Interaction between amount and pattern of training in the induction of intermediate- and long-term memory for sensitization in aplysia

In Aplysia, three distinct phases of memory for sensitization can be dissociated based on their temporal and molecular features. A single training trial induces short-term memory (STM, lasting <30 min), whereas five trials delivered at 15-min intervals induces both intermediate-term memory (ITM, lasting >90 min) and long-term memory (LTM, lasting >24 h). Here, we explore the interaction of amount and pattern of training in establishing ITM and LTM by examining memory for sensitization after different numbers of trials (each trial = one tail shock) and different patterns of training (massed vs. spaced). Under spaced training patterns, two trials produced STM exclusively, whereas four or five trials each produced both ITM and LTM. Three spaced trials failed to induce LTM but did produce an early decaying form of ITM (E-ITM) that was significantly shorter and weaker in magnitude than the late-decaying ITM (L-ITM) observed after four to five trials. In addition, E-ITM was induced after three trials with both massed and spaced patterns of training. However, L-ITM and LTM after four to five trials require spaced training: Four or five massed trials failed to induce LTM and produced only E-ITM. Collectively, our results indicate that in addition to three identified phases of memory for sensitization—STM, ITM, and LTM—a unique temporal profile of memory, E-ITM, is revealed by varying either the amount or pattern of training.     

Low environmental calcium blocks long-term memory formation in a freshwater pulmonate snail

The freshwater snail Lymnaea stagnalis (L.) is considered a calciphile and exhibits reduced growth and survival in environments containing less than 20 mg/l environmental calcium. Although it has no apparent effect on survival at 20 mg/l, reducing environmental calcium increases metabolic demand, and as such we consider that this level of calcium acts as a stressor on the snail. We exposed snails to acute periods of low environmental calcium and tested their ability to form intermediate-term memory (ITM) and long-term memory (LTM) following one trial operant conditioning (1TT) to reduce aerial respiratory activity in hypoxic conditions. We also assessed whether there were changes in the electrophysiological properties of a single neuron, right pedal dorsal 1 (RPeD1), which has been demonstrated to be necessary for LTM formation. Following training in high (80 mg/l) environmental calcium, L. stagnalis formed ITM and LTM lasting 24 h and demonstrated a significant reduction in all activity measured from RPeD1; however when snails were exposed to low (20 mg/l) environmental calcium they were able to form ITM but not LTM. Although no behavioral LTM was formed, a partial reduction in RPeD1 activtiy measured 24 h after training was observed, indicating a residual effect of training. The strong effect that environmental calcium concentration had on physiology and behavior in response to training to reduce aerial respiration in L. stagnalis suggests that it is an element of gastropod husbandry that needs to be carefully considered when studying other traits. This study also indicates that L. stagnalis found naturally in low calcium environments may be less able to adapt to novel stressors than populations found in harder waters.     

One-trial conditioning of aerial respiratory behaviour in Lymnaea stagnalis

Repeated spaced training sessions of contingent tactile stimulation to the pneumostome as it opens are required to cause long-term memory (LTM) formation of aerial respiratory behaviour making if difficult to determine exactly when memory forms. We have devised a single-trial aversive operant conditioning training procedure in Lymnaea to be better able to elucidate the causal mechanisms of LTM formation. Observations of baseline breathing behaviour in hypoxia were first made. Twenty-four hours later the snails were trained using the single trial procedure, by placing them in a small Petri dish containing 4 ml of 25 mM KCl for 30-35s as soon as the first pneumostome opening in hypoxia was attempted. LTM was present if (1) breathing behaviour following training was significantly less than before; and (2) breathing behaviour post-training was significantly less in experimental groups than in yoked control groups. LTM persisted for 24 h but not 48 h. Yoked controls that received an aversive stimulus not contingent with pneumostome opening had no evidence of memory. Cooling directly after, but not at any other time, blocks LTM formation. LTM formation was also prevented by removal of the cell body of the neuron RPeD1 before training.     

