Burying Behavior Test Research Articles

Prolame produces anxiolytic- and antidepressant-like effects in middle-aged female rats with less uterotrophic effects than 17β-estradiol

Estrogen hormone replacement therapy (EHRT), improving women's life quality at menopause, reduces anxiety and depression symptoms associated with ovarian hormonal decline. However, its potential adverse effects, like thromboembolism and cancer risk, limit its use. Prolame is a synthetic 17β-amino estrogen with antithrombotic actions that exerts anxiolytic- and antidepressant-like effects on young adult ovariectomized female rats. It is unknown if prolame's effects may be observed in age and endocrine conditions emulating menopause. This study aimed to identify the antidepressant- and anxiolytic-like effects of prolame and E2 (used as a reference estrogen treatment) in middle-aged female rats coursing with irregular cycles, in two different conditions: ovariectomized or gonadally intact. Results were compared with those from young adult ovariectomized rats. Prolame (60 or 120 μg/kg), 17β-estradiol (E2, 40 or 80 μg/kg), or vehicle were chronically administered, and their effects were evaluated in the elevated plus-maze, defensive burying behavior test, open field test, and forced swimming test. Uterotrophic actions were estimated by uterine weight related to body weight. Prolame and E2 produced robust anxiolytic- and antidepressant-like effects in young adult ovariectomized rats, but these effects were absent in gonadally intact middle-aged rats. Interestingly, only prolame induced anxiolytic- and antidepressant-like effects in middle-aged ovariectomized rats. Uterotrophic effects of prolame were weaker than E2 effects, notably in middle-aged females. Altogether, present data support the notion that prolame has the potential to be considered an EHRT with relevant psychoactive actions and with apparently lower adverse-side effects, especially in middle-aged populations.

Acute taurine reduced alcohol intake and preference in alcohol-experienced, but not in alcohol-näive rats by central mechanisms

Taurine is a non-essential β sulfonated amino acid involved in a plethora of biological functions in the mammalian central nervous system. Taurine is easily accessible in energy drinks for human consumption. Previous preclinical and clinical reports suggest that acute systemic administration of taurine could inhibit some of the behavioral and metabolic effects of alcohol use disorder. Overall, both in rodent and human studies, acute taurine administration reduced voluntary alcohol intake. This study aimed to assess the pharmacological effects of taurine (intracerebroventricular; i.c.v.) on ethanol intake/preference of rats either control (i.e., alcohol naïve) or forced ethanol intake (since juvenile age with a chronic intermittent access model). In addition, to explore anxiety-like behavior (through defensive burying behavior test) as pharmacological control of taurine. We found that acute (i.c.v.) taurine reduced alcohol consumption, i.e., taurine significantly decreased both alcohol intake and preference in adult male Wistar rats. Moreover, taurine elicits an anxiolytic-like effect in all administered groups independently of previous alcohol exposure.

Effect of chronic unpredictable stress in female Wistar-Kyoto rats subjected to progesterone withdrawal: Relevance for Premenstrual Dysphoric Disorder neurobiology

Premenstrual Dysphoric Disorder (PMDD) is related to an abrupt drop in progesterone and impairments in the HPA axis that cause anxiety. Suffering persons report higher daily-life stress and anxiety proneness that may contribute to developing PMDD, considered a chronic stress-related disorder. Here, we explored the effect of chronic unpredictable stress (CUS) in rats subjected to progesterone withdrawal (PW) and evaluated gene expression of HPA axis activation in the stress-vulnerable Wistar-Kyoto (WKY) rat strain that is prone to anxiety. Ovariectomized WKY rats were randomly assigned to CUS or Standard-housed conditions (SHC) for 30 days. To induce PW, animals received 2 mg/kg of progesterone on day 25th for 5 days; 24 h later, they were tested using the anxiety-like burying behavior test (BBT). After behavioral completion, rats were euthanized, and brains were extracted to measure Crh (PVN) and Nr3c1 (hippocampus) mRNA. Blood corticosterone and vasopressin levels were determined. Results showed that PW exacerbated anxiety-like behaviors through passive coping in CUS-WKY. PW decreased Crh-PVN mRNA and the Nr3c1-hippocampal mRNA expression in SHC. CUS decreased Crh-PVN mRNA compared to SHC, and no further changes were observed by PW or BBT exposure. CUS reduced Nr3c1-hippocampal gene expression compared to SHC animals, and lower Nr3c1 mRNA was detected due to BBT. The PW increased corticosterone in SHC and CUS rats; however, CUS blunted corticosterone when combined with PW+BBT and similarly occurred in vasopressin concentrations. Chronic stress blunts the response of components of the HPA axis regulation when PW and BBT (systemic and psychogenic stressors, respectively) are presented. This response may facilitate less adaptive behaviors through passive coping in stress-vulnerable subjects in a preclinical model of premenstrual anxiety.

