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Biochemical and functional characterisation of eggshell matrix proteins in hens

Published online by Cambridge University Press:  18 September 2007

Y. Nys ,
J. Gautron ,
M. D. McKee ,
J. M. Garcia-Ruiz  and
M. T. Hincke
Y. Nys
Affiliation:
Station de Recherches Avicoles, INRA, 37380 Nouzilly, France
J. Gautron
Affiliation:
Station de Recherches Avicoles, INRA, 37380 Nouzilly, France
M. D. McKee
Affiliation:
Faculty of Dentistry and Department of Anatomy and Cell Biology, McGill University, Montreal QC H3A 2B2, Canada
J. M. Garcia-Ruiz
Affiliation:
CSIC, University of Granada, 18002, Spain and
M. T. Hincke
Affiliation:
Department of Cellular and Molecular Medicine, University of Ottawa, ON, K1H 8M5, Canada
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Abstract

The eggshell of the hen is a highly ordered, mineralised structure deposited within an acellular milieu – the uterine fluid secreted by the distal oviduct. Spherulitic crystal growth is initiated by deposition of calcium carbonate on aggregates of organic material present on the outer surface of the eggshell membranes. Gel electrophoresis reveals a complex array of proteins in uterine fluid and eggshell extracts. The eggshell matrix proteins can be classified as egg white proteins (lysozyme, ovalbumin, ovotransferrin, clusterin), bone protein (osteopontin), or proteins specific to the uterus and eggshell (ovocleidins-17 and –116; ovocalyxins-32 and –36). Eggshell extracts, uterine fluid and purified fractions are able to modify the morphology of calcite crystals in vitro. In young hens the breaking strength of the eggshell is inversely related to the degree of calcite orientation. Conversely, reduced strength in eggshell from aged hens coincides with a high variability in crystallographic texture. In guinea fowl the exceptional mechanical properties of the eggshell are explained by an increase in the amount of eggshell produced and particular features of the crystallographic texture. These observations suggest that the eggshell matrix influences the process of crystal growth by controlling size, shape and orientation of calcite crystals. This structural control probably contributes in a substantial manner to the mechanical properties of eggshell.

Keywords

Calciteeggshellhenmatrix proteinsmineralisation
Type
Research Article
Information
World's Poultry Science Journal , Volume 57 , Issue 4 , December 2001 , pp. 401 - 413
Copyright
Copyright © Cambridge University Press 2001

