Gastrolith

Abstract: Despite the proclamation of Lowenstam and Weiner that crustaceans are the "champions of mineral mobilization and deposition of the animal kingdom," relatively few proteins from the two main calcification sites in these animals, i.e., the exoskeleton and the transient calcium storage organs, have been identified, sequenced, and their roles elucidated. Here, a 65-kDa protein (GAP 65) from the gastrolith of the crayfish, Cherax quadricarinatus, is fully characterized and its function in the mineralization of amorphous calcium carbonate (ACC) of the extracellular matrix is demonstrated. GAP 65 is a negatively charged glycoprotein that possesses three predicted domains: a chitin-binding domain 2, a low-density lipoprotein receptor class A domain, and a polysaccharide deacetylase domain. Expression of GAP 65 was localized to columnar epithelial cells of the gastrolith disk during premolt. In vivo administration of GAP 65 dsRNA resulted in a significant reduction of GAP 65 transcript levels in the gastrolith disk. Such gene silencing also caused dramatic structural and morphological deformities in the chitinous-ACC extracellular matrix structure. ACC deposited in these gastroliths appeared to be sparsely packed with large elongated cavities compared with the normal gastrolith, where ACC is densely compacted. ACC spherules deposited in these gastroliths are significantly larger than normal. GAP 65, moreover, inhibited calcium carbonate crystallization in vitro and stabilized synthetic ACC. Thus, GAP 65 is the first protein shown to have dual function, involved both in extracellular matrix formation and in mineral deposition during biomineralization.

Abstract: Gastroliths or ‘stomach stones’ occur frequently in some, but not all, groups of fossil and living m arine tetrapods. C om parative analysis of gastrolith distribution suggests a role in buoyancy control rather than food processing. Once accidental ingestion by bottom-feeding animals is excluded, gastroliths occur in most tetrapods which ‘fly’ underw ater with hydrofoil limbs, including plesiosaurs, penguins, and otariid pinnipeds, but not the m arine chelonians. They do not usually occur in cetaceans, ichthyosaurs, mosasaurs, and odobenid and phocid pinnipeds, which swim with a caudal fin or the equivalent. Occurrence in amphibious forms is variable; crocodilians often have gastroliths, but nothosaurs and placodonts do not. T he correlation of gastroliths and underw ater flight is corroborated by a com parative analysis which takes phylogenetic factors into account. There is no correlation with diet. Consideration of function and occurrence in terrestrial forms suggests that the use of gastroliths in digestion would not be useful, and might even be harmful, to a carnivorous m arine tetrapod. Gastroliths are more efficient than skeletal bone (as in pachyostosis) in terms of sinking force per unit of added mass or volume. As well as driftwood and ice, m arine tetrapods should be considered as a potential source of erratic stones in freshwater and marine sediments. Gastroliths may have evolved by the accidental ingestion of stones, the retention into adulthood of stones used by juveniles to process insect or plant food, or as a com pensatory replacem ent for dense bones habitually filling the stomach. Their presence or absence should be more carefully recorded and further studies should be carried out on their function.

Abstract: Mobilization of calcium during the molt cycle from the cuticle to transient calcium deposits is widely spread in crustaceans. The dynamics of calcium transport to transient calcium deposits called gastroliths and to the cu- ticle over the course of the molt cycle were studied in the crayfish Cherax quadricarinatus. In this species, calcium was deposited in the gastroliths during premolt and trans- ported back to the cuticle during postmolt, shown by digital X-ray radiograph analysis. The predominant mineral in the crayfish is amorphous calcium carbonate embedded in an organic matrix composed mainly of chitin. Scanning elec- tron micrographs of the cuticle during premolt showed that the endocuticle and parts of the exocuticle were the source of most of the labile calcium, while the epicuticle did not undergo degradation and remained mineralized throughout the molt cycle. The gastroliths are made of concentric layers of amorphous calcium carbonate intercalated between chiti- nous lamella. Measurements of pH and calcium levels dur- ing gastrolith deposition showed that calcium concentra- tions in the gastroliths, stomach, and muscle were about the same (10 to 11 mmol l 1 ). On the other hand, pH varied greatly, from 8.7 0.15 in the gastrolith cavity through 7.6 0.2 in muscle to 6.9 0.5 in the stomach.

Abstract: Polished pebbles occasionally found within skeletons of giant herbivorous sauropod dinosaurs are very likely to be gastroliths (stomach stones). Here, we show that based on feeding experiments with ostriches and comparative data for relative gastrolith mass in birds, sauropod gastroliths do not represent the remains of an avian-style gastric mill. Feeding experiments with farm ostriches showed that bird gastroliths experience fast abrasion in the gizzard and do not develop a polish. Relative gastrolith mass in sauropods (gastrolith mass much less than 0.1% of body mass) is at least an order of magnitude less than that in ostriches and other herbivorous birds (gastrolith mass approximates 1% of body mass), also arguing against the presence of a gastric mill in sauropods. Sauropod dinosaurs possibly compensated for their limited oral processing and gastric trituration capabilities by greatly increasing food retention time in the digestive system. Gastrolith clusters of some derived theropod dinosaurs (oviraptorosaurs and ornithomimosaurs) compare well with those of birds, suggesting that the gastric mill evolved in the avian stem lineage.