Anxiety-like Behavior and GABAAR/BDZ Binding Site Response to Progesterone Withdrawal in a Stress-Vulnerable Strain, the Wistar Kyoto Rats

Stress susceptibility could play a role in developing premenstrual anxiety due to abnormalities in the hypothalamus–pituitary–adrenal (HPA) axis and impairments in the GABAA receptors’ benzodiazepine (BDZ) site. Hence, we studied the stress-vulnerable Wistar Kyoto rat strain (WKY) to evaluate progesterone withdrawal (PW) effects on anxiety, HPA axis response, and to explore indicators of GABAA functionality in the BDZ site. For five days, ovariectomized WKY rats were administered 2.0 mg/kg of progesterone. Twenty-four hours after the last administration, rats were tested in the anxiety-like burying behavior test (BBT) or elevated plus maze test (EPM), and corticosterone was determined. [3H]Flunitrazepam binding autoradiography served as the BDZ binding site index of the GABAA receptor in amygdala nuclei and hippocampus’s dentate gyrus (DG). Finally, different doses of diazepam in PW-WKY rats were tested in the BBT. PW induced anxiety-like behaviors in both BBT and EPM compared with No-PW rats. PW increased corticosterone, but was blunted when combined with PW and BBT. PW increased [3H]Flunitrazepam binding in the DG and central amygdala compared with No-PW rats. Diazepam at a low dose induced an anxiogenic-like response in PW rats, suggesting a paradoxical response to benzodiazepines. Overall, PW induced anxiety-like behavior, a blunted HPA axis response, and higher GABAAR/BZD binding site sensitivity in a stress-vulnerable rat strain. These findings demonstrate the role of stress-susceptibility in GABAAR functionality in a preclinical approximation of PMDD.

Experimental protocol for detecting higher alcohol consumers from a conventional rat line based on basal anxiety

Predisposition for a high alcohol intake and the impact of alcohol-abstinence-relapse may be reliable experimentally performed in conventional adult rat lines if animals received juvenile exposure to alcohol (e.g., by forced consumption) and selecting those individuals with high basal anxiety levels during juvenile periods. Importantly, a forced alcohol consumption phase must be followed by an imposed withdrawal period to form an exposure-abstinence cycle (at least two cycles are required) which allow to obtain animals with notorious alcohol relapses. The easier way to test alcohol relapses is through voluntary ethanol intake models. On the other hand, the anxiety classification may be performance by classical paradigms such as an elevated plus maze test, defensive burying behavior test or any other. Here, we provide a step-by-step protocol description to detect higher alcohol consumers animals from male Wistar rats. This protocol should be especially useful for those interested in studying the participation of specific brain nucleus [e.g., periaqueductal gray (PAG)] and/or the neurotransmitters involved [e.g., neuropeptide Y (NPY)] in the alcohol intake phenomena if it is combined with stereotaxic surgery. However, every administration route of treatments or experimental design is appropriate; the limit is the own imagination, and the resources.

