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Chalky versus foliated: a discriminant immunogold labelling of shell microstructures in the edible oyster Crassostrea gigas

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Abstract

Mollusc shells are organic–inorganic biocomposites, arranged in a limited number of superimposed calcified layers that generally exhibit very different organization of their crystallites. Because of their attractive mechanical and crystallographic properties, these shell layers have been the focus of several physical and biochemical characterizations. In particular, recent proteomic data obtained from individual layers suggest that their protein contents are different. However, the direct visual evidence that some macromolecular components are layer-specific is rather tenuous. This paper is based on a non-conventional immunogold labelling approach to localize proteins in the shell of the edible oyster Crassostrea gigas. The shell microstructure of this model organism is predominantly composed of foliated calcite, interspersed by discontinuous pockets of ‘chalky layers’, a porous microstructure typical of bivalves of the ostreid family. By developing a polyclonal antibody (in two rats) elicited against a proteinaceous shell fraction, we obtained differential staining of the two microstructures. We assert that our labelling is microstructure discriminant. The difference in labelling of the two shell microstructures suggests either that they are formed by a variation of the secretory repertoire of the shell-forming cells of the calcifying mantle epithelium or that the chalky layer may be formed via a completely different mechanism. Our results allow a first glimpse on the subtle regulatory mechanisms that drive the process of chalky and foliated layers deposition.

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Acknowledgements

The work presented in this paper was made possible by the ENS PhD programme. The Earth and Natural Sciences Doctoral Studies Programme is funded under the Programme for Research in Third-Level Institutions Cycle-5 and co-funded under the European Regional Development Fund. The entire experimental work was performed in Dijon, via a financial support attributed to V. M. by the Irish Geological Association and the AllTech Innovation Competition. The work of F. M. was supported by INTERRVIE Program (INSU, CNRS) and OSU-Theta.

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Author notes

  1. Nathalie Guichard

    Present address: UMR CNRS 6457 SUBATECH, La Chantrerie, Nantes, France

  2. Françoise Immel

    Present address: UMR CNRS 5200 laboratoire de Biogenèse Membranaire, Université Bordeaux Segalen, Villenave d’Ornon, France

  3. Marc de Rafélis

    Present address: UMR CNRS 5563 Géosciences Environnements Toulouse (GET), Université de Toulouse III Paul Sabatier, Toulouse, France

  4. Vincent Mouchi

    Present address: UPMC Univ Paris 06, CNRS UMR 7193, ISTeP, Sorbonne Universités, 75005, Paris, France

Authors and Affiliations

  1. Department of Geology, School of Natural Sciences, Trinity College Dublin, Dublin 2, Ireland

    Vincent Mouchi & Quentin G. Crowley

  2. UPMC Univ Paris 06, CNRS UMR 7193, ISTeP, Sorbonne Universités, 75005, Paris, France

    Vincent Mouchi & Marc de Rafélis

  3. UPMC Univ Paris 06, CNRS, Laboratoire d’Ecogéochimie des Environnements Benthiques, Observatoire Océanologique de Banyuls, Sorbonne Universités, 66650, Banyuls/Mer, France

    Franck Lartaud

  4. UMR CNRS 6282 Biogéosciences, Université de Bourgogne – Franche Comté (UB-FC), Dijon, France

    Nathalie Guichard, Françoise Immel & Frédéric Marin

  5. UMR CNRS 8104 INSERM U1016 - Institut Cochin, Université Paris Descartes, Paris, France

    Cédric Broussard

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We declare that raising polyclonal antibodies from rats was performed according to ethical standards. We declare no conflict of interest.

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Mouchi, V., Lartaud, F., Guichard, N. et al. Chalky versus foliated: a discriminant immunogold labelling of shell microstructures in the edible oyster Crassostrea gigas . Mar Biol 163, 256 (2016). https://doi.org/10.1007/s00227-016-3040-6

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