Effect of ethanol on memory consolidation in mice: antagonism by the imidazobenzodiazepine Ro 15-4513 and decrement by familiarization with the environment

Three sets of experiments were carried out with CD1 mice tested in a one-trial inhibitory avoidance task. In a first set of experiments the posttraining administration of ethanol (1 or 2 g/kg) impaired, while that of the imidazobenzodiazepine Ro 15-4513 (5 or 10, but not 2.5 mg/kg) improved the retention performance of the animals. In a second set of experiments a by itself ineffective dose of Ro 15-4513 (2.5 mg/kg) antagonized the effect of ethanol (1 and 2 kg/kg). These results are discussed on the basis of the interaction of these drugs with the GABAergic system. In a third set of experiments, in which the performances of mice familiarized with the apparatus were compared with those of non-familiarized mice, ethanol was less effective in impairing memory processes of the experienced subjects. These results are discussed in terms of attenuation of emotionality, resulting in impaired retention, following posttraining ethanol administration.     

Morphine and memory in DBA/2 mice: Effects of stress and of prior experience

In a first set of experiments, immediately post-training morphine (1.0 or 2.5 but not 0.5 mg/kg) treatment, or immobilization stress (30 or 60 but not 15 min) impaired memory processes of non-pretrained DBA/2 mice tested in a passive avoidance box. The effects were naloxone-reversible and time-dependent (they were absent in mice injected with morphine, or immobilized, starting 120 min after training). No effect was evident in no-footshock groups injected with morphine (2.5 mg/kg) or immobilized (60 min), thus showing lack of proactive influence of the treatments on performance. In a second set of experiments, in which pretrained animals were used, both morphine and immobilization stress were less effective in disrupting memory processes of mice. In both sets of experiments a per se ineffective stress enhanced the effects of morphine. A number of possible hypotheses concerning the results obtained are examined. In particular the possible role of emotional factors in the effects of morphine on memory is discussed.     

Strain-dependent involvement of D1 and D2 dopamine receptors in muscarinic cholinergic influences on memory storage

These experiments examined the interaction of muscarinic and dopaminergic systems in influencing memory for one-trial inhibitory avoidance training in mice of the C57BL/6 and DBA/2 strains. In both strains, immediate post-training systemic administration of the muscarinic cholinergic agonist oxotremorine enhanced retention and the cholinergic antagonist atropine impaired retention. No effects were seen with injections 2 h post-training. Furthermore, the drugs did not affect retention performance of animals that received no footshock on the training trial. These results confirm previous findings indicating that muscarinic cholinergic drugs affect memory by influencing memory consolidation. In C57 mice, pretreatment with selective D1 or D2 dopamine (DA) receptor agonists (SKF 38393 or LY 171555, respectively) in otherwise non-effective doses (5 and 0.25 mg/kg, respectively) potentiated the effects of oxotremorine (0.04 mg/kg). Furthermore, in C57 mice pretreatment with selective D1 or D2 receptor antagonists (SCH 23390 or (-)-sulpiride) in otherwise non-effective doses (0.025 and 6 mg/kg, respectively) blocked the memory enhancing effects of oxotremorine. The memory impairing effects of atropine (3 mg/kg) were blocked by the D1 and D2 selective agonists and potentiated by the selective D1 or D2 antagonists. In contrast, in DBA mice, the D1 and D2 selective agonists antagonised the memory enhancing effects of oxotremorine (0.02 mg/kg) and potentiated the effects of atropine (2 mg/kg). Furthermore, the D1 and D2 antagonists potentiated the effects of oxotremorine and antagonised those of atropine. These findings indicate that although muscarinic cholinergic influences on memory storage are comparable in mice of these two strains, the cholinergic-dopaminergic interactions are opposite in the two strains. These results have implications for hypotheses of cholinergic and dopaminergic regulation of memory storage.     

