periaqueductal grey (PAG) of rats related to their exploratory activity in the elevated plus-maze model of anxiety. Animals were divided into the groups according to their exploratory activity in the plus-maze as follows: rats with low activity (‘anxious’), moderate activity (‘intermediate’) and high activity (‘non-anxious’). Control animals were not exposed to the elevated plus-maze. The differential expression of genes was analysed using the cDNA representational difference analysis (RDA) in combination with the sequencing and database search. Reverse transcription-polymerase chain reaction with specific primers was applied to confirm the differences found by the RDA. We established that animals displaying the different exploratory activity have also the different gene expression profiles in the PAG. Among the identified genes, we were able to confirm the increased expression of limbic system-associated membrane protein (LSAMP) in animals having the reduced exploratory activity in the elevated plus-maze. ‘Anxious’ group of rats had 1.6-fold higher expression of LSAMP gene compared to ‘non-anxious’ animals. By contrast, ‘home-cage’ control rats and ‘intermediate’ group did not differ significantly by their LSAMP gene expression level. In conclusion, it is likely that LSAMP plays a role in the regulation of exploratory behaviour of rats in the novel aversive environment.

Targeted mutation of CCK2 receptor gene modifies the behavioural effects of diazepam in female mice.

Evidence suggests that GABA and CCK have opposite roles in the regulation of anxiety. The aim of the present work was to study diazepam-induced anxiolytic-like action and impairment of motor co-ordination, and the parameters of benzodiazepine receptors in mice lacking CCK2 receptors. The action of diazepam (0.5-3 mg/kg i.p.) was studied in the elevated plus-maze model of anxiety and rotarod test using mice lacking CCK2 receptors. The parameters of benzodiazepine receptors were analysed using [3H]-flunitrazepam binding. In the plus-maze test, the exploratory activity of the homozygous (-/-) mice was significantly higher compared to their wild-type (+/+) littermates. However, the wild-type (+/+) mice displayed higher sensitivity to the anxiolytic-like action of diazepam. Even the lowest dose of diazepam (0.5 mg/kg) induced a significant increase of open arm entries in the wild-type (+/+) mice. A similar effect in the homozygous (-/-) mice was established after the administration of diazepam 1 mg/kg. The highest dose of diazepam (3 mg/kg) caused a prominent anxiolytic-like effect in the wild-type (+/+) mice, whereas in the homozygous (-/-) animals suppression of locomotor activity was evident. The performance of the homozygous (-/-) mice in the rotarod test did not differ from that of the wild-type (+/+) littermates. However, a difference between the wild-type (+/+) and homozygous (-/-) animals became evident after treatment with diazepam. Diazepam (0.5 and 3 mg/kg) induced significantly stronger impairment of motor co-ordination in the homozygous (-/-) mice compared to their wild-type (+/+) littermates. The density of benzodiazepine binding sites was increased in the cerebellum, but not in the cerebral cortex and hippocampus, of the homozygous (-/-) mice. Female mice lacking CCK2 receptors are less anxious than their wild-type (+/+) littermates. The reduced anxiety in homozygous (-/-) mice probably explains why the administration of a higher dose of diazepam is necessary to induce an anxiolytic-like action in these animals. The highest dose of diazepam (3 mg/kg) induced significantly stronger suppression of locomotor activity and impairment of motor co-ordination in the homozygous (-/-) mice compared to the wild-type (+/+) littermates. The increase in the action of diazepam is probably related to the elevated density of benzodiazepine receptors in the cerebellum of homozygous (-/-) mice. The present study seems to be in favour of increased tone of the GABAergic system in mice without CCK2 receptors.

Dose-related effects of propericiazine in rats.

