Spatial-temporal expression analysis of lineage-restricted shell matrix proteins reveals shell field regionalization and distinct cell populations in the slipper snail Crepidula atrasolea.

Molluscs are one of the most morphologically diverse clades of metazoans, exhibiting an immense diversification of calcium carbonate structures, such as the shell. Biomineralization of the calcified shell is dependent on shell matrix proteins (SMPs). While SMP diversity is hypothesized to drive molluscan shell diversity, we are just starting to unravel SMP evolutionary history and biology. Here we leveraged two complementary model mollusc systems, Crepidula fornicata and Crepidula atrasolea , to determine the lineage-specificity of 185 Crepidula SMPs. We found that 95% of the adult C. fornicata shell proteome belongs to conserved metazoan and molluscan orthogroups, with molluscan-restricted orthogroups containing half of all SMPs in the shell proteome. The low number of C. fornicata -restricted SMPs contradicts the generally-held notion that an animal’s biomineralization toolkit is dominated by mostly novel genes. Next, we selected a subset of lineage-restricted SMPs for spatial-temporal analysis using in situ hybridization chain reaction (HCR) during larval stages in C. atrasolea . We found that 12 out of 18 SMPs analyzed are expressed in the shell field. Notably, these genes are present in 5 expression patterns, which define at least three distinct cell populations within the shell field. These results represent the most comprehensive analysis of gastropod SMP evolutionary age and shell field expression patterns to date. Collectively, these data lay the foundation for future work to interrogate the molecular mechanisms and cell fate decisions underlying molluscan mantle specification and diversification.

Coping with stress in a warming Gulf: the postlarval American lobster's cellular stress response under future warming scenarios.

The Gulf of the Maine (GoM) is one of the fastest warming bodies of water in the world, posing serious physiological challenges to its marine inhabitants. Marine organisms can cope with the cellular and molecular stresses created by climate change through changes in gene expression. We used transcriptomics to examine how exposure to current summer temperatures (16 °C) or temperature regimes reflective of projected moderate and severe warming conditions (18 °C and 22 °C, respectively) during larval development alters expression of transcripts affiliated with the cellular stress response (CSR) in postlarval American lobsters (Homarus americanus). We identified 26 significantly differentially expressed (DE) transcripts annotated to CSR proteins. Specifically, transcripts for proteins affiliated with heat shock, the ubiquitin family, DNA repair, and apoptosis were significantly over-expressed in lobsters reared at higher temperatures relative to current conditions. Substantial variation in the CSR expression between postlarvae reared at 18 °C and those reared at 22 °C suggests that postlarvae reared under severe warming may have a hindered ability to cope with the physiological and molecular challenges of ocean warming. These results highlight that postlarval American lobsters may experience significant heat stress as rapid warming in the GoM continues, potentially compromising their ability to prevent cellular damage and inhibiting the reallocation of cellular energy towards other physiological functions beyond activation of the CSR. Moreover, this study establishes additional American lobster stress markers and addresses various knowledge gaps in crustacean biology, where sufficient 'omics research is lacking.

Ecologically relevant arsenic exposure alters female mate preference and anxiety-like behavior in Betta splendens.

Arsenic is a metalloid pollutant that is commonly found in surface and groundwater worldwide. Toxicological effects of arsenic are relatively well-known, but much less studied are its effects on behavioral endpoints, which may have considerable evolutionary and population-level consequences. Here we investigated the effects of exposure to environmentally relevant concentrations of arsenic (0, 10 and 100 µg/L) for 96-hours on female preference for male color (i.e. red versus blue) in Betta splendens, an increasingly popular fish model for contaminant-induced behavioral dysfunction. Further, we examined whether arsenic exposure altered anxiety-like behaviors using a standard scototaxis test (preference for light or dark), as well as measured tissue cortisol concentrations to increase our understanding of possible mechanisms driving behavioral responses. We found exposure to 100 µg/L arsenic results in a loss of female preference for red males, and arsenic exposed females showed increased anxiety-like behavior. The loss in preference for male coloration may have been driven by anxiety, as preference for red was negatively correlated with anxiety-like behavior for all fish. Interestingly, increase in anxiety-like behavior occurred without a parallel increase in cortisol. Female preference for red colored males may confer fitness benefits, and this study highlights important arsenic-induced behavioral changes that could have population level consequences.