Abstract
The pond snail Lymnaea stagnalis exhibits various forms of associative learning including (1) operant conditioning of aerial respiration where snails are trained not to open their pneumostome in a hypoxic pond water environment using a weak tactile stimulus to their pneumostome as they attempt to open it; and (2) a 24 h-lasting taste-specific learned avoidance known as the Garcia effect utilizing a lipopolysaccharide (LPS) injection just after snails eat a novel food substance (carrot). Typically, lab-inbred snails require two 0.5 h training sessions to form long-term memory (LTM) for operant conditioning of aerial respiration. However, some stressors (e.g., heat shock or predator scent) act as memory enhancers and thus a single 0.5 h training session is sufficient to enhance LTM formation lasting at least 24 h. Here, we found that snails forming a food-aversion LTM following Garcia-effect training exhibited enhanced LTM following operant condition of aerial respiration if trained in the presence of the food substance (carrot) they became averse to. Control experiments led us to conclude that carrot becomes a ‘sickness’ risk signal and acts as a stressor, sufficient to enhance LTM formation for another conditioning procedure.
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Data availability
The data that support the findings of this study are available on request from the corresponding author, [VR].
Abbreviations
- C:
-
Carrot slurry
- L:
-
Lettuce slurry
- LTM:
-
Long-term memory
- MT:
-
Memory test
- TS:
-
Training session
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Acknowledgments
We would like to thank Dr. Petra Hermann for providing us with the lab-reared population of snails. Furthermore, we would like to thank Consorzio Interuniversitario Biotecnologie (CIB, Trieste, Italy) for the research grant awarded to Dr. Rivi to work as a visiting researcher at the Hotchkiss Brain Institute (University of Calgary, AB, Canada).
Funding
The current work was funded by the Natural Sciences and Engineering Research Council of Canada and Regione Emilia Romagna L.R. N. 20/2002 PROGETTI DI RICERCA SUI METODI ALTERNATIVE ALL’UTILIZZO DI ANIMALI (NSERC-227993-2019); and FAR 2016 Department of Life Sciences, University of Modena and Reggio Emilia.
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Conceptualization: VR, AB, K.L; Methodology: VR, AB, KL; Validation: VR.; Formal analysis: VR; Investigation: VR, A.B; Resources: KL.; Data curation: VR.; Writing–original draft: VR; Writing–review & editing: AB, CB, FT, JM, CB, KL; Supervision: KL ; Project administration: KL; Funding acquisition: KL, JMCB, FT.
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Rivi, V., Batabyal, A., Benatti, C. et al. Investigating the interactions between multiple memory stores in the pond snail Lymnaea stagnalis. J Comp Physiol A 210, 91–102 (2024). https://doi.org/10.1007/s00359-023-01649-3
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DOI: https://doi.org/10.1007/s00359-023-01649-3