Strain-specific differences of the effects of stress on memory in Lymnaea.

Stress alters the ability to form, recall and maintain memory according to the Yerkes-Dodson/Hebb (YDH) law. The effects of environmentally relevant stressors, such as low environmental calcium and crowding, on learning and memory have previously been described in a laboratory-reared 'average' strain of Lymnaea stagnalis (i.e. the Dutch strain) as well as two strains of freshly collected L. stagnalis with enhanced memory formation abilities (i.e. 'smart' snails). Here, we use L. stagnalis to study the effects of other environmentally relevant stressors on memory formation in two other strains of freshly collected snails, one 'smart' and one 'average'. The stressors we examined are thermal, resource restriction combined with food odour, predator detection and, for the first time, tissue injury (shell damage). We show that the same stressor has significantly different effects on memory formation depending on whether snails are 'smart' or 'average'. Specifically, our data suggest that a stressor or a combination of stressors act to enhance memory in 'average' snails but obstruct memory formation in 'smart' snails. These results are consistent with the YDH law and our hypothesis that 'smart' snails are more easily stressed than 'average' snails.

Juveniles of Lymnaea 'smart' snails do not perseverate and have the capacity to form LTM.

Previously, it was concluded that the nervous systems of juvenile snails were not capable of mediating long-term memory (LTM). However, exposure and training of those juvenile snails in the presence of a predator cue significantly altered their ability to learn and form LTM. In addition, there are some strains of Lymnaea which have been identified as 'smart'. These snails form LTM significantly better than the lab-bred strain. Here, we show that juveniles of two smart snail strains not only are capable of associative learning but also have the capacity to form LTM following a single 0.5 h training session. We also show that freshly collected 'wild' 'average' juveniles are also not able to form LTM. Thus, the smart snail phenotype in these strains is expressed in juveniles.

Heat stress enhances LTM formation in Lymnaea: role of HSPs and DNA methylation.

Environmentally relevant stressors alter the memory-forming process in Lymnaea following operant conditioning of aerial respiration. One such stressor is heat. Previously, we found that following a 1 h heat shock, long-term memory (LTM) formation was enhanced. We also had shown that the heat stressor activates at least two heat shock proteins (HSPs): HSP40 and HSP70. Here, we tested two hypotheses: (1) the production of HSPs is necessary for enhanced LTM formation; and (2) blocking DNA methylation prevents the heat stressor-induced enhancement of LTM formation. We show here that the enhancing effect of the heat stressor on LTM formation occurs even if snails experienced the stressor 3 days previously. We further show that a flavonoid, quercetin, which inhibits HSP activation, blocks the enhancing effect of the heat stressor on LTM formation. Finally, we show that injection of a DNA methylation blocker, 5-AZA, before snails experience the heat stressor prevents enhancement of memory formation.

Environmentally relevant stressors alter memory formation in the pond snail Lymnaea.

Stress alters adaptive behaviours such as learning and memory. Stressors can either enhance or diminish learning, memory formation and/or memory recall. We focus attention here on how environmentally relevant stressors alter learning, memory and forgetting in the pond snail, Lymnaea stagnalis. Operant conditioning of aerial respiration causes associative learning that may lead to long-term memory (LTM) formation. However, individual ecologically relevant stressors, combinations of stressors, and bio-active substances can alter whether or not learning occurs or memory forms. While the behavioural memory phenotype may be similar as a result of exposure to different stressors, how each stressor alters memory formation may occur differently. In addition, when a combination of stressors are presented it is difficult to predict ahead of time what the outcome will be regarding memory formation. Thus, how combinations of stressors act is an emergent property of how the snail perceives the stressors.

What's hot: the enhancing effects of thermal stress on long-term memory formation in Lymnaea stagnalis.

The pond snail, Lymnaea stagnalis, naturally inhabits slow flowing, shallow and stagnant environments in the northern temperate zone. Consequently, it will experience wide temperature fluctuations dependent on prevailing weather conditions. We hypothesize that periods of warming act as a thermal stressor to alter memory formation. Snails were exposed to an acute 1 h period of 30°C pond water and we determined how memory formation following operant conditioning of aerial respiration was affected. In the snails used here (Dutch strain), a single 0.5 h training session (TS) results in intermediate-term (3 h) but not long-term memory (LTM). Applying the thermal stressor during training caused memory enhancement (i.e. LTM lasting 24 h). However, the breathing rate also increased in warm water, which might explain the enhanced memory. Therefore, we applied the thermal stressor (1 h at 30°C) up to 4 h before or 1 h after training. This did not alter baseline breathing rate during the period when snails would experience training. However, the thermal stressor whether experienced prior to or following the single TS, resulted in an enhanced memory that persisted up to 48h (i.e. LTM). We conclude that memory enhancement is due to the stress associated with the thermal stimulus.

Differences in neuronal activity explain differences in memory forming abilities of different populations of Lymnaea stagnalis.

The ability to learn and form long-term memory (LTM) can enhance an animal's fitness, for example, by allowing them to remember predators, food sources or conspecific interactions. Here we use the pond snail, Lymnaea stagnalis, to assess whether variability between natural populations (i.e., strains) in memory forming capabilities correlates with electrophysiological properties at the level of a single neuron, RPeD1. RPeD1 is a necessary site of LTM formation of aerial respiratory behaviour following operant conditioning. We used strains from two small, separate permanent ponds (TC1 and TC2). A comparison of the two populations showed that the TC1 strain had enhanced memory forming capabilities. Further, the behavioural phenotype of enhanced memory strain was explained, in part, by differences in the electrophysiology of RPeD1. Compared to RPeD1 from the naive TC2 strain, RPeD1 from the TC1 strain has both a decreased resistance and decreased excitability. Moreover, 24h after a single 0.5h training session, those membrane properties, as well as the firing and bursting rate, decrease further in the TC1 strain but not in the TC2 strain. The initial differences in RPeD1 properties in the TC1 strain coupled with their ability to further change these properties with a single training session suggests that RPeD1 neurons from the TC1 strain are "primed" to rapidly form memory.

Ecologically relevant stressors modify long-term memory formation in a model system.

Stress can alter adaptive behaviours, and as well either enhance or diminish learning, memory formation and/or memory recall. We focus attention on how environmentally relevant stressors (e.g. predator detection, crowding, and low concentrations of environmental Ca(++)) alter memory formation in the pond snail, Lymnaea stagnalis. We specifically look at operant conditioning of aerial respiration and whether or not long-term memory forms following the acquisition of the learned event, not performing aerial respiration. We will also examine the strain differences in Lymnaea which allow or cause isolated populations to possess different heritable cognitive capabilities, as manifested by differing abilities to form long-term memory.