Calcium transfer across the outer mantle epithelium in the Pacific oyster, Crassostrea gigas

Calcium transport is essential for bivalves to be able to build and maintain their shells. Ionized calcium (Ca 2+ ) is taken up from the environment and eventually transported through the outer mantle epithelium (OME) to the shell growth area. However, the mechanisms behind this process are poorly understood. The objective of the present study was to characterize the Ca 2+ transfer performed by the OME of the Pacific oyster, Crassostrea gigas, as well as to develop an Ussing chamber technique for the functional assessment of transport activities in epithelia of marine bivalves. Kinetic studies revealed that the Ca 2+ transfer across the OME consists of one saturable and one linear component, of which the saturable component fits best to Michaelis–Menten kinetics and is characterized by a K m of 6.2 mM and a V max of 3.3 nM min −1 . The transcellular transfer of Ca 2+ accounts for approximately 60% of the total Ca 2+ transfer across the OME of C. giga s at environmental Ca 2+ concentrations. The use of the pharmacological inhibitors: verapamil, ouabain and caloxin 1a1 revealed that voltage-gated Ca 2+ -channels, plasma-membrane Ca 2+ -ATPase and Na + /Ca 2+ -exchanger all participate in the transcellular Ca 2+ transfer across the OME and a model for this Ca 2+ transfer is presented and discussed.

Shell matrices of Recent rhynchonelliform brachiopods: microstructures and glycosylation studies

ABSTRACTLike most metazoan biomineralisations, the brachiopod shell is the end product of a biologically controlled calcification process. The main agent of the control is the extracellular matrix, which is secreted by the outer mantle epithelium. This matrix mediates the calcification process by allowing crystal nucleation and elongation in specific orientations and finally, by stopping crystal growth. The proteinaceous moiety of brachiopod shell matrices has been extensively studied. Less known are the post-translational modifications that occur in these matrices, in particular glycosylations. In this comparison of five species of Recent articulated brachiopods, the ratio of soluble to insoluble organic matrix varies between the species. Polydisperse macromolecular materials occur in each of these species with discrete proteins of 50 kDa in Notosaria nigricans, Calloria inconspicua and Neothyris lenticularis, 37 kDa in Terebratulina retusa and Gryphus vitreus and 20–25 kDa in N. nigricans. Protein mixtures from all five species respond differently to anionic stains (Stains-All and Alcian Blue). PAS staining results in a positive smear in C. inconspicua and T. retusa and highlights low molecular weight glycoproteins in C. inconspicua. The polysaccharide composition of the soluble matrix of T. retusa is different from the others due to high proportions of arabinose and low proportions of fucose. In all cases, polysaccharide composition of the insoluble matrix is dominated by glucose and glucosamine. Insoluble matrices have more glucose and xylose and less galactosamine and glucosamine than the corresponding soluble matrix. Relatively high amounts of glucosamine may suggest the presence of chitin in the shell matrix of rhynchonelliform brachiopods.