Localization and Bioreactivity of Cysteine-Rich Secretions in the Marine Gastropod Nucella lapillus

Marine biodiversity has been yielding promising novel bioproducts from venomous animals. Despite the auspices of conotoxins, which originated the paradigmatic painkiller Prialt, the biotechnological potential of gastropod venoms remains to be explored. Marine bioprospecting is expanding towards temperate species like the dogwhelk Nucella lapillus, which is suspected to secrete immobilizing agents through its salivary glands with a relaxing effect on the musculature of its preferential prey, Mytilus sp. This work focused on detecting, localizing, and testing the bioreactivity of cysteine-rich proteins and peptides, whose presence is a signature of animal venoms and poisons. The highest content of thiols was found in crude protein extracts from the digestive gland, which is associated with digestion, followed by the peribuccal mass, where the salivary glands are located. Conversely, the foot and siphon (which the gastropod uses for feeding) are not the main organs involved in toxin secretion. Ex vivo bioassays with Mytilus gill tissue disclosed the differential bioreactivity of crude protein extracts. Secretions from the digestive gland and peribuccal mass caused the most significant molecular damage, with evidence for the induction of apoptosis. These early findings indicate that salivary glands are a promising target for the extraction and characterization of bioactive cysteine-rich proteinaceous toxins from the species.

Hemocyanins of Muricidae: New ‘Insights’ Unravel an Additional Highly Hydrophilic 800 kDa Mass Within the Molecule

Hemocyanins are giant oxygen transport proteins that freely float within the hemolymph of most molluscs. The basic quaternary structure of molluscan hemocyanins is a cylindrical decamer with a diameter of 35 nm which is built of 400 kDa subunits. Previously published results, however, showed that one out of two hemocyanin subunits of Rapana venosa encompasses two polypeptides, one 300 kDa and one 100 kDa polypeptide which aggregate to typical 4 MDa and 8 MDa hemocyanin (di-)decamer molecules. It was shown that the polypeptides are bound most probably by one or more cysteine disulfide bridges but it remained open if these polypeptides were coded by one or two genes. Our here presented results clearly showed that both polypeptides are coded by one gene only and that this phenomenon can also be found in the gastropod Nucella lapillus. Thus, it can be defined as clade-specific for Muricidae, a group of the very diverse Caenogastropoda. In addition, we discovered a further deviation of this hemocyanin subunit within both species, namely a region of 340 mainly hydrophilic amino acids (especially histidines and aspartic acids) which have not been identified in any other molluscan hemocyanin, yet. Our results indicate that, within the quaternary structure, these additional amino acids most probably protrude within the inner part of didecamer cylinders, forming a large extra mass of up to 800 kDa. They presumably influence the structure of the protein and may affect the functionality. Thus, these findings reveal further insights into the evolution and structures of gastropod hemocyanins.

Declines over the last two decades of five intertidal invertebrate species in the western North Atlantic

Climate change has already altered the environmental conditions of the world’s oceans. Here we report declines in gastropod abundances and recruitment of mussels (Mytilus edulis) and barnacles (Semibalanus balanoides) over the last two decades that are correlated with changes in temperature and ocean conditions. Mussel recruitment is declining by 15.7% per year, barnacle recruitment by 5.0% per year, and abundances of three common gastropods are declining by an average of 3.1% per year (Testudinalia testudinalis, Littorina littorea, and Nucella lapillus). The declines in mussels and the common periwinkle (L. littorea) are correlated with warming sea temperatures and the declines in T. testudinalis and N. lapillus are correlated with aragonite saturation state, which affects rates of shell calcification. These species are common on shores throughout the North Atlantic and their loss is likely to lead to simplification of an important food web on rocky shores.

A decadal study of biometric and imposex indices in two gastropods

We have monitored tributyltin (TBT) pollution in Galicia (NW Spain) for more than a decade by means of assessing gastropod imposex in populations of Nucella lapillus (N ≥ 34) and Tritia reticulata (N ≥ 18) at regular intervals. Several thousand specimens were processed to obtain their shell height (SH), penis length (PL) and vas deferens sequence (VDS); imposex indices (including the VDS index, VDSI) were subsequently calculated. The regional mean SH of both females and males has not changed significantly in either species throughout the study. This also applies to the mean male PL in N. lapillus, but male T. reticulata penises surprisingly enlarged. On the contrary, the regional mean female PL (MFPL) and all imposex indices significantly decreased in both snails. Results confirm previous conclusions based on the chemical analyses of tissues and partial imposex observations. In addition, the close correlations between MFPL and VDSI show some potential applications to TBT biomonitoring.