Inhibition of mGluR1 and IP3Rs impairs long-term memory formation in young chicks.

Calcium (Ca(2+)) is involved in a myriad of cellular functions in the brain including synaptic plasticity. However, the role of intracellular Ca(2+) stores in memory processing remains poorly defined. The current study explored a role for glutamate-dependent intracellular Ca(2+) release in memory processing via blockade of metabotropic glutamate receptor subtype 1 (mGluR1) and inositol (1,4,5)-trisphosphate receptors (IP(3)Rs). Using a single-trial discrimination avoidance task developed for the young chick, administration of the specific and potent mGluR1 antagonist JNJ16259685 (500nM, immediately post-training, ic), or the IP(3)R antagonist Xestospongin C (5microM, immediately post-training, ic), impaired retention from 90min post-training. These findings are consistent with mGluR1 activating IP(3)Rs to release intracellular Ca(2+) required for long-term memory formation and have been interpreted within an LTP2 model. The consequences of different patterns of retention loss following ryanodine receptor (RyR) and IP(3)R inhibition are discussed.

Maternal hen calls modulate memory formation in the day-old chick: the role of noradrenaline.

Facilitation of memory for discriminative learning in young chicks is enhanced following exposure to a synthesized rhythmic auditory stimulus. Increased arousal, mediated by noradrenergic activation, is believed to underlie this effect. In this report we examine whether ethologically relevant rhythmic auditory stimuli produce the same noradrenaline-mediated memory enhancement in neonate chicks (Gallus gallus domesticus). Maternal hen attraction calls which contained a rhythmic pattern were found to facilitate retention. Intracerebral injection of noradrenaline or the beta(2)-adrenergic antagonist propranolol demonstrated that this enhancement is likely to be mediated by noradrenergic activation of central beta(2)-noradrenergic receptors. In contrast, a rhythmic alarm call inhibited retention. Subcutaneous injection of the alpha(1)-adrenergic antagonist prazosin revealed that this impairment may be due to higher arousal levels resulting in activation of alpha(1)-noradrenergic receptors. It is concluded that the maternal hen calls of domestic chickens can influence the memory ability of the offspring via noradrenaline release in the brain. The current data suggest that call meaning and rhythmicity interact to yield the appropriate levels of beta(2)-adrenergic activation required to facilitate retention for a discriminative task.

New perspectives on the mechanisms through which nitric oxide may affect learning and memory processes.

Nitric oxide (NO) has been well established as a molecule necessary for memory consolidation. Interestingly, the majority of research has focused on only a single mechanism through which NO acts, namely the up-regulation of guanylate cyclase (GC). However, since NO and NO-derived reactive nitrogen species are capable of interacting with a broad array of enzymes, ion channels and receptors, a singular focus on GC appears short-sighted. Although NO inhibits the action of a number of molecules there are four, in addition to GC, which are up-regulated by the direct presence of NO, or NO-derived radicals, and implicated in memory processing. They are: cyclic nucleotide-gated channels; large conductance calcium-activated potassium channels; ryanodine receptor calcium release (RyR) channels; and the enzyme mono(ADP-ribosyl) transferase. This review presents evidence that not only are these four molecules worthy of investigation as GC-independent mechanisms through which NO may act, but that behavioural evidence already exists suggesting a relationship between NO and the RyR channel.

Pharmaco-behavioural evidence indicating a complex role for ryanodine receptor calcium release channels in memory processing for a passive avoidance task.

Calcium signalling is an important process underlying neuronal function and consequently behaviour. The release of calcium from intracellular stores via the ryanodine receptor calcium release (RyR) channel has been implicated in both synaptic plasticity and to a limited extent in memory processing. While past investigations have suggested a role for RyR channels in long-term memory, the present study suggests their action is more complex. Using a single trial passive avoidance task developed for the day-old chick, it is proposed that RyR channels are necessary both prior to the expression of long-term memory and also in retrieval processes. Specifically, 5 mM dantrolene (a specific RyR channel blocker) resulted in a persistent retention loss from 40 min post-training while 10 nM dantrolene produced a transient retention loss centred at 40 min post-training. We speculate that in the context of memory formation, RyR channels may be activated by nitric oxide and in the context of memory retrieval may lead to the activation of large conductance calcium-activated potassium BK(Ca) channels which, when blocked by 50 nM iberiotoxin, also demonstrated a transient retention loss centred at 40 min post-training.