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References

Addadi, L. and Weiner, S. (1992) Control and design principles in biological mineralization. Angewandte Chemie (International Edition in English) 31: 153–169CrossRefGoogle Scholar
Ar, A., Rahn, H. and Paganelli, C.V. (1979) The avian egg: mass and strength. Condor 81: 331–337CrossRefGoogle Scholar
Arias, J.L., Fink, D.J., Xiao, S.Q., Heuer, A.H. and Caplan, A.I. (1993) Biomineralization and eggshells: cell-mediated acellular compartments of mineralized extracellular matrix. International Review of Cytology 145: 217–250CrossRefGoogle ScholarPubMed
Board, R.G. (1982) Properties of avian eggshells and their adaptative value. Biological Review 57: 1–28CrossRefGoogle Scholar
Butler, W.T., Ridall, A.L. and Mckee, M.D. (1996) Osteopontin. In: Principles of Bone Biology (Bilezikian, J., Raisz, L. and Rodan, G., Eds), Academic Press, San Diego, pp. 167–181Google Scholar
Carrino, D.A., Rodriguez, J.P. and Caplan, A.I. (1997) Dermatan sulfate proteoglycans from the mineralized matrix of the avian eggshell. Connective Tissue Research 36: 175–193CrossRefGoogle ScholarPubMed
Chowdhury, S.D. (1990) Shell membrane system in relation to lathyrogen toxicity and copper deficiency. World's Poultry Science Journal 46: 153–169CrossRefGoogle Scholar
Dennis, J.E., Xiao, S.Q., Agarval, M., Fink, D.J., Heuer, A.H. and Caplan, A.I. (1996) Microstructure of matrix and mineral components of eggshells from white leghorn chicken (Gallus gallus). Journal of Morphology 228: 287–3063.0.CO;2-#>CrossRefGoogle Scholar
Dominguez-Vera, J.M., Gautron, J., Garcia-Ruiz, J.M. and Nys, Y. (2000) The effect of avian uterine fluid on the growth behavior of calcite crystals. Poultry Science 79: 901–907CrossRefGoogle ScholarPubMed
Fernandez, M.S., Araya, M. and Arias, J.L. (1997) Eggshells are shaped by a precise spatio temporal arrangement of sequentially deposited macromolecules. Matrix Biology 16: 13–20CrossRefGoogle ScholarPubMed
Fernandez, M.S., Moya, A., Lopez, L. and Arias, J.L. (2001) Secretion pattern, ultrastructure and function of extracellular matrix molecules involved in eggshell formation. Matrix Biology 19: 793–803CrossRefGoogle Scholar
Garcia-Ruiz, J.M. and Rodriguez-Navarro, A. (1994) The mineral structure of the avian eggshell: a case of competitive crystal growth. In: Biomineralization 93, 7th International Symposium on Biomineralization. 1. Fundamentals of Biomineralization (Allemand, D. and Cuif, J.P., Eds), Bulletin de l'hstitut Oceanographique, Monaco, pp. 85–94Google Scholar
Gautron, J., Bain, M., Solomon, S. and Nys, Y. (1996) Soluble matrix of hen's eggshell extracts changes in vitro the rate of calcium carbonate precipitation and crystal morphology. British Poultry Science 37: 853–866CrossRefGoogle ScholarPubMed
Gautron, J., Hincke, M.T. and Nys, Y. (1997) Precursor matrix proteins in the uterine fluid change with stages of eggshell formation in hens. Connective Tissue Research 36: 195–210CrossRefGoogle ScholarPubMed
Gautron, J., Hincke, M.T., Panheleux, M., Garcia-Ruiz, J. and Nys, Y. (2000) Identification and characterization of matrix proteins from hen's eggshell. In: Chemistry and Biology of Mineralized Tissues. Proceeding of the Sixth International ConferenceVittel, France 1998 (Goldberg, M., Boskey, A. and Robinson, C. Eds), American Academy of Orthopaedic SurgeonsRosemont pp. 19–23Google Scholar
Gautron, J., Hincke, M.T., Panhéleux, M., Garcia-Ruiz, J., Boldicke, T. and Nys, Y. (2001a) Ovotransferrin is a matrix protein of the hen eggshell membranes and basal calcified layer. Connective Tissue Research 42: 1–13CrossRefGoogle ScholarPubMed
Gautron, J., Hincke, M.T., Mann, K., Panhéleux, M., Bain, M., McKee, M.D., Solomon, S.E. and Nys, Y. (2001b) Ovocalyxin-32, a novel chicken eggshell matrix protein: isolation, amino acid sequencing, cloning and immunocytochemical localization. Journal of Biological Chemistry 276: 39243–39252CrossRefGoogle ScholarPubMed
Hamilton, R.M.G. (1986) The microstructure of the hen's eggshell: a short review. Food Microstructure 5: 99–110Google Scholar
Hincke, M.T. (1995) Ovalbumin is a component of the chicken eggshell matrix. Connective Tissue Research 31: 227–233CrossRefGoogle ScholarPubMed
Hincke, M.T. and St Maurice, M. (2000) phosphorylation-dependent modulation of calcium carbonate precipitation by chicken eggshell matrix proteins. In: Chemistry and Biology of Mineralized Tissues (Goldberg, M., Boskey, A. and Robinson, C. Eds), American Academy of Orthopaedic Surgeons, Rosemont, pp. 13–17Google Scholar
Hincke, M.T., Bernard, A.M., Lee, E.R., Tsang, C.P. and Narbaitz, R. (1992) Soluble protein constituents of the domestic fowl's eggshell. British Poultry Science 33:505–516CrossRefGoogle ScholarPubMed
Hincke, M.T., Tsang, C.P., Courtney, M., Hill, V. and Narbaitz, R. (1995) Purification and immunochemistry of a soluble matrix protein of the chicken eggshell (ovocleidin 17). Calcified Tissue International 56: 578–583CrossRefGoogle ScholarPubMed
Hincke, M.T., Gautron, J., Tsang, C.P., McKee, M.D. and Nys, Y. (1999) Molecular cloning and ultrastructural localization of the core protein of an eggshell matrix proteoglycan, ovocleidin-116. Journal of Biological Chemistry 274: 32915–32923CrossRefGoogle ScholarPubMed
Hincke, M.T., Gautron, J., Panheleux, M., Garcia-Ruiz, J.M., McKee, M.D. and Nys, Y. (2000a) Identification and localization of lysozyme as a component of the eggshell membranes and shell matrix. Matrix Biology 19: 443–453CrossRefGoogle Scholar