NPY-Y1 receptors in dorsal periaqueductal gray modulate anxiety, alcohol intake, and relapse in Wistar rats

Neuropeptide Y (NPY) is likely the main endogenous anxiolytic neuromodulator involved in alcohol intake. NPY-Y1, a receptor for NPY, is highly expressed in the periaqueductal gray (PAG), a mesencephalic structure involved in integrating nervous activity to the performance of active and passive defensive behaviors related to fear and anxiety. Interestingly, anxiety and fear are some of the prevailing emotional negative states during alcohol abstinence. Moreover, an inverse relationship between NPY activity and alcohol consumption has been frequently reported, mainly in the extended amygdala. Nevertheless, both the roles of NPY and that of the receptor involved in these actions have been scarcely studied. Thus, the aim of this study was to analyze the pharmacological effect of NPY and NPY-Y1 receptor blockade into the dorsal periaqueductal gray (D-PAG) in an alcohol consumption and relapse paradigm in adult male Wistar rats.Ninety-six rats at postnatal day 42 (PND-42) were classified as having low and high anxiety (LA and HA), respectively, through the elevated plus maze test (EPM). Then, those animals were randomly divided into alcohol naïve (AN) and forced alcohol consumption (FAC) groups. A cannula was implanted in D-PAG to microinject vehicle (VEH), NPY, or BIBP-3226 (a selective NPY-Y1 receptor antagonist). A defensive burying behavior test (DBB) was performed to assess the anxiety-like state during withdrawal, followed by a 24-hour free choice voluntary alcohol intake test. Under our experimental conditions, NPY microinjection decreased alcohol consumption in HA rats, whereas NPY-Y1 receptor blockade in D-PAG produced a notably anxiogenic effect and higher alcohol intake and relapse. In conclusion, NPY in the D-PAG, most likely acting on NPY-Y1 receptors, induced a significant anxiolytic effect and prominently inhibited alcohol consumption and relapse in Wistar rats.

Reduction of dopaminergic transmission in the globus pallidus increases anxiety-like behavior without altering motor activity

The globus pallidus (GP) plays an important role in the flow of information between input and output structures of the basal ganglia (BG) circuit. In addition to participating in motor control, the GP may also be involved in cognitive and emotional functions related to the symptoms of patients with Parkinson’s disease (PD). Since the GP receives dopaminergic innervation from the substantia nigra pars compacta (SNc), it is important to determine whether a local dopamine (DA) deficit in the GP is related not only to motor but also to the cognitive and emotional alterations of PD. The aim of this study was to examine the effects of lesions in the GP (induced by 6-OHDA) on anxiety, depression and ambulation in rats. Such lesions are known to reduce dopaminergic innervation in this brain structure. Additionally, the effect on DA receptors in the GP was tested by local administration of the dopamine agonist PD168,077, antagonist haloperidol and psychostimulant amphetamine. Experimental anxiety was evaluated with the elevated plus maze (EPM), burying behavior test (BBT) and social interaction test, while depressive-like behavior was assessed with the sucrose preference test. Rats with unilateral and bilateral lesions showed a higher level of anxiety than intact animals in both the EPM and BBT, an effect also obtained after intrapallidal injection of haloperidol. The administration of methamphetamine or PD-168.077 caused the opposite effect. The dopaminergic lesions in the GP did not affect sucrose preference, social interaction or ambulation. These results show that dopamine in the GP, acting through D2 or D4 receptors, may be involved in the manifestation of anxiety, a non-motor symptom of PD that often appears before motor symptoms.

Enhanced central histaminergic transmission attenuates compulsive-like behavior in mice

Present investigation demonstrated the effect of central histaminergic transmission on the compulsive-like marble burying and spontaneous alteration behavior (SAB) in mice. Result demonstrates that on enhancement of endogenous histaminergic transmission in mice achieved by central (i.c.v.) administration of histamine or central histamine neuronal releaser, H3 receptor antagonist or on intraperitoneal (i.p.) administration of histamine precursor, l-histidine significantly attenuated the number of marble buried in marble burying behavior (MBB) test as well as obliterated the persistent behavior induced by 5-HT1A receptor agonist, 8-OH-DPAT in T-Maze test. Furthermore, central injection of histamine H1 receptor agonist, FMPH or H2 receptors agonist, amthamine also attenuated the MBB in mice. On the other hand, prior i.c.v administration of H1 but not H2 receptor antagonist attenuated the effects exhibited in MBB test on mice by all the above agents capable of enhancing the endogenous central histaminergic transmission. Thus, the results of the present investigation delineate the attenuating effect of central histaminergic transmission predominantly via H1 receptor on compulsive-like behavior in mice.

