The initial growth of the nacreous layer is crucial for comprehending the formation of nacreous aragonite. A flat pearl method in the presence of the inner-shell film was conducted to evaluate the role of matrix proteins in the initial stages of nacre biomineralization in vivo. We examined the crystals deposited on a substrate and the expression patterns of the matrix proteins in the mantle facing the substrate. In this study, the aragonite crystals nucleated on the surface at 5 days in the inner-shell film system. In the film-free system, the calcite crystals nucleated at 5 days, a new organic film covered the calcite, and the aragonite nucleated at 10 days. This meant that the nacre lamellae appeared in the inner-shell film system 5 days earlier than that in the film-free system, timing that was consistent with the maximum level of matrix proteins during the first 20 days. In addition, matrix proteins (Nacrein, MSI60, N19, N16 and Pif80) had similar expression patterns in controlling the sequential morphologies of the nacre growth in the inner-film system, while these proteins in the film-free system also had similar patterns of expression. These results suggest that matrix proteins regulate aragonite nucleation and growth with the inner-shell film in vivo.
References
[1]
Addadi L, Weiner S (1997) Biomineralization: a pavement of pearl. Nature 389: 912–915.
[2]
Weiner S, Traub W, Parker SB (1984) Macromolecules in mollusk shells and their functions in biomineralization. Phil Trans R Soc Lond B 304: 425–434.
[3]
Lopez E, Vidal B, Berland S, Camprasse S, Camprasse G, et al. (1992) Demonstration of the capacity of nacre to induce bone formation by human osteoblasts maintained in vitro. Tissue Cell 24: 667–679.
[4]
Marin F, Luquet G, Marie B, Medakovic D (2008) Molluscan shell proteins: primary structure, origin, and evolution. Curr Top Dev Biol 80: 209–276.
[5]
Sarikaya M (1994) An introduction to biomimetics: a structural viewpoint. Microsc Res Tech 27: 360–375.
[6]
Mayer G (2005) Rigid biological systems as models for synthetic composites. Science 310: 1144–1147.
[7]
Sanchez C, Arribart H, Guille MM (2005) Biomimetism and bioinspiration as tools for the design of innovative materials and systems. Nat Mater 4: 277–288.
[8]
Addadi L, Joester D, Nudelman F, Weiner S (2006) Mollusk shell formation: a source of new concepts for understanding biomineralization processes. Chemistry 12: 980–987.
[9]
Wilt FH (2005) Developmental biology meets materials science: morphogenesis of biomineralized structures. Dev Biol 280: 15–25.
[10]
Wilt FH, Killian CE, Livingston BT (2003) Development of calcareous skeletal elements in invertebrates. Differentiation 71: 237–250.
[11]
Takeuchi T, Endo K (2006) Biphasic and dually coordinated expression of the genes encoding major shell matrix proteins in the pearl oyster Pinctada fucata. Mar Biotechnol 8: 52–61.
[12]
Zhang C, Zhang R (2006) Matrix proteins in the outer shells of molluscs. Mar Biotechnol 8: 572–586.
[13]
Belcher AM, Wu XH, Christensen RJ, Hansma PK, Stucky GD, et al. (1996) Control of crystal phase switching and orientation by soluble mollusc-shell proteins. Nature 381: 56–58.
[14]
Falini G, Albeck S, Weiner S, Addadi L (1996) Control of aragonite or calcite polymorphism by mollusk shell macromolecules. Science 271: 67–69.
[15]
Gotliv BA, Addadi L, Weiner S (2003) Mollusk shell acidic proteins: in search of individual functions. Chembiochem 4: 522–529.
[16]
Miyamoto H, Miyoshi F, Kohno J (2005) The carbonic anhydrase domain protein nacrein is expressed in the epithelial cells of the mantle and acts as a negative regulator in calcification in the mollusc Pinctada fucata. Zoolog Sci 22: 311–315.
[17]
Zhang Y, Xie L, Meng Q, Jiang T, Pu R, et al. (2003) A novel matrix protein participating in the nacre framework formation of pearl oyster, Pinctada fucata. Comp Biochem Physiol B Biochem Mol Biol 135: 565–573.
[18]
Miyamoto H, Miyashita T, Okushima M, Nakano S, Morita T, et al. (1996) A carbonic anhydrase from the nacreous layer in oyster pearls. Proc Natl Acad Sci U S A 93: 9657–9660.
[19]
Gaume B, Fouchereau-Peron M, Badou A, Helléouet MN, Huchette S, et al. (2011) Biomineralization markers during early shell formation in the European abalone Haliotis tuberculata, Linnaeus. Mar Biol 158: 341–353.
[20]
Ma Z, Huang J, Sun J, Wang G, Li C, et al. (2007) A novel extrapallial fluid protein controls the morphology of nacre lamellae in the pearl oyster, Pinctada fucata. J Biol Chem 282: 23253–23263.
[21]
Suzuki M, Saruwatari K, Kogure T, Yamamoto Y, Nishimura T, et al. (2009) An acidic matrix protein, Pif, is a key macromolecule for nacre formation. Science 325: 1388–1390.
[22]
Yano M, Nagai K, Morimoto K, Miyamoto H (2007) A novel nacre protein N19 in the pearl oyster Pinctada fucata. Biochem Biophys Res Commun 362: 158–163.
[23]
Samata T, Hayashi N, Kono M, Hasegawa K, Horita C, et al. (1999) A new matrix protein family related to the nacreous layer formation of Pinctada fucata. FEBS Lett 462: 225–229.
