Assunta Imperato

Cannabinoid CB1 receptor knockout mice fail to self-administer morphine but not other drugs of abuse.

The rewarding effects of morphine, cocaine, amphetamine and nicotine were evaluated in CB1 receptor knockout mice by means of an intravenous self-administration model. Experiments were carried out on drug-naive animals using a nose-poking response (NPR)-like as operandum. The results of the present study indicate that morphine did not induce intravenous self-administration in mutant CB1 receptor knockout mice, whereas it was significantly self-administered by the corresponding wild type mice. On the contrary, cocaine, amphetamine and nicotine were self-administered to the same extent by both wild type and CB1 receptor knockout mice. These data clearly indicate that the CB1 cannabinoid receptor is essential not only for the expression of cannabinoid reinforcing effects but also for the modulation of morphine rewarding effects. The specificity of such interaction is supported by the finding that contrary to morphine, cocaine, d-amphetamine and nicotine were self-administered by mice at the same extent either in presence or in absence of the CB1 receptor.

Enhancement of memory in cannabinoid CB1 receptor knock-out mice.

We have used cannabinoid CB knock-out mice in a two-trial object recognition test to assess the role of cannabinoid CB receptors in memory. Cannabinoid CB1 knock-out mice are able to retain memory for at least 48 h after the first trial whereas the wild-type controls lose their capacity to retain memory after 24 h. These results suggest that endogenous cannabinoid CB receptors play a crucial role in the process of memory storage and retrieval.

Complex pharmacology of natural cannabivoids: Evidence for partial agonist activity of Δ9-tetrahydrocannabinol and antagonist activity of cannabidiol on rat brain cannabinoid receptors

Delta9-tetrahydrocannabinol (delta9-THC), cannabinol and cannabidiol are three important natural cannabinoids from the Marijuana plant (Cannabis sativa). Using [35S]GTP-gamma-S binding on rat cerebellar homogenate as an index of cannabinoid receptor activation we show that: delta9-THC does not induce the maximal effect obtained by classical cannabinoid receptor agonists such as CP55940. Moreover at high concentration delta9-THC exhibits antagonist properties. Cannabinol is a weak agonist on rat cerebellar cannabinoid receptors and cannabidiol behaves as an antagonist acting in the micromolar range.

Enhanced long-term potentiation in mice lacking cannabinoid CB1 receptors.

Marijuana is known to affect learning and memory in humans, and cannabinoids block long-term potentiation in the hippocampus, a model for the synaptic changes that are believed to underlie memory at the cellular level. We have now examined the physiological properties of the Schaffer collateral-CA1 synapses in mutant mice in which the CB1 receptor gene has been invalidated and found that these animals exhibit a half-larger long-term potentiation than wild-type controls. Other properties of these synapses, such as paired-pulse facilitation, remained unchanged. This indicates that disrupting CB1 receptor-mediated neurotransmission at the genome level produces mutant mice with an enhanced capacity to strengthen synaptic connections in a brain region crucial for memory formation.

Lack of morphine-induced dopamine release in the nucleus accumbens of cannabinoid CB(1) receptor knockout mice

Morphine (10 and 20 mg/kg, s.c.) does not modify dopamine release in the nucleus accumbens of cannabinoid CB(1) knock-out mice under conditions where it dose-dependently stimulates the release of dopamine in the corresponding wild-type mice. These results demonstrate that cannabinoid CB(1) receptors, regulate mesolimbic dopaminergic transmission in brain areas known to be involved in the reinforcing effects of morphine.

Repeated stressful experiences differently affect the time-dependent responses of the mesolimbic dopamine system to the stressor

The increase in mesolimbic dopamine (DA) release observed during the first 40 min of 120 min restraint in naive rats is not evident in repeatedly stressed animals (daily 60 min restraint, for 5 days). However, repeatedly stressed rats show a significant decrease in DA release from 80 min of restraint onwards which is not observable in naive rats. These results indicate that repeated stressful experiences do not produce habituation but alter the response of mesolimbic DA system to the stressor. Moreover, they point to a possible neuronal mechanism underlying stress-induced depression.

