Histochemistry

, Volume 45, Issue 1, pp 1–21 | Cite as

Tracer and freeze-etching analysis of intra-cellular membrane-junctions in Paramecium

With a note on a new heme-nonapeptide tracer
  • Helmut Plattner
  • Doris Wolfram
  • Luis Bachmann
  • Elmar Wachter
Article

Summary

In paramecia the membranes of alveoli and trichocysts are permanently connected to the cell membrane by membrane-junctions, which consist of membrane-intercalated particles in a regular geometrical arrangement. Trichocysts contain secretory material discharged by exocytosis. In unfixed or fixed cells these two compartments were impermeable to the following tracers: To “microperoxidases”, i.e. a cytochrome c-derived heme-nonapeptide and a heme-undecapeptide (WM∼1650, 1900) applied in vivo, as well as to lanthanum and cytochrome c used during (La) or after (cytochrome c) fixation. The heme-nonapeptide was prepared by TPCK-trypsin digestion of cytochrome c and subsequent purification by Sephadex gel chromatography—a simple and inexpensive new procedure resulting in preparations of high yield and purity. Tracers entered alveoli only when the plasmalemma and the alveolar membranes ruptured upon glutardialdehyde fixation. In no case were transmembraneous channels detectable in regions containing membrane-intercalated particles; this holds true for all tracers used and for freeze-fracture replicas obtained by tantalum-tungsten evaporation. With regard to attachment sites over trichocysts our results do not support the assumptions by others according to which exocytosis would be driven by an osmotic shift via transmembraneous channels (which would be analogous to inter-cellular coupling phenomena mediated by gap-junctions), unless such channels would be assumed to operate as carriers rather than via diffusion. Tracers did not penetrate trichocysts before exocytosis occurred. The functional role of membrane-intercalated particles on trichocyst attachments remains nuclear. Despite some resemblance with gap-junctions all types of intra-cellular membrane-junctions investigated are functionally “tight” at the level of “resolution” obtained with tantalum-tungsten-shadowing and with the tracers used.

Keywords

Public Health Cell Membrane Functional Role Lanthanum Attachment Site 

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Copyright information

© Springer-Verlag 1975

Authors and Affiliations

  • Helmut Plattner
    • 1
  • Doris Wolfram
    • 1
  • Luis Bachmann
    • 2
  • Elmar Wachter
    • 3
  1. 1.Institut für ZellbiologieUniversität MünchenFed. Rep. Germany
  2. 2.Institut für Technische ChemieTechnische Universität MünchenFed. Rep. Germany
  3. 3.Institut für Physiologische Chemie und Physikalische BiochemieUniversität MünchenFed. Rep. Germany

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