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A Whole Mount In Situ Hybridization Method for the Gastropod Mollusc Lymnaea stagnalis
Published on: March 15, 2016
The shell organic matrix of the crossed lamellar queen conch shell (Strombus gigas)
Antonio Osuna-Mascaró1, Teresa Cruz-Bustos2, Sana Benhamada3
1Departamento de Estratigrafía y Paleontología, Facultad de Ciencias, Universidad de Granada, Av. Fuentenueva, S/N, 18071 Granada, Spain; Departamento de Bioquímica y Parasitología Molecular, Facultad de Ciencias, Universidad de Granada, Av. Fuentenueva, S/N, 18071 Granada, Spain; UMR CNRS 6282 Biogéosciences, Université de Bourgogne, 6 Bd. Gabriel, 21000 Dijon, France.
Abstract:
In molluscs, the shell organic matrix comprises a large set of biomineral-occluded proteins, glycoproteins and polysaccharides that are secreted by the calcifying mantle epithelium, and are supposed to display several functions related to the synthesis of the shell. In the present paper, we have characterized biochemically the shell matrix associated to the crossed-lamellar structure of the giant queen conch Strombus gigas. The acid-soluble (ASM) and acid-insoluble (AIM) matrices represent an extremely minor fraction of the shell. Both are constituted of polydisperse and of few discrete proteins among which three fractions, obtained by preparative SDS-PAGE and named 1P3, 2P3 and 3P3, are dominant and were further characterized. Compared to other matrices, the acid-soluble matrix is weakly glycosylated (3%) and among the discrete components, only 3P3 seems noticeably glycosylated. The monosaccharide composition of the ASM shows that mannose represents the main monosaccharide. To our knowledge, this is the first report of a high ratio of this sugar in a skeletal matrix. Furthermore, the ASM interacts with the in vitro crystallization of calcium carbonate, but this interaction is moderate. It differs from that of the isolated 1P3 fraction but is similar to that of the 2P3 and 3P3 fractions. At last, antibodies developed from the 3P3 fraction were used to localize this fraction within the shell by immunogold. This study is the first one aiming at characterizing the organic matrix associated to the crossed-lamellar structure of the queen conch shell.
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