Pharmaco-ethological analysis of agonistic behavior between resident and intruder mice: effects of psychotropic drugs

The present study was conducted to investigate the effects of psychotropic drugs on agonistic behavior between resident and intruder mice. The effects of four doses of the following drugs were assessed in either resident or group-housed intruder mice: chlordiazepoxide (0, 5, 10 and 20 mg/kg, i.p.), haloperidol (0, 0.25, 0.50 and 0.75 mg/kg, i.p.) and imipramine (0, 5, 10 and 20 mg/kg, i.p.). Residents and intruders were drugged on alternate test days, and all animals received different sequences of each of the drug conditions according to a random schedule. The injection-test interval was 30 min. When resident mice were treated with chlordiazepoxide the resident's aggressive episodes (sideways posture, attack bite, tail rattle) were significantly suppressed. Both haloperidol and imipramine also showed a similar suppressive effect on the resident's aggressive episodes, but haloperidol significantly suppressed locomotor activity at all doses. When intruder mice were treated with chlordiazepoxide, attack bites by untreated residents were significantly increased in a dose-dependent manner, and the frequency of defensive upright posture displayed by intruder animals were significantly decreased. Haloperidol and imipramine did not alter resident's behavior and intruder's upright posture when intruders were drugged. The results suggest that chlordiazepoxide has specific effects on both the hostility of the resident and the anxiety of the intruder, differing from haloperidol and imipramine.     

Psychotropic effects of ginseng saponins on agonistic behavior between resident and intruder mice

The psychotropic actions of crude ginseng saponins, pure ginsenoside Rb1 (GS-Rb1) and ginsenoside Rg1 (GS-Rg1) obtained from the root of Panax ginseng, were evaluated from their effects on agonistic behavior in mice. A resident-intruder test situation was used. When the resident mouse was treated with crude ginseng saponins (25, 50 and 100 mg/kg i.p.), aggressive episodes (offensive sideways posture and attack bite) were significantly suppressed in a dose-dependent manner. However, the agonistic behavior was not altered when the intruder was treated with crude ginseng saponins. GS-Rb1 (2.5, 5 ad 10 mg/kg i.p.) also significantly suppressed aggressive episodes when given to the resident, whereas GS-Rg1 (2.5, 5 and 10 mg/kg i.p.) was ineffective. Neither GS-Rb1 nor GS-Rg1 given to the intruder caused any significant changes in the behavior of the resident. Although the highest dose of crude ginseng saponins suppressed locomotion frequency, it appears that both crude ginseng saponins and GS-Rb1 possess a specific psychotropic action on agonistic behavior.     

Ethopharmacology of maternal aggression in mice: effects of diazepam and SM-3997.

The present study investigated whether there is any difference between the effects of benzodiazepine and non-benzodiazepine anxiolytics on maternal aggression in lactating mice, using an ethological technique. We used SM-3997, a 5-HT1A receptor ligand, as a non-benzodiazepine anxiolytic. Behavior towards an intruder male mouse was assessed on postpartum days 5 and 7 in female mice that had been housed alone since the end of the 4-day mating period. Acute oral administration of diazepam had a biphasic effect on the frequency of bites: 1 mg/kg diazepam significantly increased bite frequency, while 2.5 mg/kg diazepam significantly decreased it. However, 2.5 mg/kg diazepam also caused a significant decrease in locomotor activity. In contrast, SM-3997 (1, 2.5 and 5 mg/kg p.o.) significantly decreased the frequency of bites in a dose-dependent manner without causing motor dysfunction. Chronic treatment with 5 mg/kg SM-3997 significantly decreased the frequency of bites when compared with vehicle, whereas diazepam was ineffective at the doses used (0.5 and 1 mg/kg p.o.). The findings suggest that the proaggressive effect is specific to benzodiazepines, and that 5-HT1A receptors may be involved in the suppression of maternal aggression in mice.     

Anticonflict action of tandospirone in a modified Geller-Seifter conflict test in rats.