The effects of acute restraint stress and dexamethasone on retrieval of long-term memory in rats: an interaction with opiate system

This study investigated whether application of acute restraint stress or dexamethasone, as a glucocorticoid receptor agonist, impaired retrieval of long-term memory and if pretreatment with opiate antagonist naloxone blocked their effects on memory retrieval. Young adult male rats were trained in one trial inhibitory avoidance task (1 mA, 1.5 s footshock). On retention test given 48 h after training, the latency to re-enter dark compartment of the apparatus was recorded. Thirty minutes before retention test, the rats were exposed to a 10 min of restraint stress in a Plexiglass tube or were injected with dexamethasone (1 mg/kg) with or without prior treatment of naloxone (1 or 2 mg/kg). The results showed that both acute restraint stress and dexamethasone impaired retention performance. Both doses of naloxone were effective in blocking the impairing effect of stress, but only higher dose of naloxone blocked dexamethasone-induced impairment. The applied stress increased circulating corticosterone levels as assessed immediately after the retention test, indicating that stress-induced impairment of memory retrieval is mediated, in part, by increased plasma levels of glucocorticoids. These findings further indicate that acute restraint stress and glucocorticoids impair retrieval of long-term memory, and provide evidence for the existence of an interaction between glucocortioids and opiate system on this process.     

Sildenafil, a selective phosphodiesterase type 5 inhibitor, enhances memory reconsolidation of an inhibitory avoidance task in mice

Intracellular levels of the second messengers cAMP and cGMP are maintained through a balance between production, carried out by adenyl cyclase (AC) and guanylyl cyclase (GC), and degradation, carried out by phosphodiesterases (PDEs). Recently, PDEs have gained increased attention as potential new targets for cognition enhancement, with particular reference to phosphodiesterase type 5 (PDE5A). It is accepted that once consolidation is completed memory becomes permanent, but it has also been suggested that reactivation (memory retrieval) of the original memory makes it sensitive to the same treatments that affect memory consolidation when given after training. This new period of sensitivity coined the term reconsolidation. Sildenafil (1, 3, and 10mg/kg, ip), a cGMP-PDE5 inhibitor, facilitated retention performance of a one-trial step-through inhibitory avoidance task, when administered to CF-1 male mice immediately after retrieval. The effects of sildenafil (1mg/kg, ip) were time-dependent, long-lasting and inversely correlated with memory age. The administration of sildenafil (1mg/kg, ip) 30 min prior to the 2nd retention test did not affect retention of mice given post-retrieval injections of either vehicle or sildenafil (1mg/kg, ip). Finally, an enhancement of retention was also observed in CF-1 female mice receiving sildenafil (1mg/kg, ip) immediately, but not 180 min after retrieval. In the present paper we reported for the first time that systemic administration of sildenafil after memory reactivation enhances retention performance of the original learning. Our results indirectly point out cGMP, a component of the NO/cGMP/PKG pathway, as a necessary factor for memory reconsolidation.     

Effects of kappa opiate agonists on palatable food consumption in non-deprived rats,with and without food preloads

There is increasing evidence to suggest that kappa opiate receptors may be importantly involved in the mediation of feeding responses in the rat. A series of experiments is reported in which the effects of four kappa receptor agonists (ethylketocyclazocine, U-50,488H, tifluadom, bremazocine) on the consumption of a highly palatable diet were investigated. Under one condition, non-deprived male rats were administered drug treatments before a 30 min feeding test. Bremazocine (0.1 mg/kg) and ethylketocyclazocine (3.0 mg/kg) both significantly decreased the level of food consumption. In contrast, U-50,488H and tifluadom each produced significant increases in food intake. In a second condition, non-deprived male rats were first allowed to consume some of the palatable diet to achieve partial satiation, prior to the administration of the drug treatments. In this case, evidence for hyperphagic effects of all four kappa agonists was obtained, within the first 30 min access to the palatable diet. Thus, hyperphagia occurred with 0.01 mg/kg bremazocine and 0.1 mg/kg ethylketocyclazocine. We conclude that some kappa agonists have mixed stimulant/inhibitory effects on food intake, whereas others are more consistent in producing hyperphagia. In neither condition did morphine (0.3–10.0 mg/kg) show any hyperphagic effect. Our data support an involvement of kappa opiate receptors in mechanisms which control palatable food consumption in non-deprived rats.     

