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Characterization of a Non-fibrillar-Related Collagen in the Mollusc Haliotis tuberculata and its Biological Activity on Human Dermal Fibroblasts

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Abstract

In invertebrates, members of the collagen family have been found in various phyla. Surprisingly, in mollusc, little is known about such molecules. In this study, we characterize the full-length abalone type IV collagen and we analysed its biological effects on human fibroblast in order to gain insights about this molecule in molluscs and particularly clues about its roles. We screened a cDNA library of Haliotis tuberculata hemocytes. The expression pattern of the transcript is determined using real-time polymerase chain reaction and in situ hybridization. The close identity between α1(IV) C-terminal domain and the vertebrate homologue led us to produce, purify and test in vitro a recombinant protein corresponding to this region using human dermal fibroblasts cell culture. The biological effects were evaluated on proliferation and on differentiation. We found that the 5,334-bp open reading frame transcript encodes a protein of 1,777 amino acids, including an interrupted 1,502-residue collagenous domain and a 232-residue C-terminal non-collagenous domain. The expression pattern of this transcript is mainly found in the mantle and hemocytes. The recombinant protein corresponding α1(IV) C-terminal domain increased fibroblast proliferation by 69% and doubled collagen synthesis produced in primary cultures. This work provides the first complete primary structure of a mollusc non-fibrillar collagen chain and the biological effects of its C-terminal domain on human cells. In this study, we prove that the NC1 domain from a molluscan collagen can improve human fibroblast proliferation as well as differentiation.

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Abbreviations

BM:

Basement membranes

ECM:

Extracellular matrix

NC1:

Non-collagenous domain 1

NC1 FL:

Non-collagenous domain 1 full length

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Acknowledgments

The work of C. Fleury was financially supported by a CIFRE fellowship from the BiotechMarine company (Zone Industrielle—BP65, 22260 Pontrieux, France, www.biotechmarine.com). The authors would like to thank Dr. Isabella Vlisidou and Miss Zoë Freeman (university of Bath) for helpful comments and/or improvement of the English language. The authors thank the technical staff of the CREC (Centre de Recherche en Environnement Côtier-Luc/Mer) for their assistance in animal care.

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Authors and Affiliations

  1. UMR 100 IFREMER “Physiologie et Ecophysiologie des Mollusques Marins”—IFR 146 ICORE—IBFA—Esplanade de la Paix, Université de Caen Basse-normandie, 14032, Caen, France

    Christophe Fleury, Antoine Serpentini & Jean-Marc Lebel

  2. Laboratoire Matrice Extracellulaire et Pathologie, EA 3214, IFR 146 ICORE, Faculté de Médecine, Université de Caen Basse-Normandie, CHU niveau 3, avenue de la Côte de nacre, 14032, Caen, France

    Magdalini Kypriotou, Emmanuelle Renard & Philippe Galéra

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  1. Christophe Fleury
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  2. Antoine Serpentini
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  3. Magdalini Kypriotou
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  4. Emmanuelle Renard
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  5. Philippe Galéra
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  6. Jean-Marc Lebel
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Correspondence to Jean-Marc Lebel.

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Fleury, C., Serpentini, A., Kypriotou, M. et al. Characterization of a Non-fibrillar-Related Collagen in the Mollusc Haliotis tuberculata and its Biological Activity on Human Dermal Fibroblasts. Mar Biotechnol 13, 1003–1016 (2011). https://doi.org/10.1007/s10126-011-9364-9

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  • DOI: https://doi.org/10.1007/s10126-011-9364-9

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