Tandospirone is a novel non-benzodiazepine compound possessing potent anxiolytic properties in a water lick conflict paradigm in rats and a high affinity for central 5-HT1A receptors. In the present study, tandospirone was evaluated for anxiolytic activity in a modified Geller-Seifter conflict paradigm in rats. Tandospirone produced significant increases in the punished responding at doses of 1.25, 2.5 and 5.0 mg/kg, i.p. or 20 mg/kg, p.o., although it decreased unpunished responding at doses of 2.5 and 5.0 mg/kg, i.p. or 20 mg/kg, p.o. Likewise, diazepam was also effective after i.p.-administration in this test, and its minimum effective dose was slightly higher than that of tandospirone. This suggests that tandospirone might be as effective in the treatment of anxiety as diazepam. The anticonflict action of tandospirone was not inhibited by Ro-15-1788, a benzodiazepine antagonist, although that of diazepam was completely inhibited. 8-OH-DPAT, a full agonist of 5-HT1A receptors, was also effective in this test with a high potency. Therefore, the possibility exists that the anticonflict action of tandospirone is related to its agonist action on 5-HT1A receptors, not on benzodiazepine receptors.     

Alcohol and chlordiazepoxide increase suppressed aggression in mice

The purpose of the present experiments was to investigate conditions under which alcohol and chlordiazepoxide (CDP) enhance aggression. Alcohol (300, 600, and 1200 mg/kg orally) and CDP (5, 10, and 20 mg/kg) failed to alter attack behavior in male mice confronting intruder mice in their home cages. When confrontations between attack-experienced mice and group-housed mice occurred in a neutral cage, attack and threat behavior was suppressed to about 50% of that in the resident's home cage. A low dose of alcohol (300 mg/kg) more than doubled the frequency of attacks and threats in the neutral cage. Higher doses of both drugs decreased attacks in the neutral cage. Combined administration of low doses of alcohol (150 and 300 mg/kg) and CDP (2.5 and 5.0 mg/kg) increased attack and threat frequency in the neutral cage to a larger extent than when either drug was given itself. The aggression-enhancing effects of alcohol and CDP in the neutral cage situation are consistent with the view that both drugs can lead to behavioral disinhibition. Alternatively, the results could reflect simply the ratedependency of alcohol effects on attack behavior, that is, low rates are increased whereas high rates remain unaffected. The present results also provide evidence for an additive interaction of both drugs in their depressant effects and, also, in their aggressionheightening effects.     

Lack of effect of the 5-HT(1A) receptor antagonist WAY-100635 on murine agonistic behaviour.

The present study examined the influences of the selective 5-HT1A receptor antagonist, WAY-100635, on the social and agonistic behavior exhibited by male resident mice during encounters with unfamiliar intruder conspecifics. Acute administration of WAY-100635 (0.01-1.0 mg/kg sc) dose dependently enhanced the duration of resident maintenance behavior, reaching statistical significance at 1.0 mg/kg. The duration of resident attend/approach behavior was reduced at 0.01 mg/kg. Drug-free intruder animals showed a reduction in the frequency and duration of attend/approach behavior when the resident mice were treated with 0.01 mg/kg WAY-100635. No other significant effects on behavior were detected for WAY-100635. A previous investigation reported that WAY-100635 induced anxiolytic-like effects in the mouse light/dark box test. In the present study, however, the level of defensive behavior of the saline-treated resident mice was too low for any further anxiolytic-like attenuation of this behavior to be observed. Therefore, no conclusions regarding the potential anxiolytic activity of WAY-100635 may be drawn from the data presented here. Current results are consistent with data for the lack of effect of WAY-100635 on rat agonistic behavior but contrast with findings for the effects of the 5-HT1A receptor antagonists (+)-WAY-100135 and SDZ 216-525 on mouse agonistic behavior.     