Effects of flunitrazepam on passive avoidance behaviour in mice subjected to immobilization stress or familiarized with the testing apparatus

The effects of flunitrazepam on passive avoidance behaviour were investigated in DBA/2 mice. In a first set of experiments retention performance impairment was observed in mice injected with the drug immediately but not 120 min after training. In a second set of experiments, immobilization stress enhanced, while familiarization with the apparatus decreased, the effects of flunitrazepam, suggesting involvement of emotional factors. All the effects observed were antagonized by naltrexone, showing involvement of opioid receptors.     

Effects of concurrent manipulations of cholinergic and noradrenergic function on learning and retention in mice

Interactions between the neuromodulators acetylcholine and norepinephrine (NE) have been reported in both developmental neural plasticity and learning and memory. In a test of the generality of this phenomenon, we assessed the amnestic effects of the muscarinic antagonist scopolamine in normal and NE-depleted mice. Pretraining administration of scopolamine impaired 24-h retention of inhibitory (passive) avoidance training (at doses of 0.1, 0.3 and 1.0 mg/kg) and the acquisition of place-training in a water maze (at a dose of 1.0 mg/kg). NE depletion resulting from systemic administration of DSP-4 did not affect performance on these tasks and did not significantly alter the effects of scopolamine. NE depletion did, however, impair the retention of place learning when mice were retested 16 days after initial training; and this impairment in the retest was additive with one observed in mice originally trained under scopolamine. Normal acquisition but rapid forgetting has also been reported in aged rodents, who display deterioration of the noradrenergic system. Thus, observation of a similar pattern of performance consequent to experimental NE depletion suggests a role for noradrenergic dysfunction in age-related memory decline.     

Alpha2-adrenoceptor antagonist SL 84.0418 reduces the expression of neocortical spike-and-wave spindling episodes in DBA/2J mice

The present work was undertaken to study the effects of alpha(2)-adrenoceptor antagonist 2-(4,5-dihydro-1H-imidazol-2-yl)-1,2,4,5-tetrahydro-2-propyl-pyrrolo[3,2,1-hi]-indole hydrochloride (SL 84.0418) on the neocortical spike-and-wave spindling episodes (S and W) in the cortical electroencephalogram (ECoG) of DBA/2J mice. Our data indicate that SL 84.0418 (0.1-1.0-10 mg/kg ip) dose-dependently reduces the S and W of DBA/2J mice. This effect appears 30 min after drug administration and lasts for the duration of the recording period (240 min). The reducing effect of the SL 84.0418 on the S and W of mice was comparable to that induced by tolazoline (10-20-40 mg/kg ip), a well-known alpha(2)-adrenoceptor antagonist, whereas the alpha(2)-adrenoceptor agonist clonidine (0.02-0.1-0.5 mg/kg ip) significantly increased it. The present results indicate that SL 84.0418 administration induces significant reduction on S and W of DBA/2J mice suggesting a possible involvement of noradrenergic system in the development of S and W of DBA/2J mice.     

Central and peripheral actions of amphetamine on memory storage

These experiments investigated the effects of central (intracerebroventricular) and peripheral (i.p.) posttraining administration ofd-amphetamine on rat's retention of a one-trial inhibitory avoidance response. While retention was enhanced by i.p. administration (1.0 mg/kg) the central administration (dose range 50–500 μg) did not affect retention. In rats given peripheral 6-OHDA 24 h prior to training a lower dose (i.p.) of amphetamine (0.25 mg/kg) was most effective in enhancing retention. These findings suggest that the memory enhancing effects ofd-amphetamine are mediated at least in part through peripheral systems.     

Enhancement of contextual fear-conditioning by putative (±)-α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor modulators and N-methyl-

