Cell and Tissue Research

, Volume 250, Issue 1, pp 125–134 | Cite as

Studies on microplasmodia of Physarum polycephalum

VI. Functional analysis of a cortical and fibrillar actin system by use of fluorescent-analog cytochemistry
  • Jörg Kukulies
  • Klaudia Brix
  • Wilhelm Stockem
Article
Accepted:

Summary

Fluorescently labeled actin (TRITC-G-actin) and heavy meromyosin (TRITC-HMM) derived from skeletal muscle and injected into microplasmodia of the acellular slime mold Physarum polycephalum were used to analyze the function of a cortical and fibrillar actin system in living specimens. The plasma membrane-attached cortical system can be labeled with TRITC-G-actin as well as with TRITC-HMM and visualized as a continuous sheath along the entire cell surface. Long-term experiments over time periods of several hours in conjunction with digital grey-value evaluations revealed that changes in the intensity of the fluorescent signal, as caused by alternative contraction and relaxation cycles of the cortical system, are distinctly correlated with periodic changes in the volume and shuttle streaming activity of the microplasmodia. The fibrillar actin system extending through the cytoplasmic matrix can be labeled only with TRITC-HMM. Formation and disappearance of fibrils were found to take place during relaxation and contraction of the cortical system, respectively. Results of the present paper indicate that the cortical actin system is mainly involved in motive force generation for alterations in cell surface morphology and locomotion activity, whereas the fibrillar actin system rather appears to maintain the mechanical stability of microplasmodia.

Key words

Cortical and fibrillar actin system Dynamic activity Cell adhesion Fluorescent-analog cytochemistry Physarum polycephalum 

Abbreviations

ATP

adenosine-5'-triphosphate

BSA

bovine serum 'albumin

DTE

1,4-dithioerythrit

EGTA

ethyleneglycol-bis-(β-amino-ethylether)-N,N,N′,N′,-tetraacetic acid

HMM

heavy meromyosin

PIPES

l,4-piperazine-N,N′-bis-(2-ethanesulfonic acid)

Rh

rhodamine

TRIS

Tris-(hydroxylmethyl)-aminomethane

TRITC

tetramethyl rhodamine isothiocyanate

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Copyright information

© Springer-Verlag 1987

Authors and Affiliations

  • Jörg Kukulies
    • 1
  • Klaudia Brix
    • 1
  • Wilhelm Stockem
    • 1
  1. 1.Institute for Cytology, University of BonnBonnFederal Republic of Germany

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