ATOMOXETINE FOR HOARDING DISORDER: A PRECLINICAL AND CLINICAL INVESTIGATION

Purpose of the study Hoarding disorder is a mental disorder that has been newly included in the DSM-5 into the obsessive-compulsive and related disorders chapter. To date only two open studies investigated the pharmacological management of hoarding disorder and its management remains controversial. Despite some studies suggested that childhood attention-deficit hyperactivity disorder (ADHD) and inattention symptoms may be related to hoarding [1], only a small case series study investigated the effectiveness of ADHD medications (metilphenidate) in hoarding disorder [2]. The aim of the present study was to evaluate the preclinical and clinical effectiveness of atomoxetine, a noradrenaline reuptake inhibitor approved for childhood and adulthood ADHD, in an animal model of compulsive-like behaviors (marble burying test) and in two patients with a primary diagnosis of hoarding disorder. Methods We performed a preclinical investigation assessing the effects of atomoxetine on the marble burying behavior test in mice. During the test, mice were placed individually in a clear plastic box containing 24 black marbles (1 cm diameter), evenly spaced on 5 cm sawdust. The number of marbles buried (to at least 60% of the depth of the sawdust) within 30 min in vehicle and atomoxetine treated groups was measured. Atomoxetine (10, 30 and 60 mg kg-1) was administered per os 30 min before the test. Subsequently, we conducted a clinical investigation on two patients fulfilling the DSM-5 criteria for hoarding disorder. These patients were treated with atomoxetine 40-100 mg for 12 weeks. A history of adult or childhood ADHD was excluded through the diagnostic interview DIVA 2.0. No changes of the ongoing medications or psychotherapy were allowed during the treatment period. To measure the severity of hoarding, the Saving Inventory-Revised (SI-R) and the UCLA Hoarding Severity Scale (UHSS) were administered before and after the treatment. Response was defined as a more of 30% decrease in UHSS and SI-R scores and at least “much improved” on the Clinical Global Impression/Improvement (CGI-I) scale. Results Atomoxetine significantly reduced the number of buried marbles in a dose dependent manner in comparison to control mice, without affecting the locomotor activity. Atomoxetine (30 and 60 mg kg-1) significantly reduced the number of buried marbles in a dose dependent manner (12.3 ± 1.0 and 10.0 ± 1.8, respectively) in comparison to control mice (19.3 ± 1.0), without affecting the locomotor activity. Atomoxetine 10 mg kg-1 was not effective (21.3 ± 0.9). Atomoxetine-treated patients were both classified as treatment responders after 12 weeks of treatment. Atomoxetine was well tolerated during all the treatment period. Furthermore, both patients as were still classified responders at the 6 months follow-up visits. Conclusions These preclinical and clinical data suggest that atomoxetine may be considered as a potentially effective compound for hoarding disorder. Therefore, atomoxetine should be considered for future controlled trials in hoarding disorder.

Anxiolytic- and antidepressant-like effects of an aqueous extract of Tanacetum parthenium L. Schultz-Bip (Asteraceae) in mice

AimTanacetum parthenium L. Schultz-Bip (Asteraceae) is widely used worldwide in traditional medicine for the treatment of convulsions and culture-bound syndromes such as susto (fear). The aim of this work was to evaluate the anxiolytic- and antidepressant-like effects of an aqueous extract of T. parthenium in behavioral paradigms in mice. The effects of T. parthenium were compared with those produced by anxiolytic and antidepressant drugs. We carried out the chemical characterization of the main constituents of T. parthenium. The involvement with the GABAergic and serotoninergic neurotransmitter systems were explored be means of synergic and antagonist experiments. Materials and methodsThe anxiolytic-like effect was evaluated using the Burying Behavior Test (BBT) and the Elevated Plus-Maze Test (PMT). The antidepressant-like effect was evaluated in the Forced Swimming Test (FST), and ambulatory activity was assessed in the Open Field Test (OFT). Employing the behavioral tests, synergism and antagonism experiments with Alprazolam, Muscimol, and Picrotoxin were carried out in the PMT. In a series of independent experiments, concomitant administration of T. parthenium and Alprazolam, Fluoxetine, or p-chlorophenylalanine were conducted in the FST.For chemical characterization, High-Performance Liquid Chromatography-Electro Spray Ionization-Mass Spectrometry (HPLC-ESI-MS) analysis was performed. ResultsT. parthenium exerts clear anxiolytic- and antidepressant-like effects in mice, without affecting the ambulatory activity of the experimental subjects. ConclusionsAnxiolytic- and antidepressant-like T. parthenium effects result, at least part from the involvement of the GABAergic system. Our results support the use of Tanacetum parthenium in traditional medicine and suggest its therapeutic potential in the comorbid anxiety and depression.