Noradrenaline involvement in the memory-enhancing effects of exposure to a complex rhythm stimulus following discriminated passive avoidance training in the young chick.

Previous research in our laboratory has demonstrated a significant memory-enhancing effect of exposure to a complex rhythm stimulus following weakly-reinforced passive avoidance learning in chicks. The aim of this study was to explore whether noradrenaline mediates this process. Chicks were trained on a strongly-reinforced single-trial passive avoidance task involving discrimination between two coloured beads. Intracerebral administration of the protein synthesis blocker, anisomycin, revealed that a phase of memory formation sensitive to arousal levels was extended by approximately 35 min following exposure to the complex rhythm stimulus. Administration of 2,4-dinitrophenol showed that this extension occurred during phase B of intermediate-term memory. Finally, a higher dose of the beta-adrenergic receptor antagonist, propranolol, was required to inhibit long-term memory in the presence of the auditory stimulus than in its absence. These findings suggest that the memory-enhancing effects of the complex rhythm stimulus may be mediated by noradrenaline, possibly via an increase in physiological arousal.

A scavenger of peroxynitrite prevents long-term memory formation using a single trial passive avoidance task for the day-old chick.

The important role of nitric oxide (NO) in memory processing has been recognised for some time. However, the mechanisms through which NO may act are only partially understood. One highly reactive radical brought about by the reaction of NO and superoxide ions is peroxynitrite. The current study investigated the effect of peroxynitrite scavenging on retention for a single trial passive avoidance task developed for the day-old chick. Administration of a range of concentrations of the peroxynitrite scavenger Trolox (0.1 microM-1.2 mM) yielded a bimodal retention curve. This dose-response curve had nadirs at 300 and 800 microM. A time of administration study was conducted for each optimal concentration of Trolox and in both studies, the effective range of administration times extended from at least 10 min before training to 20 min post-training. Finally, a retention function was conducted for each optimum concentration of Trolox and in both studies a persistent retention loss was observed from 40 min post-training until the conclusion of the experiment 24 h post-training. The findings suggest that physiological levels of peroxynitrite may be required for the consolidation of long-term memory in this model of memory formation. Interestingly, the effective times of administration and time of retention loss onset are consistent with previous studies which blocked NO synthesis. Therefore it may be suggested that NO acts to facilitate long-term memory formation through the production of peroxynitrite.

Variations in intensity and frequency moderate the facilitative effects of a complex rhythm stimulus on long-term memory consolidation in the day-old chick (Callus gallus).

The authors have previously shown that exposure to 1 min of a complex, but not an isochronous, rhythm stimulus facilitates long-term memory consolidation in chicks (Gallus gallus) trained on a passive-avoidance task (S. R. Toukhsati & N. S. Rickard, 2001). The acoustic parameters of this stimulus were explored further in the current study. Retention was found to be best facilitated when the complex rhythm stimulus was presented at intensities between 5 and 15 dBA above background laboratory noise levels and at a frequency of 1 kHz. Removal of an accent from the stimulus did not moderate the effect. These findings provide confirmation that memory in an avian species can be facilitated by exposure to a complex rhythm stimulus and suggest that pattern repetition may be an important feature of this effect.

Effects of nitric oxide inhibition on avoidance learning in the chick are lateralized and localized.