Context extinction and associative learning in Lymnaea

Aerial respiratory behavior in the pond snail Lymnaea was operantly conditioned so that snails learned not to perform aerial respiration in a hypoxic environment. Snails were trained in either the standard context (no food odorant) or a carrot (food-odorant) context. An operant training procedure of two 45-min training sessions with a 1-h interval between the sessions followed by a third 45-min training session 18 h later was sufficient to produce associative learning and long-term memory (LTM) that persisted for at least 5 days. If, however, following the third operant training session snails received three 45-min extinction training sessions, with each extinction session separated by at least a 1-h interval, LTM was not observed when tested the following day. That is, the memory was extinguished. Extinction, however, did not occur if the context of the extinction training was different from the context of the associative training. That is, in the snails trained in the standard context, extinction did not occur if the extinction training sessions were performed in the food-odorant context and vice versa.     

Operant conditioning in Lymnaea: evidence for intermediate- and long-term memory

Aerial respiration of the pond snail, Lymnaea stagnalis, can be operantly conditioned; however, the parameters necessary to produce long-term (LTM) or intermediate term memory (ITM) have not previously been investigated. We conducted training using procedures that varied in the duration of the training session, the number of training sessions per day or the amount of time between subsequent training sessions (SI). We found that by varying the duration and frequency of the training session learning could be differentially produced. Furthermore, the ability to form LTM was dependent not only on the duration of the training session was also the interval between training sessions, the SI. Thus it was possible to produce ITM, which persists for up to 3 hr, and not form LTM, which persists at least 18 hr. Learning, ITM, and LTM can be differentially produced by altering the SI, the duration of the training session, or the number of training sessions per day. These findings may allow us to begin to elucidate the underlying neural mechanisms of learning, ITM, and LTM.     

Reconsolidation and memory infidelity in Lymnaea

Lymnaea stagnalis were operantly conditioned to not perform aerial respiratory behaviour in a specific context (i.e. context-1). The memory for this learned response was reactivated 3 days later in context-1. During the 1 h reconsolidation period following memory reactivation, randomly picked snails were either maintained in context-1 or exposed to a new context (i.e. context-2). One hour later in the post-reconsolidation period, snails in context-1 were placed for 1 h in context-2 and vice-versa. In neither the hypoxic reconsolidation nor the post reconsolidation periods did snails receive a reinforcing stimulus when they opened their pneumostome. All snails were blindly tested for memory 24 h later period in context-2. Only those snails that had been exposed to context-2 during the reconsolidation period exhibited 'memory' for context-2. That is, memory infidelity was observed. Snails exposed to context-2 in only the post-reconsolidation period did not show memory for context-2. The immediate cooling of snails after their exposure to the new context in the reconsolidation period blocked the formation the implanted memory. Snails trained in context-1 and exposed to context-2 in the consolidation period only, also did not have memory for context-2. However, the memory for context-1 could still be recalled following successful implantation of the 'new' memory. All data presented here are consistent with the notion that during the reconsolidation process memory can be updated.     

Modulation of working, short- and long-term memory by nicotinic receptors in the basolateral amygdala in rats

Male Wistar rats were exposed to one-trial step-down inhibitory avoidance training using a 0.5 mA footshock. Through bilaterally implanted indwelling cannulae, they received bilateral 0.5 microL infusions of saline, mecamylamine (1.0 or 10.0 microg/side), or nicotine (0.6 or 3.0 microg/side) into the basolateral complex of the amygdaloid nucleus (BLA). Infusions were either 10 min before training (Experiment 1) or 4 min after training (Experiment 2). In Experiment 1, the animals were tested three times: first for working memory (WM) 5 s after training, then for short-term memory (STM) 90 min later, and finally for long-term memory (LTM) 24 h later. Mecamylamine depressed and nicotine enhanced WM, STM, and LTM. In Experiment 2, the treatments were given after WM was presumably over. Again, mecamylamine inhibited and nicotine enhanced STM and LTM. The results indicate that nAChRs in BLA participate in the regulation of WM formation and STM and LTM acquisition and consolidation.     

