Sensory input from osphradium is involved in fluoride detection that alters feeding and memory phenotype in Lymnaea stagnalis.

Fluoride (F-) exposure in organisms remains a significant concern due to its widespread presence and potential health implications. Investigating its detection and subsequent effects on behaviour in aquatic organisms like Lymnaea stagnalis provides valuable insights. Our study focused on elucidating the sensory pathways involved in F- detection and its impact on feeding and memory formation. We explored two potential detection mechanisms: direct flow across the integument onto neurons; and sensory input to the central nervous system (CNS) via the osphradium-osphradial ganglion-osphradial nerve pathway (snails use this system for olfactory sensation of multiple compounds). Injection of F- into snails did not alter feeding behaviour or central neuronal activity, suggesting that internalization might not be the primary detection mode. In contrast, severing the osphradial nerve abolished F-'s suppressive effects on feeding and memory formation, implicating the osphradial pathway in F- sensing and behavioural changes. This finding supports the idea that osphradial nerve signaling mediates the behavioural effects of F-. Our study underscores the importance of sensory pathways in F- detection and behavioural modulation in L. stagnalis. Understanding these mechanisms could provide critical insights into how organisms respond to and adapt to environmental chemical stressors like F-.

Fluoride alters feeding in lab-bred pond snails but not in wild snails or their progeny.

Increasing concentrations of fluoride in natural bodies of water due to anthropogenic activities can lead to potentially detrimental effects on residing species. Here we investigated the differences in fluoride exposure on feeding behaviour between freshly collected (i.e., wild) and lab-bred Lymnaea stagnalis and if developmental exposure plays a key role in fluoride tolerance. We show that wild snails that reside in naturally fluoridated waters and their fluoride naïve lab-reared progeny have a fluoride tolerance that does not suppress feeding when introduced to a fluoridated food stimulus. These results were also seen in our lab-bred snails who were exposed to artificially fluoridated pond water (at similar concentrations to natural levels) throughout development. However, lab-bred snails that have never been exposed to fluoride, or only exposed during the egg stage demonstrated suppression of feeding in the fluoridated food stimulus condition compared to an unfluoridated food stimulus. Genetic diversity and phenotypic plasticity are suspected to be two key underlying mechanisms for fluoride ion tolerance. These results are critical in understanding how parental and developmental exposure can influence a phenotypic tolerance to a potential chemical pollutant.

Fluoride affects memory by altering the transcriptional activity in the central nervous system of Lymnaea stagnalis.

Fluoride (F-), has been found to affect learning and memory in several species. In this study, we exposed an F--naïve, inbred strain of Lymnaea stagnalis to a concentration of F- similar to that naturally occurring in wild ponds. We found that the exposure to F- before the configural learning procedure obstructs the memory formation and blocks the configural learning-induced upregulation of CREB1, GRIN1, and HSP70 in snails' central ring ganglia. Along with altering the mRNA levels of these key genes for memory formation, a single acute F- exposure also upregulates Cytochrome c Oxidase, a major regulatory enzyme of the electron transport chain, which plays direct or indirect roles in reactive oxygen species production. As the central nervous system is sensitive to oxidative stress and consistent with previous studies from mammals, our results suggest a potential role of oxidative stress in memory impairment. To our knowledge, this is the first study investigating the neuronal mechanism of memory impairment in an invertebrate species that is exposed to natural F- levels.

Fluoride alters feeding and memory in Lymnaea stagnalis.

Fluoride occurs naturally in the terrestrial and aquatic environment and is a major component in tea. Prolonged fluoride exposure alters metabolic activity in several aquatic invertebrates. For the first time, we investigated the effects of fluoride on cognition in the pond snail Lymnaea stagnalis as it is capable of a higher form of associative learning called configural learning. We first showed suppressive effects of black tea and fluoride on feeding (i.e., rasping) behavior. We then investigated how fluoride may alter cognition by introducing fluoride (1.86 mg/L) before, during, after, a day before and a week before the snails underwent the configural learning training procedure. Our results show that any 45-min exposure to fluoride (before, during or after a configural learning training procedure) blocks configural learning memory formation in Lymnaea and these effects are long-lasting. One week after a fluoride exposure, snails are still unable to form a configural learning memory and this result is upheld when the snails are exposed to a lower concentration of fluoride, one which is naturally occurring in ponds that a wild strain of Lymnaea can be found (0.3 mg/L). Thus, fluoride obstructs configural learning memory formation in a fluoride-naïve, inbred strain of Lymnaea.