Involvement of the dorsal hippocampus in mediation of the antianxiety action of tandospirone, a 5-hydroxytryptamine1A agonistic anxiolytic

The effect of tandospirone, a 5-hydroxytryptamine (5-HT)1A agonist/anxiolytic, injected directly into dorsal hippocampus, on Vogel-type conflict behavior in rats was investigated and the findings were compared with the effects of diazepam and zopiclone. Tandospirone (30 micrograms/2 microliters and 60 micrograms/2 microliters) and diazepam (40 micrograms/2 microliters) but not zopiclone (20 micrograms/2 microliters), produced a potent anticonflict action in rats. The anticonflict action of tandospirone (30 micrograms/2 microliters), injected into the dorsal hippocampus, was significantly blocked by (-)-propranolol (5 mg/kg i.p.). The present findings provide evidence that suggests that tandospirone has an antianxiety action, presumably by stimulating 5-HT1A receptors in the dorsal hippocampus.     

Abecarnil enhances recovery from diazepam tolerance.

Treatment with diazepam (25 mg/kg; p.o., twice-daily for 17 days) induced tolerance to the anticonvulsant effect of diazepam against bicuculline-induced convulsions in mice. Cross-tolerance was observed to the anticonvulsant action of clonazepam, imidazenil but not abecarnil. While substitution of clonazepam (12 mg/kg; p.o., twice-daily for 15 days) for diazepam did not affect tolerance to diazepam, substitution of imidazenil (17 mg/kg; p.o., twice-daily for 15 days) for diazepam significantly increased sensitivity to the anticonvulsant effect of diazepam, although tolerance was not abolished. Tolerance to diazepam progressively decreased either after suspension of diazepam administration or replacement treatment with abecarnil (20 mg/kg; p.o., twice-daily). Complete recovery of diazepam efficacy was detected after 8 and 15 days of administration of abecarnil and vehicle, respectively. Binding experiments using [3H]-flumazenil showed that Kd values did not differ among treatment groups. A significant decrease in Bmax (-42%) was observed in the cortex of diazepam-tolerant mice whether or not also treated with imidazenil and clonazepam. Conversely, chronically diazepam-treated mice, that further received abecarnil for either 8 or 15 days or vehicle for 15 days showed Bmax values similar to those of vehicle-treated mice never exposed to diazepam. Results suggest that repeated abecarnil administration to diazepam-tolerant mice can facilitate re-adaptation of receptors to the diazepam-free state. It is proposed that replacement therapy with abecarnil after long-term treatment with conventional benzodiazepines (BDZs) may provide a novel approach for reducing tolerance to their anticonvulsant effects.     

Pharmaco-ethological analysis of agonistic behavior between resident and intruder mice: effect of anticholinergic drugs

The resident-intruder paradigm was employed in order to evoke an agonistic behavior in mice. In this situation a resident male mouse has been cohabiting with a female for 5 weeks, and an intruder male mouse is introduced into the resident's home cage. A species-specific pattern of agonistic behavior was observed in all mice. The significance of cholinergic mechanisms in the mediation of the agonistic behavior was evaluated by pharmacological manipulations. Drugs were administered to resident mice. Scopolamine hydrobromide (0.25, 0.50 and 0.75 mg/kg, i.p.) significantly suppressed the resident's aggressive episodes (offensive sideways posture, tail rattling and attack biting) in a dose-dependent manner, whereas the peripheral anticholinergic drug methylscopolamine nitrate (0.25, 0.50 and 0.75 mg/kg, i.p.) was ineffective. On the other hand, the resident's locomotor activity and rearing response were significantly increased after the administration of scopolamine hydrobromide. The evidence suggests that brain cholinoceptive mechanisms may participate in the regulation of intraspecies aggressive behavior. However, it appears that other nonspecific behavioral effects of scopolamine cannot be ruled out.     