Fritz M, Belcher AM, Radmacher M, Walters DA, Hansma PK, et al. (1994) Flat pearls from biofabrication of organized composites on inorganic substrates. Nature 371: 49–51.
[26]
Lin AY, Chen PY, Meyers MA (2008) The growth of nacre in the abalone shell. Acta Biomater 4: 131–138.
[27]
Bezares J, Asaro RJ, Hawley M (2008) Macromolecular structure of the organic framework of nacre in Haliotis rufescens: implications for growth and mechanical behavior. J Struct Biol 163: 61–75.
[28]
Bezares J, Asaro RJ, Hawley M (2010) Macromolecular structure of the organic framework of nacre in Haliotis rufescens: implications for mechanical response. J Struct Biol 170: 484–500.
[29]
Checa AG, Cartwright JH, Willinger MG (2009) The key role of the surface membrane in why gastropod nacre grows in towers. Proc Natl Acad Sci U S A 106: 38–43.
[30]
Shen X, Belcher AM, Hansma PK, Stucky GD, Morse DE (1997) Molecular cloning and characterization of lustrin A, a matrix protein from shell and pearl nacre of Haliotis rufescens. J Biol Chem 272: 32472–32481.
[31]
Liu X, Li J, Xiang L, Sun J, Zheng G, et al. (2012) The role of matrix proteins in the control of nacreous layer deposition during pearl formation. Proc Biol Sci 279: 1000–1007.
[32]
Yan Z, Ma Z, Zheng G, Feng Q, Wang H, et al. (2008) The inner-shell film: an immediate structure participating in pearl oyster shell formation. Chembiochem 9: 1093–1099.
[33]
Sch?ffer TE, Ionescu-Zanetti C, Proksch R, Fritz M, Walters DA, et al. (1997) Does abalone nacre form by heteroepitaxial nucleation or by growth through mineral bridges? Chem Mater 9: 1731–1740.
[34]
Zaremba CM, Belcher AM, Fritz M, Li Y, Mann S, et al. (1996) Critical transitions in the biofabrication of abalone shells and flat pearls. Chem Mater 8: 679–690.
[35]
Saruwatari K, Matsui T, Mukai H, Nagasawa H, Kogure T (2009) Nucleation and growth of aragonite crystals at the growth front of nacres in pearl oyster, Pinctada fucata. Biomaterials 30: 3028–3034.
[36]
Inoue N, Ishibashi R, Ishikawa T, Atsumi T, Aoki H, et al. (2011) Can the quality of pearls from the Japanese pearl oyster (Pinctada fucata) be explained by the gene expression patterns of the major shell matrix proteins in the pearl sac? Mar Biotechnol 13: 48–55.
[37]
Kinoshita S, Wang N, Inoue H, Maeyama K, Okamoto K, et al. (2011) Deep sequencing ESTs from nacreous and prismatic layer producing tissues and a screen for novel shell formation-related genes in the pearl oyster. PLoS One 6: e21238.
[38]
Dauphin Y, Ball AD, Cotte M, Cuif JP, Meibom A, et al. (2008) Structure and composition of the nacre-prisms transition in the shell of Pinctada margaritifera (Mollusca, Bivalvia). Anal Bioanal Chem 390: 1659–1669.
[39]
Yan Z, Meng W, Liu Z, Yang S, Liu X, et al. (2011) In vivo and in vitro biomineralization in the presence of the inner-shell film of pearl oyster. Acta Oceanol Sin 30: 87–93.
[40]
Gries K, Heinemann F, Gummich M, Ziegler A, Rosenauer A, et al. (2011) Influence of the insoluble and soluble matrix of abalone nacre on the growth of calcium carbonate crystals. Cryst Growth Des 11: 729–734.
[41]
Heinemann F, Treccani L, Fritz M (2006) Abalone nacre insoluble matrix induces growth of flat and oriented aragonite crystals. Biochem Biophys Res Commun 344: 45–49.
[42]
Wang N, Kinoshita S, Nomura N, Riho C, Meayama K, et al. (2012) The mining of pearl formation genes in pearl oyster Pinctada fucata by cDNA suppression subtractive hybridization. Mar Biotechnol 14: 177–188.
[43]
Miyazaki Y, Nishida T, Aoki H, Samata T (2010) Expression of genes responsible for biomineralization of Pinctada fucata during development. Comp Biochem Physiol B Biochem Mol Biol 155: 241–248.
[44]
Wang N, Kinoshita S, Riho C, Maeyama K, Nagai K, et al. (2009) Quantitative expression analysis of nacreous shell matrix protein genes in the process of pearl biogenesis. Comp Biochem Physiol B Biochem Mol Biol 154: 346–350.
[45]
Masaoka T, Samata T, Nogawa C, Baba H, Aoki H, et al. (2013) Shell matrix protein genes derived from donor expressed in pearl sac of Akoya pearl oysters (Pinctada fucata) under pearl culture. Aquaculture 384–387: 56–65.
[46]
Fang D, Pan C, Lin H, Lin Y, Xu G, et al. (2012) Ubiquitylation functions in the calcium carbonate biomineralization in the extracellular matrix. PLoS One 7: e35715.
[47]
Fang D, Xu G, Hu Y, Pan C, Xie L, et al. (2011) Identification of genes directly involved in shell formation and their functions in pearl oyster, Pinctada fucata. PLoS One 6: e21860.
[48]
Mount AS, Wheeler AP, Paradkar RP, Snider D (2004) Hemocyte-mediated shell mineralization in the eastern oyster. Science 304: 297–300.