Environmental enrichment prevents anxiety-like behavior induced by progesterone withdrawal in two strains of rats

Stress vulnerability could influence the treatment response to anxiety associated with abrupt hormonal suppression. The present study explored the effects of different treatments on experimental anxiety induced by progesterone withdrawal (PW) in a stress-sensitive rat strain, Wistar Kyoto (WKY), in the burying behavior test (BBT). The following experimental series was conducted using independent groups of Wistar (control strain) and WKY ovariectomized rats: Experiment 1: Rats were treated for 5days with oil, a constant dose of progesterone (0.5mg/rat, s.c) or a combination of progesterone (0.5mg/rat, s.c) plus fluoxetine (10 mg/kg, i.p); on day 6, all rats were subjected to BBT. Experiment 2: Rats received corn oil or decreasing doses of progesterone (0.84, 0.67, 0.5, 0.33 and 0.17mg/rat; one dose daily); on day 6, the rats were subjected to BBT. Experiment 3: Rats were divided into two groups that were subjected to 30days of standard conditions or environmental enrichment (EE); from days 25 to 30, all rats received a fixed dose of progesterone (0.5mg/rat, s.c.) or vehicle. On day 31, the rats were tested with BBT. Results showed that PW increased anxiety in both strains, and fluoxetine prevented anxiety in WKY rats. In contrast, a gradual reduction of progesterone prevents the anxiety in Wistar but not in WKY. EE was preventive against the anxiety induced by PW in both strains of rats. Thus, the results suggest that anxiety induced by PW is prevented by EE while the anxiolytic effect of pharmacological treatments depends on stress vulnerability.

GABAA/benzodiazepine receptor complex mediates the anxiolytic-like effect of Montanoa tomentosa

Ethnopharmacological relevanceMontanoa tomentosa also named Cihuapatli is a native plant of Mexico that has been used in traditional medicine for the last five centuries mainly as a remedy for reproductive impairments. However, there are reports indicating that this plant was also consumed by Mexican ancient people for its relaxing properties. In order to corroborate this information, the present study was conducted to evaluate the effect of Montanoa tomentosa lyophilisate (MT) on rat׳s anxiety-like behavior and to analyze its mechanism of action. Materials and methodsThe anxiolytic-like action of MT (1.5, 3.0, 6.0 and 12.0mg/kg) was investigated in male Wistar rats tested in three animal models of anxiety: the burying behavior, the elevated plus maze and the hole-board tests. As a positive control, the anti-anxiety effects of different doses of the selective GABAA receptor agonist muscimol were also analyzed. In order to evaluate the participation of the GABAA and oxytocin receptors in the anxiolytic-like actions of MT, the GABAA receptors blockers picrotoxin (0.25 and 0.50mg/kg), bicuculline (2.0mg/kg) and flumazenil (5.00 and 10.0mg/kg), the neurosteroid inhibitor finasteride (50.0 and 100mg/kg) and the oxytocin receptor antagonist atosiban (0.25µg) were used. Finally, to evaluate general activity, and motor coordination, the open field and rota-rod tests were used. ResultsMT at 3.0mg/kg showed anxiolytic-like effects in the three anxiety paradigms without affecting reactivity, general motor activity or motor coordination; however, at higher doses sedative effects were observed. Picrotoxin (0.25 and 0.50mg/kg), flumazenil (10.0mg/kg) and finasteride (100mg/kg) antagonized the anxiolytic-like actions of MT in the burying behavior test. In the plus maze and hole-board tests bicuculline (2.0mg/kg) blocked the effects of the plant as well. Atosiban (0.25µg) did not antagonize the anxiolytic-like actions of MT. ConclusionsThe results corroborate the anxiolytic-like actions of Montanoa tomentosa and suggest that this effect is mediated through GABAA receptors but not oxytocin receptors.