Perinatal diazepam exposure: behavioral and neurochemical consequences

In the present study the effects of perinatal diazepam (DZP) exposure on behavior and benzodiazepine binding site characteristics in offspring were investigated. Pregnant F344 rats were treated during the last trimester of gestation with vehicle or diazepam (3 mg/kg, 5 mg/kg, or 10 mg/kg). Lactating dams were similarly treated on postnatal days 1-7. Both prenatal and postnatal exposure to diazepam resulted in significant effects on the acquisition and extinction of active avoidance behavior measured postweaning. The number of trials to extinction of one-way active avoidance behavior was significantly greater in offspring exposed gestationally to 3 mg/kg and 10 mg/kg diazepam and postnatally to 10 mg/kg diazepam. The mean response latencies for all diazepam treated groups were significantly shorter than those of the vehicle treated rats during the first 15 trials under extinction conditions. In contrast, neither gestational nor postnatal diazepam exposure significantly affected either acquisition or retention of a passive avoidance task. In addition, the binding affinity between the benzodiazepine type I selective ligand, CL218,872, and cortical membranes, as well as the degree to which GABA potentiated 3H-flunitrazepam were significantly altered by perinatal diazepam exposure. No treatment altered the approximate number of benzodiazepine binding sites in either the cortex, hippocampus, or cerebellum. The results of this study further support diazepam as a behavioral teratogen and give new evidence for neurochemical effects following gestational as well as postnatal diazepam exposure.     

Benzodiazepine-receptor mediated inhibition of isolation-induced aggression in mice

The effects of a benzodiazepine receptor agonist (diazepam), antagonist (Ro 15-1788), and an "active" antagonist [inverse agonist] (3-carboethoxy-beta-carboline) were examined in an isolation-induced model of aggression. Diazepam (4 mg/kg) and 3-carboethoxy-beta-carboline (10 mg/kg), but not Ro 15-1788, significantly inhibited aggressive behavior in this model. Ro 15-1788 (10 mg/kg) reduced the anti-aggressive actions of both diazepam and 3-carboethoxy-beta-carboline, while mice treated with a combination of diazepam and 3-carboethoxy-beta-carboline had aggression scores increased to values not significantly different from vehicle treated mice. These findings suggest that both diazepam and 3-carboethoxy-beta-carboline have anti-aggressive properties in the isolation-induced model of aggression which are mediated through CNS benzodiazepine receptors.     

No tolerance to antiaggressive effect of d-amphetamine in mice

Agonistic, locomotor, and stereotyped behavior were measured in male Swiss-Webster mice in their home cage, normally shared with a female, while confronting an intruder mouse. Acute administration of d-amphetamine (2, 4, 8 mg/kg, IP) to resident mice decreased the frequency of attacks toward an untreated intruder, increased the resident's locomotor activity, and induced a small amount of stereotyped behavior. Redetermination of dose-effect functions during chronic treatment (8 or 16 mg/kg/day) indicated that tolerance did not develop to the antiaggressive effect of d-amphetamine. By contrast, the chronically treated mice showed sensitization to amphetamine-induced stereotypies and a diminished sensitivity to the drug's enhancement of locomotor activity. Subsequent tests with cocaine indicated no differences between amphetamine-maintained and saline control animals, providing no evidence for cross-tolerance or cross-sensitization between cocaine's and amphetamine's effects on attack, locomotion, and stereotypies.     

Naltrexone blocks amphetamine-induced hyperactivity,but not disruption of social and agonistic behavior in mice and squirrel monkeys