When administered systemically, glucose attenuates deficits in memory produced by several classes of drugs, including cholinergic antagonists and opiate agonists. Glucose also enhances memory in aged rats, mice, and humans. In addition, glucose ameliorates age-related reductions in paradoxical sleep. Because deficits in paradoxical sleep are most marked in those individual aged rats that also have deficits in memory, treatments which improve one of these functions may similarly improve the other. The present experiments show that glucose attenuates deficits in paradoxical sleep and memory after atropine administration, with similar dose-response curves for both actions. In the first experiment, rats received saline, atropine (1 mg/kg), glucose (100 mg/kg) or combinations of atropine + glucose (10, 100, 250, and 500 mg/kg) 30 min before assessment on a spontaneous alternation task. In the second experiment, 3-h EEGs were assessed for spontaneous daytime sleep in rats administered saline, atropine (1 mg/kg), glucose (100 mg/kg) or combinations of atropine + glucose (10, 100 and 250 mg/kg). In both experiments, glucose significantly attenuated deficits at an optimal dose of 100 mg/kg. A third experiment assessed blood glucose levels after injections of atropine + glucose (100 mg/kg) and determined that blood glucose levels were similar to those produced by other treatments which enhance memory. These results are consistent with the view that paradoxical sleep and at least one test of memory are similarly influenced by atropine and glucose.     

Glucose attenuation of atropine-induced deficits in paradoxical sleep and memory

When administered systemically, glucose attenuates deficits in memory produced by several classes of drugs, including cholinergic antagonists and opiate agonists. Glucose also enhances memory in aged rats, mice, and humans. In addition, glucose ameliorates age-related reductions in paradoxical sleep. Because deficits in paradoxical sleep are most marked in those individual aged rats that also have deficits in memory, treatments which improve one of these functions may similarly improve the other. The present experiments show that glucose attenuates deficits in paradoxical sleep and memory after atropine administration, with similar dose-response curves for both actions. In the first experiment, rats received saline, atropine (1 mg/kg), glucose (100 mg/kg) or combinations of atropine + glucose (10, 100, 250, and 500 mg/kg) 30 min before assessment on a spontaneous alternation task. In the second experiment, 3-h EEGs were assessed for spontaneous daytime sleep in rats administered saline, atropine (1 mg/kg), glucose (100 mg/kg) or combinations of atropine + glucose (10, 100 and 250 mg/kg). In both experiments, glucose significantly attenuated deficits at an optimal dose of 100 mg/kg. A third experiment assessed blood glucose levels after injections of atropine + glucose (100 mg/kg) and determined that blood glucose levels were similar to those produced by other treatments which enhance memory. These results are consistent with the view that paradoxical sleep and at least one test of memory are similarly influenced by atropine and glucose.     

Amygdala and dorsal hippocampus lesions block the effects of GABAergic drugs on memory storage

These experiments examined the effects of posttraining systemic administration of the GABAergic agonist muscimol and the GABAergic antagonist bicuculline on retention in mice with bilateral lesions of the amygdala, dorsal hippocampus or caudate nucleus. Unoperated male CD1 mice and mice with either sham lesions or electrolytically induced lesions of these 3 brain regions were trained in a one-trial inhibitory avoidance task and, immediately after training, received i.p. injections of either muscimol, (1.0, 2.0 or 3.0 mg/kg), bicuculline, (0.25, 0.5 or 1.0 mg/kg), or control solutions. Retention was tested 24 h after training. Lesions of the 3 brain regions produced comparable impairment of retention. In the unoperated controls and sham controls muscimol and bicuculline produced dose-dependent impairment and enhancement, respectively, of retention. The drug effects on retention were blocked by lesions of the amygdala and hippocampus, but were not blocked by lesions of the caudate nucleus. These findings are consistent with other recent evidence suggesting that the amygdala and hippocampus are involved in mediating posttraining neuromodulatory influences on memory storage.     

Effect of Leu-enkephalin on the memory reactivation produced by blockade of the benzodiazepine/GABA-chloride ionophore receptor complex

This investigation sought to characterize the interaction between GABA/benzodiazepine and opioid systems in memory retrieval deficit induced by detaining an animal in the training apparatus after acquisition. Mice pretreated with saline or Leu-enkephalin (0.2 mg/kg) were trained in one-trial passive avoidance test with following detention. Pre-testing administration of bicuculline (1 mg/kg), picrotoxin (1 mg/kg), or flumazenil (10 mg/kg) produced the memory-enhancing effect in the saline-pretreated mice. Pretraining treatment with Leu-enkephalin blocked the reactivation of memory produced by bicuculline and picrotoxin, but not flumazenil. The present investigation suggest that both benzodiazepine/GABA and opioid systems are important modulators of memory retrieval and that a specific interaction between these systems is responsible for the observed recovery of impaired memory trace.     