Experimental evidence for involvement of nitric oxide in low frequency magnetic field induced obsessive compulsive disorder-like behavior

It is well documented that extremely low frequency magnetic field (ELF MF) produced effects on the function of nervous system in humans and laboratory animals. Dopaminergic and serotonergic pathways have been implicated in obsessive compulsive disorder (OCD). Recently involvement of nitric oxide (NO) in OCD-like behavior is suggested. Hence, the present study was carried out to understand the involvement of dopamine, serotonin and NO in ELF MF induced OCD-like behavior. Swiss albino mice were exposed to ELF MF (50Hz, 10G) for 8h/day for 7, 30, 60, 90 and 120days by subjecting them to Helmholtz coils. OCD-like behavior was assessed in terms of marble burying behavior (MBB) test. Results revealed that ELF MF induced time dependant MBB, on 7th, 30th, 60th, 90th, and 120th exposure day.Further, levels of dopamine, serotonin and NO after 120days of ELF MF exposure were determined in the regions of the brain. The neurohumoral studies revealed that exposure to ELF MF increased NO levels in cortex, hippocampus and hypothalamus, and levels of dopamine and serotonin remain unaffected. As OCD-like behavior after ELF MF exposure was associated with higher levels of NO with no significant change in serotonin and dopamine. The effect of such exposure was studied in groups concurrently treated with NO modulators, NO precursor, l-ARG (400mg/kg) or NOS inhibitor, l-NAME (15.0mg/kg) or 7-NI (10.0mg/kg). These treatments revealed that NO precursor exacerbated and NOS inhibitors attenuated ELF MF induced OCD-like behavior with corresponding changes in the levels of NO.

Long-Term Behavioral Consequences of Prenatal Binge Toluene Exposure in Adolescent Rats

The continued abuse of inhaled organic solvents, especially among women of childbearing age, raises the risk of long-term behavioral effects of maternal toluene abuse. In this study, the effects of short-term exposures to high toluene concentrations (i.e., “binges”) were tested in independent groups of adolescent rats with different toluene treatments: (a) acute: 30-day-old animals exposed for 30min to air (A) or 6,000 ppm toluene (T); (b) prenatal and postnatal: rats exposed to T or A from gestation days 8–20 and re-exposed to T or A from postnatal day (PN) 22 to PN30 (A/A, T/A, A/T, and T/T, resp.). On PN30, animals were evaluated in different tests. Postnatal toluene exposure produced anxiolytic-like effects in the burying behavior test, and the T/T group received the highest number of electrical shocks. Antinociception was observed in the T, A/T, and T/T groups in the hotplate test. All toluene treatments impaired short-term memory in the object recognition test, but only postnatal exposure impaired long-term memory in the passive avoidance test. Sensitization occurred in the T/T group in locomotor activity. These results indicate that prenatal exposure to a concentration of toluene that does not produce evident malformations can modify behavioral toluene’s effects in adolescent rats.

Environmental enrichment increases doublecortin-associated new neurons and decreases neuronal death without modifying anxiety-like behavior in mice chronically exposed to toluene

Toluene misuse is a health problem worldwide with broad effects at the level of the central nervous system; however, therapeutic alternatives for inhalant abusers are limited. Chronic use of volatile substances is associated with different neurological and cognitive alterations, being anxiety a psychiatric condition with high prevalence. At cellular level toluene reduces neurogenesis and induces neuronal death. On the other hand, environmental enrichment has demonstrated to produce positive effects at behavioral and neuronal levels. Thus, the aim of the present work was to model alterations occasioned after repeated exposure to toluene (anxiety, reduction in neurogenesis – measured as doublecortin-labeled cells – and neuronal death). Subsequently, the influence of environmental enrichment on these effects was evaluated. Adolescent mice were exposed to toluene vapors from 1 to 4 weeks. Effects on anxiety were evaluated with the burying behavior test, whereas neurogenesis and hippocampal cell death were analyzed with immunohistochemistry, using anti-doublecortin or anti-active-Caspase-3 antibodies, respectively. Results showed that chronic toluene exposure increased anxiety in the burying behavior test; additionally, toluene decreased neurogenesis and enhanced neuronal death. Environmental enrichment (EE) enhanced the anxiety like response in air-exposed mice but did not modify the toluene anxiety response. Additionally, EE enhanced neurogenesis in toluene-pretreated animals at the same level to that found in animals unexposed to toluene and decreased neuronal death. Overall, the present study showed that environmental enrichment positively impacts some effects produced by repeated exposure to toluene.