Significant anatomical overlap of opioid and dopamine receptors as well as reciprocity of control over synthesis, metabolism, and release of opioid peptides and dopamine in brain suggests functional interactions between the two systems. In the first of two studies, the behavioral effects of amphetamine and naltrexone alone, and in combination were studied in established groups of socially interacting squirrel monkeys. Naltrexone (0.1–10.0 mg/kg, IM) increased locomotion and marking behavior in subordinate monkeys. The frequency of social initiatives directed at treated subordinate monkeys by untreated members of the group was also increased. The behavior of dominant monkeys was relatively unaffected, except at the highest dose when autonomic distress was also evident. The frequency of walking bouts by both dominant and subordinate monkeys was increased by amphetamine (0.1–0.6 mg/kg, IM), and the social behavior of dominant monkeys was disrupted by drug treatment. Naltrexone (0.1 mg/kg, IM) significantly antagonized amphetamine's effects on motor behavior, and enhanced or did not affect amphetamine's effects on social behavior. In a second study, the interaction of amphetamine (0.63–10.0 mg/kg, IP) and naltrexone (0.1–10.0 mg/kg, IP) on the behavior of resident male mice during confrontations with a male intruder was studied. Naltrexone selectively reduced the frequency of attack at the highest dose tested. Amphetamine increased locomotor activity and decreased attack and threat behavior in resident mice. A low dose of naltrexone (1.0 mg/kg, IP) blocked amphetamine's effects on locomotion and enhanced the disruption of aggressive behavior. The amphetamine-naltrexone interaction on locomotor activity in mice and monkeys is consistent with opioid receptor modulation of dopamine mediated functions. However, this mechanism does not appear to account for the interaction of naltrexone and amphetamine on social behavior.     

Effects of social defeat and of diazepam on behavior in a resident-intruder test in male DBA/2 mice

Social stress induces robust behavioral and physiological changes, some of which may alter the responsiveness to pharmacological agents, including diazepam (DZP). We used a resident-intruder paradigm to (1) develop a comprehensive ethogram of behavioral changes following social defeat (SD) in the socially reactive strain, DBA/2 male mice, (2) determine whether acute exposure of DBA/2 mice to low-dose DZP would induce flight or aggressive behavior, both of which have been observed in other rodent models and (3) to test whether prior social stress affects responses to DZP. Behavioral responses to a nonaggressive intruder (NAI) mouse 24 h post-SD were measured in resident subject mice exposed to DZP (0, 0.5, 2.0 mg/kg, ip) either prior to the resident-intruder test (Experiment 1) or immediately post-SD (Experiment 2); control mice were not defeated (NOSD). In general, SD mice displayed increased passive and active avoidance, defense, immobility, and risk assessment relative to NOSD mice. In Experiment 1, mice treated acutely with 0.5 mg/kg DZP had more approach and flight behavior, while those treated with 2.0 mg/kg DZP had more avoidance than vehicle-treated mice, independent of SD. In Experiment 2, acute DZP (2 mg/kg) induced effects 24 h later, possibly secondary to withdrawal. In a nonsocial context (Experiment 3), DZP increased exploratory activity.     

Effects of triazolam on conditioned behavior in rats (author's transl)]

Effects of triazolam on various types of conditioned behavior were investigated and compared mainly with diaepam in rats. The active conditioned avoidance response of the rat in a Shuttle box was inhibited by triazolam and diazepam only at large doses. The passive avoidance response in a step-down method was not affected by either triazolam or diazepam, but was markedly suppressed by chlorpromazine. The low rate response of hypothalamic self-stimulation behavior was markedly increased by triazolam at doses ranging from 2 to 40 mg/kg p.o., but was suppressed at doses over 80 mg/kg p.o. The high rate response was unaffected by triazolam even at doses of 40 approximately 180 mg/kg p.o. The low rate response was increased by diazepam at doses of 1 approximately 10 mg/kg p.o. and was suppressed at 80 mg/kg p.o. The high rate response was reduced by diazepam at 180 mg/kg p.o. In the conflict situation of the rat subjected to food reward and foot-shock punishment, the lever press response in the unpunished period was reduced by triazolam at doses of 1 approximately 5 mg/kg p.o., whereas that in the punished period was markedly increased. Similar effects were observed with diazepam at doses of 15 approximately 20 mg/kg p.o. Triazolam appeared to be 10 approximately 15 times more potent than diazepam in this anticonfluct effect. Thus, triazolam appears to be a potent antianxiety agent.     