The memory-modulatory effects of glucocorticoids depend on an intact stria terminalis

This study examined the effects of stria terminalis (ST) lesions on glucocorticoid-induced modulation of memory formation for inhibitory avoidance training and spatial learning in a water maze. Systemic (s.c.) posttraining injections of the glucocorticoid receptor agonist dexamethasone (0.3 or 1.0 mg/kg) enhanced memory for inhibitory avoidance training in rats with sham ST lesions. Removal of the adrenal glands (adrenalectomy; ADX) significantly impaired spatial memory in a water maze, and immediate posttraining injections of dexamethasone (0.3 mg/kg) attenuated the memory impairment. Bilateral lesions of the ST did not significantly affect retention of these two tasks. However, ST lesions did block the effects of short-term ADX and dexamethasone administration on memory for both tasks. These results are similar to those of previous experiments examining the effects of lesions of the basolateral nucleus of the amygdala on the glucocorticoid-induced modulation of memory for both tasks. These findings suggest that the integrity of the ST, which connects the amygdala with other brain structures, is essential for the modulating effects of glucocorticoids on memory storage.     

Opposite strain-dependent effects of post-training corticosterone in a passive avoidance task in mice: role of dopamine

Post-training administration of corticosterone (0.1–1 mg/kg) dose-dependently improves retention of an inhibitory avoidance response in C57BL/6 mice, whilst impairing it in the DBA/2 strain. The effects on retention performance induced by the hormone in C57BL/6 and DBA/2 mice appear to be due to an effect on memory consolidation. In fact, they were observed when the drug was given at short, but not long, periods of time after training, i.e., when the memory trace is susceptible to modulation. In the absence of pharmacological manipulations, the two strains showed a significant increase of plasma corticosterone levels 15 min after passive avoidance training that disappeared within 30 min, and similar step-through latencies on the test day. However, although no strain differences were observed for sensitivity to shock thresholds, the increase in plasma corticosterone levels elicited by passive avoidance training was more pronounced in mice of the DBA/2 strain (+ 160%) than in C57BL/6 mice (+ 52%). Moreover, DBA/2 mice were characterised by a higher number of either Type I or Type Il corticosteroid receptors in the hippocampus in comparison with C57BL/6 mice. Finally, the strain-dependent effects of an intermediate dose of corticosterone were enhanced by pretreatment with either the selective D₁ or D₂ dopamine (DA) receptor agonists SKF 38393 and LY 171555 and reversed by pretreatment with either selective D₁ or D₂ DA receptor antagonists SCH 23390 and (−)-sulpiride administered at per se non-effective doses. The present results indicate that studies in inbred strains of mice can dissect opposite effects of corticosterone on memory consolidation possibly due to its action at different steps or components of the multiphasic pathway of memory consolidation. Moreover, they suggest that some of these steps involve an interaction between the hormone and brain DA system.     

Concurrent muscarinic and beta-adrenergic blockade in rats impairs place-learning in a water maze and retention of inhibitory avoidance

These experiments examined the effects of separate and concurrent muscarinic cholinergic and beta-adrenergic blockade on inhibitory (passive) avoidance performance and spatial learning in the Morris water maze. Pretraining systemic administration of either scopolamine (0.3 or 1.0 mg/kg) or propranolol (3.0 or 10.0 mg/kg) had no significant effect on one-day retention of step-through inhibitory avoidance training. Similarly, pretraining administration of either 0.3 mg/kg scopolamine or 10 mg/kg propranolol did not affect spatial learning in the Morris water maze. However, combined administration of scopolamine and 10.0 mg/kg of propranolol impaired performance on these tasks. These findings further support a role for interactions between norepinephrine and acetylcholine in the modulation of learning and memory and implicate the participation of beta-adrenergic mechanisms in this interaction. Because cholinergic and noradrenergic deterioration is found in aging and Alzheimer's disease, these results also have implications regarding the role of age-related noradrenergic and cholinergic dysfunction in cognitive decline.     

