Cell and Tissue Research

, Volume 269, Issue 3, pp 411–420 | Cite as

Electron microscopy of a double helical tubular filament in keyhole limpet (Megathura crenulata) hemolymph

  • J. Robin Harris
  • Jürgen Markl
Article

Summary

A ≈25 nm hollow double helical filament has been detected ultrastructurally in the cell-free supernatant from hemolymph of the keyhole limpet Megathura crenulata (Gastropoda: Prosobranchia: Fissurellidae). Subsequently, much higher concentrations of this material were found in the cell pellet from hemolymph. Both negative staining and thin sectioning have been performed in an attempt to obtain a preliminary structural characterization of this new filament. It is proposed that the filaments are released or secreted from blood hemocytes in response to bleeding, but it has not been possible to define absolutely an intracellular organelle containing this material. It is shown that the hollow filaments, which have fine fibrillar surface extensions, are readily distinguishable from collagen fibrils and from tubular (multi-decameric) keyhole limpet hemocyanin (KLH).

Key words

Hemolymph Filaments Hemocyanin Hemocytes Megathura crenulata (Mollusca) 

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References

  1. Bang FB (1961) Reaction of injury in the oyster (Crassostrea virginica). Biol Bull 121:57–68Google Scholar
  2. Bayer ME (1964) An electron microscopical examination of urinary mucoprotein and its interaction with influenza virus. J Cell Biol 21:265–274Google Scholar
  3. Harris JR, Agutter PS (1970) A negative staining study of human erythrocyte ghosts and rat liver nuclear membranes. J Ultrastruct Res 33:219–232Google Scholar
  4. Harris JR, Horne RW (1991) Negative staining. In: Harris JR (ed) Electron microscopy in biology. IRL Press, Oxford, pp 203–228Google Scholar
  5. Harris JR, Cejka Z, Wegener-Strake A, Gebauer W, Markl J (1992) Two-dimensional crystallization, transmission electron microscopy and image processing of keyhole limpet haemocyanin (KLH). Micron 23: in pressGoogle Scholar
  6. Horne RW, Pasquali-Ronchetti I (1974) A negative staining-carbon film technique for studying viruses. I.Preparative procedure for studying icosahedral and filamentous viruses. J Ultrastruct Res 47:361–383Google Scholar
  7. Markl J, Savel-Niemann A, Wegener-Strake A, Süling M, Schneider A, Gebauer W, Harris JR (1991) The role of two distinct subunit types in the architecture of keyhole limpet hemocyanin (KLH). Naturwissenschaften 78:512–514Google Scholar
  8. Ratcliffe NA, Rowley AF (1979) A comparative synopsis of the structure and function of the blood cells of insects and other invertebrates. Dev Comp Immunol 3:189–243Google Scholar
  9. Reynolds ES (1963) The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J Cell Biol 17:208–212Google Scholar
  10. Rifkin E, Cheng TC, Hohl HR (1969) An electron microscope study of the constituent of encapsulating cysts in Crassostrea virginica formed in response to Tylocephalum metacestodes. J Invertebr Pathol 14:211–226Google Scholar
  11. Ruddell CL (1971) The fine structure of the granular amebocytes of the Pacific oyster, Crassostrea gigas. J Invertebr Pathol 18:269–275Google Scholar
  12. Sminia T, Knaap WPW van der (1987) Cells and molecules in molluscan immunology. Dev Comp Immunol 11:17–28Google Scholar
  13. Stern W, Dales S (1976) Biogenesis of vaccinia: Isolation and characterization of a surface component that elicits antibody suppressing infectivity and cell-cell fusion. Virology 75:232–241Google Scholar
  14. Suzuki T, Yoshinaka R, Mizuta S, Funakoshi S, Wada W (1991) Extracellular matrix formation by amebocytes during epithelial regeneration in the pearl oyster Pinctada fucata. Cell Tissue Res 266:75–82Google Scholar
  15. Toh Y, Mizutani A, Tokunaga F, Muta T, Iwanaga S (1991) Morphology of the granular hemocytes of the Japanese horseshoe crab Tachypleus tridentatus and immunocytochemical localization of clotting factors and antimicrobial substances. Cell Tissue Res 266:137–147Google Scholar
  16. Watson ML (1958) Staining of tissue sections for electron microscopy with heavy metals. J Biophys Biochem Cytol 4:475–478Google Scholar

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • J. Robin Harris
    • 1
  • Jürgen Markl
    • 2
  1. 1.Institut für Zell- und TumorbiologicDeutsches KrebsforschungszentrumHeidelbergFederal Republic of Germany
  2. 2.Institut für Zoologie der UniversitätMainzFederal Republic of Germany

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