Maternal aggression in mice: effects of treatments with PCPA, 5-HTP and 5-HT receptor antagonists

Drug treatments which influence brain serotonergic systems were administered to lactating female mice during the early postpartum period, and their effects on aggressive behavior, locomotor activity and brain monoamines were examined. P-chlorophenylalanine (200 and 400 mg/kg) and 5-hydroxytryptophan (100 mg/kg) inhibited fighting behavior of postpartum mice toward unfamiliar male intruder mice. These drug-treated postpartum females showed increased latencies to attack male intruders and also reduced frequencies of attack. In addition, postpartum mice treated with the serotonin receptor antagonists, mianserin (2 and 4 mg/kg), methysergide (4 mg/kg) and methiothepin (0.25 and 0.5 mg/kg), displayed significantly less aggressive behavior than control mice, as measured by reduced number of attacks. Whole brain monoamine and monoamine metabolite levels were measured after drug treatments. The behavioral results are discussed in terms of drug-induced changes in brain chemistry and indicate a possible role for serotonin in the mediation of maternal aggressive behavior of mice.     

Oral drug self-administration in the home cage of mice: alcohol-heightened aggression and inhibition by the 5-HT1B agonist anpirtoline

RATIONALE: In order to model heightened aggression after alcohol consumption and to study the inhibitory influence of 5-HT1B receptors on drinking and fighting, an experimental procedure should enable self-administration of precise amounts of alcohol in a limited period of time before an aggressive confrontation. OBJECTIVES: To design a new device that can reinforce operant responding by the delivery of sweet alcohol in the resident mouse home cage, where aggressive behavior toward an intruder can subsequently be examined, and to demonstrate inhibition of alcohol-heightened aggression by 5-HT1B receptor agonist treatment. METHODS: Within one experimental session, all singly housed CFW male mice (n=26) performed a nose-poke response that was reinforced by 0.05 ml sucrose. Using the sucrose fading technique, eventually the mice consumed a 6% ethanol/4% sucrose solution after each fifth nose poke during daily 15-min experimental sessions. The number of ethanol reinforcements was adjusted so that 0.6, 1.0, 1.7, and 3.0-g/kg doses were consumed in 15 min or less. Assays confirmed blood alcohol levels at 68.1 mg/dl for intake of 1.0 g/kg. After consuming a specific dose of ethanol in the form of a fixed number of response-dependent deliveries, the response panel was removed from the home cage and, 15 min later, the resident confronted a male intruder. Anpirtoline was administered either before alcohol self-administration or before the aggressive confrontation. RESULTS: After being reinforced with 1.0 g/kg or 1.7 g/kg sweet ethanol, the mice significantly increased attack and threat behavior relative to their aggressive behavior following sucrose or water consumption only. Treatment with the 5-HT1B receptor agonist anpirtoline (0.125, 0.25, 0.5 mg/kg, i.p.) before the confrontation decreased alcohol-heightened aggression and species-typical aggression in the absence of changes in other elements of the behavioral repertoire. Anpirtoline affected ethanol-reinforced behavior only at doses that were 5-10 times higher than those producing anti-aggressive effects. CONCLUSIONS: Self-administration of alcohol in the home cage of mice is readily accomplished with the aid of a simple, removable panel. The effective inhibition of high levels of aggressive behavior due to alcohol consumption after anpirtoline treatment confirm the 5-HT1B receptor as a critical site in the termination of aggression.     

Aggression during morphine withdrawal: Effects of method of withdrawal,fighting experience,and social role

Offensive and defensive components of aggressive behavior were determined in resident and intruder mice. Withdrawal aggression was measured after the removal of a subcutaneous morphine pellet or after precipitation by naloxone in naive mice and after removal of a morphine pellet in mice with prior fighting experience. In naive mice, removal of a morphine pellet led to increases in attack bites and threats but naloxone-precipitated withdrawal led to decreases in these behaviors and to increases in defensive posturing, escape attempts and vocalizations. Prior fighting experience abolished the enhanced attack behaviors of resident mice following morphine pellet removal, but led to heightened defensive behavior in intruder mice. The behavior of intruder mice appeared more sensitive to naloxone administration than the behavior of resident mice; naloxone influenced not only intruder defensive behavior, but also other non-aggressive behaviors. The social role of the drug recipient and his prior history of aggressive behavior are important determinants of morphine and naloxone effects on aggression.