Evaluation of emotional behaviors in young offspring of C57BL/6J mice after gestational and/or perinatal exposure to nicotine in six different time-windows

Nicotine replacement treatments are being alternatively applied as an aid to smoking cessation during pregnancy. However, the effects of nicotine exposed at the prenatal stage on the emotional behaviors in offspring are not well understood due to the lack of systematic investigations. The current study has therefore initially aimed to evaluate emotional behaviors in young mouse offspring (postnatal day 28–36) which experienced gestational and/or perinatal nicotine exposure (GPNE) in six different time-windows. Pregnant C57BL/6J mice were exposed to nicotine via sweetened (2% sucrose) drinking water during 6 different time-windows including gestational day 0–day 13 (G0–G13), G14–perinatal day 0 (P0), G0–P0, G14–P7, G0–P7, and P0–P7. During P28–P36 days, both male and female offspring were given a battery of behavioral tests including light and dark box test, marble burying behavior test, novelty-suppressed feeding test, sociability and social novelty preference test, social avoidance tube test, and elevated plus maze test. GPNE during G0–P0, G14–P0, G14–P7, and G0–P7 induced abnormal behaviors in male and female offspring to different extent. Results indicated that nicotine at any time points of gestational and/or perinatal period impairs emotional behaviors in offspring, and suggested certain time-windows for further neurochemical or molecular studies in relation with GPNE-induced emotional abnormalities.

Sex differences in the burying behavior test in middle-aged rats: Effects of diazepam

The full behavioral profile displayed during the burying behavior test was studied in middle aged (11–14months) males, females with irregular estrous cycles, and females in persistent diestrus, with and without diazepam (0.5–2.0mg/kg). Ambulation and motor coordination were also tested to discern behavioral changes from general motor alterations. Without diazepam treatment, middle-aged males showed longer burying behavior latencies, more prod explorations and less freezing than both groups of females. Untreated middle aged males also showed less cumulative burying and more immobility compared to females with irregular cycles. None of the parameters showed any difference between the female groups. Diazepam (0.5 and 1.0mg/kg) increased burying behavior latency in females, but had no effect on any parameter in middle aged males. However, a higher dose (2.0mg/kg) of diazepam increased immobility, freezing and the number of prod shocks and decreased prod explorations and groomings, but impaired motor coordination in males. In contrast with young males and females, diazepam at any dose reduced cumulative burying. Data are discussed on the bases of (1) sex and age differences in burying behavior and on (2) the anxiolytic-like action of diazepam and its side effects.

Evaluation of the anxiolytic-like effects of clomipramine in two rat strains with different anxiety vulnerability (Wistar and Wistar-Kyoto rats)

The main aim of this study was to evaluate the anxiolytic-like effects of the serotonergic antidepressant, clomipramine (0.6-5.0 mg/kg), and the 5-HT1A agonist, (±)8-hydroxy-2-(di-n)-propil-aminotetraline hydrobromide (8-OH-DPAT; 0.01-0.5 mg/kg), in two strains of rat with different anxiety vulnerability: Wistar-Kyoto (WKY; with trait anxiety) and Wistar rats (control strain). The anxiety model used was the burying behavior test; decreases in burying, grooming of the snout, and freezing were interpreted as a reduction of anxiety-like levels. A second objective was to explore the participation of 5-HT1A receptors in the effects of clomipramine and 8-OH-DPAT. Behavior in the burying behavior test was strain dependent. In addition to the burying behavior, WKY rats showed high levels of freezing and grooming of the snout. Clomipramine and 8-OH-DPAT decreased the burying behavior in both strains of rats through a direct interaction with the 5-HT1A receptor. 8-OH-DPAT decreased freezing behavior in both strains through a mechanism that was not related to 5-HT1A receptors. Finally, clomipramine was able to block freezing and grooming behaviors only in WKY rats. In conclusion, strains with different anxiety vulnerability express different behavioral responses toward the same aversive stimulus, and the anxiolytic-like effects of clomipramine and 8-OH-DPAT are both behavior and strain dependent.

