Molecular and Cellular Biochemistry

, Volume 243, Issue 1–2, pp 9–14

Metabolism of ovorubin, the major egg lipoprotein from the apple snail

  • Marcos S. Dreon
  • Horacio Heras
  • Ricardo J. Pollero
Article

Abstract

The site of synthesis of molluscs lipoproteins is little known and was investigated for the egg lipoprotein perivitellin 1 (PV1) or ovorubin in the freshwater snail Pomacea canaliculata. Tissues (albumen gland, gonad–digestive gland complex and muscle) of vitellogenic females were incubated in vitro at 25°C for 12 h with 14C Leucine. After that, soluble proteins from tissue homogenates and medium samples were analysed for de novo protein synthesis by electrophoresis and HPLC, and radiolabelled proteins quantified by liquid scintillation. Gonad–digestive gland complex did not synthesise ovorubin, in spite its high protein synthesis levels. Three albumen gland radiolabelled proteins (35, 32 and 28 kDa) comigrated with the subunits of ovorubin and represented 1.3% of the total labelled protein of that tissue. Western blot analysis with polyclonal antibodies confirmed that these were ovorubin subunits. In vivo experiments where vitellogenic females were injected with 3H Leucine, revealed that ovorubin was not present in hemolymph. ELISA analysis confirmed ovorubin presence only in albumen gland and developing eggs with levels of 800 and 582 mg/g protein, which represent 30.3 and 28.4 mg ovorubin/g of tissue, respectively. Therefore, albumen gland is the single site of ovorubin synthesis as no extragland synthesis, circulation or accumulation could be detected in the apple snail.

