SUMMARYThe 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 48 h (i.e. LTM). We conclude that memory enhancement is due to the stress associated with the thermal stimulus.

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.