Bilateral administration of nitric oxide synthase inhibitors into the intermediate medial hyperstriatal (IMHV) region of the chick brain impairs memory formation for an avoidance task. The aim of the current study was to determine whether this effect was restricted to a particular location in the brain, and whether inhibition was equally effective in both hemispheres. White Leghorn x black Australorp chicks were administered 0.5 mM N(omega)-Nitro-L-arginine methyl ester bilaterally into the lobus parolfactorius (LPO), or unilaterally into the IMHV. Injections into the LPO between 5 min pre-training and 40 min post-training had no effect on retention. In contrast, unilateral injections into the IMHV impaired retention and memory loss occurred from 40 min post-training. The effective administration time was hemisphere-dependent, requiring left hemisphere administration around the time of training and right hemisphere administration between 15 and 25 min post-training. These data suggest that localized nitric oxide activity in each hemisphere of the chick brain is necessary for the consolidation of memory for this task.

Inhibition of monoADP-ribosylation prevents long-term memory consolidation of a single-trial passive avoidance task in the day-old chick.

The cytosolic posttranslational protein-modifying mechanism of monoADP-ribosylation has been implicated in long-term potentiation, a synaptic model of memory formation. The current study investigated the effect of inhibiting mono(ADP-ribosyl) transferase on memory for the passive avoidance task in day-old chicks (white Leghorn-black Australorp). Various doses of novobiocin or menadione sodium bisulfite were administered intracranially at different times before or after training. Control chicks were administered saline at matched times. Novobiocin (650 microM) or menadione sodium bisulfite (250 microM) administered between 5.0 min pretraining and 2.5 min posttraining was found to cause a persistent loss of retention from 120 min posttraining. These data provide the first demonstration that monoADP-ribosylation is required for the maintenance of long-term memory. Furthermore, the temporal characteristics of the memory loss caused by monoADP-ribosylation inhibition appears to exclude this mechanism as a downstream effect of the well-established nitric oxide activity previously shown to occur within 40 min of passive avoidance training.

Inhibition of guanylate cyclase and protein kinase G impairs retention for the passive avoidance task in the day-old chick.

Nitric oxide (NO) is a highly labile chemical messenger which has previously been implicated in memory processes in a variety of learning paradigms and species. However, there is only limited evidence to suggest which enzymes are acted upon by NO during the formation of memory. The present study investigates the role of guanylate cyclase (GC) and protein kinase G (PKG) in a form of passive avoidance learning known to be dependent on nitric oxide activity. It was determined that in vivo pharmacological inhibition of GC using either 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one or 6-anilino-5,8-quinolinedione resulted in two transitory memory retention deficits centred around 40 and 120 min posttraining, respectively. In contrast, inhibition of PKG with N-[2-(methylamino)ehtyl]-5-isoquinoline-sulfornamide hydrochloride (H-8) resulted in a single temporary retention loss centered at 120 min posttraining. These temporary retention losses appear to be specific to memory since they were dose-dependent and could not be explained by nonspecific performance effects. Further, these results suggest that these agents inhibit memory retrieval rather than formation, since memory is subsequently available. The current findings indicate that guanylyl cyclase mediates two memory retrieval processes, the latter of which appears to be PKG-dependent. In contrast, since inhibition of NO results in a permanent retention loss, it is suggested that NO is required for memory formation through GC-independent processes.

Effect of the Ginkgo biloba extract, EGb 761, on memory formation in day-old chicks.

Previous studies indicate that the Ginkgo biloba extract, EGb 761, has a facilitative effect on deficient memory. The temporal parameters of this effect, however, have not been clearly defined or distinguished from the effect on normal memory. The aim in the current study was to investigate the effects of EGb 761 on memory using a well-controlled animal model. Day-old chicks were trained on either a weakly or strongly reinforced version of a passive avoidance task. Long-term memory formation of the weakly reinforced version of the task was improved significantly by EGb 761 (3 mg/ml) when administered between 10 and 30 min after training. However, the same dose of EGb 761 impaired retention when administered prior to strongly reinforced training. These data provide convincing evidence that posttraining administration of EGb 761 initiates long-term memory in chicks with only short-term memory, but that the same dose-administered pretraining can be deleterious for normal retention. This dual effect has important implications for the clinical use of Ginkgo biloba extracts.

Exposure to a rhythmic auditory stimulus facilitates memory formation for the passive avoidance task in the day-old chick.

