A change in taste: The role of MicroRNAs in altering hedonic value

The mechanisms associated with neophobia, and anhedonia remain largely unknown. Neuropsychological disorders such as depression and schizophrenia are associated with excessive fear and anhedonia and have been linked to microRNA 137. We hypothesized that microRNAs (miRNAs) in the snail Lymnaea stagnalis are important for regulating feeding behaviour through either preventing neophobia or establishing hedonic value. To test these hypotheses, we used an injection of Poly-L-Lysine (PLL) to inhibit miRNA biogenesis and observed its effects on feeding behaviour. We repeated these experiments with pre-exposure to novel stimuli capable of eliciting neophobia to disentangle the processes predicted to regulate feeding behaviour. Next, we exposed snails to food stimuli of high hedonic value after PLL injection to reset their hedonic value for that food. Finally, we consolidated our results with previous research by examining the effect of PLL injection on a one trial appetitive classical conditioning procedure (1TT) to induce long term memory (LTM). We found that miRNAs are likely not required for preventing neophobia. Moreover, we discovered that snails experienced anhedonia in response to inhibition of miRNA biogenesis, resulting in diminished feeding behaviour for food stimuli with a previously high hedonic value. Snails showed diminished feeding behaviour for multiple food stimuli of high hedonic value post 1TT with PLL injection. This finding suggested that PLL causes anhedonia rather than an impairment of LTM formation following the 1TT procedure. This is the first evidence suggesting that inhibiting the biogenesis of miRNAs contributes to anhedonia in Lymnaea.

Comparative Analysis of Neuropeptides in Homologous Interneurons and Prohormone Annotation in Nudipleuran Sea Slugs

Despite substantial research on neuronal circuits in nudipleuran gastropods, few peptides have been implicated in nudipleuran behavior. In this study, we expanded the understanding of peptides in this clade, using three species with well-studied nervous systems, Hermissenda crassicornis, Melibe leonina, and Pleurobranchaea californica. For each species, we performed sequence homology analysis of de novo transcriptome predictions to identify homologs to 34 of 36 prohormones previously characterized in the gastropods Aplysia californica and Lymnaea stagnalis. We then used single-cell mass spectrometry to characterize peptide profiles in homologous feeding interneurons: the multifunctional ventral white cell (VWC) in P. californica and the small cardioactive peptide B large buccal (SLB) cells in H. crassicornis and M. leonina. The neurons produced overlapping, but not identical, peptide profiles. The H. crassicornis SLB cells expressed peptides from homologs to the FMRFamide (FMRFa), small cardioactive peptide (SCP), LFRFamide (LFRFa), and feeding circuit activating peptides prohormones. The M. leonina SLB cells expressed peptides from homologs to the FMRFa, SCP, LFRFa, and MIP-related peptides prohormones. The VWC, previously shown to express peptides from the FMRFa and QNFLa (a homolog of A. californica pedal peptide 4) prohormones, was shown to also contain SCP peptides. Thus, each neuron expressed peptides from the FMRFa and SCP families, the H. crassicornis and M. leonina SLB cells expressed peptides from the LFRFa family, and each neuron contained peptides from a prohormone not found in the others. These data suggest each neuron performs complex co-transmission, which potentially facilitates a multifunctional role in feeding. Additionally, the unique feeding characteristics of each species may relate, in part, to differences in the peptide profiles of these neurons. These data add chemical insight to enhance our understanding of the neuronal basis of behavior in nudipleurans and other gastropods.