Gender- and morphine dose-linked expression of spontaneous somatic opiate withdrawal in mice

The opiate withdrawal syndrome powerfully motivates opiate seeking and abuse. Development of effective medications for opiate withdrawal symptoms is thus a primary research goal and heavily relies on improved experimental models. This study was carried out to establish a clinically relevant paradigm to assess somatic opiate withdrawal in mice. Female and male C57BL/6J mice were treated with saline or increasing morphine doses (10-50mg/kg or 20-100mg/kg) during 6 consecutive days and tested for the spontaneous expression of somatic opiate withdrawal signs 8, 32, 56 and 80 h after last drug administration. Contrary to opioid receptor antagonist-precipitated procedures, the spontaneous opiate withdrawal paradigm used here revealed interesting gender- and morphine dose-linked differences. In particular, 56 h after last morphine administration elevated global opiate withdrawal scores were still evident in female but not in male mice treated with 20-100mg/kg. The severity of somatic opiate withdrawal directly correlated with the prior cumulative morphine exposure. Timing of expression of somatic opiate withdrawal signs also varied as a function of both gender and morphine dose. For example, expression of paw tremors and wet dog shakes was earlier in opiate-withdrawn male than in female mice. Overall, these findings highlight the possibility to detect gender- and opiate dose-linked differences in the expression and duration of somatic opiate withdrawal using a clinically relevant research model. The behavioral paradigm described here may represent a more appropriate tool to investigate the neurobiological bases of opiate withdrawal as opposed to opioid receptor antagonist-precipitated opiate withdrawal procedures.     

Cholinergic regulation of neocortical spindling in DBA/2 mice

Brief episodes of high-amplitude, bilaterally synchronous, seven-cycles-per-second spindles appear in the EEG of DBA/2 inbred mice during active waking, quiet waking, and slow-wave sleep. They do not occur during waking in C57BL/6 mice. This difference might result from differences in acetylcholine-mediated arousal as nicotine powerfully blocks brief spindle episodes in awake DBA/2 mice. The following results are reported. (i) Physostigmine (0.1 and 0.3 mg/kg, i.p.) desynchronized the EEG and produced behavioral immobility, but did not block brief spindle episodes in free-moving DBA/2 mice. (ii) Atropine (1.0 mg/kg, i.p.) reduced arousal and provoked slow waves without facilitating brief spindle episodes. (iii) Mecamylamine (1.0 mg/kg, i.p.) weakly activated spindles without producing any noticeable behavioral alterations. Because these treatments had little effect on spindle occurrence, the action of nicotine in brain stem-transected DBA/2 mice was investigated. Nicotine (1.0 mg/kg, i.p.) had no effect on brief spindle episodes released by rostropontine transection but powerfully blocked those provoked by pentylenetetrazol (20 mg/kg, i.p.) in midpontine-transected mice. Hence nicotine's antispindling action may be mediated in the rostral pons. As both nicotine and physostigmine produce behavioral immobility and EEG activation in free-moving DBA/2 mice, but only nicotine inhibits cortical spindling, the mechanisms that produce EEG desynchronization are probably not identical to those that prevent spindling. They may, though, be linked, parallel processes that are somehow dissociated in DBA/2 mice.     

Interactions between angiotensin II and baclofen in shuttle-box and passive avoidance performance

In experiments on male rats, we established that angiotensin-II (AT II) at a dose of 0.1 micrograms injected intracerebroventricularly immediately after training improved memory when retention tests (active and passive avoidance) were given 24 hours later. Baclofen at doses of 2, 5 and 10 mg/kg injected intraperitoneally immediately after training also improved retention in both active and passive avoidance tasks. Baclofen at a dose of 20 mg/kg was without effect on active avoidance performance. Combination of AT II and baclofen (2, 5 and 10 mg/kg) facilitated memory in active avoidance as compared to controls, but impaired retention as compared to the AT II-treated group. The impairment of the AT II-improved retention was stronger when the dose of baclofen in the combination was 20 ng/kg. Combination of AT II and baclofen (10 mg/kg) did not impair retention in passive avoidance. These data favor the view that GABA receptors may interfere with the AT II effects on memory consolidation or retention and that interactions of GABA (GABAA and GABAB) receptors with AT II receptors are of importance for memory processes.