Long-term memory in pigeons: I. The role of discrimination problem difficulty assessed by reacquisition measures II. The role of stimulus modality assessed by generalization slope

Pigeons learned either an easy or a difficult line angle discrimination (Experiment 1) or wavelength discrimination (Experiment 2), and then they were given a reacquisition test of retention after delays of 1 min, 1 day, or 1 week. Both percentage of responses to the S+ in the initial 10-trial block and number of blocks to criterion showed a progressive memory loss which was greater for the difficult problem. These results extend recent findings by using a free operant rather than a discrete trial task and by varying problem difficulty by altering the dimensional separation between training stimuli. In Experiment 3, pigeons were given variable interval training with either a wavelength or a line angle stimulus, and then they were tested for generalization in extinction after delays of 1 min, 1 day, and 1 week. With both dimensions, the relative gradients became progressively flatter with increasing delay intervals. This replicates earlier findings and extends them to the line angle dimension. The evidence of substantial forgetting in the first 24 h in all three experiments suggests that operant free-response procedures are more sensitive to forgetting effects than are discrete trial tasks.     

Context learning and the effect of context on memory retrieval in Lymnaea

Aerial respiratory behavior in Lymnaea was operantly conditioned so that the animals perform aerial respiration significantly less often. Using the standard training procedure (pond water made hypoxic by bubbling N₂ through it) both food-deprived and fed animals learned and exhibited long-term memory (LTM). However, food-deprived animals exhibited neither learning nor memory when trained under a condition in which the hypoxic pond water also contained a food odorant (carrot, the food-odorant procedure). Fed animals, however, learned and exhibited LTM with the food-odorant procedure. Thus, the presence of the food odorant per se did not prevent learning or the establishment of LTM. Further experimentation, however, revealed that the ability of the snails to have recall (i.e., memory) for the learned behavior was dependent on the context in which memory was tested. That is, if animals were trained with the food-odorant procedure they could only exhibit recall if tested in the food-odorant context and vice versa with the standard training procedure. Thus, although fed animals could learn and show LTM with either training and testing procedure, LTM could only be seen when they were tested in the context in which they were trained.     

不同刺激强度训练后一日龄小鸡的记忆形成

一日龄小鸡,分别接受不同浓度的回避性刺激物邻氨基苯甲酸甲酯（ＭｅＡ）训练,间隔一定时间后测试记忆保持。主要结果为：５％和２０％ＭｅＡ训练,分别使动物记忆保持１５分钟和５０分钟左右,相当于记忆形成多阶段模型中的ＳＴＭ和ＩＴＭ阶段。４０％ＭｅＡ训练,记忆保持到训练后８－１０小时,相当于不依赖新糖蛋白合成的ＬＴＭ形成的早期阶段。只有６０％以上浓度的ＭｅＡ训练才使记忆保持至少２４小时。上述结果提示,利用弱化训练的方法,不仅可以将ＳＴＭ,ＩＴＭ和ＬＴＭ分离开,而且可把ＬＴＭ形成的早期阶段分离出来独立研究,增加了对ＬＴＭ新的理解,为进一步探讨记忆形成机制提供了一条新的途径。     

Participation of hippocampal cholinergic system in memory persistence for inhibitory avoidance in rats

Memory persistence needs a new event of consolidation 12h after the acquisition. We investigated the role of the cholinergic activity on the persistence of memory. For this purpose, we performed the treatments 9 or 12h after acquisition and the memory tested 2 or 7 days after inhibitory avoidance (IA) training. Here we report that activity of medial septum, by transitorily inactivating this structure with lidocaine 12h after IA training, is essential for memory persistence at the 7th day, but not for the formation at the 2nd day. We also report that muscarinic and nicotinic cholinergic receptors of CA1 area are engaged on memory persistence. Since scopolamine (mAChRs antagonist) and mecamylamine (nAChRs blocker) infusions, 12h post-training, demonstrated impairment on long term memory (LTM), persistence on the 7th day but no effect on LTM formation was found on the 2nd day in the IA test. The same effects were found with pirenzepine, an M1 antagonist. No effects on the formation and persistence of memory on the 2nd and 7th days were demonstrated after DHβE infusions (nAChRs subtype antagonist α4β2, α3β2). These findings suggest that mAChR and nAChR at the CA1 area, and also MS activation, are required for the persistence of memory.     

