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.

Silver nanoparticles alter learning and memory formation in an aquatic organism, Lymnaea stagnalis.

We tested the effect of silver nanoparticles (AgNPs) on the ability of the pond snail, Lymnaea stagnalis, to learn and form long-term memory (LTM) following operant conditioning of aerial respiration. We hypothesized that the AgNPs would act as a stressor and prevent learning and LTM formation. We tested snails exposed for either 72 h or only during training and testing for memory (i.e. 0.5 h) and found no difference between those treatments. We found that at a low concentration of AgNPs (5 µg/L) neither learning and nor memory formation were altered. When we increased the concentration of AgNPs (10 µg/L) we found that memory formation was enhanced. Finally, at a higher concentration (50 µg/L) memory formation was blocked. To determine if the disassociation of Ag+ from the AgNPs caused the effects on memory we performed similar experiments with AgNO3 and found similar concentration-dependent results. Finally, we found that snails perceive the AgNPs differently from Ag+ as there was context specific memory. That is, snails trained in AgNPs did not show memory when tested in Ag+ and vice-versa. We believe that changes in memory formation may be a more sensitive determination of AgNPs on aquatic organisms than the determination of a LC50.

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.