Previous research regarding the beneficial effects of auditory stimuli on learning and memory in humans has been inconsistent. In the current study, day-old chicks were used to reduce the impact of individual differences on responses. Chicks were trained on a passive avoidance task and exposed to various auditory stimuli. Exposure to a complex rhythmic sequence for 1 min strongly facilitated chicks' long-term memory. The optimal time of presentation of the stimulus was between 10 min before and 20 min after training. Moreover, the enhancing effect was not generalized to the other auditory stimuli tested. It is suggested that this effect may be due to arousal because arousal hormones are critical to long-term memory formation. This study indicates that the temporal characteristics and type of stimulus may be important considerations when investigating the effects of auditory stimuli on cognitive functioning.

Inhibition of the endothelial isoform of nitric oxide synthase impairs long-term memory formation in the chick.

Previous studies with general inhibitors of nitric oxide synthase have yielded variable and contradictory results with respect to their effects on memory. This may have been due to differential effects of blocking the various isoforms of this enzyme. We show that day-old chicks trained on a single-trial passive-avoidance task suffered significant memory loss from approximately 40 min post-training following post-training intracranial administration of a potent inhibitor of eNOS. Administration of a specific nNOS or iNOS inhibitor at the same time had no effect on retention, although a role for either of these isoforms when administered at a different time after learning has yet to be fully investigated. The onset of memory loss following eNOS inhibition is the same as observed following general NOS inhibition, which suggests that amnestic effects observed in previous studies using nonspecific inhibitors may be attributable to blocking the function of eNOS. The findings indicate that eNOS may play a role in memory formation for this task, which is at least distinct from any role that may be played by nNOS.

Further support for nitric oxide-dependent memory processing in the day-old chick.

There is considerable evidence that nitric oxide activity is essential for memory formation, particularly from studies using inhibitors of nitric oxide synthase. The particular stage of memory formation requiring nitric oxide activity has not, however, been systematically investigated. In the current experiments, day-old black Australorp-white Leghorn chicks were trained on a passive avoidance task. Intracranial injections of the nitric oxide synthase inhibitor, l-NG-nitroarginine methylester (0.5 mM), were found to inhibit memory formation shortly after training, when injected pre- or posttraining. This effect was replicated with a second inhibitor, l-NG-nitroarginine (1 mM), and counteracted by the NO donor, sodium nitroprusside (150 microM). These findings provide covergent evidence that nitric oxide activity plays a critical role in the consolidation of memory in the day-old chick.

Complex roles of glutamate in the Gibbs-Ng model of one-trial aversive learning in the new-born chick.

Glutamate is the most widespread excitatory transmitter in the CNS and is probably involved in LTP, a neural phenomenon which may be associated with learning and memory formation. Intracerebral injection of large amounts of glutamate between 5 min and 2.5 min after passive avoidance learning in young chicks inhibits short-term memory, which occurs between 0 and 10 min post-learning in a three-stage model of memory formation first established by Gibbs and Ng(25) [Physiol. Behav. 23:369-375; 1979]. This effect may be attributed to non-specific excitation. Blockade of glutamate uptake by L-aspartic and beta-hydroxamate also abolishes this stage of memory, provided the drug is administered within 2.5 min of learning. Interference with either production of percursors for transmitter glutamate in astrocytes or with glutamate receptors is also detrimental to memory formation, but the effects appear much later. After its release from glutamatergic neurons, glutamate is, to a large extent, accumulated into astrocytes where it is converted to glutamine, which can be returned to glutamatergic neurons and reutilized for synthesis of transmitter glutamate, and partly oxidized as a metabolic substrate. The latter process leads to a net loss of transmitter glutamate which can be compensated for by de novo synthesis of a glutamate precursor alpha-ketoglutarate (alpha KG) in astrocytes, a process which is inhibited by the astrocyte-specific toxin fluoroacetate (R. A. Swanson, personal communication). Intracerebral injection of this toxin abolishes memory during an intermediate stage of memory processing occurring between 20 and 30 min post-training (50) [Cog. Brain Res, 2:93-102; 1994]. Injection of methionine sulfoximine (MSO), a specific inhibitor of glutamine synthetase, which interferes with the re-supply of transmitter glutamate to neurons by inhibition of glutamine synthesis in astrocytes, has a similar effect. This effect of MSO is prevented by intracerebral injection of glutamate, glutamine, or a combination and alpha KG and alanine. MSO must be administered before learning, but does not interfere with acquisition since short-term memory remains intact. Administration of either the NMDA antagonist AP5, the AMPA antagonist DNQX, or the metabotropic receptor antagonist MCPF, also induces amnesia. Memory loss in each case does not occur until after 70 min post-training, during a protein synthesis-dependent long-term memory stage which begins at 60 min following learning. However, to be effective, AP5 must be administered within 60 s following learning, MCPG before 15 min post-learning, and DNQX between 15 and 25 min after learning. Together, these findings suggest that learning results in an immediate release of glutamate, followed by a secondary release of this transmitter at later stages of processing of the memory trace, and that one or both of these increases in extracellular glutamate concentration are essential for the consolidation of long-term memory. Since both fluoroacetate and MSO act exclusively on glial cells, the findings also show that neuronal-glial interactions are necessary during the establishment of memory.