ovorubin lipoprotein snail glycolipocarotenoprotein in vitro synthesis carotenoprotein 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Wallace RA, Walker SL, Hauschka PV: Crustacean lipovitellin. Isolation and characterization of the major high-density lipoprotein from the eggs of decapods. Biochemistry 6: 1582-1590, 1967Google Scholar
  2. 2.
    Fainzilber M, Tom M, Shafir S, Applebaum SW, Lubzens E: Is there extraovarian synthesis of vitellogenin in Penaeid shrimp? Biol Bull 183: 233-241, 1992Google Scholar
  3. 3.
    Kanost MR, Kawooya JK, Law JH, Ryan RO, Van Heusden MC, Ziegler R: Insect hemolymph proteins. In: P.D. Evans and V.B. Wigglesworth (eds). Advances in Insect Physiology, Vol 22. Academic Press, London, 1990, pp 299-396Google Scholar
  4. 4.
    De Jong-Brink M, Boer HH, Joosse J: Reproductive biology of invertebrates. In: K.G. Adiyodi and R.G. Adiyodi (eds). Mollusca, Vol 1. John Wiley and Sons, New York, 1983, pp 297-355Google Scholar
  5. 5.
    Bride M, Petraca B, Faivre D: The synthesis of vitellogenins by the digestive gland of Helix aspersa: Evidence from cell-free translation of mRNA. Cell Mol Biol 38: 181-187, 1992Google Scholar
  6. 6.
    Barre P, Bride M, Beliard R, Petracca B: Localization of yolk proteins and their possible precursors using polyclonal and monoclonal antibodies in Helix aspersa. Cell Mol Biol 37: 639-650, 1991Google Scholar
  7. 7.
    Miksys S, Saleuddin ASM: Ferritin as an exogenously derived yolk protein in Helisoma duryi (Mollusca: Pulmonata). Can J Zool 64: 2678-2682, 1986Google Scholar
  8. 8.
    Bottke W: Immunolocalization of ferritin polypeptides in oocytes and somatic tissue of the freshwater snails Lymnaea stagnalis L. and Planorbarius corneus L. Cell Tissue Res 243: 397-404, 1986Google Scholar
  9. 9.
    Garín CF, Heras H, Pollero RJ: Lipoproteins of the egg perivitelline fluid of the snail Pomacea canaliculata (Mollusca: Gastropoda). J Exp Zool 276: 307-314, 1996Google Scholar
  10. 10.
    Cheesman DF: Ovorubin, a chromoprotein from the eggs of the gastropod mollusk Pomacea canaliculata. Proc R Soc Lond 149: 571-587, 1958Google Scholar
  11. 11.
    Dreon MS, Lavarias S, Garín CF, Heras H, Pollero RJ: Synthesis, distribution and levels of an egg lipoprotein from the apple snail Pomacea canaliculata (Mollusca Gastropoda). J Exp Zool 292: 323-330, 2003Google Scholar
  12. 12.
    Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72: 248-254, 1976Google Scholar
  13. 13.
    Laemmli VK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685, 1970Google Scholar
  14. 14.
    Towbin H, Staehelin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76: 4350-4354, 1977Google Scholar
  15. 15.
    Engwall E, Perlmann P: Enzyme-linked immunosorbent assay, ELISA III. Quantitation of specific antibodies by enzyme-labelled anti-immunoglobulin in antigen-coated tubes. J Immunol 109: 129-135, 1972Google Scholar
  16. 16.
    Kater SB, Mattson MP: Extrinsic and intrinsic regulators of neurite outgrowth and synaptogenesis in isolated, identified Helisoma neurons in culture. In: D.J. Beadle, G. Lees, S.B. Kater (eds). Cell Culture Approaches to Invertebrate Neuroscience. Associated Press, London, 1988, pp 1-32Google Scholar
  17. 17.
    Yano I, Chizei Y: Ovary is the site of vitellogenin synthesis in kuruma prawn, Penaeus japonicus. Comp Biochem Physiol 86B: 213-218, 1987Google Scholar
  18. 18.
    Meusy JJ: Vitellogenin, the extraovarian precursor of the protein yolk in Crustacea: A review. Reprod Nutr Develop 20: 1-21, 1980Google Scholar
  19. 19.
    Yepis-Plascencia G, Vargas-Albores F, Higuera-Ciapara I: Penaeid shrimp hemolymph lipoproteins. Aquaculture 191: 177-189, 2000Google Scholar
  20. 20.
    Lee RF: Lipoproteins from the hemolymph and ovaries of marine invertebrates. In: R. Gilles (ed). Advances in Comparative and Environmental Physiology, Vol 7. Springer-Verlag, London, 1991, pp 187-208Google Scholar
  21. 21.
    Hinsch GH, Vermeire PE: Histochemistry and ultrastructure of the capsule gland duct of the prosobranch gastropod Pomacea paludosa. Invertebr Reprod Dev 17: 203-201, 1990Google Scholar
  22. 22.
    Morishita F, Mukai ST, Saleuddin ASM: Release of proteins and polysaccharides from the albumen gland of the freshwater snail Helisoma duryi: Effect of cAMP and brain extracts. J Comp Physiol 182: 817-825, 1998Google Scholar
  23. 23.
    Heras H, Garin CF, Pollero RJ: Biochemical composition and energy sources during embryo development and in early juveniles of the snail Pomacea canaliculata (Mollusca: Gastropoda). J Exp Zool 280: 375-393, 1998Google Scholar
  24. 24.
    Cheesman DF, Lee WL, Zagalsky PF: Carotenoproteins in invertebrates. Biol Rev 42: 132-160, 1967Google Scholar
  25. 25.
    Zagalsky PF: Comparative studies on the aminoacid compositions of some carotenoid-containing glycolipoproteins and glycoproteins from the eggs and ovaries of certain aquatic invertebrates. Comp Biochem Physiol 41B: 385-395, 1972Google Scholar

Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • Marcos S. Dreon
    • 1
  • Horacio Heras
    • 2
  • Ricardo J. Pollero
    • 1
  1. 1.CONICET-UNLPInstituto de Investigaciones Bioquímicas de La Plata (INIBIOLP)La PlataArgentina
  2. 2.Instituto de Investigaciones Bioquímicas de La Plata (INIBIOLP)La PlataArgentina

Personalised recommendations