The influence of acoustic and semantic similarity on long-term memory for word sequences

It has been shown that short-term memory (STM) for word sequences is grossly impaired when acoustically similar words are used, but is relatively unaffected by semantic similarity. This study tests the hypothesis that long-term memory (LTM) will be similarly affected. In Experiment I subjects attempted to learn one of four lists of 10 words. The lists comprised either acoustically or semantically similar words (A and C) or control words of equal frequency (B and D). Lists were learned for four trials, after which subjects spent 20 min. on a task involving immediate memory for digits. They were then asked to recall the word list. The acoustically similar list was learned relatively slowly, but unlike the other three lists showed no forgetting. Experiment II showed that this latter paradox can be explained by assuming the learning score to depend on both LTM and STM, whereas the subsequent retest depends only on LTM. Experiment III repeats Experiment I but attempts to minimize the effects of STM during learning by interposing a task to prevent rehearsal between the presentation and testing of the word sequences. Unlike STM, LTM proved to be impaired by semantic similarity but not by acoustic similarity. It is concluded that STM and LTM employ different coding systems.     

Long-term memory survives nerve injury and the subsequent regeneration process

A three-neuron network (a central pattern generator [CPG]) is both sufficient and necessary to generate aerial respiratory behavior in the pond snail, Lymnaea stagnalis. Aerial respiratory behavior is abolished following a specific nerve crush that results in axotomy to one of the three CPG neurons, RPeD1. Functional regeneration of the crushed neurite occurs within 10 days, allowing aerial respiratory behavior to be restored. Functional regeneration does not occur if the connective is cut rather than crushed. In unaxotomized snails, aerial respiratory behavior can be operantly conditioned, and following memory consolidation, long-term memory (LTM) persists for at least 2 weeks. We used the Lymnaea model system to determine (1) If in naive animals axotomy and the subsequent regeneration result in a nervous system that is competent to mediate associative learning and LTM, and (2) if LTM survives RPeD1 axotomy and the subsequent regenerative process. We show here that (1) A regenerated nervous system is competent to mediate associative memory and LTM, and (2) LTM survives axotomy and the subsequent regenerative process.     

Multiple Memory Processes Following Training That a Food Is Inedible in Aplysia

In many organisms, memory after training can be separated into a number of processes. We now report that separable memory processes are also initiated by a training procedure affecting Aplysia feeding behavior, a model system for examining the neural mechanisms underlying the regulation of a complex behavior. Four distinct memory process were identified: (1) a very short-term memory that declines within 15 min, (2) a short-term memory that persists for 0.5–1.0 hr, (3) an intermediate-term memory, observed 4 hr after training, and (4) a long-term memory that is seen only after a 12- to 24-hr delay. The four memory processes can be distinguished by the different training procedures that are required to elicit them. A single 5-min training session is sufficient to elicit the very short-term memory. However, a longer training session that continues until the animal stops responding to food is needed to elicit short-term memory. Intermediate-term memory is observed only after a spaced training procedure (three 5-min training sessions separated by 30-min intervals). A single 5-min training session that does not cause either short-term or intermediate-term memory is sufficient to induce long-term memory, indicating that short- and long-term memory are independent, parallel processes. Short- and long-term memory can also be separated by the effects of a post-training experience. Long-term, but not short-term, memory can be attenuated by cooling animals immediately after training. Cooling before the training does not affect either the training or the subsequent short- or long-term memory.     

d-Cycloserine: effects on long-term retention of a conditioned response and on memory for contextual attributes.

d-Cycloserine (DCS), a partial agonist of the glycine recognition site of the N-methyl d-aspartate receptor, has beneficial effects on learning and memory. In order to investigate its potential to influence learning and memory of both the response and the stimulus attributes of training, male Sprague-Dawley albino rats were trained in a one-trial inhibitory avoidance task following an acute intraperitoneal injection of DCS (3 mg/kg) or an equal volume of saline. In order to measure memory for stimulus attributes, testing involved a context shift paradigm, in which subjects are tested in either the environment of training or a different one. Good memory for the contextual attributes of training is indicated by poor performance in the alternate context. Retention was assessed either 1, 7, or 14 days after training. At 1 day, Saline subjects were affected by a change in context, while DCS subjects were not. In subjects tested 1 week following training, Saline subjects were no longer affected by a change in context, in that they performed the avoidance response in both contexts. This indicates the forgetting of stimulus attributes in Saline subjects. DCS subjects did show the context shift effect at 1 week, indicating the retention of stimulus attributes. Finally, Saline subjects demonstrated the context shift rebound at 14 days, while DCS subjects performed equivalently in both contexts. Taken together, these data suggest that DCS may enhance retention of fear and slow the forgetting of stimulus attributes.