Blockade of metabotropic glutamate receptors prevents long-term memory consolidation.

Metabotropic glutamate receptor activation has been shown to be essential for establishment of long-term potentiation, a phenomenon increasingly thought to be associated with the laying down of permanent memory. However, these receptors may also play a part in the initiation of protein kinase C activity, which has been demonstrated to be involved in prelong-term memory processes. Blockade of the metabotropic glutamate receptors by the specific antagonist, (RS)-alpha-Methyl-4-carboxyphenylglycine (500 microM) is shown to induce amnesia during a long-term memory stage in day-old chicks trained on a passive avoidance task, and to have no effect on prelong-term stages. The results suggest a specific role for these receptors in a possibly LTP associated mechanism of memory processing.

Both non-NMDA and NMDA glutamate receptors are necessary for memory consolidation in the day-old chick.

Day-old chicks (black Australorp-white Leghorn) trained to avoid an aversive stimulus will usually retain memory for this event indefinitely. The passive avoidance task used involves a period of pretraining where chicks peck freely at two differently colored glass beads, a single training trial where one of the beads is coated in a chemical aversant eliciting typical disgust reactions from the chicks, and a test trial where both beads are presented dry, and discrimination memory is demonstrated by avoidance of the previously aversive bead with continued pecking of the nonaversive bead. Intracranial administration of a N-methyl-D-aspartate glutamate receptor antagonist (50 microM 2-amino-5-phosphopentanoate) immediately after or prior to learning, or a non-N-methyl-D-aspartate glutamate receptor antagonist (100 microM 6,7-dinitro-quinoxaline-2,3-dione) between 10 and 25 min after learning, resulted in amnesia for this task at 80 to 90 min post-training. These data indicate that processes dependent on N-methyl-D-aspartate and non-N-methyl-D-aspartate glutamate receptor activation are necessary for memory consolidation of a passive avoidance task in the day-old chick. Since these agents must be administered during the earlier stages of memory formation to cause amnesia, the receptors are probably activated close to the time of learning. The delayed effect of these antagonists, however, suggests that memory is independent of these receptors until quite late in the memory consolidation process.

A nitric oxide agonist stimulates consolidation of long-term memory in the 1-day-old chick.

Chicks, age 1 to 2 days, that have been trained on a passive avoidance task with a strongly reinforced training trial yield a memory trace that is composed of 3 behaviorally and pharmacologically distinguishable stages, with the final long-term memory stage being dependent on protein synthesis. In contrast, chicks trained with a weakly reinforced learning trial typically do not demonstrate this final stage of memory. Sodium nitroprusside 150 microM intracranially administered immediately after a weak training trial promoted the formation of long-term memory, whereas saline did not. The results suggest that nitric oxide synthesis is either itself critical or stimulates other processes that are critical for the consolidation of long-term memory.