Wilhelm Roux's archives of developmental biology

, Volume 194, Issue 8, pp 453–461 | Cite as

A new biochemical marker for foot-specific cell differentiation in hydra

  • Sabine Hoffmeister
  • H. Chica Schaller
Article

Summary

Mucous cells in the basal disk of hydra contain a peroxidase-like enzyme allowing specific staining of these cells with substrates for peroxidases. The peroxidase activity provides an excellent marker for foot mucous cell, differentiation and was used to follow the reappearance of footspecific cells during foot regeneration after amputation. By choosing the appropriate either soluble or precipitable substrate the peroxidase reaction was used both for a qualitative and for a quantitative evaluation of foot-specific differentiation in hydra. For histological studies diaminobenzidien was found to be a suitable substrate which forms a dark brown precipitate within the cells containing the peroxidase activity. For a quantitative evaluation of foot regeneration the soluble substrate 2,2-azino-di(3-ethyl-benzthiazoline-sulfonic acid-6) ammonium salt was used which after reaction with the enzyme gives rise to a diffusible green reaction product the concentration of which can be measured by its specific absorption at 415 nm. Based on the diffusible enzyme product a new quantitative assay for foot regenration was developed and applied to confirm the effect and specificity of morphogenetic substances which either inhibit or activate foot or head regeneration in hydra.

Key words

Hydra Peroxidase activity Foot cell differentiation Foot activator Foot inhibitor Head inhibitor 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Berking S (1977) Bud formation in hydra: Inhibition by an endogenous morphogen. Wilhelm Roux's Arch 186:189–210Google Scholar
  2. Berking S (1983) The fractionation of a hydra-derived inhibitor into head and foot inhibitor may be an artefact. Wilhelm Roux's Arch 192:327–332Google Scholar
  3. Birr C, Zachmann B, Bodenmüller H, Schaller HC (1981) Synthesis of a new neuropeptide, the head activator from hydra. FEBS Lett 131:317–322PubMedGoogle Scholar
  4. Bodenmüller H, Zachmann B (1983) A radioimmunoassay for the Hydra head activator. FEBS Lett 159:237–240Google Scholar
  5. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Analytical. Biochemistry 72:248–254CrossRefPubMedGoogle Scholar
  6. Burnett AL (1966) A model of growth and differentiation in hydra. Am Naturalist 100:165–190Google Scholar
  7. Davis LE (1973) Histological and ultrastructural studies of the basal disk of hydra. Z Zellforsch 139:1–45PubMedGoogle Scholar
  8. Grimmelikhuijzen CJP, Schaller HC (1977) Isolation of a substance activating foot formation in hydra. Cell Differ 6:297–305PubMedGoogle Scholar
  9. Grimmelikhuijzen CJP, Schaller HC (1979) Hydra as a model organism for the study of morphogenesis. Trends Biochem Sci 12:265–267Google Scholar
  10. Kemmner W (1984) A model of head regeneration in hydra. Differentiation 26:83–90Google Scholar
  11. Kemmner W, Schaller HC (1984) Actions of head activator and head inhibitor during regeneration in hydra. Differentiation 26:91–96Google Scholar
  12. Lentz TL (1966) The cell biology of hydra. North-Holland Publishing Co, AmsterdamGoogle Scholar
  13. Schaller HC (1973) Isolation and characterisation of a lowmolecular-weight substance activating head and bud formation in hydra. J Embryol Exp Morphol 29:27–38PubMedGoogle Scholar
  14. Schaller HC (1981) Morphogenetic substances in hydra, Fortschritte der Zoologie 26: 153–162. Gustav Fischer Verlag, Stuttgart, New YorkGoogle Scholar
  15. Schaller HC (1984) The head and the foot inhibitor from hydra are not Dowex artefacts Wilhelm Roux's Arch 193:117–118Google Scholar
  16. Schaller HC, Bodenmüller H (1981) Isolation and amino acid sequence of a morphogenetic peptide from hydra. Proc Natl Acad Sci [USA] 78:7000–7004Google Scholar
  17. Schaller HC, Bodenmüller H, Zachmann B, Schilling E (1984) Enzyme-linked immunosorbent assay for the neuropeptide head activator. Eur J Biochem 138:365–371PubMedGoogle Scholar
  18. Schaller HC, Schmidt T, Grimmelikhuijzen CJP (1979) Spearation and specificity of action of four morphogens from hydra. Wilhelm Roux's Arch 186:138–149Google Scholar
  19. Schmidt T, Schaller HC (1976) Evidence for a foot-inhibiting substance in hydra. Cell Differ 5:151–159PubMedGoogle Scholar
  20. Schmidt T, Schaller HC (1980) Properties of the foot inhibitor from hydra. Wilhelm Roux's Arch 188:133–139Google Scholar
  21. Sersig BL, Lesh-Laurie GE (1981) Bud separation inHydra oligactis. Biol Bull 160:431–437Google Scholar
  22. Tardent P (1978) Cnidaria in Seidel F Morphogenese der Tiere: 68-415 VEB. Gustav Fischer Verlag, JenaGoogle Scholar
  23. Wilby OK, Webster G (1970) Experimental studies on axial polarity in hydra. J Embryol Exp Morphol, 24:595–613PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • Sabine Hoffmeister
    • 1
  • H. Chica Schaller
    • 1
  1. 1.Department of BiophysicsMax-Planck-Institute for Medical ResearchHeidelbergGermany

Personalised recommendations