Hincke, M.T., St Maurice, M., Nys, Y., Gautron, J., Panheleux, M., Tsang, C.P.W., Bain, M., Solomon, S. and McKee, M.D. (2000b) Eggshell matrix proteins and shell strength: molecular biology of eggshell matrix proteins and industrial applications. In: Egg Nutrition and Biotechnology (Sim, J.S., Nakai, S. and Guenter, W., Eds), CAB International, Wallingford, pp. 447–461Google Scholar
Hunter, G.K. and Goldberg, H.A. (1994) Modulation of crystal formation by bone phosphoproteins: role of glutamic acid-rich sequences in the nucleation of hydroxyapatite by bone sialoprotein. Biochemical Journal 302: 175–179CrossRefGoogle ScholarPubMed
Krampitz, G. and Graser, G. (1988) Molecular mechanisms of biomineralization in the formation of calcified shells. Angewandte Chemie (International Edition in English) 27: 1145–1156CrossRefGoogle Scholar
Lavelin, L., Yarden, N., Ben-Bassat, S., Bar, A. and Pines, M. (1998) Regulation of osteopontin gene expression during egg shell formation in the laying hen by mechanical strain. Matrix Biology 17: 615–623CrossRefGoogle ScholarPubMed
Mann, K. (1999) Isolation of a glycosylated form of the chicken eggshell protein ovocleidin and determination of the glycosylation site. Alternative glycosylation/phosphorylation at an N-glycosylation sequon. FEBS Letters 463: 12–14CrossRefGoogle ScholarPubMed
Mann, K. and Siedler, F. (1999) The amino acid sequence of ovocleidin 17, a major protein of the avian eggshell calcified layer. Biochemistry & Molecular Biology International 47: 997–1007Google Scholar
McKee, M.D. and Nanci, A. (1996) Osteopontin: an interfacial extracellular matrix protein in mineralized tissues. Connective Tissue Research 35: 197–205CrossRefGoogle ScholarPubMed
Nys, Y. (1987) Progesterone and testosterone elicit increases in the duration of shell formation in domestic hens. British Poultty Science 28: 57–68CrossRefGoogle ScholarPubMed
Nys, Y. (1993) Regulation of 1, 25 (OH)2D3, of osteocalcin and of intestinal and uterine calbindin in hens. In: Avian Endocrinology (Sharp, P.J., Ed), Journal of Endocrinology, Bristol, pp. 345–357Google Scholar
Nys, Y., Zawadzki, J., Gautron, J. and Mills, A.D. (1991) Whitening of brown-shelled eggs: mineral composition of uterine fluid and rate of protoporphyrin deposition. Poultry Science 70: 1236–1245CrossRefGoogle ScholarPubMed
Nys, Y., Hincke, M.T., Arias, J.L., Garcia-Ruiz, J.M. and Solomon, S.E. (1999) Avian eggshell mineralization. Poultry & Avian Biology Reviews 10: 143–166Google Scholar
Panheleux, M., Bain, M., Fernandez, M.S., Morales, I., Gautron, J., Arias, J.L., Solomon, S.E., Hincke, M. and Nys, Y. (1999a) Organic matrix composition and ultrastructure of eggshell: a comparative study. British Poultry Science 40: 240–252CrossRefGoogle ScholarPubMed
Panheleux, M., Kalin, O., Gautron, J. and Nys, Y. (1999b) Features of eggshell formation in guinea fowl: kinetics of shell deposition, uterine protein secretion and uterine histology. British Poultry Science 40: 632–643CrossRefGoogle ScholarPubMed
Panheleux, M., Nys, Y., Williams, J., Gautron, J., Boldicke, T. and Hincke, M.T. (2000) Extraction and quantification by ELISA of eggshell organic matrix proteins (ovocleidin-17, ovalbumin, ovotransferrin) in shell from young and old hens. Poultry Science 79: 580–588CrossRefGoogle Scholar
Pilon, M., Gautron, J., Nys, Y. and Hincke, M.T. (2000) Cloning of a hen protein related to mammalian LPS-binding and lipid transport proteins. XXI World's Poult y Congress, Montreal, CD S3.6.03Google Scholar
Pines, M., Knopov, V. and Bar, A. (1995) Involvement of osteopontin in egg shell formation in the laying chicken. Matrix Biology 14: 765–771CrossRefGoogle ScholarPubMed
Quintana, C. and Sandoz, D. (1978) Coquille de l'oeuf de caille: étude ultrastructurale et cristallographique. Calcified Tissue Research 25: 145–149CrossRefGoogle Scholar
Rodriguez-Navarro, A., Messier, C., Jimenez-Lopez, C. and Garcia-Ruiz, J.M. (2000) Importance of electrostatic interactions between proteins and calcite surfaces. In: Mineralization in Natural and Synthetic Biomaterials (Calvert, P., Levy, R., Kokubo, T. and Scheid, C., Eds), Material Research Society Symposium Proceedings pp. 353–359Google Scholar
Rodriguez-Navarro, A., Kalin, O., Nys, Y. and Garcia-Ruiz, J.M. (2001) Influence of the microstructure and crystallographic texture on the shell strength of eggs laid by hens of different ages. British Poultry Science (submitted)CrossRefGoogle Scholar
Romanoff, A.L. and Romanoff, A.J. (1949) The Avian Egg, John Wiley & Sons, Inc, New YorkGoogle Scholar
Sharp, R.M. and Silyn-Roberts, H. (1984) Development of preferred orientation in the eggshell of the domestic fowl. Biophysical Journal 46: 175–179CrossRefGoogle ScholarPubMed
Tullet, S.G. (1987) Egg shell formation and quality. In: Egg Quality Current Problems and Recent Advances (Wells, R.G. and Belyavin, C.G., Eds), Butterworth, London, pp. 123–146Google Scholar
Tyler, C. (1964) Wihhelm zlon Nathusius 1821–1899 on avian eggshells. University of Reading, ReadingGoogle Scholar
Wu, T.M., Rodriguez, J.P., Fink, D.J., Carrino, D.A., Blackwell, J., Caplan, A.I. and Heuer, A.H. (1995) Crystallization studies on avian eggshell membranes: implications for the molecular factors controlling eggshell formation. Matrix Biology 14: 507–513CrossRefGoogle ScholarPubMed