Anandamide content is increased and CB1 cannabinoid receptor blockade is protective during transient, focal cerebral ischemia.

The role of endocannabinoid signaling in the response of the brain to injury is tantalizing but not clear. In this study, transient middle cerebral artery occlusion (MCAo) was used to produce ischemia/reperfusion injury. Brain content of N-arachidonoylethanolamine (AEA) and 2-arachidonoylglycerol were determined during MCAo. Whole brain AEA content was significantly increased after 30, 60 and 120 min MCAo compared with sham-operated brain. The increase in AEA was localized to the ischemic hemisphere after 30 min MCAo, but at 60 and 120 min, was also increased in the contralateral hemisphere. 2-Arachidonoylglycerol content was unaffected by MCAo. In a second set of studies, injury was assessed 24 h after 2 h MCAo. Rats administered a single dose (3 mg/kg) of the cannabinoid receptor type 1 (CB1) receptor antagonist SR141716 prior to MCAo exhibited a 50% reduction in infarct volume and a 40% improvement in neurological function compared with vehicle control. A second CB1 receptor antagonist, LY320135 (6 mg/kg), also significantly improved neurological function. The CB1 receptor agonist, WIN 55212-2 (0.1-1 mg/kg) did not affect either infarct volume or neurological score.

The failure of some rats to acquire intravenous cocaine self-administration is attributable to conditioned place aversion.

Although cocaine administration in humans includes euphoric and anxiogenic effects, the latter are less well understood. Acute cocaine administration produces aversive effects including anxiogenic effects as well as appetitive effects in rats and mice. In the present study the self-administration and conditioned place preference paradigms were used to determine whether the failure of some rats to acquire intravenous cocaine self-administration is attributable to either an interference with learning or an aversion to cocaine. Rats were classified as self-administrators or non-self-administrators based on the mean number of cocaine self-infusions per session and whether or not rats exhibited either a stable high level of responding or a stable low level of responding. Intravenously administered cocaine produced place preference for the self-administrators, while intravenously administered cocaine produced place aversion for the non-self-administrators. The fact that the non-self-administrators showed place aversion is inconsistent with the interpretation that the failure of these rats to readily self-administer is attributable to cocaine-mediated interference of learning. This is the first study in which both the self-administration and the conditioned place preference paradigms have been used in the same animals to demonstrate that the effects of cocaine are appetitive for some rats and aversive for others, and are not an artifact of cocaine's interference with learning.

The effects of amperozide on cocaine-induced social withdrawal in rats.

Cocaine decreases social interactions in both humans and animals, but it is not known whether the drugged animal withdraws from an undrugged conspecific, the undrugged conspecific avoids the drugged animal, or a combination of these two factors occurs. In the first experiment, the source of cocaine-induced social withdrawal was determined using a tether paradigm, in which the movement of one of the rats was restricted to one half of the observation chamber, such that the freely moving rat had the option of escaping social interactions. There were decreases in social interactions in the condition in which both rats were freely moving, and in the condition in which the undrugged rat was tethered, but not when the drugged rat was tethered and could not escape social contact. A second experiment was conducted to test the efficacy of the potent serotonin receptor antagonist, amperozide, in attenuating cocaine-induced social withdrawal using the condition in which the drugged rat was freely moving. Either amperozide (1.0, 3.0, and 5.0 mg/kg) or saline vehicle was injected into rats 1 h before receiving a 30.0 mg/kg cocaine dose. Cocaine decreased social interactions. Amperozide restored social interactions to near control levels and elevated social interactions in the animals treated with saline vehicle.

Short-term exposure to dietary Pb and/or DMSA affects dopamine and dopamine metabolite levels in the medulla, optic tectum, and cerebellum of rainbow trout (Oncorhynchus mykiss).

Rainbow trout (Oncorhynchus mykiss) were randomly assigned to one of the following dietary exposure conditions: lead (Pb) solvent (2% nitric acid), meso-2,3-dimercaptosuccinic acid (DMSA) solvent (0.1 N NaOH), Pb, DMSA, Pb followed by Pb solvent, or Pb followed by DMSA. Medulla, cerebellum, and optic tectum homogenates were analyzed for dopamine (DA), homovanillic acid (HVA), and 3,4-dihydroxyphenylacetic acid (DOPAC). DA levels in all brain regions tended to be highest for trout exposed to dietary Pb followed by dietary DMSA. DA levels were elevated for trout exposed to dietary DMSA and Pb followed by Pb solvent. DA levels were below control levels for trout exposed to Pb only. HVA levels varied across brain regions. However, HVA levels in all brain regions tended to be elevated for trout exposed to dietary DMSA and Pb followed by Pb solvent. DOPAC levels across all brain regions were below control levels for trout dietary exposed to DMSA, Pb only, Pb followed by Pb solvent, and Pb followed by DMSA. These data indicate that Pb and/or DMSA have the potential of altering DA, HVA, and DOPAC levels in the medulla, cerebellum, and optic tectum. The animal model of short-term dietary exposure to Pb and DMSA, both alone and sequentially, to mimic dietary exposure to Pb and the oral delivery of DMSA, that our laboratory has developed, may be useful in future studies aimed at characterizing the neurobiological mechanisms by which Pb and/or DMSA alter neurotransmitter levels and behavior.

The rewarding properties of NMDA and MK-801 (dizocilpine) as indexed by the conditioned place preference paradigm.

