Molecular characterization and evolution of haemocyanin from the two freshwater shrimps Caridina multidentata (Stimpson, 1860) and Atyopsis moluccensis (De Haan, 1849).

Haemocyanin (Hc) is a copper-containing respiratory protein, floating freely dissolved in the hemolymph of many arthropod species. A typical haemocyanin is a hexamer or oligohexamer of six identical or similar subunits, with a molecular mass around 75 kDa each. In the crustaceans, the haemocyanins appear to be restricted to the remipedes and the malacostracans. We have investigated the haemocyanins of two freshwater shrimps, the Amano shrimp Caridina multidentata and the bamboo shrimp Atyopsis moluccensis. We obtained three full-length and one partial cDNA sequences of haemocyanin subunits from the Amano shrimp, which were assigned to the α- and γ-types of decapod haemocyanin subunits. Three complete and two partial haemocyanin cDNA sequences were obtained from the bamboo shrimp, which represent subunit types α, ß and γ. This is the first time that sequences of all three subunit types of the decapod haemocyanins were obtained from a single species. However, mass spectrometry analyses identified only α- and γ-type subunits, suggesting that a ß-subunit is not a major component of the native haemocyanin of the bamboo shrimp. Phylogenetic and molecular clock analyses showed that malacostracan haemocyanins commenced to diversify into distinct subunit types already ~515 million years ago. ß-subunits diverged first, followed by α- and γ-type subunits ~396 million years ago. The haemocyanins of phyllocarids and peracarids form distinct clades within the α/γ-cluster. Within the Caridea, an early divergence of distinct α-type subunits occurred ~200 MYA. The tree of the γ-subunits suggests a common clade of the Caridea (shrimps) and Penaeidae (prawns).

Application of synchrotron-radiation-based computer microtomography (SRICT) to selected biominerals: embryonic snails, statoliths of medusae, and human teeth.

Synchrotron-radiation-based computer microtomography (SRmicroCT) was applied to three biomineralised objects First, embryonic snails of the freshwater snail Biomphalaria glabrata, second, rhopalia (complex sense organs) of the medusa Aurelia aurita, and third, human teeth. The high absorption contrast between the soft tissue and mineralised tissues, i.e. the shell in the first case (consisting of calcium carbonate) and the statoliths in the second case (consisting of calcium sulphate hemihydrate), makes this method ideal for the study of biomineralised tissues. The objects can be non-destructively studied on a micrometre scale, and quantitative parameters like the thickness of a forming a snail shell or statolith crystal sizes can be obtained on a length scale of 1-2 mum. Using SRmicroCT, the dentin-enamel border can be clearly identified in X-ray dense teeth.