Glutamate uptake is decreased tardively in the spinal cord of FALS mice.

THIS study examined high affinity Na+-dependent uptake of glutamate in synaptosomal preparations from spinal cord in mice that express a dominant mutation of human copper/zinc superoxide dismutase (SOD1) and represent an animal model of amyotrophic lateral sclerosis (ALS). Their muscle strength was also monitored by a grip traction test throughout their lifespan. The high affinity Na+-dependent uptake of [3H]glutamate was decreased between 120 and 150 days of age. A marked and significant decrease in Vmax (−40.2%; p < 0.001) on whole spinal cord synaptosomes was observed at 150 days, with no change in Km. This significant decrease was reached a week before the animals died (157.2 ± 2.2 days) and corresponded to a considerable fall in muscle strength (25% loss between 120 and 140 days, p < 0.001). The FALS mouse model therefore reproduces the decrease in glutamate uptake reported in humans suffering from sporadic or familial ALS. These results are discussed in terms of a possible tardive involvement of glutamate uptake deficiency in human ALS.

IMMUNOHISTOCHEMICAL ANALYSIS OF PRESENILIN-1 EXPRESSION IN THE MOUSE BRAIN

At least 22 different mutations associated with early‐onset familial Alzheimers disease (AD) in various kindreds have been reported to occur in a recently identified gene on chromosome 14, presenilin 1 (PS‐1) (Sherrington et al. (1995) Nature 375, 754–760 [1] and reviewed by Van Broeckhoven (1995) Nat. Genet. 11, 230–231 [2]). In order to study the localization of PS‐1 in the brain, we raised a polyclonal antiserum specific to a fragment of the predicted protein sequence of PS‐1. PS‐1 immunostaining was found intracellularly, in the perikaria of discrete cells, mostly neurons, appearing as thick granules, resembling large‐size vesicles. These granules were located in the periphery of cell bodies and extended into dendrites and neurites. PS‐1 expression was found to be broadly distributed throughout the mouse brain, not only in structures involved in AD pathology, but also in structures unaltered by this disease.

Inhibition of hippocampal acetylcholine release by benzodiazepines: antagonism by flumazenil

Diazepam (2.5-10 mg/kg i.p.) and midazolam (2.5-10 mg/kg i.p.) decreased acetylcholine release in the hippocampus of freely moving rats. This effect was antagonized by pretreatment with flumazenil (1 mg/kg i.p.). These results show that activation of benzodiazepine receptors reduces the in vivo release of acetylcholine in the hippocampus, suggesting that the septo-hippocampal cholinergic system, which has a major role in the regulation of cognitive functions, is under inhibitory control exerted by gamma-aminobutyrate (GABA) neurons.

Rapid increase in basal acetylcholine release in the hippocampus of freely moving rats induced by withdrawal from long-term ethanol intoxication

The effect of ethanol withdrawal on hippocampal acetylcholine (ACh) release was investigated by brain microdialysis in rats rendered ethanol dependent by repeated forced administration of a 20% ethanol solution for 7 days. The behavioral signs of ethanol withdrawal were accompanied by an increase in hippocampal ACh output that was significantly 6 h after the last ethanol administration, reached a maximum (fourfold) at 12 h, and persisted for >72 h. Administration of diazepam (5 mg/kg, i.p.) or gamma-hydroxybutyrate (1 g/kg, intragastric) 12 h after the last ethanol administration completely antagonized, within 30 min, the increase in ACh output induced by ethanol withdrawal. Thus, the rapid and marked increase in ACh output might contribute to the changes in cognitive function associated with ethanol withdrawal, and the septohippocampal cholinergic system may play a major role in the response to withdrawal of addictive drugs.