A workflow to investigate the impacts of weathered multi-walled carbon nanotubes to the mud snail Lymnaea stagnalis

Although the development and application of nanomaterials is a growing industry, little data is available on the ecotoxicological effects on aquatic organisms. Therefore, we set up a workflow to address the potential uptake of weathered multi-walled carbon nanotubes (wMWCNTs) by a model organism, the pulmonary mud snail Lymnaea stagnalis (L. stagnalis), which plays an important role in the food web. It represents a suitable organism for this approach because as a grazer it potentially ingests large amounts of sedimented wMWCNTs. As food source for L. stagnalis, benthic biofilm was investigated by the use of a transmission electron microscope (TEM) and a scanning electron microscope (SEM) after exposure with wMWCNTs. In addition, isotopic labeling was applied with 14C-wMWCNTs (0.1 mg/L) to quantify fate, behavior, and enrichment of 14C-wMWCNTs in benthic biofilm and in L. stagnalis. Enrichment in benthic biofilm amounted to 529.0 µg wMWCNTs/g dry weight and in L. stagnalis to 79.6 µg wMWCNTs/g dry weight. A bioconcentration factor (BCF) for L. stagnalis was calculated (3500 L/kg). We demonstrate the accumulation of wMWCNTs (10 mg/L) in the digestive tract of L. stagnalis in an effect study. Moreover, the physiological markers glycogen and triglycerides as indicators for the physiological state, as well as the RNA/DNA ratio as growth indicator, were examined. No significant differences between exposed and control animals were analyzed for glycogen and triglycerides after 24 days of exposure, but a decreasing trend is recognizable for triglycerides. In contrast, the significant reduction in the RNA/DNA ratio of L. stagnalis indicated an inhibition of growth with a following recovery after depuration. The described workflow enables a comprehensive determination of the fate and the behavior of wMWCNTs specifically and in general all kinds of CNTs in the aquatic environment and therefore contributes to a holistic risk assessment of wMWCNTs.

Vodní měkkýši přírodní rezervace Krabonošská niva [Aquatic molluscs of the Krabonošská niva Nature Reserve]

A floodplain of the Lužnice river at the Czech-Austrian border is protected by the Krabonošská niva Nature Reserve (Southern Bohemia, Czech Republic). This floodplain has a relatively natural character and, in addition to the Lužnice river, there is a high diversity of habitats at various stages of succession such as oxbows, pools and temporary wetlands. Altogether 20 species were found at 15 sites studied in 2021. Common and widespread gastropods Lymnaea stagnalis, Segmentina nitida and Planorbarius corneus belonged among the most often recorded species. Endangered Pisidium globulare was found at two sites, and only one non-native species Physa acuta was recorded. Molluscan assemblages are very similar to the assemblages of the neighboring Horní Lužnice Nature Reserve.

SPECIFIC FEATURES OF EMBRYONIC DEVELOPMENT OF POND SNAIL (LYMNAEA STAGNALIS L.) UNDER ANTHROPOGENIC INFLUENCE OF CHLORIDE COMPOUNDS IN WATER

The article highlights the research of embryonic development of hydrobionts under the influence of anthropogenic factors of water solutions – chlorobenzene and tetrachloroethylene. To detect the toxicity of chloride pollutants in a freshwater reservoir, representatives of benthic mollusks (big pond snail Lymnaea stagnalis L.) were used as a model organism. According to the results of research, it was found that at a concentration of 0.01 mg/l of chlorobenzene solution individual anomalies of embryonic development of the pond fish are observed, but at higher concentrations its development is delayed by 5-6 stages in comparison with the control. As an integral parameter of pond fish development, we can take indicators of the biological toxicity criterion: Hatchling and its survival over a 10-day period after leaving the egg shells. Changes in the physiological parameters of pond fish embryos begin with a chlorobenzene concentration of 0.01 mg/l; the maximum permissible concentration for tetrachloroethylene for the considered biological indicators should be considered 0.05 mg/l. It is noted that in big pond snail (Lymnaea stagnalis L.) the hatch of juveniles from egg shells is the most sensitive indicator when determining the toxicity of an aqueous solution; and less sensitive indicators are the number of eggs laid and the survival of juveniles in the first 10 days after hatching. The test object used is a large pond, which has shown a high sensitivity to contamination of the water environment with chlorinated ecotoxicants and can be recommended for a comprehensive quality assessment when biotesting freshwater waters