N-Methyl-D-aspartate (NMDA) ([R]-2-[Methylamino]succinic acid) is a specific excitatory amino acid. Two experiments were conducted to determine the rewarding properties of this compound using the conditioned place preference paradigm. In the first experiment, 40 male Sprague-Dawley rats received place preference conditioning for a 4 day period. The conditioned place preference apparatus consisted of two chambers with distinct visual and tactile cues, separated by a removable door. On days 2 and 4, rats were systemically administered NMDA (1.0, 15.0, and 30.0 mg/kg) paired with one chamber. On days 3 and 5, rats were systemically administered saline paired with the other chamber. Day 6 was the test day, and the rat was allowed free run of the entire apparatus in a drug-free state. Time spent in each side of the apparatus was computer recorded. NMDA produced a significant increase in the amount of time spent on the side previously paired with drug for 15.0 and 30.0, but not 1.0 mg/kg NMDA. In the second experiment, systemic administration of NMDA (30.0 mg/kg) paired with the noncompetitive NMDA receptor antagonist, MK-801 (0.5 mg/kg), resulted in neither place preference nor place aversion.

Determination of the place conditioning and locomotor effects of amperozide in rats: a comparison with cocaine.

Amperozide promotes social interactions in rats (Rattus norvegicus) and reduces craving for cocaine and alcohol without producing adverse side effects. Amperozide administration produced a place preference in the dose range from 3.0 to 10.0 mg/kg. Locomotor behavior tended to be suppressed on the days of amperozide pairings, elevated on the days of vehicle pairings, and elevated on test days. Administration of 20.0 mg/kg but neither 2.0 nor 0.2 mg/kg cocaine produced place preference. Rats exhibited a greater place preference for the chamber paired with 2.0 mg/kg and 20.0 mg/kg but not 0.2 mg/kg cocaine compared to the chamber paired with 5.0 mg/kg amperozide. The results indicate that amperozide is appetitive. The most appetitive dose of amperozide (5.0 mg/kg) is less appetitive than the most appetitive dose of cocaine (20.0 mg/kg). More research is needed before conclusions regarding the degree of abuse potential for amperozide can be made.

Neurotransmitter levels in two populations of larval Fundulus heteroclitus after methylmercury exposure.

The effects of methylmercury (MeHg) exposure on neurotransmitter (NT) levels in larval mummichogs (Fundulus heteroclitus) obtained from a mercury-polluted site (Piles Creek (PC), NJ) and a reference site (Tuckerton (TK), NJ) were examined. Population differences between PC and TK larvae in neurochemical composition and in neurochemical changes in response to MeHg intoxication were found. Heads of untreated PC larvae (7 days posthatch (dph)) contained considerably higher levels of dopamine (DA) and its metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) than TK. However, they had comparable levels of serotonin (5-hydroxytryptamine (5-HT)) and 5-hyroxy-3-indoleacetic acid (5-HIAA)/5-HT ratios. Changes in NTs with age were noticed, especially in PC larvae. Exposure of larvae to 10 microg/l MeHg induced neurochemical alterations. A significant increase in DA and 5-HT, as well as depressed dopaminergic and serotonergic activity (i.e. decreased DOPAC/DA, HVA/DA and 5-HIAA/5-HT ratios) were seen in TK larvae. Exposure of PC larvae to 10 microg/l MeHg reduced 5-HT at 14 dph, increased serotonergic activity at 7 dph, and altered dopaminergic activity (i.e. increased DOPAC/DA ratios, but decreased HVA/DA ratios). Changes in DA levels were inconsistent over time. The DA level, which was considerably higher than the control at 7 dph, was significantly lower than the control at 14 dph. For the two populations, the level of 5-HT and serotonergic activity, as well as DOPAC and HVA levels, were correlated with previously noted spontaneous activity. The changes in NT levels after exposure to MeHg are an indication of neurological dysfunction in larvae.

Virus isolation from semen of bulls serologically positive for bluetongue virus.

Isolation of bluetongue virus was attempted from 85 semen samples taken from 3 long-term seropositive bulls and 9 short-term seropositive bulls in an artificial breeding service unit. Two types of cell cultures susceptible to bluetongue virus were used for virus isolation. Extended sonication, centrifugation of specimens, and treatment of cell cultures with dimethyl sulfoxide and diethylaminoethyl-dextran were used to enhance virus attachment and infection of cell cultures. Virus isolation results were negative on all specimens. These results indicate that at the limits of the methods used, bluetongue virus-seropositive bulls do not have long-term latent bluetongue virus in their semen.

Effect of amphetamine place conditioning on excitatory synaptic events in the basolateral amygdala ex vivo.

The basolateral amygdala (BLA) plays an important role in the formation of associations between context and drug. BLA activity and BLA-dependent drug-seeking behavior are driven by excitatory inputs. Drug-seeking behavior driven by context involves participation of the BLA, and plasticity of excitatory inputs to the BLA may contribute to this behavior. In this study, amphetamine conditioned place preference (AMPH CPP) was used to model the formation of context-drug associations. Learning-induced changes of excitatory synapses within the BLA were examined. Male Sprague-Dawley rats were assigned to one of three groups, the experimental group (AMPH CPP) or one of two control groups (saline or AMPH delayed pairing). Approximately 24 h after testing their preference, spontaneous and miniature excitatory postsynaptic currents (sEPSCs and mEPSCs, respectively) in BLA pyramidal neurons were investigated using whole-cell patch-clamp recordings. There were no between-groups differences in the amplitude or frequency of sEPSCs or mEPSCs. In a higher osmolarity solution to increase release, there was a significantly greater frequency of the mEPSCs in neurons from AMPH CPP animals compared with controls. This was observed with no change detected in the probability of glutamate release. Together, these data demonstrate no evidence for increased synaptic strength, but are consistent with an increase in the number of synapses in the BLA after AMPH CPP. These findings may underlie increased excitatory drive of the BLA after AMPH CPP, and contribute to the animals' preference for the AMPH-paired compartment.