We evaluated the effects of the neuroleptic agent propericiazine on animal models of anxiety and memory. Adult male Wistar rats (250 to 350 g) received intraperitoneal injections of propericiazine (0.05, 0.075 and 0.1 mg/kg), diazepam (1 mg/kg), saline, or diazepam vehicle (20% propylene glycol and 80% saline) 30 min prior to the experimental procedure. Animals (10-15 for each task) were tested for step-down inhibitory avoidance (0.3-mA footshock) and habituation to an open-field for memory assessment, and submitted to the elevated plus-maze to evaluate the effects of propericiazine in a model of anxiety. Animals treated with 0.075 mg/kg propericiazine showed a reduction in anxiety measures (P<0.05) similar to that observed in those treated with diazepam. Propericiazine at the doses of 0.05 and 0.1 mg/kg had no significant anxiolytic effects (P>0.05) in the elevated plus-maze model of anxiety. Memory was not affected by propericiazine in any of the tests, but was impaired by diazepam. The results indicate a dose-related, inverse U-shaped effect of propericiazine in an anxiety model, but not on memory tasks, perhaps reflecting involvement of the dopaminergic system in the mechanisms of anxiety.

Brain Penetrance, Receptor Occupancy and Antistress In Vivo Efficacy of a Small Molecule Corticotropin Releasing Factor Type I Receptor Selective Antagonist

The present studies were designed to evaluate the competitive binding properties and functional effects of a novel nonpeptide CRF1 receptor antagonist, R121919. R121919 administered in doses of 0.63 to 20 mg/kg p.o. 60 min pretest in Wistar rats dose dependently attenuated the swim stress-induced anxiogenic-like behavior in the elevated plus-maze model of anxiety. Moreover, receptor autoradiography revealed that R121919 dose-dependently occupied brain CRF1 receptors in subjects tested in the plus-maze experiment. Orally administered doses of up to 20 mg/kg R121919 also blunted basal and swim stress-induced pituitary-adrenocortical activation, produced additional anxiolytic-like behavioral actions in the defensive withdrawal and defensive burying paradigms, and functionally antagonized the locomotor stimulatory properties of exogenously administered CRF. Taken together, these results suggest that the anxiolytic-like efficacy of R121919 in attenuating the stress-, novelty-, shock-, and CRF-induced increases in behavioral arousal is correlated with competitive blockade of central CRF1 receptors.

Anti-anxiety studies on extracts of Passiflora incarnata Linneaus

Passiflora incarnata Linn. has been used to cure anxiety and insomnia since time immemorial. Despite the worldwide use of P. incarnata, the pharmacological work on this plant had been inadequate, inconclusive and wage as the earlier reports were unable to infer the mode of action of the plant as well as the phytoconstituents responsible for the much acclaimed anxiolytic and sedative effects of P. incarnata. An attempt has been made to isolate and identify the bioactive phytomoiety of P. incarnata by resorting to bioactivity directed fractionation and chromatographic procedures. A fraction derived from the methanol extract of P. incarnata has been observed to exhibit significant anxiolytic activity at a dose of 10 mg/kg in mice using elevated plus-maze model of anxiety. This fraction comprises mainly two components which are visible as blue and turquoise colored fluorescent spots at 366 nm of the UV light. The possibility of a phytoconstituent having benzoflavone nucleus as the basic moiety being responsible for the bioactivity of P. incarnata is highly anticipated.

Anxiolytic-like effects in rats produced by ventral pallidal injection of both N- and C-terminal fragments of substance P.

Prior studies have shown that the neurokinin substance P (SP) has anxiolytic-like effects when administered into the nucleus basalis (NB) area of the rat ventral pallidum. The present work was performed to examine whether the anxiolytic effects of SP in the nucleus basalis can be assigned its amino (N)- or carboxy (C)-terminal moiety. Using the elevated plus-maze model of anxiety in combination with unilateral injection of N-terminal SP(1-7) or C-terminal SP(7-11) into the NB region, we found that the treatment with either SP-fragment increased the number of entries into and time spent on the open arms as well as excursions into the end of the open arms, indicative of an anxiolytic-like profile. Furthermore, the effective doses of SP(1-7) (0.67 ng) and SP(7-11) (0.45 ng) were equimolar to the dosage of the whole SP molecule (1 ng), which was effective to reduce anxiety. Thus, the results support earlier findings that ventral pallidal injection of SP has anxiolytic-like effects and provide new evidence that fragments of SP, representing the N- and C-terminal domain of the peptide can reduce fear-parameters at a concentration similar to that of the parent peptide.