Anti‐anxiety and sedative profile evaluation of imidazo[1,2‐a]pyridine derivatives

AbstractThree imidazo[1,2‐a]pyridine‐3‐nitrosated (L‐1, L‐2, L‐3) and a 3‐formyl imidazo[1,2‐a]pyridine thiosemicarbazone (L‐4) were synthesized and evaluated for their effects in the elevated plus maze, burying behavior test, rotarod performance, the horizontal wire test, and locomotor activity. L‐2 and L‐3 increased the percent time spent in the open arms of the plus maze at doses of 1 and 2 mg/kg without modifying the number of total entries. In addition, L‐2 and L‐3 (1 mg/kg) increased the number of open arm entries indicating anxiolytic‐like activity at this dose. In the burying behavioral test, L‐1 (2–8 mg/kg), L‐2 (8 mg/kg), and L‐3 (4 and 8 mg/kg), induced a clear reduction in cumulative burying behavior, without modifying burying behavior latency, thus reducing experimental anxiety. In the rotarod test, L‐1 and L‐2 impaired rotarod performance only at the highest evaluated dose (64 mg/kg) at which reduction of motor activity was observed and thereby no conclusions about myorelaxant effects can be proposed. All compounds showed a clear sedative effect and corresponding ED50 values were obtained. Results indicate that compounds L‐1, L‐2, and L‐3 show a sedative and an anxiolytic profile. Drug Dev Res 71:371–381, 2010. © 2010 Wiley‐Liss, Inc.

Influence of pharmacologically-induced experimental anxiety on nociception and antinociception in rats

Animal studies reveal that diverse environmental stimuli that generate anxiety-like behaviors also induce antinociception; conversely, clinical data show that pain perception is reduced under anxiolysis. This study was conducted to investigate the influence of pharmacologically induced-anxiety on nociception and antinociception. Experimental anxiety levels were measured using the rat burying behavior test. Diazepam (0, 0.5, 1.0 and 2.0 mg/kg, i.p.) or yohimbine (0, 0.5 and 1.0 mg/kg, i.p.) were used as anxiolytic or anxiogenic drugs, respectively. To evaluate the influence of different experimental anxiety levels on nociception, the pain-induced functional impairment in the rat (PIFIR model) was used. Nociception was induced by an intra-articular injection of 15% uric acid into the knee joint of the right hind limb. Diazepam or yohimbine were administered 15 min before uric acid and the ability of the rat to use the injured hind limb was recorded. To analyze the influence of different levels of anxiety on the antinociceptive effects produced by acetylsalicylic acid (0, 31, 100 and 310 mg/kg, p.o.); this analgesic was administered 3.5 h after uric acid. Fifteen min before diazepam (2.0 mg/kg) or yohimbine (1.0 mg/kg) were administered. We found that, in the burying behavior test, diazepam and yohimbine produced a dose-dependent decrease or augment in the cumulative time of burying, effects denoting reduced or increased experimental anxiety, respectively. Diazepam or yohimbine, administered alone, was unable to produce nociception. The results showed an influence of anxiety on nociception since a decreased (by diazepam) or increased (by yohimbine) experimental anxiety prevented nociception. Control experiments showed that acetylsalicylic acid did not modify experimental anxiety in the burying behavior test, but effectively reversed the nociception induced by uric acid (15%) in the PIFIR model. Such antinociceptive effect was unmodified by the anxiolytic or anxiogenic actions of diazepam or yohimbine. Data are discussed on the bases of clinical- and animal-studies revealing interactions between anxiety and nociception.